Methods for treating and ameliorating angina and congestive heart failure

Abstract

In alternative embodiments, provided are methods for treating, ameliorating, lessening the symptoms of, or preventing, angina, myocardial infarction, reperfusion injury, ventricular arrhythmias (VA), pump failure due to myocardial infarction (MI) in an early ischemia / reperfusion (I / R) period, and / or congestive heart failure (CHF) comprising administering to an individual in need thereof a composition or a drug capable of suppressing hypoxia-induced SUMOylation of the cardiac sodium channel NaV1.5.

Description

[0001] METHODS FOR TREATING AND AMELIORATING ANGINA, MYOCARDIAL INFARCTION, REPERFUSION INJURY, HEART ARRHYTHMIAS AND CONGESTIVE HEART FAILURE (CHF)

[0002] RELATED APPLICATIONS

[0003] This Patent Convention Treaty (PCT) International Application claims the benefit of priority under 35 U.S.C. § 119(e) of U.S. Provisional Application No. 63 / 530,438, August 2, 2023. The aforementioned application is expressly incorporated herein by reference in its entirety and for all purposes. All publications, patents, patent applications cited herein are hereby expressly incorporated by reference for all purposes.

[0004] STATEMENT AS TO FEDERALLY SPONSORED RESEARCH

[0005] This invention was made with government support under R01HL10549 awarded by the National Institutes of Health (NIH). The government has certain rights in the invention.

[0006] REFERENCE TO ELECTRONIC SEQUENCE LISTING

[0007] The application contains a Sequence Listing which has been submitted electronically in .XML format and is hereby incorporated by reference in its entirety. Said .XML copy, created on August 2, 2024, is named“5825.153163PCT.xml” and is 11,563 bytes in size. The sequence listing contained in this .XML file is part of the specification and is hereby incorporated by reference herein in its entirety.

[0008] TECHNICAL FIELD

[0009] This invention generally relates to biology and medicine. In alternative embodiments, provided are methods for treating, ameliorating, lessening the symptoms of, or preventing, angina, myocardial infarction, reperfusion injury, a ventricular arrhythmia (VA), cardiac pump failure due to myocardial infarction (MI) (optionally cardiac pump failure due to myocardial infarction (MI) in an early ischemia / reperfusion (I / R) period), congestive heart failure (CHF), hypovolemia, acidosis, hyperkalemia, hypokalemia, a cardiac poisoning or cardiopathology due to a toxin or poisoning, cardiac tamponade, tension pneumothorax, coronary thrombosis, sepsis, shock, pulmonary thrombosis, pulmonary embolism, respiratory decompensation, arrythmia a cardiac event caused by hypoperfusion and / or cardiac arrest, comprising administering to an individual in need thereof comprising suppressing hypoxia-induced SUMOylation of the cardiac sodium channel NaV1.5, a post-translational modification (PTM) that we have shown produces life-threatening elevations in the late Na+ current.

[0010] BACKGROUND

[0011] SUMOylation is a process in which SUMO proteins are covalently attached to specific lysine residues in target proteins, thereby regulating various aspects of protein function. SUMO (Small Ubiquitin-like Modifier) proteins are a family of small proteins that are covalently attached to and detached from other proteins in cells to modify their function. SUMO proteins are similar to ubiquitin and are considered members of the ubiquitin-like protein family. SUMOylation is directed by an enzymatic cascade analogous to that involved in ubiquitination. In contrast to ubiquitin, SUMO is not used to tag proteins for degradation. Mature SUMO is produced when the last four amino acids of the C-terminus have been cleaved off to allow formation of an isopeptide bond between the C-terminal glycine residue of SUMO and an acceptor lysine on the target protein. Human SUMO proteins include SUMO1, SUMO2, SUMO3 and SUMO4.

[0012] SUMO attachment to its target is similar to that of ubiquitin; however, the SUMO precursor has some extra amino acids that need to be removed, therefore a C- terminal peptide is cleaved from the SUMO precursor by a protease (in human these are the SENP proteases) to reveal a di-glycine motif. The obtained SUMO then becomes bound to an El enzyme (SUMO Activating Enzyme 1 (SAE)) which is a heterodimer (subunits SAE1 and SAE2). It is then passed to an E2 enzyme, which is a conjugating enzyme (Ubc9). In some cases, one of a small number of E3 ligating proteins (or enzymes) speeds or makes more specific the conjugation process to select targets.

[0013] SUMMARY

[0014] In alternative embodiments, provided are methods for treating, ameliorating, lessening the symptoms of, or preventing: angina, myocardial infarction, reperfusion injury, a ventricular arrhythmia (VA), cardiac pump failure due to myocardial infarction (MI) (optionally cardiac pump failure due to myocardial infarction (MI) in an early ischemia / reperfusion (I / R) period), congestive heart failure (CHF), hypovolemia, acidosis, hyperkalemia, hypokalemia, a cardiac poisoning or cardiopathology due to a toxin or poisoning, cardiac tamponade, tension pneumothorax, coronary thrombosis, sepsis, shock, pulmonary thrombosis, pulmonary embolism, respiratory decompensation, arrythmia a cardiac event caused by hypoperfusion and / or cardiac arrest, comprising administering to an individual in need thereof a compound, a composition or anucleic acid capable of modifying a NaV1.5 K442 residue to another residue that cannot be SUMOylated, thereby preventing or inhibiting or lessening the amount or severity of hypoxia-induced post-translational modification (PTM) of cardiac sodium channel NaV 1.5, and preventing, inhibiting or lessening pathologic elevations in intracellular Na+ current (ILATE), or treating, ameliorating, lessening the symptoms of, or preventing: angina, myocardial infarction, reperfusion injury, a ventricular arrhythmia (VA), cardiac pump failure due to myocardial infarction (MI) (optionally cardiac pump failure due to myocardial infarction (MI) in an early ischemia / reperfusion (I / R) period), congestive heart failure (CHF), hypovolemia, acidosis, hyperkalemia, hypokalemia, a cardiac poisoning or cardiopathology due to a toxin or poisoning, cardiac tamponade, tension pneumothorax, coronary thrombosis, sepsis, shock, pulmonary thrombosis, pulmonary embolism, respiratory decompensation, arrythmia a cardiac event caused by hypoperfusion and / or cardiac arrest.

[0015] In alternative embodiments of methods as provided herein:

[0016] - the drug or composition capable of suppressing hypoxia-induced SUMOylation of a cardiac sodium channel NaV1.5 comprises:

[0017] (a) subasumstat, or CAS No. 1858276-04-6, or a compound having the formula or structure (also called TAK-981):

[0018] (b) a compound having the formula or structure:

[0019] (also called TAK-243);

[0020] (c) a compound having CAS No. 144707-18-6, or the formula or structure: (also called 2-D08);

[0021] (d) ginkgolic acid or ginkgolic acid derivatives thereof, or a compound having the structure or formula:

[0022] (e) anacardic acid or anacardic acid derivatives thereof, or 2-Hydroxy-6- pentadecylbenzoic acid;

[0023] (f) kerriamycin B or kerriamycin B derivatives thereof, or 9-[4-[5-(4.5-dihvdroxy-6- methyloxan-2-yl)oxy-6-methyloxan-2-yl]oxy-5-hydroxy-6-methyloxan-2-yl] -3,4a,8- trihydroxy-12b-(5-hydroxy-6-methyloxan-2-yl)oxy-3-methyl-2,4-dihydrobenzo[a]anthracene- 1,7, 12 -trione;

[0024] (g) davidiin or davidiin derivatives thereof, or [(1S,19R,2OR.21S.22R)- 6,7,8,11.12,13-hexahydroxy-3,16-dioxo-21,22-bis[(3,4,5-trihydroxybenzoyl)oxy]-2,17,23- trioxatetracyclo[17.3.1.04,9.010,15]tricosa-4,6,8,10,12,14-hexaen-20-yl] 3,4,5- trihydroxybenzoate; (h) tannic, or gallotannic acid, or 1, 2,3,4, 6-penta-O-{3,4-dihydroxy-5-[(3, 4,5- trihydroxybenzoyl)oxy]benzoyl}-D-glucopyranose, or a compound having the structure or formula: (i) a SUMO adenylate (AMSN) or a tetrahedral SUMO intermediate (AVSN) analogues;

[0025] (j) MLN4924, or pevonedistat (CAS no. 905579-51-3), or a compound having the structure or formula: (k) Dimethyl 1-(1-anilinobut-3-enyl)-7-oxabicyclo[2.2.1]hepta-2,5-diene-2,3- dicarboxylate (or 1-(1-Phenylamino-but-3-enyl)-7-oxa-bicyclo[2.2.1]hepta-2,5-diene- 2, 3-dicarboxylic acid dimethyl ester), or PubChem CID no. 9549553;

[0026] (1) compound COHOOO, or CAS No. 1534358-79-6, or a compound having the structure or formula:

[0027]

[0028] (m) ML-792, or CAS No. 1644342-14-2, or a compound having the structure or formula or formula: (n) ML-93, or a compound having the structure or formula or formula:

[0029] (o) McM025044, or a compound having the structure or formula or formula:

[0030] (p) glycyrrhizin (or glycyrrhizic acid or glycyrrhizinic acid) I, or a compound having the structure or formula:

[0031]

[0032] (q) spectomycin B1, or a compound having the structure:

[0033] (r) chaetochromin A, or PubChem CID no. 6712966, or (2R,3R)-5,6,8- trihydroxy-2,3-dimethyl-9-[(2R,3R)-5,6,8-trihydroxy-2,3-dimethyl-4-oxo-2,3- dihydrobenzo[g]chromen-9-yl]-2,3-dihydrobenzo[g]chromen-4-one, or a compound having the structure:

[0034] (s) viomellein, or PubChem CID no. 3033108. or (3R)-8-[(3R)-9,10- dihydroxy-7-methoxy-3-methyl-1-oxo-3,4-dihydrobenzo[g]isochromen-8-yl]-10- hydroxy-7-methoxy-3-methyl-3,4-dihydrobenzo[g]isochromene-1,6,9-trione, or a compound having the structure:

[0035]

[0036] (t) a compound having the structure or formula:

[0037] (u) GSK145A. or CAS No. 1609945-27-8, or a compound having the structure or formula:

[0038] (v) topotecan, or HYCAMTIN™ or POTACTASOL™, or a compound having the structure or formula: (w) nocardione A, or a compound having the structure or formula:

[0039] (x) 33-DINOR-dunnione, or a compound having the structure or formula: 33-DINOR-dehydrodunnione or a compound having the structure or formula:

[0040] (y) [β-lapachone, or 3,4-Dihydro-2.2-dimethyl-2H-naphtho[1,2-b]pyran-5.6- dione, or CAS No. 4707-32-8, , or a compound having the structure or formula:

[0041] (z) macrophilone A and its methoxy derivative, or a compound having the structure or formula:

[0042] (aa) a Ubc9 inhibitor having the structure or formula:

[0043]

[0044] (bb) an SENP inhibitor (or an aza-epoxide SENP inhibitor) having the structure or formula: (also called momordin Ic),

[0045] (also called Streptonigrin),

[0046] (also called NSC45384), (also called vialinin A), (also called atromentin).

[0047]

[0048] (cc) a compound as set forth in U.S. patent application publication no. US20170002032A9, and described in WO2016004136, as set forth in further detail, below; and / or

[0049] (dd) an isomer, deuterated isoform, optical isomer or stereoisomer, a racemate or racemic mixture, an enantiomer, an individual diastereomer or a diastereomeric mixture, an analog, a crystalline product or a crystalline intermediate, a pharmaceutically acceptable salt thereof, a prodrug or a bioisostere of any of (a) to (cc); and / or

[0050] (ee) a SUMO protease or a SENP protein, or a catalytic domain of a SUMO protease or a SENP protein, or a nucleic acid encoding the SUMO protease or SENP protein, or the catalytic domain of the SUMO protease or SENP protein, wherein optionally the SUMO protease or a SENP protein, or a catalytic domain of a SUMO protease or a SENP protein, comprises a:

[0051] (i) SENP1, sentrin-specific protease 1 Homo sapiens, optionally a sentrin- specific protease 1 having an amino acid (aa) sequence SEQ ID NO: 3, or NCBI Reference Sequence NP_001254524.1, or a SENP1 catalytic domain comprising aa residues from 419 to 644 of SEQ ID NO:3;

[0052] (ii) SENP2. sentrin-specific protease 2 Homo sapiens, optionally a SENP2, sentrin-specific protease 2 having an amino acid (aa) sequence SEQ ID NO:4, or NCBI Reference Sequence NP_067640.2, or a SENP2 catalytic domain comprising aa residues 336 to 584 of SEQ ID NO:4;

[0053] (iii) SENP3, sentrin-specific protease 3 Homo sapiens, optionally a sentrin-specific protease 3 having an amino acid (aa) sequence SEQ ID NO:5, or NCBI Reference Sequence NP_056485.2, or a SENP3 catalytic domain comprising aa residues 400 to 572 of SEQ ID NO:5; (iv) SENP5, isoform 1, sentrin-specific protease 5 isoform 1 Homo sapiens, optionally a sentrin-specific protease 5 isoform 1 having an amino acid (aa) sequence SEQ ID NO:6, or NCBI Reference Sequence NP_689912.2, or a sentrin-specific protease 5 isoform 1 catalytic domain comprising aa residues 581 to 753 of SEQ ID NO:6;

[0054] (v) SENP5, isoform 2, sentrin-specific protease 5 isoform 2 Homo sapiens, optionally an isoform 2, sentrin-specific protease 5 isoform 1 having an amino acid (aa) sequence SEQ ID NO:7, or NCBI Reference Sequence NP_001294974.1, or a sentrin-specific protease 5 isoform 2 catalytic domain comprising aa residues 581 to 707 of SEQ ID NO:7;

[0055] (vi) SENP6, sentrin-specific protease 6 isoform 1 Homo sapiens, optionally a sentrin-specific protease 6 isoform 1 having an amino acid (aa) sequence SEQ ID NO:8, or protein Accession no. NP 056386.2, or a sentrin- specific protease 6 isoform 1 catalytic domain comprising aa residues 973 to 1067 of SEQ ID NO:8;

[0056] (vii) Sentrin-specific protease 6 isoform 2 Homo sapiens, optionally a Sentrin-specific protease 6 isoform 2 having an amino acid (aa) sequence SEQ ID NO:9, or protein Accession no. NP_001093879.1, or a Sentrin-specific protease 6 isoform 2 catalytic domain comprising aa residues 966 to 1060 of SEQ ID NO:9;

[0057] (viii) Sentrin-specific protease 6 isoform 3 Homo sapiens, optionally a Sentrin-specific protease 6 isoform 3 having an amino acid (aa) sequence protein Accession no. NP_001291721.1, or a Sentrin-specific protease 6 isoform 3 catalytic domain comprising aa residues 966 to 1060 of protein Accession no. NP_001291721.1;

[0058] (ix) SENP7, sentrin-specific protease 7 isoform 1 Homo sapiens, optionally a sentrin-specific protease 7 isoform 1 having an amino acid (aa) sequence SEQ ID NO: 10, or protein Accession no. NP_065705.3, or a sentrin- specific protease 7 isoform 1 catalytic domain comprising aa residues 774 to 1011 of SEQ ID NO: 10;

[0059] (x) Sentrin-specific protease 7 isoform 2 Homo sapiens, optionally a Sentrin-specific protease 7 isoform 2 having an amino acid (aa) sequence SEQ ID NO:11, orprotein Accession no. NP OO 1070671.1, or a C -terminal catalytic domain thereof;

[0060] (xi) sentrin-specific protease 7 isoform 3 Homo sapiens, optionally a sentrin-specific protease 7 isoform 3 protein Accession no. NP 001269730.1, or a C-terminal catalytic domain thereof;

[0061] (xii) sentrin-specific protease 7 isoform 4 Homo sapiens, optionally a sentrin-specific protease 7 isoform 4 protein Accession no. NP_001269731.1, or a C-terminal catalytic domain thereof;

[0062] (xiii) sentrin-specific protease 7 isoform 6 Homo sapiens, optionally a sentrin-specific protease 7 isoform 6 protein Accession no. NP 001269733.1, or a C-terminal catalytic domain thereof; and / or

[0063] (xiv) sentrin-specific protease 7 isoform 5 Homo sapiens, optionally a sentrin-specific protease 7 isoform 5 protein Accession no. NP 001269732.1, or a C-terminal catalytic domain thereof; and / or, in alternative embodiments of methods as provided herein:

[0064] - the drug or composition capable of suppressing hypoxia-induced SUMOylation of a cardiac sodium channel NaV1.5 is formulated or fabricated to be contained in:

[0065] (a) in a liquid, a gel, a hydrogel, a vesicle, a liposome, a nanoparticle, a nanolipid particle, a powder or an aqueous or a saline formulation, or for administration in vitro or in vivo,'

[0066] (b) for enteral or parenteral administration;

[0067] (c) in or as a liposome, a nanoparticle, or a nanoliposome, wherein optionally the liposome, nanoparticle, or nanoliposome is fabricated to comprise or express moieties or molecules that target the liposome, nanoparticle, or nanoliposome to a particular tissue or organ, wherein optionally the liposome, nanoparticle, or nanoliposome is fabricated to target heart tissue, myocardial tissue, and / or a coronary artery:

[0068] (d) in or as a dendrimer, a tablet, a pill, a capsule, a gel, a hy drogel, a geltab, a liquid, a powder, an emulsion, a lotion, an aerosol, a spray, a lozenge, an aqueous or a sterile or an injectable solution, an eye drop, or an implant; or

[0069] (e) for intravenous injection, subcutaneous injection, intramuscular injection, inhalation, or intravitreal injection. In alternative embodiments, of methods as provided herein:

[0070] - the drug or composition capable of suppressing hypoxia-induced SUMOylation of a cardiac sodium channel NaV1.5 is administered after, or immediately after, or between about 5 minutes and 5 hours of, or between about 15 minutes and 15 hours of, an individual in need thereof presenting with symptoms of angina, myocardial infarction, reperfusion injury, ventricular arrhythmia (VA), pump failure due to myocardial infarction (MI) in an early ischemia / reperfusion (I / R) period, congestive heart failure (CHF), hypovolemia, acidosis, hyperkalemia, hypokalemia, cardiopathology due to a toxin, cardiac tamponade, tension pneumothorax, coronary thrombosis, sepsis, shock, pulmonary thrombosis, pulmonary embolism, respiratory decompensation, arrythmia and / or cardiac arrest;

[0071] - the drug or composition capable of suppressing hypoxia-induced SUMOylation of a cardiac sodium channel NaV1.5 is administered to the individual in need thereof intravenously (IV), and optionally is administered IV via catheter to the heart;

[0072] - the drug or composition capable of suppressing hypoxia-induced SUMOylation of a cardiac sodium channel NaV1.5 is administered to the individual in need intramuscularly (IM), or by injection directly into or approximate to heart tissue, or cardiac myocytes, or heart vasculature;

[0073] - the drug or composition capable of suppressing hypoxia-induced SUMOylation of a cardiac sodium channel NaV1.5 is administered to the individual in need thereof by implantation of an implant in the individual in need thereof, wherein the implant contains therein or comprises the drug or composition, and the drug or composition is released to tissue (optionally a heart) surrounding or approximate to the implant by elution or effusion of the drug or composition, or by controlled release of the drug or composition from the implant into the surrounding tissue, wherein optionally the implant is fabricated as a stent; and / or

[0074] - the drug or composition capable of suppressing hypoxia-induced SUMOylation of a cardiac sodium channel NaV1.5 is administered to the individual in need thereof by oral, rectal, topical routes, or by inhalation, or by vaginal, topical, nasal or pulmonary administration, or by parenteral (optionally subcutaneous, intramuscular, intravenous and intradermal) infusion;

[0075] In alternative embodiments, provided are uses of a drug or composition (optionally a nucleic acid) capable of suppressing hypoxia-induced SUMOylation of a cardiac sodium channel NaV1.5 for treating, ameliorating, lessening the symptoms of, or preventing: angina, myocardial infarction, reperfusion injury, a ventricular arrhythmia (VA), cardiac pump failure due to myocardial infarction (MI) (optionally cardiac pump failure due to myocardial infarction (MI) in an early ischemia / reperfusion (I / R) period), congestive heart failure (CHF), hypovolemia, acidosis, hyperkalemia, hypokalemia, a cardiac poisoning or cardiopathology due to a toxin or poisoning, cardiac tamponade, tension pneumothorax, coronary thrombosis, sepsis, shock, pulmonary thrombosis, pulmonary embolism, respiratory decompensation, arrythmia a cardiac event caused by hypoperfusion and / or cardiac arrest.

[0076] In alternative embodiments, provided are drugs or compositions (optionally a nucleic acid) capable of suppressing hypoxia-induced SUMOylation of a cardiac sodium channel NaV 1.5 for use in treating, ameliorating, lessening the symptoms of, or preventing: angina, myocardial infarction, reperfusion injury, a ventricular arrhythmia (VA), cardiac pump failure due to myocardial infarction (MI) (optionally cardiac pump failure due to myocardial infarction (MI) in an early ischemia / reperfusion (I / R) period), congestive heart failure (CHF), hypovolemia, acidosis, hyperkalemia, hypokalemia, a cardiac poisoning or cardiopathology due to a toxin or poisoning, cardiac tamponade, tension pneumothorax, coronary thrombosis, sepsis, shock, pulmonary thrombosis, pulmonary embolism, respiratory decompensation, arrythmia a cardiac event caused by hypoperfusion and / or cardiac arrest.

[0077] In alternative embodiments, provided are methods for treating, ameliorating, lessening the symptoms of, or preventing: angina, myocardial infarction, reperfusion injury, a ventricular arrhythmia (VA), cardiac pump failure due to myocardial infarction (MI) (optionally cardiac pump failure due to myocardial infarction (MI) in an early ischemia / reperfusion (I / R) period), congestive heart failure (CHF), hypovolemia, acidosis, hyperkalemia, hypokalemia, a cardiac poisoning or cardiopathology due to a toxin or poisoning, cardiac tamponade, tension pneumothorax, coronary thrombosis, sepsis, shock, pulmonary thrombosis, pulmonary embolism, respiratory decompensation, arry thmia a cardiac event caused by hypoperfusion and / or cardiac arrest, comprising administering to an individual in need thereof a compound, composition and / or a nucleic acid capable of modifying a NaV 1.5 K442 residue to another residue that cannot be SUMOylated, wherein optionally theNaV1.5 is ahuman NaVl.5.

[0078] In alternative embodiments of methods as provided herein:

[0079] - the nucleic acid capable of modifying a NaV 1.5 K442 residue to another residue that cannot be SUMOylated: is delivered using CRISPR technology or prime editing technology; or, is or comprises: an RNAi inhibitory nucleic acid molecule, a double-stranded RNA (dsRNA) molecule, a microRNA (mRNA), a small interfering RNA (siRNA), an antisense RNA, a short hairpin RNA (shRNA), an inhibitory ribozyme, or a CRISPR or prime editing system such as CRISPR-Cas9, wherein optionally the inhibitory nucleic acid, optionally an inhibitory RNA fragment or sequence, is based on SEQ ID NO: 1 or SEQ ID NO:2;

[0080] - the nucleic acid capable of modifying a NaV 1.5 K442 residue to another residue that cannot be SUMOylated is contained in an expression construct, plasmid, expression vehicle, virus or vector, or the nucleic acid capable of modifying a NaV 1.5 K442 residue to another residue that cannot be SUMOylated is formulated or fabricated to be contained in:

[0081] (a) in a liquid, a gel, a hydrogel, a vesicle, a liposome, an exosome, a nanoparticle, a nanolipid particle, a powder or an aqueous or a saline formulation, or for administration in vitro or in vivo,'

[0082] (b) for enteral or parenteral administration;

[0083] (c) in or as a liposome, an exosome, a nanoparticle, or a nanoliposome, wherein optionally the liposome, exosome, nanoparticle, or nanoliposome is fabricated to comprise or express moieties or molecules that target the liposome, exosome, nanoparticle, or nanoliposome to a particular tissue or organ, wherein optionally the liposome, exosome, nanoparticle, or nanoliposome is fabricated to target heart tissue, myocardial tissue, and / or a coronary' artery;

[0084] (d) in or as a tablet, a pill, a capsule, a gel, a hydrogel, a geltab, a liquid, a powder, an emulsion, a lotion, an aerosol, a spray, a lozenge, an aqueous or a sterile or an injectable solution, an eye drop, or an implant; or

[0085] (e) for intravenous injection, subcutaneous injection, intramuscular injection, inhalation, or intravitreal injection; and / or

[0086] - the nucleic acid capable of modifying a NaV1.5 K442 residue to another residue that cannot be SUMOylated is contained in an expression construct, a plasmid, an expression vehicle, a virus or a vector, and the expression construct, plasmid, expression vehicle, virus or vector is delivered or administered to the individual in need thereof, wherein optionally the expression vehicle or vector is selected from the group consisting of a herpes simplex virus, a human immunodeficiency virus (HIV), a synthetic vector, an adeno-associated virus (AAV), a lentivirus. an adenovirus and a plasmid, and optionally the AAV is selected from the group consisting of AAV1, AAV2, AAV3, AAV4. AAV5, AAV6, AAV7, AAV8, AAV9, AAV 10 AAV11, AAV12, pseudotyped AAV. a rhesus-derived AAV, AAVrh8, AAVrhlO and AAV- DJan AAV capsid mutant, an AAV hybrid serotype, an organ-tropic AAV, a cardiotropic AAV, and a cardiotropic AAVM41 mutant.

[0087] The details of one or more exemplary embodiments of the invention are set forth in the accompanying drawings and the description below. Other features, objects, and advantages of the invention is apparent from the description and drawings, and from the claims.

[0088] All publications, patents, patent applications cited herein are hereby expressly incorporated by reference in their entireties for all purposes.

[0089] DESCRIPTION OF DRAWINGS

[0090] The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) is provided by the Office upon request and payment of the necessary fee.

[0091] The drawings set forth herein are illustrative of exemplary' embodiments provided herein and are not meant to limit the scope of the invention as encompassed by the claims.

[0092] FIG. 1 schematically illustrates SUMO pathway and inhibitors: 1. Maturation; SUMO pro-peptide is cleaved at C-terminus by SENP exposing Gly-Gly; 2. Activation. SUMO-activating enzy me (El) forms a thioester to yield El-GG-SUMO; 3. Conjugation. GG-SUMO is transferred to E2 conjugating enzyme UBC9 (E2-GG- SUMO); 4. Ligation. SUMO is transferred to target Lys (K442 in Nav1.5); for some targets, E3 ligases speed SUMOylation or increase specificity. 5. SENP deSUMOylases remove SUMO from targets. Site of inhibition: TAK-981 via El, 2- DO8 via E2, as described in detail in Example 1, below.

[0093] FIG. 2 schematically illustrates how hypoxia induces NaV1.5 SUMOylation on K442 so channels reopen late in the cardiac AP when normally inactive, raising ILATEto approximately 4% of peak as seen with VA and sudden cardiac death (1), as described in detail in Example 1, below.

[0094] FIG. 3A-C illustrate thatNaV1.5-K442 is required for hypoxic SUMOylation and ILATE: ILATEwas studied at -30 mV, 50 and 100 ms after the peak current and is presented as mean % of the peak. The time course for hypoxic modulation was studied with steps from -100 mV to -30 mV every' 10 s. Single-channel currents were studied in cell-attached mode and elicited every 5 s by a 50-ms depolarizing pulse to - 30 mV from a holding potential of -120 mV. Data were recorded at filter and sampling frequencies of 5 and 50 kHz, respectively, and processed offline using a 1.2- kHz Bessel filter for display. For each cell, sweeps with no channel activity (null) were averaged offline, and subtracted from the data sweeps before analysis:

[0095] FIG. 3A Left image graphically illustrates time course for changes in ILATEwith hypoxia for WT (closed circles), NaV1.5-K442Q (open circles) with control solution (black), SUMO1 (red), or SENP1 (blue) in the pipette;

[0096] FIG. 3A right image illustrates bars indicating mean ILATEas a % of the peak in 21% 02. after 200 s in 1.5% 02, and then 200 s in 21% O2;

[0097] FIG. 3B top image illustrates depolarization with single WT channels open, inactivate rapidly, and do not reopen unless exposed to 1.5% O2. Null traces in 21% and 1.5% 02 were approximately 55%±5% (n=20 patches; 2,000 sw eeps):

[0098] FIG. 3B bottom image illustrates ensemble (n=200- 220 sweeps); and

[0099] FIG. 3C Top image illustrates single NaV1.5-K442Q channels do not reopen in 21% or 1.5% O2and null traces were approximately 51% ± 3% (n=20 patches; 2,000 sweeps); and

[0100] FIG. 3C bottom image illustrates ensemble (n=200- 225 sweeps). Data mean ± SEM for 8 - 12 cells per group, as described in detail in Example 1, below. FIG. 4A-E illustrates that hypoxia recruits one SUMO1 to each NaV1.5 on the PM at the same rate as ILATEincreases. Single mTFP-NaV1.5 channels (blue) and SUMO1 tagged with mCherry (m- SUMO1, red) were studied in CH0-K1 cells by TIRFm. Stoichiometric (photobleaching) and pixel- by -pixel analysis for subunit density and co-localization were performed and Manders’ coefficients for colocalization assessed post hoc for 3-5 regions per cell; co-localization was defined as the presence of both fluorophores at more than 30% of the maximum fluorescence level recorded in that stack (overlap is represented in images as white pixels). The time course of hypoxic modulation of NaV1.5 ILATEwas studied as in Fig 3:

[0101] FIG. 4A left image illustrates single co-localized particles of mCherry and TFP at the surface of cells expressing NaV1.5 (with NaV01) and SUMO1. The time courses for bleaching of the fluorophores’ revealed complexes have one subunit of each type;

[0102] FIG. 4B graphically illustrates a histogram of photobleaching showing that hypoxia increased single m-SUMOl (red) subunits at the PM co-localized with mTFP-NaV1.5 (blue), without a change in subunit stoichiometry;

[0103] FIG. 4C Left image shows picture of cells where in ambient O2the surface density of m-SUMOl (top) is low compared to NaV1.5 (middle) with little colocalization (bottom); and Right image illustrates hypoxia recruits m- SUMO1 to the PM within 100 s (top), where it is co-localized withNaV1.5 (bottom); surface levels of NaV1.5 did not change (middle). Scale bar = 10 mm;

[0104] FIG. 4D graphically illustrates a histogram of surface density for 6 cells studied as in (A). In ambient O2, the density of pixels per mm2 with SUMO1 alone (red) was 3±1, and 305±21 for NaV1.5 (blue). The density7of pixels with both fluorophores (green) was 33±3 per mm2. Hypoxia increased co-localization to 301±13 pixels per mm2 and decreased the density of free NaV1.5 channels (37±4) without altering the density of free SUMO1 (6±2); and

[0105] FIG. 4E graphically illustrates a time course for co-localization of mTFP- NaV1.5 and m-SUMOl (Manders’ coefficient, black circle) and the magnitude of ILATE. as a % of the peak cunent (open circle) were coincident. A mean Manders’ coefficient of 0.10±0.01 in ambient O2increased to 0.9I±0.03 after 100 s hypoxia. Mean ILATErose from 0.46% ± 0.1% to 4.4% ± 0.8%. Data represent 5-8 cells; biophysical parameters and particle statistics as reported, as described in detail in Example 1, below.

[0106] FIG. 5 illustrates images of WT SENP2-mCherry or SENP2- dell8-mCherry with NaV1.5 and Navpi in HEK293T cells. Differential interference contrast (DIC) (left). Epifluorescence (right) 24 h after transfection. Full-length SENP2 observed at the nuclear membrane. SENP2-dell8 was throughout the cytosol, as described in detail in Example 1, below.

[0107] FIG. 6A-C graphically illustrates that hypoxia induces ILATEin hiPS-CMs in a SUMO- dependent manner:

[0108] FIG. 6A Left: graphically illustrates sample traces showing ILATEat -30 mV increased when 21% 02 (black) changed to 1.5% 02 (black dash). Inset: increase in ILATEwith hypoxia;

[0109] FIG. 6A Right: graphically illustrates time course for changes in ILATEin response to hypoxia. Scale bars = 500 pA and 10 ms.;

[0110] FIG. 6B Left graphically illustrates: SUMO1 in the pipette (red) increased ILATE. NO further change in ILATEwas observed when the perfusate was exchanged for 1.5% O2(red dash);

[0111] FIG. 6B Right graphically illustrates: ILATEwas not observed with SENP1 in the pipette at 21% (blue) or 1.5% O2(blue dash). In each case, dashed lines overlap with the solid lines; and

[0112] FIG. 6C graphically illustrates a histogram for mean ILATEas % of peak current at 21% O2(solid color), after 200 s in 1.5% O2(hatched color), and after 200 s at 21% O2(solid color) with control solution (black), 1 nM SUMO1 (red), or 1 nM SENP1 (blue) in the pipette. Data are mean ± SEM for 8 to 10 cells per group, as described in detail in Example 1, below.

[0113] FIG. 7 A-E illustrates hypoxia-induced, SUMO-dependent APD prolongation in hiPS-CMs:

[0114] FIG. 7A graphically illustrates exemplar spontaneous AP in hiPS-CMs in 21% and 1.5% O2+ / - ranolazine. APD50 increased with 1.5% O2by 22%, 663±2ms to 810±2 ms, and ranolazine restored the APD50 to 689±2 ms;

[0115] FIG. 7B graphically illustrates SUMO1 in the pipette APD50 was 822 ± 2 ms and increased to 870±2 ms with 1.5% 02; ranolazine + hypoxia led to an APD50 of 699±3ms as observed without SUMO1 in the pipette at 21% O2; FIG. 7C graphically illustrates APD50 was 663±2 ms for cells with SENP1 in the pipette, and the APD50 increased only to 698±2 ms with 1.5% O2and returned to 663±4 ms with ranolazine;

[0116] FIG. 7D graphically illustrates data showing a 5-fold increase in ILATE(half the observed with hypoxia or SUMOylation), in the O’Hara-Rudy model, increased APD50 by 27%, from 205 ms to 260 ms;

[0117] FIG. 7E graphically illustrate histograms mean APD50 in (A)-(C). Scale bars = 25 mV. 250 ms in (A)-(C) and 30 mV, 150 ms in (D). Data are mean ± SEM for 6- 9 cells per group (*p < 0.01. two-way paired t test). as described in detail in Example 1, below.

[0118] FIG. 8 illustrates an image showing that SUMO-Q94P improves yield of SUMO- RanGAP (arrow). RanGAP (R) - / + GFP-HA-SUMO1 (WT) or GFP-HA- SUMO1-Q94P (P) in HEK293T cells. At 24 h cells were lysed with RIP A, PMSF, Na+ orthovanadate, and protease inhibitors + / - 40 mM NEM. Protein quantified by BCA. SDS-PAGE, transfer to PVDF, stain with (top) anti-HA (Invitrogen 26183) or (lower) anti-P-actin (Santa Cruz 47778) and alkphos- 20 (Promega S3721) + NBT / BCIP solution (Thermo), as described in detail in Example 1, below.

[0119] FIG. 9A-I illustrate images showing that there are no significant changes in the systolic or diastolic function in the homozygous Scn5AK442Q / WT (KI) mice compared to WT animals:

[0120] FIG. 9A illustrates representative 2D echocardiographic images, taken at parasternal short axis. Blood flow through the mitral valve (MV) was assessed using pulse-wave Doppler to capture diastolic function, as seen in the representative images, with E and A waves (lower panels);

[0121] FIG. 9B graphically illustrates data showing that there were no significant differences in heart rate in KI mice compared to WT animals;

[0122] FIG. 9C graphically illustrates data showing that there were no significant differences in the left ventricular (LV) dimensions in KI mice compared to WT animals including LV anterior and posterior wall dimensions in diastole (d) or systole (s) (LVAW;d; LVAW;s and LVPW;d; LVPW;s) or LV internal dimension in diastole and systole (LVlD;d and LVID;s);

[0123] FIG. 9D-G graphically illustrates data showing that there were no significant differences in corrected LV mass (FIG. 9D), LV volume in diastole (FIG. 9E), LV volume in systole (FIG. 9R), or fractional shortening (FS) (FIG. 9G). The diastolic function was normal in KI mice compared to WT animals;

[0124] FIG. 9H graphically illustrates data showing quantitatively, isovolumic relaxation time (IVRT); and

[0125] FIG. 91 graphically illustrates data showing that MV E / A ratio were not significantly different in the KI mice compared to WT. Numbers of animals are shown within the bar graphs, as described in detail in Example 1, below.

[0126] FIG. 10A-E illustrate data showing a significant decrease in VA in KI mice during 45 min ischemia and reperfusion period:

[0127] FIG. 10A illustrates representative ECGs at baseline; and

[0128] FIG. 10B-E graphically illustrate and summarize data for heart rate. P duration, RR intervals. PR intervals. QRS durations, and QTc intervals. as described in detail in Example 1, below.

[0129] FIG. 11 A-G illustrate data showing that Navi .5-K442Q KI mice show better pump function compared to WT in response to LAD ligation:

[0130] FIG. 11 A illustrates 2D echocardiographic images, taken at parasternal short axis, left images wild type with (upper image) control (sham) and (lower image) reperfusion I / R, the right images SCN5AK442Q(upper image) control (sham) and (lower image) reperfusion I / R; and

[0131] FIG. 11B-G illustrate summary data for heart rate (FIG. 1 IB). LV mass (FIG. 11C). ejection fraction (FIG. 1 ID), fractional shortening (FIG. HE), LV volume in diastole (FIG. 11 F) and systole (FIG. 11 G). KI mice show a significant preservation of LVEF and FS compared to WT animals at 2 weeks after I / R injury. Animal numbers are shown in the bar graphs, as described in detail in Example 1, below.

[0132] FIG. 12A-E illustrate data showing that hemodynamic monitoring shows better pump function in KI mice than WT in response to LAD ligation:

[0133] FIG. 12A illustrates representative developed pressure (P), volume and dP / dt traces in WT sham (control) and SCN5AK442Qmice and wild type and wild type I / R and SCN5AK442QI / R KI mice; and

[0134] FIG. 12B-E graphically illustrate summary data for peak developed pressure (FIG. 12B), end diastolic volume (EDV) (FIG. 12C), end systolic volume (ESV) (FIG. 12D) and dP / dt in WT (FIG. 12E) compared to KI mice. KI mice show the preservation of developed pressure and dP / dt compared to WT animals at 2 weeks after I / R injury , as described in detail in Example 1, below.

[0135] FIG. 13A-E illustrate data showing a significant decrease in VA burdens using SUMO specific inhibitors (TAK-981 and 2-D08) in the WT animals:

[0136] FIG. 13A illustrates representative ECG recordings during ischemia and reperfusion in WT mice treated with vehicle alone compared to SUMO-specific inhibitors; and

[0137] FIG. 13B-D graphically illustrates summary data showing a significant decrease in arrhythmic events in SUMO-specific inhibitors treated mice compared to controls (*p<0.05, n = 3-5 animals per group), as described in detail in Example 1, below.

[0138] FIG. 14A-C illustrate data showing that treatment with SUMO-specific inhibitors ameliorates LV dysfunction after I / R injury in WT animals:

[0139] FIG. 14A illustrates 2D echocardiographic images, taken at parasternal short axis, with upper image control, middle image TAK-981, and lower image 2-D08; and

[0140] FIG. 14B-C illustrate summary data for fractional shortening (FIG. 14B) and ejection fraction (FIG. 14C) at 2 weeks post I / R in WT animals. Animal numbers are shown in the bar graphs, as described in detail in Example 1, below.

[0141] FIG. 15 (Left panel) illustrates ambulatory ECG recordings showing heart rate response to isoproterenol (ISO) in a WT mouse, where mice were allowed to fully recover after telemetry implantation before a 24-h recording; top panels show representative ECG tracings in basal condition, and after 0-adrenergic stimulation with ISO, and lower panel shows average HR over 20-min in WT mice (n = 8) demonstrating diurnal variations; FIG. 15 (Right panel) illustrates in vivo electrophysiologic studies showing inducible ventricular arrhythmia using programmed stimulation in a mouse model with chronic pressure- induced cardiac hypertrophy, as described in detail in Example 1, below.

[0142] FIG. 16 graphically illustrates data from an AP-clamp: an example sequential dissection of multiple currents in a rabbit ventricular myocyte, as described in detail in Example 1, below. FIG. 17A-B (Left Panels) illustrate simultaneous measurement of transients (CaT) and (right panels) sarcomere shortening, the images are representative traces for sarcomere shortening and CaT measured using Fura-2 ratio (RFURA) from WT cardiomyocytes, and summary data for percentages of sarcomere shortening and CaT; and

[0143] FIG. 17C-D illustrates assessment of the SR Ca2+ load; the myocytes were first paced to steady state; the pacing was stopped for 15 seconds (s) followed by a rapid application of 20 mM caffeine to induce SR Ca2+release, FIG. 17D graphically illustrates SRL load, as described in detail in Example 1, below.

[0144] FIG. 18A-B illustrates Ca2+sparks recorded in myocytes using confocal line scan; representative two dimensional (2D) (FIG. 18 A) and three dimensional (3D) (FIG. 18B) images; and

[0145] FIG. 18C illustrates quantification of spark amplitudes as well as FWHM (full-width at half maximum and FDHM (full-duration at half maximum). as described in detail in Example 1, below.

[0146] FIG. 19A schematically illustrates a diagram showing the quantification of sarcomere shortening during relaxed and contracted states from SHG recordings;

[0147] FIG. 19B illustrates images of example SHG recording and corresponding Ca2+ transients from a ventricular myocyte; and

[0148] FIG. 19C graphically illustrates quantification of rise time (trise) and half decay time (t 1 / 2) of a Ca2+ transient, as described in detail in Example 1, below.

[0149] FIG. 20 illustrates Table 1 and Table 2 showing data that human NaV1.5 in HEK293T cells with NaVpi, whole cell recording as in Fig 3. Data are mean ± SEM, 4-8 cells per group. TAK-981 was applied at 15 uM and 2-D08 at 20 uM:

[0150] Table 1 shows + / - 10 nM SUMO1 in the pipette after inhibitor pre-treatment for various times; and

[0151] Table 2 shows pre-treatment for 4 h with hypoxia (1.5% O2), as described in detail in Example 1, below.

[0152] FIG. 21A-B illustrates how NaV1.5 is SUMOylated: previously shown using FRET and TIRF microscopy (and functional studies with wild type and Nav1.5- K424Q to ablate SUMOylation by removing the site for attachment) we show here human NaV1.5 channels expressed with NaVpi and HA-tagged SUMO in HEK293 cells subjected to immunoprecipitation (IP) using anti-NaV1.5 (FIG. 21A, left) or anti-SUMOl (FIG. 21 B) antibodies and separation on SDS-PAGE gels. With both IP antibodies, SUMOylation of Nav1.5 is demonstrated by immunoblotting (IB) because SUMO1 and NaV1.5 are co-purified. When HEK293 cells were pre-treated by 15 pM TAK981 for 4 hrs prior to IP, NaV1.5 SUMOylation was decreased indicating its ability to suppress SUMOylation. Demonstrating specificity, IP with non-immune IgG isolated neither SUMO norNaV1.5. WCL, whole cell lysate.

[0153] FIG. 22 illustrates data showing myocardial ischemia and reperfusion (MI / R) leads to SUMOylation of NaV1.5 in vivo that is suppressed by intravenous administration of TAK-981. Protection previously shown using functional studies with wild ty pe and Navl.5-K424Q knock-in mice to ablate SUMOylation (removing the site for attachment), and TAK-981, is supplemented here by showing the effectiveness of TAK-981 in vivo by direct visualization of the channel-SUMO complex isolated from mouse heart. Wild type mice were subjected to MI / R (45-min ligation of the left anterior descending artery followed by 30-min of reperfusion), + / - TAK-981 at 7.5 mg / kg IV at the time of ligation. With MI / R, SUMOylated NaV1.5 was isolated from the free wall (frank infarct) and septum (spared) of heart (red arrow, mouse 17) that was decreased when mice were treated with TAK-981 (red arrow, mouse 12). The results were confirmed in repeat studies on two other pairs of treated and untreated mice.

[0154] FIG. 23A-C illustrate data showing that NaV1.5-K442Q knock-in (KI) mice removes the channel site of SUMOylation and protect mice from MI / R -induced tissue damage. KI mice that cannot be SUMOylated on K424 show less cardiac troponin release 24 hours after MI / R (FIG. 23 A) and less collagen deposition 2 weeks post MI / R (FIG. 23B-C) compared to WT animals. This validates the claim of protection afforded by suppression of SUMOylation.

[0155] FIG. 24A-B illustrate data showing that MI / R-induced late current (NaV1.5 channel ILATE) is suppressed inNaV1.5-K442Q KI mice (that are not SUMOylated) and wild type mice (WT) treated with TAK-981. A significant increase in ILATEis recorded in myocytes isolated from the left ventncular anterior wall of WT mice immediately after MI / R (45 min ischemia and 30 min reperfusion) compared to sham (surgery but no ligation). The increase in ILATEis not seen in KI mice or WT mice treated with TAK-981 given at the time of MI / R. FIG. 24A exhibits representative traces of cells from WT and KI post MI / R or sham, with or without TAK-981 treatment. The statistical analysis of ILAIE in the 5 different groups is summarized in FIG. 24B. This directly proves operation of the proposed mechanism of protection in cells from mouse heart.

[0156] FIG. 25A-E illustrate data showing that NaV1.5-K442Q knock-in (KI) mice are protected from MI / R-induced altered calcium flux and decreased shortening. Validating that suppressing SUMOylation and ILATEdecreases downstream pro- arrhythmic effect of alter calcium flux in vivo. WT and NaV1.5-K442Q KI mice were studied by simultaneous measurement of Ca2+ transients (CaT) using Fura-2 dual- wavelength ratiometric method and sarcomere shortening by IONOPTIX™ detection (lonOptix Co) and fast Fourier transform (FFT) analysis. A significant decrease in sarcomere shortening (FIG. 25A-B) and CaT amplitude (FIG. 25C-D) was observed in WT after MI / R (45 min ischemia and 30 min reperfusion). The changes were abrogated in KI mice (FIG. 25A-D). There is a trend towards a delay in CaT decay in WT after I / R, but the data were not significant (FIG. 25E).

[0157] Like reference symbols in the various drawings indicate like elements.

[0158] DETAILED DESCRIPTION

[0159] In alternative embodiments, provided are methods for treating, ameliorating, lessening the symptoms of, or preventing: angina, myocardial infarction, reperfusion injury, a ventricular arrhythmia (VA), cardiac pump failure due to myocardial infarction (MI) (optionally cardiac pump failure due to myocardial infarction (MI) in an early ischemia / reperfusion (I / R) period), congestive heart failure (CHF), hypovolemia, acidosis, hyperkalemia, hypokalemia, a cardiac poisoning or cardiopathology due to a toxin or poisoning, cardiac tamponade, tension pneumothorax, coronary thrombosis, sepsis, shock, pulmonary thrombosis, pulmonary embolism, respiratory decompensation, arrythmia a cardiac event caused by hypoperfusion and / or cardiac arrest, comprising administering to an individual in need thereof comprising suppressing hypoxia-induced SUMOylation of the cardiac sodium channel NaV1.5, a post-translational modification (PTM) that we have shown produces life-threatening elevations in the late Na+ current. We have demonstrated in mice a novel strategy to subdue ventricular arrhythmias (VA) and pump failure due to myocardial infarction (MI) in the early ischemia / reperfusion (I / R) period (events associated with humans’ sudden death). The method suppresses hypoxia-induced SUMOylation of the cardiac sodium channel NaV1.5, a post-translational modification (PTM) that we have shown produces life- threatening elevations in the late Na+ current (ILATE).

[0160] While the invention is not limited by any particular mechanism of action, the rationale for methods as provided herein is five-fold. First, ILATEis associated with death in 25% to 50% of patients with MI and I / R due to arrhythmias including ventricular tachycardia (VT), ventricular fibrillation (VF), and torsade de points (TdP). Second, we have demonstrated that hypoxia produces pathologic levels of ILATEin under 1-min due to SUMOylation of NaV1.5 channels on a single residue, lysine 442 (K442). and that ILATEis eliminated by mutation of the site so it is no longer subject to SUMOylation (NaV1.5-K442Q), or by application of a deSUMOylating enzyme. Third, the preliminary results shown here include a knock- in (KI) mouse strain we created, NaV1.5-K442Q, that ablates NaV1.5 SUMOylation MI. 45-min left anterior descending coronary artery (LAD) ligation, manifest VA, whereas the KI mice show decreased VA and left ventricular (LV) dysfunction compared to the WT mice. Fifth, we show- that tw o SUMO pathway inhibitors, one in phase I / II cancer trials as a transcriptional regulator, suppress Navi.5 SUMOylation, ILATE, VA, and ameliorate LV dysfunction in WT mice subjected to LAD ligation. Based on these findings, inhibiting NaV1.5 SUMOylation is a viable strategy to reduce VA and LV dysfunction in acute and post-acute phases of ML

[0161] Angina and heart attacks come from decreased blood flow' (ischemia) to the heart: the decrease in oxygen delivery (hypoxia) to the heart interferes with normal electric activity and normal pumping blood to the body. The earliest change is excess sodium going into the muscle cells and this is caused by the heart sodium channel (Nav1.5) opening abnormally late in the heartbeat when it is supposed to be closed (late sodium current). We have demonstrated that the late current is produced within a minute of hypoxia because enzymes act to link SUMO protein to the channel on one site (K442) and that no late current develops with hypoxia if the channel linkage site is mutated or the SUMO cannot attach or is cut off the channel. As described herein, we have studied the effects in wild type and knock-in mice and shown physiological effects that demonstrate new therapies to protect humans.

[0162] If the channel has a mutation at the site so it cannot be SUMOylated (K442 is changed) the mice are protected from cardiac rhythm disturbances and pump failure in response to preclinical model for heart attack (occluding the major vessel the feeds the heart, the left anterior descending artery). Moreover, we showed that giving compounds, TAK-981 or 2-D08, that block the enzyme from attaching SUMO to K442 protect wild type mice (TAK-981 has been used in phase I / II drug trials to treat cancer because the SUMO pathway has been known for controlling the expression of genes in the nucleus over days rather than the rapid effects we study of the SUMO enzymes at the cell surface) . Because these findings were generated in an art- accepted animal model for equivalent human cardiopathies, these data demonstrate that humans with angina, heart attack and heart failure can benefit from drug (for example, TAK-981 or 2-D08) or genetic therapies to stop SUMO attachment to the Nav1.5 channel at K442, as this stops the pathological late sodium current.

[0163] In alternative embodiments, treatments as provided herein can be by genetic methods, and can include: viral vectors or protein delivery via nanoparticles to alter the genetic code of NaVl .5 K442 to another residue so it cannot be SUMOylated, for example, by modifying the SCN5A gene or transcripts using CRISPR, prime editing, or other technologies.

[0164] Further, other causes of myocardial ischemia / hypoxia are therefore amenable to improvement via this approach (i.e., drug (for example, TAK-981 or 2-D08) or genetic therapies to stop SUMO attachment to the Navi .5 channel at K442); these include, but are not limited to, hypovolemia, acidosis, hyperkalemia, hypokalemia, cardiopathology due to a toxin, cardiac tamponade, tension pneumothorax, coronary thrombosis, sepsis, shock, pulmonary thrombosis, pulmonary embolism, respiratory decompensation, arrythmia and other causes of cardiac hypoperfusions and cardiac arrest.

[0165] We have showed that:

[0166] - the SUMO pathway can act at the cell surface, modifying ion channels to change their electrical activity.

[0167] - hypoxia regulates the SUMO effect on ion channels first in the CNS channel Navi.2 (for example in stroke), - hypoxia leads to SUMOylation of cardiac Navi .5 explaining late sodium current and the initial events (sodium loading) associated with hypoxia as seen with heart attacks.

[0168] As described herein, we tested the prediction for knock-in mice that changing the K442 site would protect the mice from the negative effects of left anterior descending (LAD) coronary artery occlusion for 45 min (to simulate a heart attack) and that treatment with compounds given as intraperitoneal injection that inhibit SUMOylation would also protect.

[0169] In alternative embodiments, the composition or a drug capable of suppressing hypoxia-induced SUMOylation of the cardiac sodium channel NaV1.5 comprises any compound capable of inhibiting SUMOylation at El (i.e., blocking formation of an El-SUMO intermediate, where El is El enzyme, or SUMO Activating Enzyme 1 (SAE)), E2 (i.e., blocking formation of an E2-SUMO intermediate (where E2 is an E2 conjugating enzyme), E3 (i.e., blocking formation of an E3-SUMO intermediate (E3 is a ligating protein)) or an unidentified site (i.e., blocking formation of any SUMO intermediate), for example, a drug or composition used in methods as provided herein, and to suppress hypoxia-induced SUMOylation of the cardiac sodium channel

[0170] NaV1.5, comprises any one or several of:

[0171] 1. compounds inhibiting SUMO El: 2. compounds inhibiting SUMO E2;

[0172] 3. compounds inhibiting SUMOylation with unknown sites:

[0173] El enzymes activate ubiquitin (Ub) and ubiquitin-like (Ubl) proteins in two steps by carboxy -terminal adenylation and thioester bond formation to a conserv ed catalytic cysteine in the El Cys domain. Ubiquitin (Ub) and ubiquitin-like (Ubl) proteins attached to their target proteins and modulating the activities of those targets in various ways. Three types of evolutionarily conserved enzymes are: El activating enzymes; E2 conjugating enzy mes; and, E3 ligase enzymes, and they act sequentially through parallel yet distinct pathways to conjugate ubiquitin and Ubl proteins, such as SUMO and NEDD8. to their targets. The El enzyme uses the adenosine triphosphate (ATP) and magnesium to adenylate the C -terminal Ub / Ubl glycine, releasing pyrophosphate and resulting in adenosine monophosphate (AMP).

[0174] References

[0175] 1. Fukuda, I. et al. Ginkgolic acid inhibits protein SUMOylation by blocking formation of the El-SUMO intermediate. Chem. Biol., 2009, 76(2), 133-140.

[0176] 2. Brackett, M et al. Synthesis and Evaluation of Ginkgolic Acid Derivatives as SUMOylation Inhibitors. ACS Medicinal Chemistry Letters 2020 11 (11), 2221-2226. 3. Fukuda, I. et al. Kerriamycin B inhibits protein SUMOylation. J. Antibiot. (Tokyo), 2009, 62(4), 221-224.

[0177] 4. Takemoto, M. et al. Inhibition of protein SUMOylation by davidiin, an ellagitannin from Davidia involucrata. J. Antibiot. (Tokyo), 2014. 67(4), 335-338.

[0178] 5. Suzawa, M. et al. A gene-expression screen identifies a non-toxic sumoylation inhibitor that mimics SUMO-less human LRH-1 in liver. eLife, 2015, 4, e09003.

[0179] 6. Lu, X. et al. Designed semisynthetic protein inhibitors of Ub / Ubl El activating enzymes. J. Am. Chem. Soc., 2010, 732(6), 1748-1749.

[0180] 7. Soucy, T. A. et al. An inhibitor ofNEDD8-activating enzyme as anew approach to treat cancer Nature 2009, 458 (7239) 732- 736.

[0181] 8. Kumar, A. et al. Identification of sumoylation activating enzyme 1 inhibitors by structure-based virtual screening. J. Chem. Inf. Model., 2013, 53(4), 809-820.

[0182] 9. Ashutosh Kumar et al, Identification of quinazolinyloxy biaryl urea as a new class of SUMO activating enzyme 1 inhibitors, Bioorganic & Medicinal Chemistry Letters, 2013, https: / / doi.Org / 10.1016 / j.bmcl.2013.07.022.

[0183] 10. Ashutosh Kumar et al, Identification of new SUMO activating enzy me 1 inhibitors using virtual screening and scaffold hopping, Bioorganic & Medicinal Chemistry Letters, 2016, https: / / doi.Org / 10.1016 / i.bmcl.2016.01.030.

[0184] 11. Li, Y.J. et al. Allosteric inhibition of ubiquitin-like modifications by a class of inhibitor of SUMO-activating enzy me. Cell Chem. Biol., 2019, 26(2), 278-288. e6.

[0185] 12. Lv, Z. et al. Molecular mechanism of a covalent allosteric inhibitor of SUMO El activating enzyme. Nat. Commun., 2018. 9(1). 5145.

[0186] 13. Chen, Y. et al. Bicyclic and tricyclic inhibitors of SUMOylation enzymes and methods for their use. US patent 20130245032 Al, 2013.

[0187] 14. He, X. et al. Probing the roles of SUMOylation in cancer cell biology' by using a selective SAE inhibitor. Nat Chem Biol 13, 1164-1171 (2017).

[0188] 15. Biederstadt A etal. SUMO pathway inhibition targets an aggressive pancreatic cancer subtype. Gut 2020; 69:1472-1482.

[0189] 16. Duffey, M.O. et al. Heteroaryl compounds useful as inhibitors of SUMO activating enzyme. WO Patent 2015002994 A2, 2015.

[0190] 17. Yannick D. Benoit et al. Targeting SUMOylation dependency in human cancer stem cells through a unique SAE2 motif revealed by chemical genomics, Cell Chemical Biology', Volume 28, Issue 10, 2021, Pages 1394-1406. elO, 18. Bentz GL, Lowrey AJ, Home DC, et al. Using glycyrrhizic acid to target sumoylation processes during Epstein-Barr virus latency. PLoS ONE. 2019; 14(5): e0217578.

[0191] 19. Hirohama. M. et al. Spectomycin Bl as a novel SUMOylation inhibitor that directly binds to SUMO E2. ACS Chem. Biol., 2013, 5(12), 2635-2642.

[0192] 20. Kim, Y.S. et al. An electrophoretic mobility shift assay identifies a mechanistically unique inhibitor of protein sumoy lation. Chem. Biol., 2013, 20(4), 604-613.

[0193] 21. Zlotkowski, K. et al. A small-molecule microarray approach for the identification of E2 enzyme inhibitors in ubiquitin-like conjugation pathways. SLAB Discov., 2017, 22(6), 760-766.

[0194] 22. Brandt, M. et al Development of a high-throughput screen to detect inhibitors ofTRPSl sumoylation. Assay Drug Dev. Technol., 2013, 77(5), 308-325.

[0195] 23. Leyva, M.J. et al. Synthetic derivatives of the SUMO consensus sequence provide a basis for improved substrate recognition. Bioorg. Med. Chem. Lett., 2015, 25(10), 2146-2151.

[0196] 24. Kumar, A.; Ito, A.; Hirohama, M.; Yoshida, M.; Zhang, K.Y. Identification of sumoylation inhibitors targeting a predicted pocket in Ubc9. J. Chem. Inf. Model., 2014, 54(10), 2784-2793.

[0197] 25. Cai S etal. Selective targeting of NaV1.7 via inhibition of the CRMP2-Ubc9 interaction reduces pain in rodents. Sei. Transl. Med.13.eabhl 314(2021)

[0198] 26. Patent US10441586 (CRMP2 SUMOylation inhibitors and uses thereof).

[0199] 27. Gomez K et al. Identification and targeting of a unique Navi .7 domain driving chronic pain. Proc Natl Acad Sci USA. 2023 Aug 8;120(32):e2217800120.

[0200] 28. Brackett, M. Christopher, Blagg, S.J. Brian, Current Status of SUMOylation Inhibitors, Current Medicinal Chemistry, 2021, Doi 10.2174 / 0929867327666200810135039

[0201] Formulations and pharmaceutical compositions

[0202] In alternative embodiments, provided are compounds and compositions, including formulations and pharmaceutical compositions, for use in in vivo, in vitro or ex vivo methods for treating, ameliorating, preventing or reversing: a cancer, for example, a breast cancer or a thyroid cancer by administering a drug or composition also capable of suppressing hypoxia-induced SUMOylation of a cardiac sodium channel NaV1.5.

[0203] In alternative embodiments, the pharmaceutical compositions as provided herein or as used in methods as provided herein can be administered enterally or parenterally, topically, orally or by local administration, such as by aerosol or trans dermally. In alternative embodiments, pharmaceutical compositions can be prepared in various forms, such as granules, tablets, pills, capsules, suspensions, taken orally, suppositories and salves, lotions and the like. Pharmaceutical formulations as provided herein may comprise one or more diluents, emulsifiers, preservatives, buffers, excipients, etc. and may be provided in such forms as liquids, powders, emulsions, lyophilized powders, sprays, creams, lotions, controlled release formulations, tablets, pills, lozenges, gels, geltabs, on patches, in implants, etc. In practicing embodiments as provided herein, the pharmaceutical compounds can be delivered by transdermally, by atopical route, formulated as applicator sticks, solutions, suspensions, emulsions, gels, creams, ointments, pastes, jellies, paints, powders, and aerosols. Oral carriers can be elixirs, syrups, capsules, tablets, pills, geltabs and the like.

[0204] In alternative embodiments, provided are pharmaceutically acceptable salts of compounds as provided herein or as used in methods as provided herein, including pharmaceutically acceptable non-toxic bases or acids including inorganic or organic bases and inorganic or organic acids. In alternative embodiments, salts are derived from inorganic bases such as aluminum, ammonium, calcium, copper, ferric, ferrous, lithium, magnesium, manganic salts, manganese, potassium, sodium, zinc, and the like; or, salts can be in a solid form, or in a crystal structure, or the form of hydrates. In alternative embodiments, salts are pharmaceutically acceptable organic non-toxic bases including salts of primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins, such as arginine, betaine, caffeine, choline, N,N'- dibenzylethylenediamine. diethylamine, 2-diethylaminoethanol, 2-dimethyl aminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydrabamine, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purines, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, and the like. In alternative embodiments, e.g., if a compound provided herein is basic, salts are prepared from pharmaceutically acceptable non-toxic acids, including inorganic and organic acids. Such acids include acetic, benzenesulfonic, benzoic, camphorsulfonic, carbonic, citric, ethanesulfonic, fumaric, gluconic, glutamic, hydrobromic, hydrochloric, isethionic, lactic, maleic, malic, mandelic, methanesulfonic, mucic, nitric, pamoic, pantothenic, phosphoric, succinic, sulfuric, tartaric, p-toluenesulfonic acid, and the like.

[0205] In alternative embodiments, pharmaceutically acceptable salts include hemisalts of non-toxic acids or bases, or hemihydrates.

[0206] In alternative embodiments, compounds and compositions as provided herein, or a compound used to practice methods as provided herein, are delivered orally, e.g., as pharmaceutical formulations for oral administration, and can be formulated using pharmaceutically acceptable carriers well known in the art in appropriate and suitable dosages. Such carriers enable the pharmaceuticals to be formulated in unit dosage forms as tablets, pills, powder, dragees, capsules, liquids, lozenges, gels, syrups, slurries, suspensions, etc., suitable for ingestion by the patient. Pharmaceutical preparations for oral use can be formulated as a solid excipient, optionally grinding a resulting mixture, and processing the mixture of granules, after adding suitable additional compounds, if desired, to obtain tablets or dragee cores. Suitable solid excipients can be carbohydrate or protein fillers, e.g., sugars, including lactose, sucrose, mannitol, or sorbitol; starch from com, wheat, rice, potato, or other plants: cellulose such as methyl cellulose, hydroxypropylmethyl-cellulose. or sodium carboxymethyl cellulose; and gums including arabic and tragacanth; and proteins, e.g., gelatin and collagen. Disintegrating or solubilizing agents may be added, such as the cross-linked polyvinyl pyrrolidone, agar, alginic acid, or a salt thereof, such as sodium alginate.

[0207] In alternative embodiments, liquid earners are used to manufacture or formulate compounds as provided herein, or a composition used to practice the methods as provided herein, including carriers for preparing solutions, suspensions, emulsions, syrups, elixirs and pressurized compounds. The active ingredient (e g., a composition as provided herein or as used in methods as provided herein) can be dissolved or suspended in a pharmaceutically acceptable liquid carrier such as water, an organic solvent, a mixture of both or pharmaceutically acceptable oils or fats. The liquid carrier can comprise other suitable pharmaceutical additives such as solubilizers, emulsifiers, buffers, preservatives, sweeteners, flavoring agents, suspending agents, thickening agents, colors, viscosity' regulators, stabilizers or osmoregulators.

[0208] In alternative embodiments, solid carriers are used to manufacture or formulate compounds as provided herein, or a composition used to practice the methods as provided herein, including solid carriers comprising substances such as lactose, starch, glucose, methyl-cellulose, magnesium stearate, dicalcium phosphate, mannitol and the like. A solid earner can further include one or more substances acting as flavoring agents, lubricants, solubilizers, suspending agents, fillers, glidants, compression aids, binders or tablet-disintegrating agents; it can also be an encapsulating material. In powders, the carrier can be a finely divided solid which is in admixture with the finely divided active compound. In tablets, the active compound is mixed with a carrier having the necessary compression properties in suitable proportions and compacted in the shape and size desired. Suitable solid carriers include, for example, calcium phosphate, magnesium stearate, talc, sugars, lactose, dextrin, starch, gelatin, cellulose, poly vinylpyrrolidine, low melting waxes and ion exchange resins. A tablet may be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared by compressing in a suitable machine the active ingredient in a free flowing form such as a powder or granules, optionally mixed with a binder (e g., povidone, gelatin, hydroxypropylmethyl cellulose), lubricant, inert diluent, preservative, disintegrant (e.g., sodium starch glycolate, cross-linked povidone, cross-linked sodium carboxymethyl cellulose) surface active or dispersing agent. Molded tablets may be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent. The tablets may optionally be coated or scored and may be formulated so as to provide slow or controlled release of the active ingredient therein using, for example, hydroxypropyl methylcellulose in vary ing proportions to provide the desired release profile. Tablets may optionally be provided with an enteric coating, to provide release in parts of the gut other than the stomach.

[0209] In alternative embodiments, concentrations of therapeutically active compound in a formulation can be from between about 0.1% to about 100% by weight. In alternative embodiments, therapeutic formulations are prepared by any method well known in the art, e.g., as described by Brunton et al., eds., Goodman and Gilman's: The Pharmacological Bases of Therapeutics , 12th ed., McGraw-Hill, 2011; Remington: The Science and Practice of Pharmacy, Mack Publishing Co., 20th ed., 2000; Avis et al., eds., Pharmaceutical Dosage Forms: Parenteral Medications, published by Marcel Dekker, Inc., N.Y., 1993; Lieberman et al., eds., Pharmaceutical Dosage Forms: Tablets, published by Marcel Dekker, Inc., N.Y., 1990; and Lieberman et al., eds., Pharmaceutical Dosage Forms: Disperse Systems, published by Marcel Dekker, Inc.. N.Y.. 1990.

[0210] In alternative embodiments, therapeutic formulations are delivered by any effective means appropriated for a particular treatment. For example, depending on the specific antitumor agent to be administered, the suitable means include oral, rectal, topical, by inhalation, vaginal, nasal, pulmonary administration, or parenteral (including subcutaneous, intramuscular, intravenous and intradermal) infusion into the bloodstream. For parenteral administration, antitumor agents as provided herein may be formulated in a variety of ways. Aqueous solutions of the modulators can be encapsulated in polymeric beads, liposomes, exosome, nanoparticles or other injectable depot formulations known to those of skill in the art. In alternative embodiments, compounds and compositions as provided herein, or a compound used to practice methods as provided herein, are administered encapsulated in liposomes (see below). In alternative embodiments, depending upon solubility , compositions are present both in an aqueous layer and in a lipidic layer, e.g., a liposomic suspension. In alternative embodiments, a hydrophobic layer comprises phospholipids such as lecithin and sphingomyelin, steroids such as cholesterol, more or less ionic surfactants such a diacetylphosphate, stearylamine, or phosphatidic acid, and / or other materials of a hydrophobic nature.

[0211] The pharmaceutical compositions can be formulated in any way and can be administered in a variety of unit dosage forms depending upon the condition or disease and the degree of illness, the general medical condition of each patient, the resulting preferred method of administration and the like. Details on techniques for formulation and administration are well described in the scientific and patent literature, see, e.g., the latest edition of Remington's Pharmaceutical Sciences, Maack Publishing Co., Easton PA (“Remington’s”). For example, in alternative embodiments, compounds and compositions as provided herein, or a compound used to practice methods as provided herein, are formulated in a buffer, in a saline solution, in a powder, an emulsion, in a vesicle, in a liposome, an exosome, in a nanoparticle, in a nanolipoparticle and the like. In alternative embodiments, the compositions can be formulated in any way and can be applied in a variety of concentrations and forms depending on the desired in vivo, in vitro or ex vivo conditions, a desired in vivo, in vitro or ex vivo method of administration and the like. Details on techniques for in vivo, in vitro or ex vivo formulations and administrations are well described in the scientific and patent literature. Formulations and / or carriers used to practice embodiments as provided herein can be in forms such as tablets, pills, powders, capsules, liquids, gels, syrups, slurries, suspensions, etc., suitable for in vivo, in vitro or ex vivo applications.

[0212] In practicing embodiments as provided herein, the compounds (e.g., formulations) as provided herein or as used in methods as provided herein can comprise a solution of compositions disposed in or dissolved in a pharmaceutically acceptable carrier, e.g., acceptable vehicles and solvents that can be employed include water and Ringer's solution, an isotonic sodium chloride. In addition, sterile fixed oils can be employed as a solvent or suspending medium. For this purpose any fixed oil can be employed including synthetic mono- or diglycerides, or fatty acids such as oleic acid. In one embodiment, solutions and formulations used to practice embodiments as provided herein are sterile and can be manufactured to be generally free of undesirable matter. In one embodiment, these solutions and formulations are sterilized by conventional, well known sterilization techniques.

[0213] The solutions and formulations used to practice methods as provided herein can comprise auxiliary substances as required to approximate physiological conditions such as pH adjusting and buffering agents, toxicity adjusting agents, e.g., sodium acetate, sodium chloride, potassium chloride, calcium chloride, sodium lactate and the like. The concentration of active agent in these formulations can vary widely, and can be selected primarily based on fluid volumes, viscosities and the like, in accordance with the particular mode of in vivo, in vitro or ex vivo administration selected and the desired results.

[0214] The compounds and compositions as provided herein, or a compound used to practice methods as provided herein, can be delivered by the use of a liposome, an exosome, a nanoparticle, or a nanoliposome. In alternative embodiments, by using a liposome, an exosome, a nanoparticle, or a nanoliposome, particularly where the a liposome, an exosome, a nanoparticle, or a nanoliposome surface carries moieties and / or ligands specific for targeting (binding specifically to) a particular cell, tissue or organs, or are otherwise fabricated to be directed to a specific cell, tissue or organ type (for example, directed to preferentially bind to, or target, a heart, a coronary artery or heart myocytes) the practitioner can focus the delivery of the active agent (a drug or composition capable of suppressing hypoxia-induced SUMOylation of a cardiac sodium channel NaV1.5) into a target cell, tissue and / or organ (for example, a heart, a coronary artery or heart myocytes) in an in vivo, in vitro or ex vivo application.

[0215] The compounds and compositions as provided herein, or a compound used to practice methods as provided herein, can be directly administered, e.g.. under sterile conditions, to an individual in need thereof (e.g., a patient) to be treated. The modulators can be administered alone or as the active ingredient of a pharmaceutical composition. Compositions and formulations as provided herein can be combined with or used in association with other therapeutic agents. For example, an individual may be treated concurrently with conventional therapeutic agents.

[0216] Nanoparticles. Nanolipoparticles and Liposomes

[0217] Provided are liposome, particles, nanoparticles, nanolipoparticles, exosomes, vesicles and liposomal membranes comprising compounds and compositions used to practice the methods and embodiments as provided herein. Provided are multilayered particles, nanoparticles, nanolipoparticles, exosomes, vesicles and liposomal membranes comprising compounds used to practice embodiments as provided herein, e.g., as described in Park, et al., U.S. Pat. Pub. No. 20070082042. The multilayered particles, nanoparticles, nanolipoparticles, exosomes, vesicles and liposomal membranes can be prepared using a mixture of oil-phase components comprising squalane, sterols, ceramides, neutral lipids or oils, fatty acids and lecithins, to about 200 to 5000 nm in particle size, to entrap a composition used to practice embodiments as provided herein.

[0218] Liposomes, particles, nanoparticles, nanolipoparticles. exosomes, vesicles and liposomal membranes can be made using any method, e.g., as described in Park, et al., U.S. Pat. Pub. No. 20070042031, including the method of producing a liposome by encapsulating an active agent (e.g., compounds and compositions as provided herein, or a compound used to practice methods as provided herein), the method comprising providing an aqueous solution in a first reservoir; providing an organic lipid solution in a second reservoir, and then mixing the aqueous solution with the organic lipid solution in a first mixing region to produce a liposome solution, where the organic lipid solution mixes with the aqueous solution to substantially instantaneously produce a liposome encapsulating the active agent; and immediately then mixing the liposome solution with a buffer solution to produce a diluted liposome solution.

[0219] In one embodiment, liposome, particle, nanoparticle, nanolipoparticle, exosome, vesicle and liposomal membrane compositions used to practice embodiments as provided herein comprise a substituted ammonium and / or polyanions, e.g., for targeting delivery of a compound as provided herein, or a compound used to practice methods as provided herein, to a desired cell type or organ, e.g., brain, as described e.g., in U.S. Pat. Pub. No. 20070110798.

[0220] Provided are particles, nanoparticles, nanolipoparticles, exosomes, vesicles and liposomal membranes comprising compounds as provided herein, e.g., used to practice methods as provided herein in the form of active agent-containing particles, nanoparticles, nanolipoparticles, exosomes, vesicles and liposomal membranes (e.g., a secondary nanoparticle), as described, e.g., in U.S. Pat. Pub. No. 20070077286. In one embodiment, provided are particles, nanoparticles, nanolipoparticles, exosomes, vesicles and liposomal membranes comprising a fat-soluble active agent used to practice embodiments as provided herein, or a fat-solubilized water-soluble active agent to act with a bivalent or trivalent metal salt.

[0221] In one embodiment, solid lipid suspensions can be used to formulate and to deliver compositions used to practice embodiments as provided herein to mammalian cells in vivo, in vitro or ex vivo, as described, e.g.. in U.S. Pat. Pub. No. 20050136121.

[0222] Delivery vehicles

[0223] In alternative embodiments, any delivery vehicle can be used to practice the methods as provided herein, e.g., to deliver compounds and compositions as provided herein, or a compound used to practice methods as provided herein, to mammalian cells, e.g., in vivo, in vitro or ex vivo. For example, delivery' vehicles comprising poly cations, cationic polymers and / or cationic peptides, such as polyethyleneimine derivatives, can be used e.g. as described, e.g., in U.S. Pat. Pub. No. 20060083737. In one embodiment, a dried polypeptide-surfactant complex is used to formulate compounds and compositions as provided herein, or a compound used to practice embodiments as provided herein, e.g. as described, e.g., in U.S. Pat. Pub. No. 20040151766.

[0224] In one embodiment, compounds and compositions as provided herein, or a compound used to practice methods as provided herein, can be applied to cells using vehicles with cell membrane-permeant peptide conjugates, e.g., as described in U.S. Patent Nos. 7,306,783; 6,589,503. In one aspect, the composition to be delivered is conjugated to a cell membrane-permeant peptide. In one embodiment, the composition to be delivered and / or the delivery vehicle are conjugated to a transport- mediating peptide, e.g., as described in U.S. Patent No. 5,846,743, describing transport-mediating peptides that are highly basic and bind to poly-phosphoinositides.

[0225] In one embodiment, electro-permeabilization is used as a primary or adjunctive means to deliver the composition to a cell, e.g., using any electroporation system as described e.g. in U.S. Patent Nos. 7,109,034; 6,261,815; 5,874,268.

[0226] In alternative embodiments, provided are drugs (for example, a drug or composition capable of suppressing hypoxia-induced SUMOylation of a cardiac sodium channel NaV1.5) incorporated into a stent used to open coronary arteries after a myocardial infarction (MI), and the drug or drugs elute at the highest levels right into the tissue that has been damaged in the post-MI period.

[0227] Dosaging

[0228] The pharmaceutical compositions and formulations as provided herein or as used in methods as provided herein can be administered for prophylactic and / or therapeutic treatments. In therapeutic applications, compositions are administered to a subject, e.g., a human in need thereof, in an amount of the agent sufficient to cure, alleviate or partially arrest the clinical manifestations and / or its complications (a“therapeutically effective amount”), for example, of angina, myocardial infarction, reperfusion injury, ventricular arrhythmias (VA), pump failure due to myocardial infarction (MI) in an early ischemia / reperfusion (I / R) period, and / or congestive heart failure (CHF), hypovolemia, acidosis, hyperkalemia, hypokalemia, cardiopathology due to a toxin, cardiac tamponade, tension pneumothorax, coronary thrombosis, sepsis, shock, pulmonary thrombosis, pulmonary embolism, respiratory decompensation, arrythmia and other causes of cardiac hypoperfusions and cardiac arrest..

[0229] The amount of pharmaceutical composition adequate to accomplish this is defined as a "therapeutically effective dose." The dosage schedule and amounts effective for this use, i.e., the “dosing regimen,” will depend upon a variety of factors, including the stage of the disease or condition, the severity of the disease or condition, the general state of the patient's health, the patient’s phy sical status, age and the like. Dosage levels may range from about 0.01 mg per kilogram to about 100 mg per kilogram of body weight. In calculating the dosage regimen for a patient, the mode of administration also is taken into consideration.

[0230] The dosage regimen also takes into consideration pharmacokinetics parameters well known in the art, i.e., the active agents' rate of absorption, bioavailability, metabolism, clearance, and the like (see, e.g., Hidalgo-Aragones (1996) J. Steroid Biochem. Mol. Biol. 58:611-617; Groning (1996) Pharmazie 51:337-341; Fotherby (1996) Contraception 54:59-69; Johnson (1995) J. Pharm. Sci. 84:1144-1146; Rohatagi (1995) Pharmazie 50:610-613; Brophy (1983) Eur. J. Clin. Pharmacol. 24: 103-108; the latest Remington’s, supra). The state of the art allows the clinician to determine the dosage regimen for each individual patient, active agent and disease or condition treated. Guidelines provided for similar compositions used as pharmaceuticals can be used as guidance to determine the dosage regiment, i.e., dose schedule and dosage levels, administered practicing the methods as provided herein are correct and appropriate.

[0231] Chemical synthesis

[0232] In alternative embodiment, methods as provided herein comprise administration of a drug or composition capable of suppressing hypoxia-induced SUMOylation of a cardiac sodium channel NaV1.5, which can comprise:

[0233] (a) a compound having the formula: (also called TAK-981), or

[0234] (b) a compound having the formula. (also called 2-D08, or 2’,3’,4'-trihydroxyflavone) or an isomer, deuterated isoform, optical isomer or stereoisomer, a racemate or racemic mixture, an enantiomer, an individual diastereomer or a diastereomeric mixture, an analog, a crystalline product or a crystalline intermediate, a pharmaceutically acceptable salt thereof, a prodrug or a bioisostere of TAK-981 or 2- D08, or 2’, 3 ’,4’ -trihydroxyflavone.

[0235] Any chemical synthesis protocol known in the art can be used to synthesize these compounds, for example, for TAK-981 as described by Langston, et al, J. Med. Chem. 2021, 64, 2501-2520; or. for 2’,3’,4'-trihydroxyflavone, as described by Kirn et al, Bioorg Med Chem Lett. 2014 Feb 15; 24(4): 1094-1097.

[0236] In alternative embodiment, the invention also provides bioisosteres of TAK- 981 or 2-D08. Bioisosteres of the invention are compounds comprising one or more substituent and / or group replacements with a substituent and / or group having substantially similar physical or chemical properties which produce substantially similar biological properties to TAK-981 or 2-D08, or racemer or isomer thereof. In one embodiment, the purpose of exchanging one bioisostere for another is to enhance the desired biological or phy sical properties of a compound without making significant changes in chemical structures.

[0237] For example, in one embodiment, one or more hydrogen atom(s) is replaced with one or more fluorine and / or deuterium atom(s), e.g., at a site of metabolic oxidation; this may prevent metabolism (catabolism) from taking place. Because the fluorine or deuterium atom is similar in size to the hydrogen atom the overall topology of the molecule is not significantly affected, leaving the desired biological activity unaffected. However, with a blocked pathway for metabolism, the TAK-981 or 2-D08 molecule may have a longer half-life or be less toxic, and the like.

[0238] In alternative embodiments, compounds used in methods as provided herein, TAK-981 or 2-D08, or a pharmaceutically acceptable salt or solvate thereof, or optical isomer thereof, or racemic mixture or enantiomer thereof; or, a pharmaceutical composition or formulation comprising TAK-981 or 2-D08, include or comprise their respective bioisosteres. In alternative embodiments, bioisosteres used to practice methods as provided herein comprise one or more substituent and / or group replacements with a substituent and / or group having substantially similar physical or chemical properties which produce substantially similar biological properties to a compound, or stereoisomer, racemer or isomer thereof. In one embodiment, the purpose of exchanging one bioisostere for another is to enhance the desired biological or physical properties of a compound without making significant changes in chemical structures.

[0239] Gene Therapy and Gene Delivery Vehicles

[0240] In alternative embodiments, provided are methods for treating, ameliorating, lessening the symptoms of, or preventing: angina, myocardial infarction, reperfusion injury, a ventricular arrhythmia (VA), cardiac pump failure due to myocardial infarction (MI) (optionally cardiac pump failure due to myocardial infarction (MI) in an early ischemia / reperfusion (I / R) period), congestive heart failure (CHF), hypovolemia, acidosis, hyperkalemia, hypokalemia, a cardiac poisoning or cardiopathology due to a toxin or poisoning, cardiac tamponade, tension pneumothorax, coronary thrombosis, sepsis, shock, pulmonary thrombosis, pulmonary embolism, respiratory decompensation, arrythmia a cardiac event caused by hypoperfusion and / or cardiac arrest, comprising administering to an individual in need thereof a nucleic acid capable of modifying a NaV1.5 K442 residue to another residue that cannot be SUMOylated.

[0241] In alternative embodiments, the nucleic acid capable of modifying a NaV1.5 K.442 residue to another residue that cannot be SUMOylated is delivered in vivo using a gene therapy delivery vehicle.

[0242] In alternative embodiments, expression vehicle, vector, recombinant virus, or equivalents used to practice methods provided herein are or comprise: an adeno- associated virus (AAV), a lentiviral vector or an adenovirus vector; an AAV serotype AAV5, AAV6, AAV8 or AAV9; a rhesus-derived AAV, or the rhesus-derived AAV AAVrh.10hCLN2; an organ-tropic AAV, or a neurotropic AAV; and / or an AAV capsid mutant or AAV hybrid serotype. In alternative embodiments, the AAV is engineered to increase efficiency in targeting a specific cell type that is non- permissive to a wild type (wt) AAV and / or to improve efficacy in infecting only a cell type of interest. In alternative embodiments, the hybrid AAV is retargeted or engineered as a hybrid serotype by one or more modifications comprising: 1) a transcapsidation, 2) adsorption of a bi-specific antibody to a capsid surface, 3) engineering a mosaic capsid, and / or 4) engineering a chimeric capsid. It is well known in the art how to engineer an adeno-associated virus (AAV) capsid in order to increase efficiency in targeting specific cell types that are non-permissive to wild type (wt) viruses and to improve efficacy in infecting only the cell type of interest; see for example, Wu et al., Mol. Ther. 2006 Sep;14(3):316-27. Epub 2006 Jul 7; Choi, et al., Curr. Gene Ther. 2005 Jun;5(3):299-310.

[0243] For example, in alternative embodiments, serotypes AAV-8, AAV-9, AAV-DJ or AAV-DJ / 8™ (Cell Biolabs, Inc., San Diego, CA), which have increased uptake in brain tissue in vivo, are used to deliver a nucleic acid payload for expression in the CNS. In alternative embodiments, the following serotypes, or variants thereof, are used for targeting a specific tissue:

[0244] In alternative embodiments, the rhesus-derived AAV AAVrh.10hCLN2 or equivalents thereof can be used, wherein the rhesus-derived AAV may not be inhibited by any pre-existing immunity in a human; see for example, Sondhi, et al., Hum Gene Ther. Methods. 2012 Oct;23(5): 324-35, Epub 2012 Nov 6; Sondhi, et al., Hum Gene Ther. Methods. 2012 Oct 17: teaching that direct administration of AAVrh.10hCLN2 to the CNS of rats and non-human primates at doses scalable to humans has an acceptable safety profile and mediates significant payload expression in the CNS.

[0245] Because adeno-associated viruses (AAVs) are common infective agents of primates, and as such, healthy primates carry a large pool of AAV-specific neutralizing antibodies (NAbs) which inhibit AAV-mediated gene transfer therapeutic strategies, methods provided herein can comprise screening of patient candidates for AAV-specific NAbs prior to treatment, especially with the frequently used AAV8 capsid component, to facilitate individualized treatment design and enhance therapeutic efficacy; see, for example, Sun, et al., J. Immunol. Methods. 2013 Jan 31;387(1-2): 114-20, Epub 2012 Oct 11.

[0246] In alternative embodiments, the NaVl 5 K442 residue modifying nucleic acid is delivered in vivo using methods as provided herein can be in the form of, or comprise, an RNA, for example, or miRNA or an mRNA, which can be formulated in a lipid formulation or a liposome (or in particles, nanoparticles, nanolipoparticles, exosomes, vesicles or liposomal membranes) and injected for example intramuscularly (IM), for example using formulations and methods as described in U.S. patent application no. US 20210046173 Al, which describes delivering to a subject (for example, via intramuscular administration) the NaVl .5 K442 residue modifying nucleic acid that comprises a RNA (for example. miRNA or mRNA) that comprises an open reading frame (ORF) that comprises (or consists of, or consists essentially of) or encodes for the protein; wherein optionally the RNA (or the DNA- carrying expression vehicle) is formulated in a liposome, or a lipid nanoparticle (LNP), or nanoliposome, that comprises: non-cationic lipids comprise a mixture of cholesterol and l,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), or aPEG-lipid. or PEG-modified lipid, or LNP, or an ionizable cationic lipid; or a mixture of (13Z,16Z)-N,N-dimethyl-2-nonylhenicosa-l 2, 15-dien-l -amine, cholesterol, DSPC, and PEG-2000 DMG. In alternative embodiments, the PEG-lipid is 1,2-Dimyristoyl- sn-glycerol methoxypolyethylene glycol (PEG-DMG), PEG-disteryl glycerol (PEG- DSG), PEG-dipalmetoleyl, PEG-dioleyl, PEG-distearyl, PEG-diacylglycamide (PEGDAG), PEG-dipalmitoyl phosphatidylethanolamine (PEG-DPPE), or PEG- 1,2- dimyristyloxlpropyl-3-amine (PEG-c-DMA), or, the PEG-lipid is PEG coupled to dimyristoylglycerol (PEG-DMG). In alternative embodiments, the LNP comprises 20-99.8 mole % ionizable cationic lipids, 0.1-65 mole % non-cationic lipids, and 0.1- 20 mole % PEG-lipid. In alternative embodiments, the LNP comprises an ionizable cationic lipid selected from the group consisting of (2S)-1-({6-[(3))-cholest-5-en-3- yloxy]hexyl}oxy)-N,N-dimethyl-3-[(9 Z)-octadec-9-en-1-yloxy]propan-2-amine; ( 13Z.16Z)-N,N-dimethy 1-3 -nonyldocosa- 13 , 16-dien- 1 -amine; and N,N -dimethyl- 1 - [(lS,2R)-2-octylcyclopropyl]heptadecan-8-amine; or a pharmaceutically acceptable salt thereof, or a stereoisomer of any of the foregoing. In alternative embodiments, the PEG modified lipid comprises a PEG-modified phosphatidylethanolamine, a PEG- modified phosphatidic acid, a PEG-modified ceramide, a PEG-modified dialkylamine, a PEG-modified diacylglycerol, a PEG-modified dialkylglycerol, and mixtures thereof. In alternative embodiments, the ionizable cationic lipid comprises: 2,2- dilinoleyl-4-dimethylaminoethyl-[l,3]-dioxolane (DLin-KC2-DMA). dilinoleyl- methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), di((Z)-non-2-en-l-yl) 9-((4- (dimethylamino)butanoyl)oxy) heptadecanedi oate (L319), (13Z,16Z)-N,N-dimethyl- 3-nonyldocosa-13,16-dien-1-amine, (12Z,15Z)-N,N-dimethyl-2-nonylhenicosa-12,15- dien-1-amine, and N,N-dimethyl-1-[(lS,2R)-2-octylcyclopropyl]heptadecan-8-amine. In one embodiment, the lipid is (13Z,16Z)-N,N-dimethyl-3-nonyldocosa-13,16-dien- 1 -amine or N,N-dimethyl-1-[(lS,2R)-2-octylcyclopropyl]heptadecan-8-amine, each of which are described in PCT / US2011 / 052328, the entire contents of which are hereby incorporated by reference. In some embodiments, a non-cationic lipid of the disclosure comprises l,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1.2- dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 1 ,2-dilinoleoyl-sn-glycero-3- phosphocholine (DLPC), 1 ,2-dimyristoyl-sn-gly cero-phosphocholine (DMPC), 1 ,2- dioleoyl-sn-glycero-3-phosphocholine (DOPC), l,2-dipalmitoyl-sn-glycero-3- phosphocholine (DPPC). 1,2-di undecanoy 1-sn-gly cero-phosphocholine (DUPC), 1- palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1,2-di-O-octadecenyl-sn- glycero-3-phosphocholine (18:0 Diether PC), 1-oleoyl-2 cholesterylhemisuccinoyl-sn- glycero-3-phosphocholine (OChemsPC), 1 -hexadecy 1-sn-gly cero-3-phosphocholine (C16 Lyso PC), 1,2-dilinolenoyl-sn-glycero-3-phosphocholine, 1.2-diarachidonoyl- sn-glycero-3-phosphocholine, 1,2-didocosahexaenoyl-sn-glycero-3-phosphocholine,

[0247] 1,2-diphytanoyl-sn-glycero-3-phosphoethanolamine (ME 16.0 PE), 1,2-distearoyl-sn- glycero-3-phosphoethanolamine, 1,2-dilinoleoyl-sn-glycero-3-phosphoethanolamine,

[0248] 1,2-dilinolenoyl-sn-glycero-3 -phosphoethanolamine, 1,2-diarachidonoy 1-sn-gly cero- 3 -phosphoethanolamine. 1,2-didocosahexaenoyl-sn-glycero-3-phosphoethanolamine,

[0249] 1,2-dioleoyl-sn-glycero-3-phospho-rac-(1-glycerol) sodium salt (DOPG), sphingomyelin, or mixtures thereof.

[0250] CRISPR and Prime Editing gene therapy

[0251] In alternative embodiments, NaV1.5 K442 residue modifying nucleic acids are delivered in vivo or ex vivo or nucleic acids are altered in vivo or ex vivo using a CRISPR system such as CRISPR-Cas9 or any variants of CRISPR-Cas9. and / or a prime editing system (for example, as described in Chen, P.J., Liu, D R. Prime editing for precise and highly versatile genome manipulation. Nat Rev Genet 24, 161-177 (2023)). Similar to CRISPR, prime editing requires the presence of a Cas endonuclease and a single guide (sg) RNA. However, as the premise of prime editing is to edit sequences without generating a double-stranded break, both components are slightly modified. Instead of traditional Cas9, this method utilizes Cas9 nickase — a variant of Cas9 that nicks the DNA rather than generating double-strand breaks — fused to a reverse transcriptase. This Cas9 fusion is referred to as a prime editor (PE).

[0252] In alternative embodiments, prime editing comprises use of materials and processes as described, for example, in U.S. patent application publication nos. US 2023 / 0220374 Al; US2023 / 0357766 Al; and / or U.S patentnos. US 11,840,685 B2, US 11 ,795,452 B2; and / or as described in WO / 2021 / 072328 Al .

[0253] In alternative embodiments, prime editing comprises use of the following components: a prime editing guide RNA (pegRNA), capable of (i) identifying the target nucleotide sequence to be edited, and (ii) encoding new genetic information that replaces the targeted sequence. The pegRNA consists of an extended single guide RNA (sgRNA) containing a primer binding site (PBS) and a reverse transcriptase (RT) template sequence. During genome editing, the primer binding site allows the 3’ end of the nicked DNA strand to hybridize to the pegRNA, while the RT template serves as a template for the synthesis of edited genetic information;

[0254] - a fusion protein consisting of a Cas9 H840A nickase fused to a Moloney Murine Eeukemia Virus (M-MEV) reverse transcriptase;

[0255] Cas9 H840A nickase: the Cas9 enzyme contains two nuclease domains that can cleave DNA sequences, a RuvC domain that cleaves the non-target strand and a HNH domain that cleaves the target strand. The introduction of a H840A substitution in Cas9, through which the 840th amino acid histidine is replaced by an alanine, inactivates the HNH domain. With only the RuvC functioning domain, the catalytically impaired Cas9 introduces a single strand nick, hence the name nickase;

[0256] M-MLV reverse transcriptase: an enzyme that synthesizes DNA from a single-stranded RNA template; and a single guide RNA (sgRNA) that directs the Cas9 H840A nickase portion of the fusion protein to nick the non-edited DNA strand.

[0257] In alternative embodiments, genetic engineering ofNaV1.5 by NaVl.5 residue-modifying nucleic acids is performed in conjunction with administration of a drug or composition capable of suppressing hypoxia-induced SUMOylation of a cardiac sodium channel NaV1.5, such as a drug or composition capable of suppressing hypoxia-induced SUMOylation of a cardiac sodium channel NaV1.5 as provided herein.

[0258] In alternative embodiment the drug or composition capable of suppressing hypoxia-induced SUMOylation of a cardiac sodium channel NaV1.5 is administered first, followed by genetic engineering of NaV1.5 by NaV1.5 residue-modifying nucleic acids (for example, using CRISPR).

[0259] In alternative embodiment the drug or composition capable of suppressing hypoxia-induced SUMOylation of a cardiac sodium channel NaV 1.5 is administered together (or, substantially at the same time, or substantially simultaneously) with genetic engineering of NaV1.5 by NaV1.5 residue-modifying nucleic acids (for example, using CRISPR).

[0260] In alternative embodiment the drug or composition capable of suppressing hypoxia-induced SUMOylation of a cardiac sodium channel NaV1.5 is administered after genetic engineering of NaV1.5 by NaV1.5 residue-modify ing nucleic acids (for example, using CRISPR).

[0261] In alternative embodiments, NaV1.5 K442 residue modify ing exogenous nucleic acids, Cas9, sgRNA, and associated complexes are delivered into cells such as T cells, and any vehicle for delivery can be used, for example, viral and / or non-viral systems can be used; also, electroporation of DNA, RNA, or ribonucleocomplexes can be sued; or chemical transfection techniques utilizing lipids and peptides (particularly to introduce sgRNAs can be used, optionally also in complex with Cas9 into cells); nanoparticle-based delivery for transfection, and the like can be used.

[0262] In alternative embodiments, lenti virus (LVs), adenovirus (AdV), and adeno- associated virus (AAV) are used to deliver NaV 1.5 residue-modifying nucleic acids; some categories of cells are more difficult to transfect, including stem cells, neurons, and hematopoietic cells, and these require more efficient delivery systems, such as those based on lentivirus (LVs), adenovirus (AdV), and adeno-associated virus (AAV).

[0263] In alternative embodiments, variants of CRISPR-Cas9 are used to allow gene activation or genome editing with an external trigger such as light or small molecules: including photoactivatable CRISPR systems developed by fusing light-responsive protein partners with an activator domain and a dCas9 for gene activation, or by fusing similar light-responsive domains with two constructs of split-Cas9, or by incorporating caged unnatural amino acids into Cas9, or by modifying the guide RNAs with photocleavable complements for genome editing.

[0264] In alternative embodiments, "dead" versions of Cas9 (dCas9) are used to eliminate CRISPR's DNA-cutting ability while preserving its ability to target desirable sequences. Various regulatory factors can be added to dCas9s, enabling turning any gene on or off or adjust its level of activity. Like RNAi, CRISPR interference (CRISPRi) can turn off genes in a reversible fashion by targeting, but not cutting a site. The targeted site is methylated, epigenetically modify ing the gene. This modification inhibits transcription. These precisely placed modifications may then be used to regulate the effects on gene expression (for example, HMGB2) and DNA dynamics after the inhibition of certain genome sequences within DNA.

[0265] In alternative embodiments, CRISPR-Casl3 fused to deaminases is used to direct mRNA editing; for example, Cas7-11, is better suited for therapeutic RNA editing than Casl3, and enables sufficiently targeted cuts.

[0266] In alternative embodiments, any CRISPR system can be used to practice methods as provided herein, for example, as described in US 20220387560 Al, which describes methods of treating and / or correcting ocular disease in vivo using an Adeno-associated virus (AAV) system, where the AAV system employs a nucleic acid encoding a CRISPR-Cas9 system for targeted gene disruption or correction; or US 20220389398 Al that describes using engineered CRISPR / Cas effector enzymes, such as Casl3 (Casl3d, Casl3e, or Casl3f) that maintain guide-sequence-specific endonuclease activity and lack guide-sequence-independent collateral endonuclease activity; or US 2023 0029506 Al, which describes therapeutic applications of the crispr-cas systems and compositions for genome editing; or, US 2020 0340012 Al, which describes a modular CRISPR-Cas9 architecture that allows better delivery, specificity and selectivity of gene editing; or US 8,771,945, which describes CRISPR- Cas systems and methods for altering expression of gene products; or WO 2023 283420 A2 which describes therapeutic gene silencing with crispr-casl3.

[0267] In alternative embodiments, design of a nucleic acid (for example, an inhibitory RNA, or an iRNA) capable of modifying aNaVl.5 K442 residue to another residue that cannot be SUMOylated (for example, using CRISPR) can be based on the nucleic acid sequence of the transcript of human NaV 1.5 (the codon encoding the K442 residue is underlined), SEQ ID NO:1:

[0268] In alternative embodiments, design of a nucleic acid capable of modifying a

[0269] NaV 1.5 K442 residue to another residue that cannot be SUMOylated (for example, using CRISPR) can be based on the amino acid sequence of human NaV1.5 protein (the K442 residue is underlined). SEQ ID NO:2:

[0270] SENPs. sentrin-specific proteases. SUMO desumoylases

[0271] In alternative embodiments, compositions used in methods as provided herein (including compositions capable of suppressing hypoxia-induced SUMOylation of a cardiac sodium channel NaV1.5) comprise SUMO proteases or SENP proteins: in alternative embodiments, SUMO proteases or their C-terminal catalytic domains, and / or nucleic acids encoding them, are administered to individuals in need thereof.

[0272] In alternative embodiments, any one or a mixture of six SUMO proteases in humans, designated SENP1-3 and SENP 5-7, and / or their isoforms, are administered. All SUMO proteases have a conserved C-terminal catalytic domain, and in alternative embodiments, either the full length SENP protein, or only the C-terminal catalytic domain.

[0273] In alternative embodiments, a tag such as an affinity tag such as His6, or an identification tag such as Flag or eGFP, is attached to the administered SUMO proteases or SENP proteins for purification or localization. In alternative embodiments, the tag can be removed from the protein before it is administered.

[0274] In alternative embodiments, a SENP1 or SENP 2 catalytic domain is administered. The administered catalytic domain can comprise amino acid (aa) residue from 419 to 644 in SENP1, and in SENP2 aa residues 336 to 584. In alternative embodiments, nucleic acids encoding catalytic domains are administered.

[0275] In alternative embodiments, any one or more of the following proteins, or nucleic acids encoding these proteins, are administered to individuals in need thereof to practice methods as provided herein:

[0276] SENP1, sentrin-specific protease 1 Homo sapiens, NCBI Reference Sequence: NP_001254524.1, the SENP1 catalytic domain comprises amino acid from 419 to 644.

[0277] SENP2. sentrin-specific protease 2 Homo sapiens, NCBI Reference Sequence: NP_067640.2, Catalytic domain is residues 336 to 584

[0278] SENP3, sentrin-specific protease 3 Homo sapiens, NCBI Reference Sequence: NP 056485.2, The catalytic domain is residues 400 to 572

[0279] SENP5. isoform 1, sentrin-specific protease 5 isoform 1 Homo sapiens, NCBI Reference Sequence: NP_689912.2, The catalytic domain is residues 581 to 753

[0280] SENP5. isoform 2, sentrin-specific protease 5 isoform 2 Homo sapiens,

[0281] NCBI Reference Sequence: NP_001294974.1, The catalytic domain is residues 581 to 707

[0282] SENP6. sentrin-specific protease 6 isoform 1 Homo sapiens, 1112 aa protein, catalytic domain residues 973 to 1067, Accession: NP 056386.2

[0283] Sentrin-specific protease 6 isoform 2 Homo sapiens, 1105 aa protein, catalytic domain residues 966 to 1060, Accession: NP 001093879.1. GI: 156105703

[0284] Sentrin-specific protease 6 isoform 3 Homo sapiens, 685 aa protein, catalytic domain residues 966 to 1060, Accession: NP_001291721.1

[0285] SENP7. sentrin-specific protease 7 isoform 1 [Homo sapiensl

[0286] 1050 aa protein, Catalytic domain 774 to 1011, Accession: NP_065705.3

[0287] Sentrin-specific protease 7 isoform 2 Homo sapiens, 985 aa protein, Accession: NP_001070671.1 sentrin-specific protease 7 isoform 3 Homo sapiens

[0288] 984 aa protein, Accession: NP_001269730.1 sentrin-specific protease 7 isoform 4 Homo sapiens

[0289] 1017 aa protein, Accession: NP_001269731.1 sentrin-specific protease 7 isoform 6 Homo sapiens

[0290] 169 aa protein, Accession: NP_001269733.1 sentrin-specific protease 7 isoform 5 Homo sapiens

[0291] 886 aa protein, Accession: NP_001269732.1

[0292] Dosaging

[0293] In alternative embodiments, provided are pharmaceutical formulations or compositions comprising NaV1.5 K442 residue modifying small molecules, proteins and / or nucleic acids. In alternative embodiments, provided are compositions (for example, small molecules, proteins and / or nucleic acids), including products of manufacture and kits, and methods, for treating, ameliorating, lessening the symptoms of, or preventing: angina, myocardial infarction, reperfusion injury, a ventricular arrhythmia (VA), cardiac pump failure due to myocardial infarction (MI) (optionally cardiac pump failure due to myocardial infarction (MI) in an early ischemia / reperfusion (I / R) period), congestive heart failure (CHF), hypovolemia, acidosis, hyperkalemia, hypokalemia, a cardiac poisoning or cardiopathology due to a toxin or poisoning, cardiac tamponade, tension pneumothorax, coronary thrombosis, sepsis, shock, pulmonary thrombosis, pulmonary embolism, respiratory' decompensation, arrythmia a cardiac event caused by hypoperfusion and / or cardiac arrest, which can be used by administering to an individual in need thereof a drug or composition capable of suppressing hypoxia-induced SUMOylation of a cardiac sodium channel NaV1.5

[0294] The amount of pharmaceutical composition adequate to accomplish these defined as a "therapeutically effective dose." The dosage schedule and amounts effective for this use. i.e., the "dosing regimen." will depend upon a variety of factors, including the stage of the disease or condition, the severity of the disease or condition, the general state of the patient's health, the patient’s physical status, age and the like. In calculating the dosage regimen for a patient, the mode of administration also is taken into consideration.

[0295] In alternative embodiments, viral vectors such as adenovirus or AAV vectors are administered to an individual in need therein (for example, wherein the vectors contain therein nucleic acids encoding inhibitory nucleic acids as provided herein, or nucleic acids expressing SUMO proteases or SENP proteins as provided herein, and in alternative embodiment the dosage administered to a human comprises: a dose of about 2 x 1012vector genomes per kg body weight (vg / kg), or between about 1010and 1014vector genomes per kg body weight (vg / kg), or about 109, 1010, 1011, 1012, 1013, 1014, 1015, or more vg / kg, which can be administered as a single dosage or in multiple dosages, as needed. In alternative embodiments, these dosages are administered intravitreally, orally, IM, IV, or intrathecally. In alternative embodiments, the vectors are delivered as formulations or pharmaceutical preparations, for example, where the vectors are contained in a nanoparticle, a particle, a micelle or a liposome or lipoplex, a polymersome, a polyplex or a dendrimer.

[0296] In alternative embodiments, these dosages are administered once a day, once a week, or any variation thereof as needed to maintain in vivo expression levels of NaV1.5 K442 residue modifying nucleic acids, which can be monitored by measuring actually expression of NaV1.5 or by monitoring of therapeutic effect. The dosage regimen also takes into consideration pharmacokinetics parameters well known in the art, i.e.. the active agents’ rate of absorption, bioavailability, metabolism, clearance, and the like (see. for example. Hidalgo-Aragones (1996) J. Steroid Biochem. Mol. Biol. 58:611-617; Groning (1996) Pharmazie 51:337-341; Fotherby (1996) Contraception 54:59-69; Johnson (1995) J. Pharm. Sci. 84:1144-1146; Rohatagi (1995) Pharmazie 50:610-613; Brophy (1983) Eur. J. Clin. Pharmacol. 24:103-108; the latest Remington’s, supra). The state of the art allows the clinician to determine the dosage regimen for each individual patient, active agent and disease or condition treated. Guidelines provided for similar compositions used as pharmaceuticals can be used as guidance to determine the dosage regiment, i.e., dose schedule and dosage levels, administered practicing the methods as provided herein are correct and appropriate.

[0297] Single or multiple administrations of formulations can be given depending on the dosage and frequency as required and tolerated by the patient. The formulations should provide a sufficient quantity of active agent to effectively treat, prevent or ameliorate a conditions, diseases or symptoms as described herein. For example, alternative exemplary pharmaceutical formulations for oral administration of compositions used to practice methods as provided herein are in a daily amount of between about 0.1 to 0.5 to about 20, 50, 100 or 1000 or more Mg per kilogram of body weight per day. In an alternative embodiment, dosages are from about 1 mg to about 4 mg per kg of body weight per patient per day are used. Lower dosages can be used, in contrast to administration orally, into the blood stream, into a body cavity or into a lumen of an organ. Substantially higher dosages can be used in topical or oral administration or administering by powders, spray or inhalation. Actual methods for preparing parenterally or non-parenterally admimstrable formulations is known or apparent to those skilled in the art and are described in more detail in such publications as Remington's, supra. The methods as provided herein can further comprise co-administration with other drugs or pharmaceuticals, for example, compositions for treating any neurological or neuromuscular disease, condition, infection or injury, including related inflammatory and autoimmune diseases and conditions, and the like. For example, the methods and / or compositions and formulations as provided herein can be co-formulated with and / or co-administered with, fluids, antibiotics, cytokines, immunoregulatory agents, anti-inflammatory agents, pain alleviating compounds, complement activating agents, such as peptides or proteins comprising collagen-like domains or fibrinogen-like domains (for example, a ficolin), carbohydrate-binding domains, and the like and combinations thereof.

[0298] Compounds of US20170002032A9. WO2016004136

[0299] In alternative embodiments, compounds and structures used in methods as provided herein and compounds formulated as drugs used in methods as provided herein comprise compounds as set forth in U.S. patent application publication no. US20170002032A9, and described in W02016004136: wherein in alternative embodiments, compounds and structures comprise: a compound or pharmaceutically acceptable salt of formula (I): wherein stereochemical configurations depicted at asterisked positions indicate absolute stereochemistry;

[0300] Y is — O— , — CH2— , or — N(H)— ;

[0301] Rais hydrogen, fluoro, — NH2, or hydroxyl;

[0302] Rais hydrogen or fluoro, provided that when Rais — NH2or hydroxyl, Ra' is hydrogen;

[0303] Rbis hydrogen or, together with the oxygen to which it is attached, forms a prodrug; Rcis hydrogen or C1-4alkyl;

[0304] Rdis hydrogen, halogen, — CF3, or C1-4alkyl;

[0305] X1is C(H), C(F), orN;

[0306] X2is S or O;

[0307] X3is C(Rx3) or N;

[0308] Rx3is hydrogen, methyl, or halogen;

[0309] Z1is hydrogen, halogen, cyano, Rz3, — S — Rz3, — S(O) — Rz3, or — S(O)2— Rz3;

[0310] Rz3is an optionally substituted phenyl, an optionally substituted 5- to 7-membered cycloaliphatic, an optionally substituted 5- to 7-membered heterocyclyl, or an optionally substituted C1-4aliphatic; wherein Z1is not hydrogen, halogen, methyl, or cyano if Z2is hydrogen or methyl; and

[0311] (a) Z2is a ring system having an optionally substituted 5- to 7-membered heterocyclyl with 1-2 heteroatoms or an optionally substituted 5- to 7-membered cycloaliphatic fused to

[0312] (i) an optionally substituted 5-membered heteroaryl or an optionally substituted 6- membered aryl or heteroaryl to form a bicyclic group; or

[0313] (ii) an optionally substituted 9-membered heteroaryl or an optionally substituted 10- membered aryl or heteroaryl to form a tricyclic group; or

[0314] (b) Z2is L-Rewherein L is -L1-, — V1-L2-, or -L1-V1-L2-;

[0315] L1is a C1-3alkylene chain wherein 1 or 2 saturated carbon atoms are optionally substituted by (Rf)(Rf) and in which there are optionally one or two degrees of unsaturation; each Rfis independently hydrogen; hydroxyl; — N(Rh)(Rh); C1-4aliphatic optionally- substituted with hydroxyl, — OCH3, or cyclopropyl; — O — C1-4aliphatic optionally substituted with hydroxyl, — OCH3, or cyclopropyl; or, together with Rfand the carbon atom to which they are attached, form C=CH2, or a 3- to 6-membered carbocycle or 4- to 6-membered heterocycle comprising a heteroatom chosen fromN (which may be protonated or C1-4alkylated). O, or S, the heteroatom optionally located immediately adjacent to the quaternary carbon of the heterocycle; each Rfis independently hydrogen; C1-4aliphatic optionally substituted with hydroxyl, — OCH3, or cyclopropyl; — O — C1-4aliphatic optionally substituted with hydroxyl, — OCH3, or cyclopropyl; or, together with R1and the carbon atom to which they are attached, form C=CH2, or a 3- to 6-membered carbocycle or 4- to 6- membered heterocycle comprising a heteroatom chosen from N (which may be protonated or C1-4alkylated). O, or S, the heteroatom optionally located immediately adjacent to the quaternary carbon of the heterocycle; wherein if Rfis hydroxyl, Rfis not — O — C1-4aliphatic optionally substituted with hydroxyl, — OCH3, or cyclopropyl;

[0316] Rhand Rh’ are each independently hydrogen or C1-4alkyl;

[0317] Vi is — S— , — O— , — S(O)— , — S(O)2— , — C(O)— or — N(RS)— ;

[0318] L2is a C0-2 alkylene chain wherein one saturated carbon atom is optionally substituted by (Rf)(Rf);

[0319] R is hydrogen or C1-4alkyl; and either (i) Reis hydrogen, hydroxyl, halogen. — CF3, or an optionally substituted C1-4aliphatic, with the proviso that Reis not hydrogen if Rfand Rfare present and form a ring; or

[0320] (ii) Reis a ring chosen from optionally substituted 6-membered aryl, optionally substituted 5- to 6-membered heteroaryl, optionally substituted 3- to 7-membered cycloaliphatic, or optionally substituted 4- to 7-membered heterocyclyl, which is optionally fused to a second optionally substituted 6-membered aryl, optionally substituted 5- to 6-membered heteroaryl, optionally substituted 3- to 7-membered cycloaliphatic, or optionally substituted 4- to 7-membered heterocyclyl; or

[0321] (c) Z2is hydrogen.

[0322] In alternative embodiments;

[0323] (a) Z2is a ring system having a 5- to 7-membered heterocyclyl with 1-2 heteroatoms or a 5- to 7-membered cycloaliphatic fused to

[0324] (i) a 5-membered heteroaryl or a 6-membered aryl or heteroaryl to form a bicyclic group; or

[0325] (ii) a 9-membered heteroaryl or a 10-membered aryl or heteroaryl to form a tricyclic group; wherein the ring system is optionally substituted by 1-3 independent occurrences of halogen, hydroxyl, cyano, C1-4aliphatic, C1-4fluoroaliphatic, C1-4alkoxy, C1- 4 fluoroalkoxy, — S — C1-4aliphatic, — S — C1-4fluoroaliphatic, — N(Rz7)2, — C(O)Rz8, S(O)Rz8, S(O)2RZ8, C(O)2RZ7, C(O)N(RZ7)2, S(O)2N(RZ7)2,

[0326] OC(O)N(RZ7)2, — N(RZ7)C(O)RZ8, — N(RZ7)SO2RZ8, — N(RZ7)C(O)ORZ8, T2-RZ9, a 5- to 6-membered heteroaryl, a 6-membered aryl, a 3- to 6-membered cycloaliphatic, or a 4- to 6-membered heterocyclyl; and the ring system is optionally substituted at one saturated carbon with oxo, a spirocyclic 3- to 6-membered carbocycle, or a spirocyclic 4- to 6-membered heterocycle; each occurrence of Rz7is independently hydrogen or C1-4alkyl; each occurrence of Rz8is independently C1-4alkyl;

[0327] T2is a C1-2alkylene chain;

[0328] Rz 9is cyano, N(Rz7)2, ORz7, C(O)Rz8, C(O)2Rz7, or C(O)N(Rz7)2; or

[0329] (b) Z2is L-Rewherein either:

[0330] (i) Reis hydrogen, hydroxyl, halogen, — CF3, or C1-4aliphatic optionally substituted with one or more hydroxyl, halogen, or C1-4aliphatic, with the proviso that Reis not hydrogen if Rfand Rfare present and form a ring; or

[0331] (ii) Reis a ring chosen from 6-membered aryl, 5- to 6-membered heteroaryl, 3- to 7- membered cycloaliphatic, or 4- to 7-membered heterocyclyl, which is optionally fused to a second 6-membered aryl, 5- to 6-membered heteroaryl, 3- to 7-membered cycloaliphatic, or 4- to 7-membered heterocyclyl, Rebeing optionally substituted by 1-3 independent occurrences of halogen, hydroxyl, cyano, C1-4aliphatic, C1-

[0332] 4fluoroaliphatic, C1-4alkoxy, C1-4fluoroalkoxy, S — C1-4aliphatic, S — C1-

[0333] 4fluoroaliphatic, — N(Rz7)2, — C(O)Rz8, S(O)Rz8, S(O)2Rz8, C(O)2Rz7, —

[0334] C(O)N(Rz7)2, — S(O)2N(RZ7)2, — OC(O)N(RZ7)2, — N(RZ7)C(O)RZ8, — N(RZ7)SO2RZ8. — N(RZ7)C(O)ORZ8, T2-RZ9, a 5- to 6-membered heteroaryl, a 6-membered aryl, a 3- to 6-membered cycloaliphatic, or a 4- to 6-membered heterocyclyl; and which is optionally substituted at one saturated carbon with oxo, a spirocyclic 3- to 6- membered carbocycle, or a spirocyclic 4- to 6-membered heterocycle; each occurrence of Rz7is independently hydrogen or C1-4alkyl; each occurrence of Rz8is independently C1-4alkyl;

[0335] T2is a C1-C2alkylene chain; and

[0336] Rz9is cyano, NO2, N(Rz7)2, ORz7, C(O)Rz8, C(O)2Rz7, or C(O)N(Rz7)2. In alternative embodiments: L is — C(R*)(Rf) — , — S — , — S(O) — , — S(O)2— , — C(O) , C(=CH2) , C(Rf)(Rf) C(=CH2) , — C(Rf)(Rf)— C=C— , C(Rf)(Rf)— O— . — C(Rf)(Rf)— S— , — C(Rf)(Rf)— N(Rg)— , — C(Rf)(Rf)— N(Rg)— CH2— . — C(Rf)(Rf)— CH2— , — C(Rf)(Rf)— CH2— CH2— . or — C(O)— C(Rf)(Rf)— .

[0337] In alternative embodiments: Z1is hydrogen, halogen, cyano, Rz3, — S(O) — Rz3, or — S(O)2— Rz3; Rz3is a phenyl, 5- to 7-membered cycloaliphatic, 5- to 7- membered heterocyclyl, or C1-4aliphatic, any of which may be substituted with one or more independently selected Rz4; Rz4is hydroxyl, halogen, cyano, C1-4aliphatic, C1-4fluoroaliphatic, C1-4alkoxy, C1-4fluoroalkoxy, — N(Rz5)2, — C(O)Rz6, — C(O)2Rz5, 5- or 6-membered cycloaliphatic or heterocyclyl, or a phenyl optionally substituted with one or more independently selected halogens; each occurrence of Rz5is independently hydrogen or C1-4alkyl; and each occurrence of Rz6is independently C1-4alkyl.

[0338] In alternative embodiments: Z1is hydrogen; halogen; cyano; phenyl optionally substituted with one or more independently selected halogens; 5- to 7-membered cycloaliphatic or heterocyclyl optionally fused to a 6-membered aryl, wherein the 5- to 7-membered cycloaliphatic or heterocyclyl optionally fused to a 6-membered aryl is optionally substituted with one or more independently selected halogens; C1-4 fluoroaliphatic; or a C1-4aliphatic group optionally substituted with one or more hydroxyl, C1-4alkoxy, phenyl optionally substituted with one more independently selected halogens, 5- or 6-membered cycloaliphatic, 5- or 6-membered heterocyclyl, or — N(RZ5)2.

[0339] In alternative embodiments: Z2is a ring system having a 5- to 7-membered heterocyclyl with 1-2 heteroatoms or a 5- to 7-membered cycloaliphatic fused to

[0340] (i) a 5-membered heteroaryl or 6-membered aryl or heteroaiyl to form a bicyclic group; or

[0341] (ii) a 9-membered heteroaryl or 10-membered aryl or heteroaryl to form a tricyclic group; wherein the ring system is optionally substituted by 1-3 independent occurrences of halogen, hydroxyl, cyano, C1-4aliphatic, C1-4fluoroaliphatic, C1-4alkoxy, C1-

[0342] 4 fluoroalkoxy, S — C1-4aliphatic, S — C1-4fluoroaliphatic, — N(Rz7)2, — C(O)Rz8, — S(O)Rz8, — S(O)2RZ8, — C(O)2RZ7, — C(O)N(RZ7)2, — S(O)2N(RZ7)2, — OC(O)N(RZ7)2, — N(RZ7)C(O)RZ8, — N(RZ7)SO2RZ8, — N(RZ7)C(O)ORZ8, T2-RZ9, 5- to 6-membered heteroaiy 1, 6-membered aryl, 3- to 6-membered cycloaliphatic or heterocyclyl; and the ring system is optionally substituted at one saturated carbon with oxo, a spirocyclic 3- to 6-membered carbocycle, or a spirocyclic 4- to 6-membered heterocycle; each occurrence of Rz7is independently hydrogen or C1-4alkyl; each occurrence of Rz8is independently C1-4alkyl;

[0343] T2is a C1-2alkylene chain; and

[0344] Rz9is cyano, — N(Rz7)2, — ORz7, — C(O)Rz8, — C(O)2Rz7, or — C(O)N(Rz7)2.

[0345] In alternative embodiments: Z2is a 5- to 7-membered heterocyclyl with 1-2 heteroatoms or a 5- to 7-membered cycloaliphatic fused to a 5-membered heteroaryl or 6-membered aryl or heteroaryl ring to form a bicyclic group, wherein the ring system is optionally substituted as described above.

[0346] In alternative embodiments: Z2is a 6-membered heterocyclyl, the heterocyclyl containing 1 N or O atom, fused to a 6-membered aryl or heteroaryl ring to form a bicyclic group, wherein the ring system is optionally substituted as defined herein.

[0347] In alternative embodiments: a compound or pharmaceutically acceptable salt of formula (V) is: wherein: m is 0, 1, or 2;

[0348] X4is S, O, orN(Rn4);

[0349] X5is O, C(O), or C(RX5)(RX5’), wherein Xs is not O if X4is N(Rn4) or S;

[0350] Rn4is hydrogen or C1-4alkyl;

[0351] Rx5is hydrogen, fluoro, hydroxyl, or C1-4alkyl;

[0352] Rx5is hydrogen, fluoro, or C1-4alkyl, wherein Rx5is not fluoro if Rx5is hydroxyl; or Rx5and Rx5, taken together with the carbon atom to which they are attached, form a spirocyclic 3- to 6-membered carbocycle or a spirocyclic to 4- to 6-membered heterocycle comprising only one heteroatom, the heteroatom being chosen from O, N, or S; dashed lines indicate single or double bonds;

[0353] Ring A is a fused 5-membered heteroaryl or 6-membered aryl or heteroaryl optionally substituted with 1-3 independent occurrences of halogen, hydroxyl, cyano, C1-4 aliphatic, C1-4fluoroaliphatic, C1-4alkoxy, C1-4fluoroalkoxy, — S — C1-4aliphatic, — S—C1-4fluoroaliphatic, — N(Rz7)2, — C(O)Rz8, — S(O)Rz8, — S(O)2Rz8, — C(O)2Rz7, — C(O)N(RZ7)2, — S(O)2N(RZ7)2. — OC(O)N(RZ7)2, — N(RZ7)C(O)RZ8, — N(RZ7)SO2RZ8, — N(RZ7)C(O)ORZ8, T2-RZ9, a 5- to 6-membered heteroaryl, a 6- membered aryl, a 3- to 6-membered cycloaliphatic, or a 4- to 6-membered heterocyclyl;

[0354] T2is a C1-C2alkylene chain;

[0355] Rz9is cyano, — N(Rz7)2, — ORz7. — C(O)Rz7, — C(O)2Rz8. or — C(O)N(Rz7)2; and Rkis hydrogen or methyl.

[0356] In alternative embodiments, a compound or pharmaceutically acceptable salt wherein: m is 1 or 2;

[0357] X6is N or C(Rx6);

[0358] X6'is N or C(Rx6); and each of Rx6, Rx6, RJand Rmis independently hydrogen, halogen, hydroxyl, cyano, C1-4 aliphatic, C1-4fluoroaliphatic. C1-4alkoxy, C1-4fluoroalkoxy. — N(Rz7)2. — C(O)2Rz7, — C(O)N(RZ7)2, — S(O)2N(RZ7)2, — CH2— ORZ7, — CH2NRZ7, a 3- to 6-membered cycloaliphatic, or a 4- to 6-membered heterocyclyl; wherein at least one of Rx6, Rx6, Rj, and Rmis hydrogen. In alternative embodiments: m is 1;

[0359] X5is CH2;

[0360] Rn4is hydrogen or methyl; and each of Rx6, Rx6, R>, and Rmis independently hydrogen, chloro, fluoro, bromo, iodo, methyl, ethyl, isopropyl, cyano, cyclopropyl, CF3, — OCH3, — OCH2CH3, or — C=CH: wherein at least one of Rx6, Rx7, R' and Rmis hydrogen.

[0361] In alternative embodiments:

[0362] X i is O or N(H);

[0363] X6is N or C(H);

[0364] X6'is C(H);

[0365] Rmis hydrogen, fluoro or chloro; and

[0366] Rjis methyl, ethyl, isopropyl, hydrogen, fluoro, chloro, bromo, cyclopropyl, — C=CH or — CF3.

[0367] In alternative embodiments: a compound or pharmaceutically acceptable salt of formula (Via) or (VIb):

[0368] In alternative embodiments: Z2is

[0369] X4is O or (Rn4);

[0370] X5is C(RX5)(RX5');

[0371] X6is N or C(Rx6); X7is O or S;

[0372] X8is S or N(H); and

[0373] Rx6and Rjare each independently hydrogen, halogen, hydroxyl, cyano, C1-4aliphatic. C1-4fluoroaliphatic, C1-4alkoxy, C1-4fluoroalkoxy, — N(Rz7)2. — C(O)2Rz7, — C(O)N(RZ7)2, — S(O)2N(RZ7)2, — CH2— ORZ7, — CH2N(RZ7)2, a 3- to 6-membered cycloaliphatic, or a 4- to 6-membered heterocyclyl.

[0374] In alternative embodiments: Z2is

[0375] X4is O or N(Rn4);

[0376] X5is C(Rx5)(Rx5);

[0377] X6is N or C(Rx6);

[0378] X6'is N or C(Rx6); and each Rx6, Rj, and Rmis independently hydrogen, halogen, hydroxyl, cyano, C1-

[0379] 4aliphatic, C1-4fluoroaliphatic. C1-4alkoxy, C1-4fluoroalkoxy. — N(Rz7)2, — C(O)2Rz7, — C(O)N(RZ7)2, — S(O)2N(RZ7)2, — CH2— ORZ7, — CH2N(RZ7)2, a 3- to 6-membered cycloaliphatic, or a 4- to 6-membered heterocyclyl, wherein at least one Rx6, Rj, or Rmis hydrogen.

[0380] In alternative embodiments: Z2is

[0381] X5is C(RX5)(RX5);

[0382] Rx5and Rx5are independently hydrogen or fluoro; or, together with the carbon to which they are attached, form a cyclopropyl ring; Rjis hydrogen, chloro, fluoro, bromo, methyl, ethyl, isopropyl, cyano, cyclopropyl,

[0383] CF3, — OCH3, OCH2CH3, or C=CH: and

[0384] Rmis hydrogen, fluoro, or chloro.

[0385] In alternative embodiments: Z2is

[0386] X4is O or N(Rn4);

[0387] X6is N or C(Rx6);

[0388] X6'is N or C(Rx6);

[0389] Rn4is hydrogen or C 1-4 alkyl; and each of Rx6, Rjor Rmis independently halogen, hydroxyl, cyano, C1-4aliphatic, C1-4fluoroaliphatic, C1-4alkoxy, C1-4fluoroalkoxy, — N(Rz7)2, — C(O)2Rz7, — C(O)N(RZ7)2. — S(O)2N(RZ7)2, — CH2— ORZ7, — CH2NRZ7, a 3- to 6-membered cycloaliphatic, or a 4- to 6-membered heterocyclyl.

[0390] In alternative embodiments: Z2is L-Re.

[0391] In alternative embodiments: L is — C(Rf)(Rf) — , S— , C(=O) , C(Rf)(Rf)

[0392] O— , — C(Rf)(Rf)— S— , — C(Rf)(Rf)— N(Rg)— , -C(Rf)(Rf)— CH2— or — C(Rf)(Rf)— C=C— .

[0393] In alternative embodiments: a compound or pharmaceutically acceptable salt of formula (VIII):

[0394] In alternative embodiments: R6is a ring chosen from 3- to 7-membered cycloaliphatic or 4- to 7-membered heterocyclyl, which is optionally fused to a second 6-membered aryl, 5- to 6-membered heteroaryl, 3- to 7-membered cycloaliphatic, or 4- to 7- membered heterocyclyl, which is optionally substituted by 1-3 independent occurrences of halogen, hydroxyl, cyano, C1-4aliphatic, C1-4fluoroaliphatic, C1- 4 alkoxy, C1-4fluoroalkoxy, S — C1-4aliphatic, S — C1-4fluoroaliphatic, — N(Rz7)2, — C(O)Rz8, S(O)Rz8, S(O)2RZ8, C(O)2RZ7, — C(O)N(RZ7)2, — S(O)2N(RZ7)2, — OC(O)N(RZ7)2, — N(RZ7)C(O)RZ8, — N(RZ7)SO2RZ8, — N(RZ7)C(O)ORZ8, T2-RZ9, a 5- to 6-membered heteroaryl, a 6-membered aryl, a 3- to 6-membered cycloaliphatic, or a 4- to 6-membered heterocyclyl; and which is optionally substituted at one saturated carbon with oxo, a spirocyclic 3- to 6-membered carbocycle, or a spirocyclic 4- to 6- membered heterocycle; each occurrence of Rz7is independently hydrogen or C1-4alkyl; each occurrence of Rz8is independently C1-4alkyl;

[0395] T2is a C1-C2alky dene chain; and

[0396] Rz9is cyano, N(Rz7)2, ORz7, C(O)Rz7, C(O)2Rz7, or C(O)N(Rz7)2.

[0397] In alternative embodiments: each Rfis independently hydrogen, hydroxyl, N(Rh)(Rh), C1-4alkoxy, cyclopropyl, or C1-4alkyl optionally substituted with hydroxyl or —

[0398] OCH3; each Rfis independently hydrogen, cyclopropyl, or C1-4alkyd optionally substituted with hydroxyl or — OCH3; and

[0399] Reis a 5- to 7-membered cycloaliphatic ring or a 5- to 7-membered heterocyclyl having only one heteroatom, wherein the ring is optionally substituted as defined in any one of claims 18 to 21.

[0400] In alternative embodiments: Reis and wherein dashes indicate single or double bonds;

[0401] E1is N or C(H);

[0402] E2is O. S, or CH2;

[0403] E3is O, S, N(Re3), or C(H)(Re3); E3' is O, N(Re3) or C(H)(Re3);

[0404] Re1and Re1are each independently hydrogen or fluoro;

[0405] Re2is hydrogen or methyl; and

[0406] Re3is hydrogen or methyl.

[0407] In alternative embodiments: Rfand Rfare each independently hydrogen, C1-4alkyd, or cyclopropyl; or are taken together to form =CH2; and

[0408] Reis a ring chosen from 6-membered aryl or 5- to 6-membered heteroaryl, which is optionally fused to a second 6-membered aryl, 5- to 6-membered heteroaryl, 3- to 7-membered cycloaliphatic, or 4- to 7-membered heterocyclyl, and is optionally substituted with 1-3 independent occurrences of halogen, hydroxyl, cyano, C1-4aliphatic, C1-4fluoroaliphatic, C1-4alkoxy, C1-4fluoroalkoxy, — N(Rz7)2, — C(O)2Rz7, — C(O)N(RZ7)2, — S(O)2N(RZ7)2. — CH2— ORZ7, — CH2NRz7or a 3- to 6-membered cycloaliphatic or 4- to 6-membered heterocyclyl.

[0409] In alternative embodiments: Rfis hydrogen, hydroxyl, N(Rh)(Rh), — OCH3, cyclopropyl, or C1-4aliphatic optionally substituted with hydroxyl or — OCH3;

[0410] Rfis hydrogen, cyclopropyl, or C1-4aliphatic optionally substituted with hydroxyl or — OCH3; or. together with the carbon atom to which they are attached. Rfand Rfform a 4- to 6-membered heterocycle comprising a heteroatom chosen from N (which may be protonated or C1-4alkylated), O, or S, the heteroatom optionally located immediately adjacent to the quaternary carbon of the heterocycle; wherein at least one of Rfand Rfcomprises at least one heteroatom; and

[0411] Reis a ring chosen from 6-membered aryl or 5- to 6-membered heteroaryl, which is optionally fused to a second 6-membered aryl, 5- to 6-membered heteroaryl, 3- to 7-membered cycloaliphatic, or 4- to 7-membered heterocyclyl, and is optionally substituted with one or more halogen, C1-4aliphatic optionally substituted with 1-3 independent occurrences of halogen, hydroxyl, cyano, C1-4aliphatic, C1-4fluoroaliphatic, C1-4alkoxy, C1-4fluoroalkoxy, — N(Rz7)2, — C(O)2Rz7, — C(O)N(RZ7)2, — S(O)2N(RZ7)2, — CH2— ORZ7, — CH2NRZ7, a 3- to 6-membered cycloaliphatic, or a 4- to 6-membered heterocyclyl.

[0412] In alternative embodiments: Reis a ring chosen from 6-membered aryl or 5- to 6-membered heteroaryl, which is optionally substituted with 1-3 independent occurrences of chloro, fluoro, bromo, iodo, methyl, ethyl, cyano, cyclopropyl, CF3, — OCH3, OCH2CH3, or C=CH. In alternative embodiments: Reis:

[0413] E4is or O;

[0414] E5is N or C(Re5);

[0415] E6is N or C(H);

[0416] Re4is hydrogen, methyl, chloro, fluoro, bromo, iodo, cyano, or — CF3; and Re5is hydrogen or halogen.

[0417] In alternative embodiments: Reis

[0418] E5is N or C(Re5);

[0419] E6is N or C(H); each of Re5, Re6, Re7, and Re8is independently hydrogen, halogen, methyl, ethyl, isopropyl, — OCFh, — CFs, or — C=CH; and at least two of Re5, Re6, Re7, and Re8are hydrogen.

[0420] In alternative embodiments: Reis

[0421] E5is N or C(Re5);

[0422] E6is N or C(H);

[0423] Re5is hydrogen, halogen, methyl, — OCH3, — CF3, or — C=CH;

[0424] Re6is hydrogen, fluoro, or chloro;

[0425] Re7is hydrogen, fluoro, or chloro; and

[0426] Re8is hydrogen, halogen, methyl, — OCHs, or cyano; wherein at least one of Re6, Re7, and Re8is hydrogen.

[0427] In alternative embodiments, a compound or pharmaceutically acceptable salt of formula (Villa) or (Vlllb):

[0428]

[0429] In alternative embodiments: Reis hydrogen, hydroxyl, halogen, — CF3, or C1-4alkyl optionally substituted with one or more hydroxyl, halogen, or C1-4alkyl.

[0430] In alternative embodiments: Z1is hydrogen, halogen, cyano, or C1-4aliphatic optionally substituted with one or more hydroxyl, C1-4alkoxy, — N(Rz5)2, or phenyl optionally substituted with one more independently selected halogens; and each occurrence of Rz5is independently hydrogen or C1-4alkyl.

[0431] In alternative embodiments: wherein Z1is hydrogen, chloro, or methyl; or Z2is hydrogen.

[0432] In alternative embodiments: wherein Rbis hydrogen, or Rbis — C(O) — Rbx, Rbxis C1-4 alkyl, — CH(Rby) — NH2, pyrrolidinyl, or -Lb-OPO3H2, Rbyis C1-4alkyl optionally substituted with hydroxyl, phenyl, phenolyl. imidazolyl, carboxyl, amino, guanidino. — SCH3, — C(O)NH2, or indolyl. Lb is a bivalent linker chosen from C1-4alkylene, — (CH2)n1-phenylene-(CH2)n2— where nl is 0 or 1 and n2 is 1 or 2.

[0433] In alternative embodiments: Y is — O — ; Rais hydrogen; Rais hydrogen; Rcis hydrogen; X1is N; Rdis hydrogen; or X3is C(H).

[0434] In alternative embodiments: a compound used in methods as provided herein is:

[0435] [(1R,2R,3S,4R)-4-{[5-({4-[(1S)-1-(6-bromopyridin-2-yl)-1-hydroxyethyl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2,3-dihydroxyeyelopentyl]methyl sulfamate;

[0436] [(1R,2R,3S,4R)-4-{[5-({4-[(1R)-1-(6-bromopyridin-2-yl)-1-hydroxyethyl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2,3-dihydroxycyclopentyl]methyl sulfamate;

[0437] [(1R,2S,4R)-4-{[5-({5-chloro-4-[(.S)-(6-chloropyridin-2-yl)(hydroxy)methyl]-2- thienyl } carbonyl)pyrimidin-4-yl] amino } -2-hydroxycyclopentyl] methyl sulfamate ; [(1R,2S,4R)-4-{[5-({5-chloro-4-[(R)-(6-chloropyridin-2-ylXhydroxy)methyl]-2- thienyl} carbonyl )pyrimidin-4-yl ]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0438] [(1R,2S,4R)-4- { [5-({4-[(1R)-1-(3-bromophenyl)-1-hydroxyethyl]-2-thienyl}carbonyl)pyrimidin-4- yl] amino }-2-hydroxycyclopentyl]methyl sulfamate;

[0439] [(1R,2S,4R)-4-{[5-({4-[(1S')-1-(3-bromophenyl)-1-hydroxyethyl]-2-thienyl}carbonyl)pyrimidin-4- yl]amino}-2-hydroxycyclopcntyl]methyl sulfamate;

[0440] [(1R,2S,4R)-4-{[5-({4-[(R)-(3-chlorophenyl)(hydroxy)methyl]-2-thienyl}carbonyl)pyrimidm-4- yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0441] [(1R,2 S,4R)-4- { [5-( {4-[(S)-(3-chlorophenyl)(hy droxy)methyl]-2-thienyl} carbonyl)pyrimidin-4- yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0442] [(1R,2S,4R)-4-{[5-({4-[(1R)-1-(3-chlorophenyl)-1-hydroxyetliyl]-2-thienyl}carbonyl)pyrimidin-4- yl]amino}-2-hydroxycyclopentylJmethyl sulfamate;

[0443] [(1R,2S,4R)-4-{[5-({4-[(1S)-1-(3-chlorophenyl)-1-hydroxyethyl]-2-thienyl}carbonyl)pyrimidm-4- yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0444] [(1R,2S,4R)-4- { [5-({4-[(R)-amino(3-chlorophenyl)methyl]-5-methyl-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0445] [(1R,2S,4R)-4-{[5-({4-[(S')-amino(3-chlorophenyllmethyl]-5-methyl-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0446] [(1R,2S,4R')-4-{[5-({4-[(R)-(3-chlorophenyl)(hydroxy)methyl]-5-(methoxymethyl)-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0447] [(1R,2S,4R')-4-{[5-({4-[(S)-(3-chlorophenyl)(hydroxy)methyl]-5-(methoxymethyl)-2- thienyl} carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0448] [(1R,2R,3S,4R)-4-{[5-({4-[(1S)-1-ammo-1-(3-chlorophenyl)ethyl]-2-thienyl}carbonyl)pyrimidin- 4-yl] amino} -2, 3-dihydroxycyclopentyl]methyl sulfamate;

[0449] [(1R,2R,3S,4R)-4- { [5-( { 4- [(1R)-1-amino-1-(3-chlorophenyl)ethyl] -2-thienyl } carbonyl)pyrimidin- 4-yl]amino}-2,3-dihydroxycyclopentyl]methyl sulfamate;

[0450] [(1R,2S,4R)-4-{[5-({5-chloro-4-[(R)-(3-chlorophenylXhydroxy)methyl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0451] [(1R,2S,4R)-4-{[5-({5-chloro-4-[(S)-(3-chlorophenyl)(hydroxy)methyl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0452] [(1R,2R,3S,4R)-4-{[5-({4-[(R)-amino(6-bromopyridin-2-yl)methyl]-5-chloro-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2,3-dihydroxycyclopentyl]methyl sulfamate;

[0453] [(1R,2R,3S,4R')-4-{[5-({4-[(S)-amino(6-bromopyridin-2-yl)methyl]-5-chloro-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2,3-dihydroxycyclopentyl]methyl sulfamate;

[0454] [(1R,2S,4R)-4-{[5-({4-[(R)-(3-chlorophenyl)(hydroxy)methyl]-5-methyl-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0455] [(1R,2S,4R)-4-{[5-({4-[(S)-(3-chlorophenyl)(hydroxy)methyl]-5-methyl-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0456] [(1R,2S,4R)-4-{[5-({4-[(1S)-1-amino-1-(3-chlorophenyl)ethyl]-2-thienyl}carbonyl)pyrimidin-4- yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0457] [(1R,2S,4R)-4-{[5-({4-[(1R)-1-amino-1-(3-clilorophenyl)ethyl]-2-thienyl}carbonyl)pyrimidin-4- yl]amino}-2-hydroxyeyelopentyl]methyl sulfamate;

[0458] [(1R,2S,4R)-4-{[5-({4-[(R)-(3-chlorophenyl)(hydroxy)methyl]-5-(hydroxymethyl)-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0459] [(1R,2S,4R)-4-{[5-({4-[fS)-(3-chlorophenyl)(hydroxy)methyl]-5-(hydroxymetliyl)-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0460] [(1R,2R,3S,4R)-4- { [5-({4-[(R)-amino(3-chlorophenyl)methyl]-5-chloro-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2,3-dihydroxycyclopentyl]methyl sulfamate; [(1R,2R,3S,4R')-4-{[5-({4-[(S)-amino(3-chlorophenyl)methyl]-5-chloro-2- thienyl} carbonyl )pyrimidin-4-yl ]amino} -2, 3-dihydroxy cyclopentyl ]methyl sulfamate;

[0461] [(1R,2S,4R)-4-{[5-({4-[(R)-amino(3-chlorophenyl)methyl]-5-chloro-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0462] [(1R,2S,4R')-4-{[5-({4-[(S)-amino(3-chlorophenyl)metliyl]-5-chloro-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopcntyl]methyl sulfamate;

[0463] [(1R,2S,4R')-4-{[5-({4-[(S)-(3-bromophenyl)(hydroxy)methyl]-5-chloro-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0464] [(1R,2S,4R)-4-{[5-({4-[(R)-(3-bromophenyl)(hydroxy)methyl]-5-chloro-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0465] [(1R,2S,4R)-4-{[5-({4-[(R)-(3-bromophenyl)(hydroxy)methyl]-2--thienyl}carbonyllpyrimidin-4- ylJamino}-2-hydroxycyclopentylJmethyl sulfamate;

[0466] [(1R,2S,4R)-4-{[5-({4-[(S)-(3-bromophenyl)(hydroxy)methyl]-2-thienyl}carbonyl)pyrimidm-4- yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0467] [(1R,2S,4R)-4-{[5-({4-[(R)-amino(6-bromopyridin-2-yl)methyl]-5-chloro-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0468] [(1R,2S,4R)-4-{[5-({4-[(S')-amino(6-bromopyridin-2-yl)methyl]-5-chloro-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0469] [(1R,2S,4R)-4-{[5-({4-[(R)-(5-bromo-2-fluorophenyll(hydroxy)methyl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0470] [(1R,2S,4R)-4-{[5-({4-[(S)-(5-bromo-2-fluorophenyl)(hydroxy)methyl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0471] [(1R,2S,4R)-4-{[5-({4-[(2S)-2-(3-chlorophenyl)tetrahydrofuran-2-yl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0472] [(1R,2S,4R)-4-[[5-({4-[(2R)-2-(3-chlorophenyl)tetrahydrofuran-2-yl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0473] [(1R,2S,4R)-4-{[5-({4-[(S)-(5-chloro-2-furyl)(hydroxylmetliyl]-2-thienyl}carbonyl)pyrimidin-4- yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0474] [(1R,2S,4R)-4-{[5-({4-[(R)-(5-chloro-2-furyl)(hydroxy)methyl]-2-thienyl}carbonyl)pyrimidin-4- yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0475] [( 1R,2S,4R)-4- { [5-({4-[(R)-amino(3-bromophenyl)methyl]-5-chloro-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0476] [(1R,2S,4R)-4-{[5-({4-[(S)-amino(3-bromophenyl)methyl]-5-chloro-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0477] [(1R,2R,3S,4R)-4-{[5-({4-[(R)-amino(3-bromophenyl)methyl]-5-chloro-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2;3-dihydroxycyclopentyl]methyl sulfamate;

[0478] [(1R,2R,3S,4R)-4-{[5-({4-[(S)-amino(3-bromophenyl)methyl]-5-chloro-2- thienyl} carbonyl)pyrimidin-4-yl]amino} -2, 3-dihydroxy cyclopentyl]methyl sulfamate;

[0479] [( 1 R,2S,4R )-4- { [ 5-( {4-[( S )-amino(6-chloropyridin-2-yl)m ethyl ]-5-chloro-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0480] [(1R,2S,4R)-4- {[5-({4-[(R)-amino(6-chloropyridin-2-yl)methyl]-5-chloro-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxyeyelopentyl]methyl sulfamate;

[0481] {(1R.2R.3S.4R)-4-[(5-{[4-(3-bromobenzyl)-2-thienyl]carbonyl}pyrimidin-4-yl)amino]-2,3- dihydroxy cyclopentyl} methyl sulfamate;

[0482] [(1R,2R,3S,4R)-4-{[5-({4-[(1S)-1-amino-1-(3-bromophenyl)ethyl]-2-thienyl}carbonyl)pyrimidin-

[0483] 4-yl]amino}-2,3-dihydroxycyclopentyl]methyl sulfamate:

[0484] [(1R,2R,3S,4R)-4-{[5-({4-[(1R)-1-amino-1-(3-bromophenyl)ethyl]-2-thienyl}carbonyl)pyrimidin-

[0485] 4-yl]amino}-2,3-dihydroxycyclopentyl]methyl sulfamate; [(1R,2S,4R)4-{[5-({4-[(R)-(3-chlorophenyl)sulfmyl]-5-methyl-2-thienyl}carbonyl)pyrimidin-4- yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0486] [(1R,2S,4R)-4-{[5-({4-[(S)-(3-chlorophenyl)sulfinyl]-5-methyl-2-thienyl}carbonyl)pyrimidin-4- yl] amino} -2-hy droxy cyclopentyl]methyl sulfamate;

[0487] [(1R,2S,4R)-4- {[5-({4-[(S)-(6-chloropyridin-2-yl)(hydroxy)methyl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopcntyl]methyl sulfamate;

[0488] [(1R,2S,4R)-4-{[5-({4-[(R)-(6-chloropyridin-2-yl)(hydroxy)methyl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0489] {(1R,2S,4R)-4-[(5-{4-[(1S)-1-(3-chlorophcny1)-1-hydroxyethyl]-2-furoyl}pyrimidin-4-yl)amino]- 2-hydroxycyclopentyl}methyl sulfamate;

[0490] {(1R,2S,4R)-4-[(5-{4-[(1R)-1-(3-chlorophenyl)-1-hydroxyethyl]-2-furoyl}pyriinidin-4-yl)amino]- 2-hydroxycyclopentyl}methyl sulfamate;

[0491] [(1R,2R,3S,4R)-4-{[5-({4-[(2S)-2-(3-chlorophenyl)pyrrolidin-2-yl]-2-thienyl}carbonyl)pyrimidin-

[0492] 4-yl]ammo}-2,3-dihydroxycyclopentyl]methyl sulfamate:

[0493] [(1R,2R,3S,4R)-4-{[5-({4-[(2R)-2-(3-chlorophenyl)pyrrolidin-2-yl]-2-thienyl}carbonyl)pyrimidin- 4-yl]amino}-2,3-dihydroxycyclopentyl]methyl sulfamate;

[0494] [(1R,2S,4R)-2-hydroxy-4-{[5-({4-[(2S)-2-phenyltetrahydrofuran-2-yl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}cyclopentyl]methyl sulfamate;

[0495] [(1R,2S,4R)-2-hydroxy-4-{[5-({4-[(2R)-2-phenyltetrahydrofuran-2-yl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}cyclopentyl]methyl sulfamate;

[0496] [(1R,2S,4R)-4-({5-[4-(3-chlorobenzyl)-5-methyl-2-furoyl]pyrimidin-4-yl]ammo)-2- hydroxy cyclopentyl} methyl sulfamate;

[0497] [(1R,2S,4R)-4-({5-[4-(3-bromobenzyl)-5-metliyl-2-furoyl]pyrimidin-4-yl}amino)-2- hydroxycyclopentyl]methyl sulfamate;

[0498] {(1R,2S,4R)-4-[(5-{[4-(3-chlorobenzyl)-5-(hydroxymethyl)-2-thienyl]carbonyl}pyrimidin-4- yl)amino] -2-hy droxy cyclopentyl } methyl sulfamate ;

[0499] [(1R,2S,4R)-4-{[5-({4-[(R)-amino(3-chlorophenyl)methyl]-2-thienyl}carbonyl)pyrimidin-4- yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0500] [(1R,2S,4R)-4-{[5-({4-[(S)-amino(3-chlorophenyl)methyl]-2-thienyl}carbonyl)pyrimidin-4- yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0501] [(1R,2S,4R)-2-hydroxy-4-{[5-({4-[(4-iodo-1H-pyrazol-1-yl)methyl]-5-methyl-2- thienyl}carbonyl)pyrimidin-4-yl]amino}cyclopentyl]methyl sulfamate:

[0502] [(1R,2S,4R)-4-{[5-({4-[(2S)-2-(3-chlorophenyl)pyrrolidin-2-yl]-2-thienyl}carbonyl)pyrimidin-4- yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0503] [(1R,2S,4R)-4-{[5-({4-[(2R)-2-(3-chlorophenyl)pyrrolidin-2-yl]-2-thienyl}carbonyl)pyrimidin-4- yl] amino} -2-hy droxy cyclopentyl]methyl sulfamate;

[0504] [(1R,2S,4R)-4-{[5-({4-[(4-bromo-1H-pyrazol-1-yl)methyl]-5-methyl-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0505] [( 1 R,2 S,4R)-4- { [5-( { 5-chloro-4- [(R)-(3 -chloro-2-fhiorophenyl)(hydroxy )methyl] -2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0506] [(1R,2S,4R)-4- { [5-({5-chloro-4-[(S)-(3-chloro-2-fluorophenyl)(hydroxy)methyl]-2- thienyl} carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0507] [(1R,2S,4R)-4-{[5-({4-(3-chlorobenzyl)-5-[(2S)-tetrahydrofuran-2-yl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0508] [(1R,2S,4R)-4-{[5-({4-(3-chlorobenzyl)-5-[(2R)-tetrahydrofuran-2-yl]-2- thienyl } carbonyl)pyrimidin-4-yl] amino } -2-hydroxycyclopentyl]methyl sulfamate ;

[0509] [(1R,2S,4R)-4-[(5-{4-[(R)-(3-chlorophenyl)(hydroxy)methyl]-2-furoyl}pyrimidin-4-yl)amino] -2- hydroxycyclopentyl]methyl sulfamate: {(1R,2S,4R)-4-[(5-{4-[(S)-(.3-chlorophenyl)(hydroxy)methyl]-2-furoyl}pyrimidin-4-yl)amino]-2- hydroxy cyclopentyl } methyl sulfamate;

[0510] {(1R2S,4R)-4-[(5-{[4-(3-chlorobenzyl)-2-thienyl]carbonyl}pyrimidin-4-yl)amino]-2- hydroxycyclopentyl}methyl sulfamate;

[0511] {(1R,2S,4R)-4-[(5-{[4-(3-chlorobenzyl)-5-(methoxymethyl)-2-thienyl]carbonyl}pyrimidin-4- yl)amino] -2-hy droxy cy clopcnty 1 } methyl sulfamate ;

[0512] {(1R,2S;4R)-2-hydroxy-4-[(5-{[5-methyl-4-(3-methylbenzyl)-2-thienyl]carbonyl}pyrimidin-4- yl)amino] cyclopentyl } methyl sulfamate ;

[0513] [(1R,2S,4R)-4-{[5-({4-[(6-bromopyridin-2-yl)methyl]-5-chloro-2-thienyl}carbonyl)pyrimidin-4- yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0514] [(1R,2S,4R)-4-{[5-({4-[(R)-(3-chlorophenyl)(methylamino)methyl]-2-thienyl}carbonyl)pyrimidin- 4-ylJamino}-2-hydroxycyclopentylJmethyl sulfamate;

[0515] [(1R,2S,4R)-4-{[5-({4-[(.S)-(3-chlorophenyl)(methylamino)methyl]-2-thienyl}carbonyl)pyrimidin- 4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0516] [(1R,2 S,4R)-4- { [5-( { 5-chloro-4- [(R)-(5-chloro-2-fluorophenyl)(hydroxy )methyl] -2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0517] [(1R,2S,4R)-4-{[5-({5-chloro-4-[(S)-(5-chloro-2-fluorophenyl)(hydroxy)methyl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0518] [(1R,2S,4R)-4-{[5-({5-chloro-4-[(1R)-1-(3-chlorophenyl)-1,3-dihydroxypropyl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0519] [(1R,2S,4R)-4-{[5-({5-chloro-4-[(lS)-1-(3-chlorophenyl)-1,3-diliydroxypropyl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0520] [(1R,2S,4R)-4-{[5-({4-[(S)-(3-chlorophenyl)(cyclopropyl)hydroxymethyl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0521] [(1R,2S,4R)-4- {[5-({4-[(R)-(3-chlorophenyl)(cyclopropyl)hydroxymethyl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0522] {(1R,2S,4R)-4-[(5-{[4-(3-chlorobenzyl)-5-methyl-2-thienyl]carbonyl}pyrimidin-4-yl)amino]-2- hydroxycyclopentyl}methyl sulfamate;

[0523] {(1R,2S,4R)-4-[(5-{[4-(3-bromobenzyl)-5-methyl-2-thienyl]carbonyl}pyrimidin-4-yl)amino]-2- hydroxy cyclopentyl}methyl sulfamate;

[0524] [(1R,2S,4R)-4-{[5-({4-[(6-bromopyridin-2-yl)methyl]-2-thienyl}carbonyl)pyrimidin-4-yl]amino}- 2-hy droxy cyclopentyl] methyl sulfamate ;

[0525] [(1R,2S,4R)-4-{[5-({4-[(l S)-1-(6-bromopyridin-2-yl)-1-hy droxy ethyl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0526] [(1R,2S,4R)-4-{[5-({4-[(1R)-1-(6-bromopyridin-2-yl)-1-hydroxyethyl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methylsulfamate;

[0527] [(1R,2S,4R)-4-{[5-({4-[(4-chloro-1H-pyrazol-1-yl)methyl]-5-methyl-2- thienyl]carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0528] [(1R, 2S,4R)-4-{[5-({5-chloro-4-[(R)-hy droxy (phenyl)methyl]-2-thienyl}carbonyl)pyrimidin-4- yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0529] [(1R,2S,4R)-4- {[5-({5-chloro-4-[(S)-hydroxy(phenyl)methyl]-2-thienyl}carbonyl)pyrimidin-4- yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0530] [(1R,2S,4R)-2-hydroxy-4-({5-[4-(3-methylbenzyl)-2-furoyl]pyrimidin-4- yl } amino)cyclopentyl]methyl sulfamate;

[0531] {(1R,2S,4R)-2-hydroxy-4-[(5-{4-[(1S)-1-hydroxy-1-phenylethyl]-2-furoyl}pyrimidin-4- yl)amino] cyclopentyl} methyl sulfamate ;

[0532] {(1R,2S,4R)-2-hydroxy-4-[(5-{4-[(1R)-1-hydroxy-1-phenylethyl]-2-furoyl}pyrimidin-4- yl)amino] cyclopentyl} methyl sulfamate ; {(1R,2S,4R)-4-[(5-{[4-(3-bromobenzyl)-2-thienyl]carbonyl}pyrimidin-4-yl)amino]-2- hydroxycyclopentyl } methyl sulfamate;

[0533] {(1R,2S,4R')-2-hydroxy-4-[(5-{[4-(3-methylbenzyl)-2-thienyl]carbonyl}pyrimidin-4- yl)amino] cy clopentyl} methyl sulfamate ;

[0534] {(1R,2S,4R)-4-[(5-{[4-(3-chloro-4-fluorobenzyl)-2-thienyl]carbonyl}pyrimidin-4-yl)amino]-2- hydroxy cyclopcntyljmethyl sulfamate;

[0535] {(1R,2S;4R)-2-hydroxy-4-[(5-{[4-(3-iodobenzyl)-2-thienyl]carbonyl}pyrimidin-4- yl)amino]cyclopentyl} methyl sulfamate;

[0536] [(1R,2S,4R)-4-{[5-({5-chloro-4-[(R)-(5-chloro-2-mcthoxyphenyl)(hydroxy)methyl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0537] [(1R,2S,4R)-4-{[5-({5-chloro-4-[(S)-(5-chloro-2-methoxyphenyl)(hydroxy)methyl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-24iydroxycyclopentyl]methyl sulfamate;

[0538] [(1R,2R,3R,4R)-4-({5-[4-(3-bromobenzyl)-5-methyl-2-furoyl]pyriniidin-4-yl}amino)-3-fluoro-2- hydroxy cy clopenty f]methyl sulfamate;

[0539] [( 1R,2R,3R,4R')-3-fluoro-2-hydroxy-4-( {5-[5-methyl-4-(3-methylbenzyl)-2-furoyl]pyrimidin-4- yl]amino)cyclopentyl}methyl sulfamate;

[0540] [(1R,2S,4R)4-{[5-({4-[(5-chloro-2-furyl)methyl]-2-thienyl}carbonyl)pyrimidin-4-yl]ammo}-2- hydroxy cy clopenty l]m ethyl sulfamate ;

[0541] [(1R,2S,4R)-4-{[5-({5-chloro-4-[(2R)-2-(3-chlorophenyl)oxetan-2-yl]-2- thienyl} carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopenty l]methyl sulfamate;

[0542] [(1R,2S,4R)-4-{[5-{[5-chloro-4-[(2S')-2-(3-chlorophenyl)oxetan-2-yl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0543] [(1R,2S,4R)-2-hydroxy-4-{[5-{[5-methyl-4-[(R)-phenylsulfinyl]-2-thienyl}carbonyl)pyrimidin-4- y l]amino} cyclopenty l]methyl sulfamate;

[0544] [(1R,2S,4R)-2-hydroxy-4-{[5-({5-methyl-4-[(S)-phenylsulfinyl]-2-thienyl}carbonyl]pyrimidin-4- yl]amino}cyclopentyl]methyl sulfamate;

[0545] [(1R,2S,4R)-2-hydroxy-4-({5-[(5-methyl-4- {(R)-[3-(trifluoromethyl)phenyl] sulfinyl} -2- thienyl)carbonyl]pyrimidin-4-yl}amino)cyclopentyl]methyl sulfamate;

[0546] [(1R,2S,4R)-2-hydroxy-4-(5-[(5-methyl-4-{(S)-[3-(trifluoromethyl)phenyl]sulfinyl}-2- thienyl)carbonyl]pyrimidin-4-yl}amino)cyclopentyl}methyl sulfamate;

[0547] {(1R,2S,4R)-4-[(5-{[4-(3-ethynylbenzyl)-2-thienyl]carbonyl}pyrimidin-4-yl)amino]-2- hydroxycyclopentyl } methyl sulfamate;

[0548] [( 1R,2 S,4R)-4- { [5-( { 4- | ( 6-chloropvndm-2-v] ;m ethyl] -2-thienyl } carbonyl)pyrimidin-4-y 1] amino } - 2-hydroxycyclopentyl]methyl sulfamate;

[0549] [(1R,2S,4R)-4-({5-[5-chloro-4-(3-chlorobenzyl)-2-furoyl]pyrimidin-4-yl}amino)-2- hydroxycy clopenty l]methyl sulfamate;

[0550] [(1R,2S,4R)-4-{[5-({4-[(R)-(3-bromophenyl)(methoxy)methyl]-2-thienyl}carbonyl)pyrimidin-4- y l]amino} -2-hydroxycy clopenty l]methyl sulfamate;

[0551] [(1R,2S,4R')-4-{[5-({4-[(S)-(3-bromophenyl)(methoxy)methyl]-2-thienyl}carbonyl)pyrimidin-4- yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0552] [(1R,2S,4R)-2-hydroxy-4-{[5-({4-[(4-methyl-1H-pyrazol-1-yl)methy]yl) thienyl} carbonyl)pyrimidin-4-yl] amino } cyclopentyl]methyl sulfamate ;

[0553] [(1R,2R,3R,4R)-4-(5-[4-(3-chlorobenzyl)-5-methyl-2-furoyl]pyrimidin-4-yl}amino)-3-fluoro-2- hydroxy cyclopentyl]methyl sulfamate;

[0554] {(1R,2R,3S,4R)-4-[(.5-{[4-(.3-bromobenzyl)-5-chloro-2-thienyl]carbonyl-4-yl)amino]-2,3- dihydroxycyclopentyl}methyl sulfamate;

[0555] [(1R,2S,4R)-4-{[5-f{4-(3-chlorobenzyl)-5-[(dimethylamino)methyl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate; [(1R,2S,4R)-2-hydroxy-4-[5-(5-methyl-4-[(4-methyl-1H-pyrazol-1-yl)methyl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}cyclopentyl]methyl sulfamate;

[0556] [(1R,2S,4R)-4-({5-[4-(3-chlorobenzyl)-2-furoyl]pyrimidm-4-yl}amino)-2- hydroxycyclopentyl]methyl sulfamate;

[0557] [(1R,2S,4R')-2-hydroxy-4-{[5-({4-[(2-methoxyphenoxy')methyl]-2-thienyl}carbonyl)pyrimidin-4- yl]amino}cyclopcntyl]methyl sulfamate;

[0558] [( 1 R,2 S,4R)-2-hydroxy-4- { [5-( {4-[3-(methylsulfanyl)benzyl]-2-thienyl } carbonyl)pyrimidin-4- yl]amino}cyclopentyl]methyl sulfamate;

[0559] [( 1 R,2S,4R)-4-( { 5-[5-(3-bromobenzyl)-2-furoyl]pyrimidin-4-yl} amino)-2- hydroxycyclopentyl]methyl sulfamate;

[0560] [(1R,2S,4R)-4-{[5-({4-[(6-chloro-2,3-diliydro-1H-indol-1-yl)methyl]-2- thienyl}carbonyl)pyrimidin-4-ylJamino}-2-hydroxycyclopentylJmethyl sulfamate;

[0561] {(1R,2S,4R)-4-[(5-{[4-(3-chloro-2-fluorobenzy1)-2-thienyl]carbonyl}pyrimidin-4-yl)amino]-2- hydroxy cyclopentyl}methyl sulfamate;

[0562] [(1R,2S,4R)-4- { [5-({5-chloro-4-[(R)-hydroxy(2-methoxyphenyl]methyl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0563] [(1R,2S,4R)-4-{[5-({5-chloro-4-[(S')-hydroxy(2-methoxyphenyl')methyl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0564] [(1R,2S,4R)-4- { [5-({ 5-chloro-4- [( S)(2-chlorophenyl')(hydroxy)methyl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0565] [(1R,2S,4R')-4-{[5-({5-chloro-4-[(R)-(2-chlorophenyl)(hydroxy)methyl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0566] [(1R,2R;3S,4R)-4-{[5-(5-benzyl-2-furoyl)pyrimidin-4-yl]amino}-2,3- dihydroxycyclopentyl]methyl sulfamate;

[0567] [(1R,2S,4R)-4-({5-[(4-benzy1-2-thienyl)carbonyl]pyrimidin-4-y1}amino)-2- hydroxycyclopentyl]methyl sulfamate;

[0568] {(1R,2S,4R)-4-[(5-{[4-(3-chlorobenzyl)-5-fluoro-2-thienyl]carbonyl}pyrimidin-4-yl)amino]-2- hydroxy cyclopentyl}methyl sulfamate;

[0569] [(1R,2S,4R)-4-{[5-({5-chloro-4-[(R)-(3-chlorophenyl)(methoxy)methyl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0570] [(1R,2S,4R)-4-{[5-({5-chloro-4-[(SX3-chlorophenyl)(methoxy)methyl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0571] {( 1R,2S,4R)-2-hydroxy-4-[15- { |4-( phenoxymethyl )-2-thienyl ]carbonyl} pyrimidin-4- yl)am ino] cyclopentyl } methyl sulfamate ;

[0572] {(1R,2S,4R)-2-hydroxy-4-[(5-{[4-(1H-pyrrolo[2,3-b]pyridin-1-ylmethyl)-2- thienyl] carbonyl} pyrimidin-4-yl)am ino] cyclopentyl } methyl sulfamate ;

[0573] [(1R,2S,4R}-4-({5-[5-(3-chlorobenzyl)-2-furoyl]pyrimidin-4-yl}amino)-2- hydroxycyclopentyl]methyl sulfamate;

[0574] [(1R,2S,4R)-4-{[5-({5-chloro-4-[(R)-(3-chlorophenyl)sulfinyl]-2-thienyl}carbonyl)pyrirnidin-4- yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0575] [(1R,2S,4R)-4- {[5-({5-chloro-4-[(S)-(3-chlorophenyl)sulfinyl]-2-tlrienyl}carbonyl)pyrimidin-4- yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0576] [(1R,2S,4R)-4-{[5-({5-chloro-4-[(R)-(3-chlorophenyl)(methylamino)methyl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0577] [(1R,2S,4R)-4-{[5-({5-chloro-4-[(S)-(3-chlorophenyl)(methylamino)methyl]-2- thienyl } carbonyl)pyrimidin-4-y l] amino } -2-hydroxy cy clopenty l]methyl sulfamate ;

[0578] [(1R,2S,4R)-4-({5-[(4-benzyl-5-chloro-2-thienyl)carbonyl]pyrimidin-4-yl}amino)-2- hydroxycyclopentyl]methyl sulfamate; {(1R,2S,4R)-4-[(5-{[4-(3-fluorobenzyl)-2-thienyl]carbonyl}pyrimidin-4-yl)amino]-2- hydroxycyclopentyl}methyl sulfamate;

[0579] [(1R,2S,4R)-4-{[5-({4-[(2-bromophenoxy)methyl]-2-thienyl}carbonyl)pyrimidin-4-yl]amino}-2- hydroxycy clopenty l]methyl sulfamate;

[0580] {(1R,2R,3R,4R)-3-fluoro-2-hydroxy-4-[(5-{[4-(3-iodobenzyl)-2-thienyl]carbonyl}pyrimidin-4- yl)amino]cyclopentyl{ methyl sulfamate;

[0581] [(1R,2S,4R)-4-{[5-({4-[1-(3-chlorophenyl)vinyl]-5-methyl-2-thienyl{carbonyl)pyrimidin-4- yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0582] {(1R,2R;3R,4R)-4-[(5-{[4-(3-bromobcnzyl)-2-thienyl]carbonyl}pyrimidin-4-yl)amino]-3-fluoro-2- hydroxycyclopentyl } methyl sulfamate ;

[0583] [(1R,2S,4R)-4-{[5-({5-chloro-4-[(1R)-1-(3-chlorophenyl)-1-hydroxyethyl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0584] [(1R,2S,4R)-4-{[5-({5-chloro-4-[(1S)-1-(3-chlorophenyl)-1-hydroxyethyl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl}methyl sulfamate;

[0585] [(1R,2S,4R)-4-{[5-({5-chloro-4-[(S)-(5-chloro-2-thienyl)(hydroxy)methyl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0586] [(1R,2S,4R>4-{[5-({5-chloro-4-[(R)-(5-chloro-2-thienylXhydroxy)methyl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0587] {(1R,2S,4R)-4-[(5-{[4-(3,4-dichlorobenzyl)-2-thienyl]carbonyl}pyrimidin-4-yl)amino]-2- hydroxy cyclopentyl {methyl sulfamate;

[0588] [(1R,2S,4R)-2-hydroxy-4-{[5-({4-[(3-methyl-1H-indol-1-yl)methyl]-2- thienyl} carbonyl)pyrimidin-4-yl] amino }cyclopenty l]methyl sulfamate;

[0589] [(1R,2S,4R)-2-hydroxy-4-{[5-({4-[(1R)-1-phenylethyl]-2-thienyl}carbonyl)pyrimidin-4- yl]amino{ cy clopenty l]methyl sulfamate;

[0590] [(1R,2S,4R)-2-hydroxy-4-{[5-({4-[(1S)-1-phenylethyl]-2-thienyl{carbonyl)pyrimidin-4- yl]amino}cyclopentyl]methyl sulfamate;

[0591] {(1R,2S,4R)-4-[(5-{[5-chloro-4-(3-chlorobenzyl)-2-thienyl]carbonyl}pyrimidin-4-yl)amino]-2- hydroxy cyclopentyljmethyl sulfamate;

[0592] {(1R,2S,4R)-2-hydroxy-4-[(5-{[4-(2-phenylethyl)-2-thienyl]carbonyl}pyrimidin-4- yl)amino] cyclopentyl} methyl sulfamate ;

[0593] {(1R,2S,4R)-2-hydroxy-4-[(5-{[5-methyl-4-(1H-pyrazol-1-ylmethyl)-2- thienyl]carbonyl}pyrimidin-4-yl)amino]cyclopentyl}methyl sulfamate;

[0594] {( 1R,2S,4R)-4-[(5- { [4-(3-chlorobenzyl)-5-(tetrahydro-2H-pyran-4-ylmethyl)-2- thienyl]carbonyl}pyrimidin-4-yl')amino]-2-hydroxycyclopentyl {methyl sulfamate;

[0595] {(1R,2S,4R)-4-[(5-{[4-(3-ethylbenzyl)-5-methyl-2-thienyl]carbonyl}pyrimidin-4-yl)amino]-2- hydroxycyclopentyl {methyl sulfamate;

[0596] {(1R,2S,4R')-4-[(5-{[4-(3-bromobenzyl)-5-chloro-2-thienyl]carbonyl}pyrimidin-4-yl)amino]-2- hydroxy cyclopentyl} methyl sulfamate;

[0597] [(1R,2S,4R')-4-{[5-({4-[3-(difluoromethoxy)benzyl]-2-thienyl}carbonyl)pyrimidin-4-yl]amino}-2- hydroxycyclopentyl]methyl sulfamate;

[0598] {(1R,2S,4R)-2-hydroxy-4-[(5-{[4-(1H-indol-1-ylmethyl)-2-thienyl]carbonyl}pyrimidin-4- yl)amino]cyclopentyl}methyl sulfamate;

[0599] [(1R,2R,3S,4R)-4-{[5-[(5-benzyl-2-thienyl)carbonyl]pyrimidin-4-yl}amino)-2,3- dihydroxycyclopentyl]methyl sulfamate;

[0600] [(1R,2S,4R)-4-{[5-({4-[(2S)-2-(eyelohex-1-en-1-yl)tetrahydrofuran-2-yl]-2- thienyl}carbonyl)pyrimidin-4-y1]amino}-2-hydroxycyclopentyl]methylsulfamate;

[0601] {(1R,2S,4R)-4-{[5-({4-[(2R)-2-(cyclohex-1-en-1-yl)tetrahydrofuran-2-yl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate; [(1R,2S,4R)-4-{[5-({4-[(1S)-1-(3-chlorophenyl)ethyl]-2-thienyl}carbonyl)pyrimidin-4-yl]amino}- 2-hydroxycyclopentyl]methyl sulfamate;

[0602] [(1R,2S,4R)-4-{[5-({4-[(1R')-1-(3-chlorophenyl)ethyl]-2-thienyl}carbonyl')pyrimidin-4-yl]ammo}-

[0603] 2-hydroxycyclopentyl}methyl sulfamate;

[0604] {(1R,2S,4R)-4-[(5-{[5-bromo-4-(3-chlorobenzyl)-2-thienyl]carbonyl}pyrimidin-4-yl)amino]-2- hydroxy cyclopcntyljmethyl sulfamate;

[0605] [(1R,2S,4R)-2-hydroxy-4-({5-[(4-{[5-(trifluoromethyl)-2-fuiyl]methyl}-2- thienyl)carbonyl]pyrimidin-4-yl}amino)cyclopentyl]methyl sulfamate;

[0606] [(1R,2S,4R)-4-{[5-({4-[(3-chlorophcnoxy)methyl]-2-thienyl}carbonyl)pyrimidin-4-yl]amino}-2- hydroxy cyclopentyl]methyl sulfamate;

[0607] [(1R,2S,4R)-2-hydroxy-4-{[5-({5-methyl-4-[(2S)-2-phenyltetrahydrofuran-2-yl]-2- thienyl}carbonyl)pyrimidin-4-ylJamino}cyclopentylJmethyl sulfamate;

[0608] [f1R,2S,4R)-2-hydroxy-4-{[5-({5-methyl-4-[(.2R)-2-phenyltetrahydrofuran-2-yl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}cyclopentyl]methyl sulfamate;

[0609] [(1R,2S,4R)-4-({5-[(5-chloro-4-{(R)-[3-(trifluoromethyl)phenyl]sulfinyl}-2- thienyl)carbonyl]pyrimidin-4-yl } amino)-2-hydroxycyclopent\'l]methyl sulfamate;

[0610] [(1R,2S,4R)-4-({5-[(5-chloro-4-{(S')-[3-(trifluoromethyl)phenyl]sulfinyl}-2- thienyl (carbonyl |pvrimidin-4-vl} amino )-2-hydroxvcvclopentyl |methvl sulfamate;

[0611] {(1R,2S,4R)-2-hydroxy-4-[(5-{[4-(3-methoxybenzyl)-2-thienyl]carbonyl}pyrimidin-4- yl]amino} cy clopentyl } methyl sulfamate ;

[0612] [(1 R,2 S,4R)-4- { [5-( {4-[(2-cyanophenoxy)methyl] -2-thienyl } carbonyl)pyrimidin-4-yl]amino} -2- hydroxycy clopenty l]methyl sulfamate;

[0613] [(1R,2S.4R)-4-{[5-({4-[(6-chloro-1H-indol-1-yl)methyl]-2-thienyl}carbonvhpvrimidm-4- yl]amino}-2-hydroxycyclopentyl]methylsulfamate;

[0614] [( 1R,2 S,4R)-2-hydroxy-4- { [5-( {4-[(6-methoxy-2,3-dihydro- 1H-indol- 1 -yl)methyl]-5-methyl-2- thienyl}carbonyl)pyrimidin-4-yl]amino}cyclopentyl]methyl sulfamate;

[0615] [(1R,2S,4R)-4-{[5-(5-benzyl-2-furoyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0616] [(1R,2S,4R)-4-{[5-({4-[(6-cyano-2,3-dihydro-1H-indol-1-yl)methyl]-5-methyl-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0617] [(1R,2S,4R)-4-{[5-({4-[(2-chlorophenoxy)methyl]-2-thienyl}carbonyl)pyrimidin-4-yl]amino}-2- hydroxy cyclopentyl]methyl sulfamate;

[0618] [(1R,2S,4R)-2-hydroxy-4-({5-[(4-{[4-(tritluoromethyl')-1H-pyrazol-1-yl]methyl}-2- thienyl (carbonyl ]pvrimidin-4-vl} amino icvclopentyl |methvl sulfamate;

[0619] [(1R,2S,4R)-2-hydroxy-4-{[5-({4-[(2-methylphenoxy)methyl]-2-thienyl}carbonyl)pyrimidin-4- yl]amino}cyclopentyl]methyl sulfamate;

[0620] [(1R,2S,4R}-2-hydroxy-4-{[5-({4-[(6-methyl-1H-indol-1-yl)methyl]-2- thienyl} carbonyl)pyrimidin-4-yl] amino }cy clopenty l]methyl sulfamate;

[0621] [(1R,2R,3 S,4R)-2,3-dihydroxy-4- { [5-(5-phenyl-2-furoyl)pyrimidin-4- yl]amino}cyclopentyl]methyl sulfamate;

[0622] [(1R,2S,4R)-4- {[5-({4-[(1R)-1-(3-chlorophenyl)ethyl]-5-methyl-2-thienyl}carbonyl)pyrimidin-4- yl]amino} -2-hydroxycyclopentyl]methyl sulfamate;

[0623] [(1R,2S,4R)-4-{[5-({4-[(1S)- 1 -( 3-chlorophenyl )ethyl ]-5-methyl-2-thienyl}carbonyl )pyrimidin-4- yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0624] [(1R,2S,4R)-4-{[5-({5-chloro-4-[(1R)-1-hydroxy-2-phenylethyl]-2-thienyl}carbonyl)pyrimidm-4- yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0625] [(1R,2S,4R)-4-{[5-({5-chloro-4-[(1S)-1-hydroxy-2-phenylethyl]-2-thienyl}carbonyl)pyrimidin-4- yl]amino}-2-hydroxycyclopentyl]methyl sulfamate; {( 1R,2S,4R)-2-hydroxy-4-[(5- { [5-methyl-4-(phenylsulfanyl)-2-thienyl]carbonyl}pyrimidin-4- yl)amino]cyclopentyl} methyl sulfamate;

[0626] [( 1 R,2 S ,4R)-4-( { 5- [(4- { [(3 -chlorophenyl)(methyl)amino]methyl } -2-thienyl)carbony 1] pyrimidin-4- yl]amino)-2-hydroxy cyclopentyl} methyl sulfamate;

[0627] [(1R,2 S,4R')-4- { [5-( {4,5-dibenzyl-2-furoyl)pyrimidin-4-yl] amino } -2-hydroxycyclopentyl]methyl sulfamate;

[0628] {(1R,2S;4R)-4-[(5-[4-(cyclohex-1-en-1-ylmethyl)-2-thienyl]carbonyl}pyrimidin-4-yl)amino]-2- hydroxycyclopentyl } methyl sulfamate ;

[0629] [(1R,2S,4R)-4-{[5-({5-chloro-4-[(1S)-1-hydroxy-2-methylprop-2-en-1-yl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0630] [(1R,2S,4R)-4-{[5-({5-chloro-4-[(1R)-1-hydroxy-2-methylprop-2-en-1-yl]-2- thienyl}carbonyl)pyrimidin-4-ylJamino}-2-hydroxycyclopentyl]methyl sulfamate;

[0631] {f1R,2S,4R)-4-[(5-{[5-(3-chlorobenzyl)-4-(hydroxymethyl)-2-thienyl]carbonyl}pyrimidin-4- yl)aminol-2-hydroxycyclopentyl}methyl sulfamate;

[0632] [(1R,2S,4R)-4-{[5-({4-[(3-chlorophenyl)sulfanyl]-5-methyl-2-thienyl}carbonyl)pyrimidin-4- yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0633] [(1R,2S,4R)4-({5-[(4,5-dibenzyl-2-thienyl)carbonyl]pyrimidin-4-yl}amino)-2- hydroxy cy clopenty l]m ethyl sulfamate ;

[0634] [(1R,2S,4R)-2-hydroxy-4-({5-[(5-methyl-4-{[3-(trifluoromethyl)phenyl]sulfanyl}-2- thienyl)carbonyl]pyrimidin-4-yl}amino)cyclopentyl]methyl sulfamate;

[0635] [(1R,2S,4R')-4-{[5-({4-[2-(3-chlorophenyl)ethyl]-2-thienyl}carbonyl)pyrimidin-4-yl]amino}-2- hydroxy cy clopenty l]methyl sulfamate;

[0636] [(1R,2S,4R')-4-({5-[(4-{[(2-chlorophenyl)sulfanyl]methyl}-2-thienyl)carbonyl]pyrimidin-4- yl}amino)-2-hydroxycyclopentyl]methyl sulfamate;

[0637] [( 1R,2 S,4R)-4- { [5-( {4-[(4-bromo-2-cyano-1H-pyrrol- 1 -yl)methyl]-5-methyl-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0638] [(1R,2S,4R)-4-{[5-({5-chloro-4-[(R)-(2,5-dichlorophenyl)(hydroxy)methyl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0639] [(1R,2S,4R)-4-{[5-({5-chloro-4-[(S)-(2,5-dichlorophenyl)(hydroxy)metliyl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0640] [( 1R,2S,4R)-4- { [5-({5-chloro-4-[(1R)-3-cyclopropyl-1-hydroxyprop-2-yn-1-yl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0641] [( 1 R,2S,4R)-4- { [5-( {5-chloro-4-[( 1 S)-3-cyclopropyl- 1 -hydroxyprop-2-yn- 1 -yl] -2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0642] [(1R,2S,4R)-4-{[5-(5-[(1R)-1-(3-chlorophenyl)-1-hydroxyethyl]-2-thienyl}carbonyl)pyrimidin-4- yl] amino }-2-hydroxycyclopentyl]methyl sulfamate;

[0643] [(1R,2S,4R)-4-{[5-(5-[(1S)-1-(3-chlorophenyl)-1-hydroxyethyl]-2-thienyl}carbonyl)pyrimidin-4- yl]amino-2-hydroxycyclopentyl]methyl sulfamate;

[0644] [(1R,2S,4R)-4-{[5-({4-[(R)-(3-chlorophenylXdimethylamino)methyl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0645] [(1R,2S,4R)-4-{[5-({4-[(S)-(3-chlorophenyl)(dimethylamino)methyl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0646] [(1R,2S,4R)-4-{[5-({4-[(2R)-2-(3-chlorophenyl)tetrahydro-2H-pyran-2-yl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0647] [(1R,2S,4R)-4-{[5-({4-[(2S)-2-(3-chlorophenyl)tetrahydro-2H-pyran-2-yl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methylsulfamate;

[0648] {(1R,2S,4R)-4-[(5-{[4-(2,3-dichlorobenzyl)-2-thienyl]carbonyl}pyrimidin-4-yl)amino]-2- hydroxycyclopentyl} methyl sulfamate; [(1R,2S,4R)-4-{[5-({4-[(2-ethoxyphenoxy)methyl]-2-thienyl}carbonyl)pyrimidin-4-yl]amino}-2- hydroxycyclopentyl]methyl sulfamate;

[0649] {(1R,2S,4R')-4-[(5-{[4-(4-chlorobenzyl')-2-thienyl]carbonyl}pyrimidin-4-yl)amino]-2- hydroxycyclopentyl}methyl sulfamate;

[0650] [(1R2R,3S,4R)-4-({5-[5-(2-chlorophenyf)-2-furoyl]pyrimidin-4-yl}amino)-2,3- dihydroxycyclopcntyl]methyl sulfamate;

[0651] [(1R,2S,4R)-4-{[5-({4-[(5-chloro-2,3-dihydro-1H-indol-1-yl)methyl]-5-methyl-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0652] [( 1R,2 S,4R)-2-hydroxy-4- { [5-( {4-[(3-methyl- 1 H-pyrazolo[3 ,4-c]pyridin- 1 -yl)methyl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}cyclopentyl]methyl sulfamate;

[0653] [(1R,2S,4R)-2-hydroxy-4-{[5-({4-[(2-iodophenoxy)methyl]-2-thienyl}carbonyl)pyrimidin-4- yl]amino}cyclopentyl]methyl sulfamate;

[0654] [( 1R,2R,3R,4R)-4-( { 5-[(4-benzyl-5-chloro-2-thienyl)carbonyl]pyrimidin-4-yl] amino )-3-fluoro-2- hydroxycyclopentyl}methyl sulfamate;

[0655] [(1R,2S,4R)-4-{[5-({4-[(4-chlorophenoxy)methyl]-2-thienyl}carbonyl)pyrimidin-4-yl]amino}-2- hydroxycyclopentyl]methyl sulfamate;

[0656] [(1R,2S,4R)4-{[5-({4-[l-(3-bromophenyl)vinyl]-2-thienyl}carbonyl)pyrimidin-4-yl]amino}-2- hydroxycyclopentyl]m ethyl sulfamate;

[0657] {(1R,2S,4R)-4-[(5-{[4-(2-chlorobenzyl)-2-thienyl]carbonyl}pyrimidin-4-yl)amino]-2- hydroxy cyclopentyl {methyl sulfamate;

[0658] [(1R,2S,4R')-4-{[5-({4-[(2S')-2-cyclohexyltetrahydrofuran-2-yl]-2-thienyl}carbonyl)pyrimidin-4- yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0659] [(1R,2S,4R -4-{[5-({4-[(2R)-2-cyclohexyltetrahydrofuran-2-yl]-2-thienyl}carbonyl)pyrimidin-4- yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0660] {(1R,2S,4R)-4-[(5-{[4-(3,4-dihydroisoquinolin-2(1H)-ylmethyl)-2-thienyl]carbonyl}pyrimidin-4- yl)amino] -2-hy droxycyclopentyl } methyl sulfamate ;

[0661] {}1R,2S,4R)-4-[(5-{[4-(6,7-dihydrothieno[3,2-c]pyridin-5(4H)-ylmethyl)-2- thienyl] carbonyl } py rimidin-4-yl)amino] -2-hy droxy cyclopentyl } methyl sulfamate ;

[0662] [(1R,2S,4R)-4-{[5-({4-[(6-cyano-1H-indol-1-yl)methyl]-2-thienyl}carbonyl)pyrimidin-4- yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0663] [( 1R,2S,4R)-2-hydroxy-4- { [5-( {4-[(3-methyl- 1 H-pyrrol- 1 -yl)methyl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}cyclopentyl]methyl sulfamate;

[0664] {(1R,2S,4R)-4-[(5-{[4-(2,3-dihydro-1H-pyrrolo[2,3-b]pyridin-1-ylmethyl)-5-methyl-2- thienyl] carbonyl }pyrimidin-4-y Ham ino] -2-hy droxy cyclopentyl {methyl sulfamate;

[0665] [(1R,2S,4R)-4-{[5-(5-chloro-4-[(R)-cyclohexyl(hydroxy)methyl]-2-thienyl}carbonyl)pyrimidin-4- yl] amino} -2-hy droxy cyclopentyl]methyl sulfamate;

[0666] [(1R,2S,4R')-4-{[5-({5-chloro-4-[(S)-cyclohexyl(hydroxy)methyl]-2-thienyl}carbonyl')pyrimidin-

[0667] 4-yl] amino} -2-hydroxycyclopentyl]methyl sulfamate;

[0668] {(1R,2S,4R)-4-[(5-{[4-(3,6-dihydro-2H-thiopyran-4-ylmethyl)-2-thienyl]carbonyl}pyrimidin-4- yl)amino] -2-hy droxy cyclopentyl } methyl sulfamate ;

[0669] [(1R,2S,4R)-4- {[5-(5-chloro-4-[(R)-hydroxy(tetrahydro-2H-pyran-4-yl)methyl]-2- thienyl} carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0670] [(1R,2S,4R)-4-{[5-({5-chloro-4-[(S)-hydroxy(tetrahydro-2II-pyran-4-yl)methyl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0671] [(1R,2S,4R)-4-{[5-({5-chloro-4-[(3-chlorophenyl)sulfanyl]-2-thienyl}carbonyl)pyrimidin-4- yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0672] [(1R,2S,4R)-2-hydroxy-4-({5-[(4- { [2-(trifluoromethoxy)phenoxy]methyl}-2- thienyl)carbonyl]pyrimidin-4-yl } amino)cyclopentyl]methyl sulfamate; [(1R,2S,4R)-2-hydroxy-4-{[5-({4-[(phenylsulfanyl)methyl]-2-thienyl}carbonyl)pyrimidin-4- yl]amino} cyclopentyl]methyl sulfamate;

[0673] [(1R,2S,4R)-4-{[5-({4-[2-(4-chlorophenyl)ethyl]-2-thienyl}carbonyl)pyrimidin-4-yl]amino}-2- hydroxycyclopentyl]methyl sulfamate;

[0674] {(1R,2S,4R)-4-[(5-{[4-(3-chlorobenzyl)-5-cyano-2-thienyl]carbony1}pyrimidin-4-yl)amino]-2- hydroxy cyclopcntyl}methyl sulfamate;

[0675] [(1R,2S,4R)-4-{[5-({4-[(2,3-dichlorophenoxy)methyl]-2-thienyl}carbonyl)pyrimidin-4-yl]amino}- 2-hydroxycyclopentyl]methyl sulfamate;

[0676] [(1R,2S,4R)-4-{[5-({4-[(3-chlorophenyl)sulfonyl]-5-methyl-2-thienyl}carbonyl)pyrimidin-4- yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0677] [(1R,2S,4R)-4-{[5-({4-[(2-ethylphenoxy)methyl]-2-thienyl}carbonyl)pyrimidin-4-yl]amino}-2- hydroxy cyclopentyljmethyl sulfamate;

[0678] [(1R,2S,4R)-2-hydroxy-4-{[5-({4-[2-(2-methoxyphenyl)ethyl]-2-thienyl}carbonyl)pyrimidin-4- yl]amino } cyclopentyl]methyl sulfamate;

[0679] [(1R, 2S,4R)- 2-hydroxy-4-({5-[(4-{[6-(trifluoromethyl)-1H-indol-1-yl]methyl}-2- thienyl)carbonyl]pyrimidin-4-yl } amino)cyclopentyl]methyl sulfamate;

[0680] {(1R,2S,4R)-4-[(5-{[4-(3-cyanobenzyl)-2-thienyl]carbonyl}pyrimidm-4-yl)amino]-2- hydroxycyclopentyl } methyl sulfamate;

[0681] {(1R,2S,4R)-2-hydroxy-4-[(5-{[4-(1H-pyrrolo[2,3-c]pyridin-1-ylmethyl)-2- thienyl] carbony 1} pyrimidin-4-yl)amino] cyclopentyl } methyl sulfamate ;

[0682] {(1R,2S,4R)-4-[(5-{[4-(l,3-dihydro-2H-isoindol-2-ylmetliyl)-2-thienyl]carbonyl}pyrimidin-4- yl)amino]-2-hydroxycyclopentyl} methyl sulfamate;

[0683] [(1R,2S,4R)-2-hydroxy-4-({5-[(4-{[2-(trifluoromethyl)phenoxy]methyl}-2- thienyl)carbonyl]pyrimidin-4-yl}amino)cyclopentyl]methyl sulfamate;

[0684] [(1R,2S,4R)-4-({5-[(5-chloro-4-{[3-(trifluoromethyl)phenyl]sulfanyl}-2- thienyl)carbonyl]pyrimidin-4-yl}amino)-2-hydroxycyclopentyl]methyl sulfamate;

[0685] {(1R,2S,4R)-4-[(5-{[4-(5,6-dihydroimidazo[1,2-a]pyrazin-7(8H)-ylmethyl)-2- thienyl] carbonyl } pyrimidin-4-yl)amino] -2-hydroxycyclopentyl {methyl sulfamate ;

[0686] [(1R,2S,4R)-2-hydroxy-4-{[5-({4-[(2-isopropylphenoxy)methyl]-2-thienyl}carbonyl)pyrimidin-4- yl]amino}cyclopentyl]methyl sulfamate;

[0687] {(1R,2S,4R)-4-[(5-{[4-(1H-benzimidazol-1-ylmethyl)-2-thienyl]carbonyl}pyrimidin-4-yl)amino]- 2-hydroxycyclopentyl}methyl sulfamate;

[0688] {(1R,2S,4R)-4-[(5-{[4-(2,5-dihydrofuran-3-ylmethyl)-2-thienyl]carbonyl}pyrimidin-4-yl)amino]- 2-hydroxycyclopentyl }methyl sulfamate;

[0689] [(1R,2S,4R)-4-{[5-({4-[(3-cyano-1H-pyrrol-1-yl)methyl]-5-methyl-2-thienyl}carbonyl)pyrimidin- 4-yl] amino} -2-hydroxycyclopentyljmethyl sulfamate;

[0690] [(1R,2S,4R)-4-{[5-({5-chloro-4-[(1R)-1-hydroxy-2-methylpropyl]-2-thienyl}carbonyl)pyrimidin- 4-yl]amino} -2-hydroxy cyclopentyljmethyl sulfamate;

[0691] [(1R,2S,4R)-4-{[5-({5-chloro-4-[(1S)-1-hydroxy-2-methylpropyl]-2-thienyl}carbonyl)pyrimidin- 4-yl] amino} -2-hydroxycyclopentyljmethyl sulfamate;

[0692] {(1R,2S,4R)-4-[(5-{[4-(3,6-dihydro-2H-pyran-4-ylmethyl)-2-thienyl]carbonyl}pyrimidin-4- yl)amino] -2-hydroxy cyclopentyl {methyl sulfamate;

[0693] {(1R,2S,4R)-2-hydroxy-4-[(5-{4-[(1S)-1-hydroxy-2-methylprop-2-en-1-yl]-2-furoyl}pyrimidin-4- yl)amino] cyclopentyl {methyl sulfamate ;

[0694] {(1R,2S,4R)-2-hydroxy-4-[(5-{4-[(1R)-1-hydroxy-2-methylprop-2-en-1-yl]-2-furoyl}pyrimidin-4- yl)amino] cyclopentyl} methyl sulfamate ;

[0695] {(1R,2S,4R)-4-[(5-{[4-(cyclohexylmethyl)-2-thienyl]carbonyl}pyrimidin-4-yl)amino]-2- hydroxycyclopentyl } methyl sulfamate; {( 1R,2S,4R)-2-hydroxy-4-[(5- { [4-(phenylsulfonyl}-2-thienyl]carbonyl}pyrimidin-4- yl)amino]cyclopentyl} methyl sulfamate;

[0696] [(1R,2S,4R)-2-hydroxy-4-{[5-({4-[(2-isopropoxyphenoxy)methyl]-2-thienyl}carbonyl)pyrimidin- 4-yl] amino } cyclopentyl]methyl sulfamate;

[0697] {(1R,2R,3S,4R)-2,3-dihydroxy-4-[(5-{[5-(2-hydroxypropan-2-yl)-2-thienyl]carbonyl}pyrimidin-4- yl)amino]cyclopentyl} methyl sulfamate;

[0698] [( 1 R,2 S,4R)-4- { [5-( { 5-chloro-4- [(3 -chlorophenyl)sulfony 1] -2-thienyl } carbonyl)pyrimidin-4- yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0699] { (1 R,2 S,4R)-4- [(5- { [4-(3 ,6-dihy dropyridin- 1 (2H)-y Imethyl)-2-thienyl] carbony 1} pyrimidin-4- yl)amino] -2-hydroxycyclopentyl } methyl sulfamate ;

[0700] {(1R,2R,3R,4R)-4-[(5-{[5-(3-chlorobenzyl)-2-thienyl]carbonyl}pyrimidin-4-yl)amino]-3-fluoro-2- hydroxycyclopentyl]methyl sulfamate;

[0701] [(1R,2S,4R)-2-hydroxy-4-({5-[5-(hydroxymethyl)-2-furoyl]pyrimidin-4- y 1] amino)cy clopentyl} methyl sulfamate ;

[0702] {(1R,2S,4R)-4-[(5-{[5-chloro-4-(3-chlorobenzoyl)-2-thienyl]carbonyl}pyrimidin-4-yl)amino]-2- hydroxycyclopentyl } methyl sulfamate;

[0703] [(1R,2S,4R)-4-{[5-({4-benzoyl-2-furoyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0704] {(1R,2S,4R)-4-[(5-{[(2S)-2-(3-chlorophenyD-2,3,4,5-tetrahydro-2,3'-bithiophen-5'- yl]carbonyl}pyrimidin-4-yl')amino]-2-hydroxycyclopentyl}methyl sulfamate;

[0705] {(1R,2S,4R)-4-[(5-[(2R)-2-(3-chlorophenyl)-2,3,4,5-tetrahydro-2,3'-bithiophen-5'- yl] carbonyl } pyrimidin-4-yl)amino] -2-hydroxycyclopentyl } methyl sulfamate ;

[0706] [(1R,2S,4R)-4-{[5-({4-[(5-chloropyridin-3-yl)methyl]-2-thienyl}carbonyl)pyrimidin-4-yl]amino}- 2-hydroxycyclopentyl]methyl sulfamate;

[0707] {(1R,2S,4R)-4-[(5-{[5-chloro-4-(hydroxymethyl)-2-thienyl]carbonyl}pyrimidin-4-yl)amino]-2- hydroxycyclopentyl}methyl sulfamate;

[0708] [(1R,2S,4R)-2-hydroxy-4-({5-[5-(methoxymethyl)-2-furoyl]pyrimidin-4- yl } amino)cyclopentyl]methyl sulfamate;

[0709] [(1R,2S,4R)-4-( {5-[(4-benzoyl-5-chloro-2-thienyl)carbonyl]pyrimidin-4-yl} amino )-2- hydroxycyclopentyl]methyl sulfamate;

[0710] [(1R,2S,4R)-2-hydroxy-4-({5-[4-(hydroxymethyl)-2-furoyl]pyrimidin-4- yl } amino {cyclopentyl ]methyl sulfamate;

[0711] {( 1R,2S,4R)-2-hydroxy-4-[(5- { [4-( methoxymethyl )-2-thienyl ]carbonyl}pyrimidin-4- yl] (amino] cyclopentyl } methyl sulfamate ;

[0712] { ( 1 R,2 S,4R)-4- [(5- { [5-chloro-4-(methoxymethyl)-2-thienyl] carbonyl } pyrimidin-4-yl)amino] -2- hydroxycyclopentyl {methyl sulfamate;

[0713] {( 1 R, 2 S,4R)-4- [(5- { [4-(2 ,5-dihydro- 1 H-pyrrol- 1 -ylmethyl)- 2-thienyl] carbony 1 } pyrimidin-4- yl)amino]-2-hydroxycyclopentyl} methyl sulfamate;

[0714] [(1R,2S,4R)-2-hydroxy-4-({5-[4-(2-hydroxypropan-2-yl)-2-furoyl]pyrimidin-4- yl}amino)cyclopentyl]methyl sulfamate;

[0715] {(1R,2S,4R)-4-[(5-{[4-({3-[(dimethylamino)methyl]-1H-indol-1-yl]methyl)-2- thienyl} carbony l}pyrimidin-4-yl)amino]-2-hydroxycyclopentyI]methyl sulfamate;

[0716] [( 1 R,2 S,4R)-4- { [5-( { 4- [(benzylamino {methyl] -2-thienyl } carbonyl)pyrimidin-4-y 1] amino-2- hydroxycyclopentyl } methyl sulfamate ;

[0717] [(1R,2S,4R)-2-hydroxy-4-({5-[4-(methoxymethyl)-2-furoyl]pyrimidin-4- yl] amino)cyclopentyl} methyl sulfamate ;

[0718] [(1R,2S,4R)-4-{[5-({4-[(3,3-difluoropiperidin-1-yl)methyl]-2-thienyl}carbonyl)pyrimidin-4- yl]amino}-2-hydroxycyclopentyl]methyl sulfamate; [(1R,2S,4R)-2-hydroxy-4-({5-[(4-{[(3R)-3-methylpiperidin-1-yl]methyl}-2- thienyl)carbonyl ]pyrimidin-4-yl ] amino ]cyclopentyl} methyl sulfamate;

[0719] [(1R,2S,4R)-2-hydroxy-4-({5-[(4-{[(3S -3-methylpiperidin-1-yl]methyl}-2- thienyl)carbonyl]pyrimidin-4-yl}amino)cyclopentyl]methyl sulfamate;

[0720] [(1R,2S,4R)-4-{[5-({5-chloro-4-[(1S)-1-hydroxyethyl]-2-thienyl}carbonyl)pyrimidin-4- yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0721] [(1R,2S,4R)-4-{[5-{[5-chloro-4-[(1R)-1-hydroxyethyl]-2-thienyl}carbonyl)pyrimidin-4- yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0722] [(1R,2S,4R)-4-{[5-({4-[(2S)-2-(3-chlorophenyl)-1-methylpyrrolidin-2-yl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0723] [(1R,2S,4R)-4-{[5-({4-[(2R)-2-(3-chlorophenyl)-1-methylpyrrolidin-2-yl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0724] [(1R,2S,4R)-4-{[5-({4-[(3,3-difluoropyrrolidin-1-yl)methyl]-2-thienyl}carbonyl)pyrimidin-4- yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0725] [(1R,2S,4R)-4-({5-[(4-acetyl-2-thienyl)carbonyl]pyrimidin-4-yl}aminol-2- hydroxycyclopentyl]methyl sulfamate;

[0726] {(1R,2S,4R)-2-hydroxy-4-1(5-{[4-(hydroxymethyl)-2-thienyl]carbonyl}pyrimidin-4- yl)amino] cyclopentyl } methyl sulfamate ;

[0727] {(1R,2S,4R)-2-hydroxy-4-[(5-{[4-(1H-imidazol-1-ylmethyl)-2-thienyl]carbonyl}pyrimidin-4- yl)amino]cyclopentyl ; methyl sulfamate;

[0728] {(1R,2S,4R)-4-[(5-{[5-(3-chlorobenzyl)-3-methyl-2-thienyl]carbonyl}pyrimidin-4-yl)amino]-2- hydroxycyclopentyl}methyl sulfamate;

[0729] [(1R,2S,4R)-4-{[5-(4-acety4-2-furoyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0730] {(1R,2S,4R)-2-hydroxy-4-[(5-{[5-(2-hydroxypropan-2-yl)-2-thienyl]carbonyl}pyrimidin-4- yl)amino] cyclopentyl} methyl sulfamate ;

[0731] [(1R,2R,3S,4R)-4-[2-chloro-5-(5-phenyl-2-furoyl)pyrimidin-4-yl]amino}-2,3- dihydroxycyclopentyl]methyl sulfamate;

[0732] [(1R,2S,4R)-2-hydroxy-4-{[5-({4-[(5-methyl-2-furyl)methyl]-2-thienyl}carbonyl)pyrimidin-4- yl]amino } cyclopentyl]methyl sulfamate;

[0733] [(1R,2S,4R)-4-{[5-({4-[(2S)-2-cyclopropyltetrahydrofuran-2-yl]-2-thienyl}carbonyl)pyrimidin-4- yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0734] [(1R,2S,4R)-4-{[5-({4-[(2R)-2-cyclopropyltetrahydrofuran-2-yl]-2-thienyl}carbonyl)pyrimidin-4- yl] amino } -2-hydroxycyclopentyl }methyl sulfamate ;

[0735] [(1R,2S,4R)-2-hydroxy-4-({5-[5-(phenylsulfonyl)-2-furoyl]pyrimidin-4- yl}amino)cyclopentyl]methyl sulfamate;

[0736] {(1R,2S,4R)-2-hydroxy-4-[(5-{[4-(2-hydroxypropan-2-yl)-2-thienyl]carbonyl}pyrimidin-4- yl)amino]cyclopentyl} methyl sulfamate;

[0737] [(1R,2S,4R)-2-hydroxy-4-{[5-({4-[(4-phenylpiperazin-1-yl)methyl]-2-thienyl}carbonyl)pyrimidin- 4-yl]amino}cyclopentyl]methyl sulfamate;

[0738] [(1R,2S,4R)-4-({5-[(4-acetyl-5-chloro-2-thienyl)carbonyl]pyrimidin-4-yl}amino)-2- hydroxy cy clopenty l]me thy 1 sulfamate ;

[0739] [(1R,2S,4R)-4-{[5-({4-[(4-bromo-lII-imidazol-1-yl)methyl]-2-thienyl}carbonyl)pyrimidm-4- yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0740] [(1R,2R,3S,4R)-4-{[5-({4-[(1R)-7-bromo-1,2,3,4-tetrahydroisoquinolin-1-yl]-5-methyl-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2,3-dihydroxycyclopentyl]methylsulfamate;

[0741] [(1R,2R,3S,4R)-4-{[5-({4-[(1S)-7-bromo-1,2,3,4-tetrahydroisoquinolin-1-yl]-5-methyl-2- thienyl}carbnyl)pyrimidin-4-yl]amino}-2,3-dihydroxycyclopentyl]methyl sulfamate; [(1R,2S,4R)-4-{[5-({4-[(1S)-7-bromo-1,2,3,4-tetrahydroisoquinolm-1-yl]-5-methyl-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0742] [(1R,2S,4R)-4-{[5-({4-[(1R)-7-bromo-1,2,3,4-tetrahydroisoquinolin-1-yl]-5-methyl-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0743] [(1R,2R,3 S,4R)-4- { [5-( { 5-chloro-4- [( 1 R)-7-chloro- 1,2,3 ,4-tetrahydroisoquinolin- 1 -yl] -2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2,3-dihydroxycyclopcntyl]methyl sulfamate;

[0744] [(1R,2R,3S,4R)-4-{[5-({5-chloro-4-[(1S)-7-chloro-1,2,3,4-tetrahydroisoquinolin-1-yl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2,3-dihydroxycyclopentyl]methyl sulfamate;

[0745] [(1R,2R,3S,4R)-4-{[5-({4-[(1R)-3,4-dihydro-1H-isochromen-1-yl]-5-methyl-2- thienyl}carbonyl)pyrimidin-4-yl]amiiio}-2,3-dihydroxycyclopentyl]methyl sulfamate;

[0746] [(1R,2R,3S,4R)-4-{[5-({4-[(1S)-3,4-dihydro-1H-isochromen-1-yl]-5-methyl-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2,3-dihydroxycyclopentyl]methyl sulfamate;

[0747] [(1R,2S,4R)-4-{[5-({4-[(1R)-7-bromo-1,2,3,4-tetrahydroisoquinolin-1-yl]-5-chloro-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0748] [(1R,2S,4R)-4-{[5-({4-[(1S)-7-bromo-1,2,3,4-tetrahydroisoquinolin-1-yl]-5-chloro-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0749] [(1R,2S,4R)-4-{[5-({4-[(1R)-7-ethynyl-3,4-dihydro-1H-isochromen-1-yl]-5-methyl-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0750] [(1R,2S,4R)-4-{[5-({4-[(1S)-7-ethynyl-3,4-dihydro-1H-isochromen-1-yl]-5-methyl-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0751] [(1R,2S,4R)-4-{[5-({5-chloro-4-[(8S')-2-chloro-5,6,7,8-tetrahydro-1,7-naphthyridin-8-yl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0752] [(1R,2S,4R)4-{[5-({5-chloro-4-[(8R)-2-chloro-5,6,7,8-tetrahydro-1,7-naphthyridin-8-yl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0753] [(1R,2S,4R)-4-{[5-({4-[(1R)-3,4-dihydro-1H-isochromen-1-yl]-2-thienyl}carbonyl)pyrimidin-4- yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0754] [(1R,2S,4R)-4-{[5-({4-[(1S)-3,4-dihydro-1H-isochromen-1-yl]-2-thienyl}carbonyl)pyrimidin-4- yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0755] [(1R,2S,4R)-4-{[5-({5-chloro-4-[(1R)-7-chloro-1,2,3,4-tetrahydroisoquinolin-1-yl]-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0756] [( 1R,2S,4R)-4- {[5-({5-chloro-4-[( 1 S)-7-chloro- 1 ,2,3,4-tetrahydroisoquinolin- 1 -yl] -2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0757] [( 1R,2S,4R)-4- { [5-( { 4- [( 1R)-3 ,4-dihydro- 1H-isochromen- 1 -yl]-5-methyl-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0758] [(1R,2S,4R)-4-{[5-({4-[(1S)-3,4-dihydro-1H-isochromen-1-yl]-5-methyl-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0759] [(1R,2 S,4R)-4- { [5-( {4- [( 1 R)-7-chloro-3 ,4-dihydro- 1H-isochromen- 1 -yl]-5-methyl-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0760] [(1R,2S,4R)-4-{[5-({4-[(lS)-7-chloro-3,4-dihydro-1H-isochromen-1-yl]-5-methyl-2- thienyl}carbonyl)pyriniidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0761] [( 1R,2 S,4R)-2-hydroxy-4- { [ 5 -( { 5-methyl-4-[(l R)-7-(trifluoromethyl)-3 ,4-dihydro- 1H- isochromen-1-yl]-2-thienyl}carbonyl)pyrimidin-4-yl]amino}cyclopentyl]methyl sulfamate;

[0762] [(1R,2S,4R)-2-hydroxy-4-{[5-({5-methyl-4-[(l)-7-(trifluoromethyl)-3,4-dihydro-HI-isochromen- 1-yl] -2-thienyl } carbonyllpyrimidin-4-yl] amino } cyclopentyl] methyl sulfamate ;

[0763] [(1R,2R,3S,4R)-4-{[5-({4-[(1R)-7-bromo-1, 2,3,4- tetrahydroisoquinolin-1-yl]-5-chloro-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2,3-dihydroxycyclopentyl]methyl sulfamate;

[0764] [(1R,2R,3 S,4R)-4- { [5-( {4- [(1S)-7-bromo- 1 ,2,3 ,4-tetrahydroisoquinolin- 1 -yl]-5-chloro-2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2,3-dihydroxycyclopentyl]methyl sulfamate; [(1R,2S,4R)-4-{[5-({4-[(8R)-2-chloro-5,5-difluoro-5,8-dihydro-6H-pyrano[3,4-b]pyridin-8-yl]-5- metliyl-2-thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0765] [(1R,2S,4RM-{[5-({4-[(8S)-2-chloro-5,5-difluoro-5,8-dihydro-6H-pyrano[3,4-b]pyridin-8-yl]-5- methyl-2-thienyl}carbonyl]pyrimidin-4-yl ]amino}-2-hydroxy cyclopentyl] methyl sulfamate;

[0766] [(1R,2S,4R)-4- { [5-( {4- [(1R)-7-chloro-3 ,4-dihydro- 1H-isochromen-1-yl] -2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopcntyl]methyl sulfamate;

[0767] [(1R,2S,4R)-4-{[5-({4-[(1S)-7-chloro-3,4-dihydro-1H-isochromen-1-yl]-2- thienyl} carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0768] [( 1R,2 S,4R)-4- { [5-( { 5-chloro-4- [(1R)-6-chloro-2, 3 -dihydro-1H-isoindol-1-yl] -2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentyl]methyl sulfamate;

[0769] [( 1R,2S,4R)-4- { [5-( { 5-chloro-4-[(1S)-6-chloro-2,3-dihydro- 1H-isoindol-1-yl] -2- thienyl}carbonyl)pyrimidin-4-yl]amino}-2-hydroxycyclopentylJmethyl sulfamate.

[0770] Products of manufacture and Kits

[0771] Provided are products of manufacture and kits for practicing methods as provided herein; and optionally, products of manufacture and kits can further comprise instructions for practicing methods as provided herein.

[0772] Any of the above aspects and embodiments can be combined with any other aspect or embodiment as disclosed here in the Summary, Figures and / or Detailed Description sections.

[0773] As used in this specification and the claims, the singular forms “a,” “an” and “the” include plural referents unless the context clearly dictates otherwise.

[0774] Unless specifically stated or obvious from context, as used herein, the term “or” is understood to be inclusive and covers both “or” and “and”.

[0775] Unless specifically stated or obvious from context, as used herein, the term “about” is understood as within a range of normal tolerance in the art, for example within 2 standard deviations of the mean. About (use of the term “about”) can be understood as within 20%. 19%. 18%. 17%. 16%. 15%. 14%. 13%. 12% 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, 0.05%, or 0.01% of the stated value. Unless otherwise clear from the context, all numerical values provided herein are modified by the term “about.”

[0776] Unless specifically stated or obvious from context, as used herein, the terms “substantially all”, “substantially most of”, “substantially all of” or “majority of” encompass at least about 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 99.5%, or more of a referenced amount of a composition.

[0777] The entirety of each patent, patent application, publication and document referenced herein hereby is incorporated by reference. Citation of the above patents, patent applications, publications and documents is not an admission that any of the foregoing is pertinent prior art, nor does it constitute any admission as to the contents or date of these publications or documents. Incorporation by reference of these documents, standing alone, should not be construed as an assertion or admission that any portion of the contents of any document is considered to be essential material for satisfying any national or regional statutory disclosure requirement for patent applications. Notwithstanding, the right is reserved for relying upon any of such documents, where appropriate, for providing material deemed essential to the claimed subject matter by an examining authority or court.

[0778] Modifications may be made to the foregoing without departing from the basic aspects of the invention. Although the invention has been described in substantial detail with reference to one or more specific embodiments, those of ordinary skill in the art will recognize that changes may be made to the embodiments specifically disclosed in this application, and yet these modifications and improvements are within the scope and spirit of the invention. The invention illustratively described herein suitably may be practiced in the absence of any element(s) not specifically disclosed herein. Thus, for example, in each instance herein any of the terms "comprising", “consisting essentially of”, and “consisting of” may be replaced with either of the other two terms. Thus, the terms and expressions which have been employed are used as terms of description and not of limitation, equivalents of the features shown and described, or portions thereof, are not excluded, and it is recognized that various modifications are possible within the scope of the invention. Embodiments of the invention are set forth in the following claims.

[0779] The invention is further described with reference to the examples described herein; however, it is to be understood that the invention is not limited to such examples.

[0780] EXAMPLES

[0781] Unless stated otherwise in the Examples, all recombinant DNA techniques are carried out according to standard protocols, for example, as described in Sambrook et al. (2012) Molecular Cloning: A Laboratory Manual, 4th Edition, Cold Spring Harbor Laboratory Press, NY and in Volumes 1 and 2 of Ausubel et al. (1994) Current Protocols in Molecular Biology, Current Protocols, USA. Other references for standard molecular biology techniques include Sambrook and Russell (2001) Molecular Cloning: A Laboratory Manual, Third Edition, Cold Spring Harbor Laboratory Press, NY, Volumes I and II of Brow n (1998) Molecular Biology LabFax, Second Edition, Academic Press (UK). Standard materials and methods for polymerase chain reactions can be found in Dieffenbach and Dveksler (1995) PCR Primer: A Laboratory Manual, Cold Spring Harbor Laboratory Press, and in McPherson at al. (2000) PCR - Basics: From Background to Bench, First Edition, Springer Verlag, Germany.

[0782] Example 1: Exemplary Methods

[0783] This example demonstrates that methods as provided can be effective for treating, ameliorating, lessening the symptoms of, or preventing: angina, myocardial infarction, reperfusion injury, a ventricular arrhythmia (VA), cardiac pump failure due to my ocardial infarction (MI) (optionally cardiac pump failure due to my ocardial infarction (MI) in an early ischemia / reperfusion (I / R) period), congestive heart failure (CHF), hypovolemia, acidosis, hyperkalemia, hypokalemia, a cardiac poisoning or cardiopathology due to a toxin or poisoning, cardiac tamponade, tension pneumothorax, coronary hrombosis, sepsis, shock, pulmonary thrombosis, pulmonary embolism, respiratory decompensation, arrythmia a cardiac event caused by hypoperfusion and / or cardiac arrest, by administering to an individual in need thereof a drug or composition capable of suppressing hypoxia-induced SUMOylation of a cardiac sodium channel NaV1.5.

[0784] Cardiac ILATEcurrent and arrhythmia. In healthy hearts, ILATEis less than 0.5% of the peak sodium current (iNa) and permeates through a small number of NaV1.5 channels that do not stay closed after inactivation (2). In patients with ischemic and failing heart, ILATEcan increase to 4%-5% of peak INa, a level we and others have also shown in subty pes of cardiac sudden infant death and long QT syndrome (3 -5). Acute hypoxia and ischemia are recognized to increase ILATEin cardiac myocytes (6-8) prior to slower processes like remodeling (18). Excess ILATEis pro-arrhythmic because it prolongs action potential duration (APD), reducing repolarization reserve, increasing susceptibility to after-depolarizations, and causing a predisposition to TdP, a ventricular dysrhythmia that is lethal when sustained, with increases in ILATEabove baseline by as little 2-fold associated with sudden cardiac death (3,5,9-11). These observations motivated studies by our group and others to investigate inhibition of late NaV1.5 re-openings with medications like mexiletine and ranolazine4,8,19 and this proposal to suppress the regulatory event that generates ILATEwith MI, hypoxia- induced SUMOylation of NaVl.5.1 SUMOylation at the PM. Among PTMs that regulate cardiac proteins (i.e., phosphorylation, acetylation, and ubiquitinoylation). Small Ubiquitin-like Modifier (SUMO) protein modification (SUMOylation) at the PM has been little studied. Present in all eukaryotic cells, the SUMO pathway, and its component enzy mes, were first identified to regulate the trafficking, activity, and lifespan of nuclear proteins, now including more than 60 transcription factors, mediating their movement between the cytosol and nucleus to control cell division. DNA repair and cellular metabolism in adaptation to environmental changes.20-22 Therefore, it was unexpected when we demonstrated the direct SUMOylation of Na+ and K+ channels in the PM of nerves and muscles to regulate excitability, showing that SUMOylation of different channel types varies (1,23-28) and that NaV1.2 in the central nervous system (CNS) neurons (28) and NaV1.5 in cardiac myocytesl show little SUMOylation at baseline but are modified within min in response to hypoxia.

[0785] Given the important role of hypoxia-induced increases in ILATEin MI morbidity and mortality, we test the hypothesis that SUMOylation of Nav1.5 channels may offer a new target for therapy. More generally, we propose to elucidate the mechanism of SUMOy lation at the PM since we have shown that SUMOylation can proceed in off-cell membrane patches, supporting the notion that the SUMO enzy me cascade is stably resident on the cytosolic face of the PM, not just at the nucleus and in the cytosol (23-25). Our studies may inform investigations of heart failure and cardiac hypertrophy where the SUMO pathway exercises its classical role in metabolic adaptation to stress (29,30). The studies is relevant to other cardiac ion channels we have shown to be SUMOylated (27) and in the CNS since we have shown SUMOylation of NaV1.2 regulates both backpropagation (31) and deleterious neuronal sodium loading with hypoxia (28).

[0786] SUMOylation of NaV1.5 is a novel therapeutic target for MI; whereas the channel has been targeted by existing medication to suppress ILATE, the drugs have significant side-effects, in some cases exacerbating arrhythmias. Here, we seek to prevent and suppress the pathology before it is fully manifest. Further, SUMOylation of PM targets to regulate excitability in health and disease is a novel strategy. The SUMO pathway has recently been identified as a therapeutic target for heart failure based on its regulation of nuclear events and cellular metabolism. Novel preliminary data. We created a KI mouse that ablates NaV1.5 SUMOylation and have preliminary data to support the hypothesis that suppressing SUMOylation of NaV 1.5 can decrease VA and, unexpectedly, ameliorate cardiac dysfunction. Moreover, we demonstrate that SUMO pathway inhibitor in phase I / II trials can inhibit Nav1.5 SUMOylation and protect WT mice subjected to LAD occlusion, therefore the study is extremely timely. The SUMO pathway and inhibitors

[0787] The SUMO pathway. Our observation that K+ and Na+ channels at the PM are subject to regulation by covalent linkage of SUMOs (1,23,24,26-28,31,34) should, perhaps, have been more readily accepted. Although the majority of vertebrate targets are nuclear transcription factors, SUMO activating, conjugating, and cleavage enzymes were first identified in the yeast X cerevisiae. including deSUMOylases (SENPs) that move near the PM at the neck between mother and daughter cells (35,36). In vertebrates, the pathway has three SUMO isoforms (SUMO -3) of approximately 100 amino acids that covalently linked to the s-amino group of specific intracellular target lysine residues to alter function (Fig 1) (37) SUMO1 is approximately 50% identical to SUMO2 and SUMO3, that differ at only 4 residues. Although SUMOylation is a covalent PTM, it is subject to reversal by SENPs 1-7, which vary in tissue distribution and roles; SENP2 / 3 are most significant to homeostasis in healthy, adult human heart (38).

[0788] SUMO pathway inhibitors. Given the role of the SUMO pathway in cellular metabolism via nuclear transcription factors, El and E2 have been targeted for the treatment of cancer. Among the emerging pharmacopeia, TAK-981, brand name SUBASUMSTAT™, selectively inhibits the El activating enzyme by forming a covalent adduct with SUMO in the catalytic site and has promising activity in preclinical models for lymphomas (2). pancreatic adenocarcinomas (3) and multiple myeloma (4), and has advanced to four phase 1 / 2 clinical trials (NCT03648372, NCT04074330, NCT04381650, and NCT04776018), showing “a manageable safety profile along with preliminary anti-tumor activity” (5,6). 2-D08 is a selective inhibitor of the E2 SUMO conjugase (Ubc9) that prevents transfer of SUMO to the target (7) and has been used to study SUMOylation in preclinical work on genome integrity (8), cancer (9,10), hepatic ischemic reperfusion injury (11), and viral replication (12,13). SUMOylation of ion channels. Since we first reported SUMO regulation of K2P 1 channels in 2005 (23) we have additionally reported SUMOylation of other K+ and Na+ channels at the PM of excitable cells, demonstrating the PTM to increase excitability by decreasing K+ flux or increasing Na+ currents, and deSUMOylation to produce the opposite effects. The SUMOylated channels we have described include K2P1; K2P1 / K2P3 and K2P1 / K2P9 (IKSO); Kv2.1; KCNQ1 / KCNE1 (IKs); NaV1.2; and, NaV1.5; in rat cerebellar granular neurons, cortical pyramidal neurons, neonatal rat cardiac myocytes, human cardiac iPS cells, and on heterologous expression (1, 23, 24. 26-28, 31, 34). SUMOylation of PM proteins has now also been described by other in the CNS,39,40 and pancreas (41).

[0789] Hypoxia-induced SUMOylation of NaV1.2 in central neurons. Seeking natural events in physiology that might regulate SUMOylation, we studied hypoxia because ground squirrels show increased SUMOylation during hibernation when tissue oxygen and glucose levels fall (42). We studied NaV1.2 in rat cerebellar granule neurons because the earliest effects of hypoxia with ischemic stroke on central neurons is an increase in sodium current (INa) and sodium loading (43-45). We observed hypoxia-induced SUMOylation of NaV1.2 channels that increased INa, and that the time-course for SUMOylation of single channels at the PM and changes in INa coincided. Mutation of a single residue (NaV1.2-Lys38) or application of a deSUMOylating enzyme prevented hypoxia-induced SUMOylation (28). The NaV1.2 channels were not SUMOylated at baseline but hypoxia led to rapid linkage of SUMO1 to almost all NaV1.2 channels at the PM, left-shifting the voltage-dependence of activation. Recently, we used CRISPR / Cas9 to create a KI mouse to replace the only SUMO- conjugation site (NaV1.2- Lys38Gln) so the channel no longer responds to hypoxia; we showed that SUMOylation of NaV1.2 in cortical pyramidal neurons exclusively controls persistent sodium current (INaP) and AP backpropagation, thereby playing a prominent role in synaptic integration and plasticity (46).

[0790] Hypoxia-induced SUMOylation of cardiac NaVl.5 and ILATE. Given the importance of hypoxia-induced ILATEin heart disease, we tested the hypothesis that SUMOylation ofNaV1.5 was an underlying mechanism. The confirmaloiy results were publishedl (Fig 2) and are summarized here briefly to rationalize the proposal and demonstrate two of the methods we would apply in this proposal. One salient feature of hypoxia- induced SUMOylation of NaV1.5 is that SUMOylation ofNaV1.5 on K442 is necessary and sufficient to explain hypoxia-induced changes in ILATEas shown by reconstitution of the response in CHO cells using whole-cell and single-channel patch clamp recording (Fig 3). This panel demonstrates an important differentiating aspect of our approach to studying SUMOylation compared to most others: we do not overexpress SUMO or SUMO pathway enzymes for days before cells are studied because these manipulations alter scores of off-target genes and secondary pathways; rather, we study live-cell responses in real-time by applying the regents to excised patches or within cells via the pipette to limit confounding effects. Thus, in Fig 3 wild-type NaV1.5 (WT) or NaV1.5- K.442Q expressed in CHO cells with NaV[31 with normoxic (21% O2) or hypoxic (1.5% O2) solution and 1 nM SUMO (red) or 1 nM SENP1 (blue) is applied via the recording pipette. In the research plan section, the same approach shows that hypoxia also induces native ILATEin hiPS-CMs in a SUMO-dependent manner.

[0791] A second key aspect of hypoxia-induced SUMOylation is that hypoxia recruits one SUMO1 to each NaV1.5 on the cell surface at the same rate as ILATEincreases; this is shown strikingly by imaging the arrival of single SUMO molecules to the PM of live cells in real-time only at sites of single NaV1.5 channels in response to hypoxia using TIRFm (Fig 4). Whole-cell current measurements show that NaV1.5 is SUMOylated in response to hypoxia at the same rate as ILATEdevelops, that SUMO is only attached to K442, as no SUMO is recruited by NaV1.5-K442Q (we also show using macroscopic FRET1), and that SUMO chains do not form, since only single SUMOs link to single WT channels).

[0792] Discovery ofTAK-981 and pharmacology in mice. Elevated levels of SUMO pathway elements are associated with poor outcomes for patients w ith some cancers.47 Agents were therefore developed to suppress SUMOylation as potential drugs. TAK-981 acts by linking covalently to the SUMO activating enzyme (SAE) component of the El enzyme when SUMO is in the catalytic site, a process called substrate-assisted inhibition. Studies ofTAK-981 with various cell types and in mice offered findings to support human trials.48, 49 Thus, treatment of pancreatic tumor cell lines for 4 h decreased SUMO conjugates in the nucleus (where the majority of SUMO conjugates reside) almost completely in a concentration dependent manner and decreased viability of the cancer cells lines; importantly, the same treatment w as well tolerated by normal VH10 fibroblasts.48 In mice, intravenous administration w as notable for decreasing SUMOylation in HCT-116 tumors after a single dose of 50 mg / kg TAK- 981 such that nuclear SUMOylation was eliminated by 4 h with SUMO-adducts measured by mass spectrometry (MS) represented approximately 80% of the El at 2 hours (h) and approximately 100% by 8 h. Of note, the pathway recovered after a single dose between 8 to 24 h in different studies consistent with lysis of the adduct or synthesis of new El. The pharmacokinetic (PK) and pharmacodynamic (PD) studies in rat were similar (where poor oral bioavailability was also demonstrated).

[0793] Conclusion: given the role of ILATEin arrhythmogenesis and our demonstration that SUMOylation of NaV1.5 is a mechanistic basis for hypoxia-induced late re- openings, we conjectured these as viable targets for therapeutic interv ention for MI- associated VA and LV dysfunction. These findings led us to create the NaV1.5- K442Q KI mouse model.

[0794] Prevention of excess ILATEbv SUMOylation inhibitors. Seeking to study SUMO inhibitors in clinically relevant manners, we first applied them in a scenario reflecting a patient with symptoms in whom vascular occlusion has not yet produced significant ischemic hypoxia nor NaV1.5 SUMOylation. Here, we applied the inhibitors for various periods (0 to 4 h) at levels we had screened to avoid toxicity prior a challenge with SUMO or hypoxia. In the first study, we raised intracellular SUMO1 to 10 nM via the recording pipette, a condition that leads to full SUMOylation of NaV1.5 within mins. As expected, we observed mean ILATEto rise from normal to pathological levels, approximately 0.4% to 3.8% (Table 1, see FIG. 20). When cells we pre-treated for 4 h to suppress El ligase function with TAK-981 (15 uM), mean ILATEdecreased to <1%. The time course for 2-D08 (20 uM) inhibition of SUMO conjugase (E2 aka Ubc9) appeared to be like TAK-981 in these preliminary studies; both agents decreased exogenous SUMO-induced ILAIE to approximately 1.5% in 2 h and this suggested that targeting both El and E2 merit study. Next, we sought to assess the inhibitors when NaV1.5 SUMOylation and excess ILATEwas induced by hypoxia (Table 2, see FIG. 20). Again, exposure to 1.5% O2led to an expected increase in ILATEfrom -0.4% to 3.8% and in these preliminary studies 4 h treatment with TAK- 981 (15 uM) or 2-D08 (20 uM) limited ILATEto approximately 2%. Because patients can also present with significant ischemia, a scenario reflecting vascular occlusion, producing NaVl .5 SUMOylation, we sought to develop an assay to study the speed of recovery' after hypoxic challenge on reperfusion. Figures 3 and 4 show that hypoxia-induced ILATEin cells in whole-cell clamp mode remains elevated when cells return to 21% O2for 3 min (the reproducible duration of such experiments being limited to approximately 10 min). We therefore confirmed that 20 min in a 1.5% O2incubator induced ILATEand the cells could be returned to 21% O2and studied for 24 h thereafter, displaying full recovery by 18 h (not shown); this leads us to propose to assess if NaV1.5 channels were deSUMOylated, if new channels were recruited to the cell surface replacing those that were SUMOylated, or both, in HEK293T cells and isolated adult murine cardiomyocytes.

[0795] The N-terminus of SENP2 and its PM location. We have shown that intracellular application of soluble SENPs leads to deSUMOylation of ion channels at the PM, 1,23, 24, 26-28, 31, 34 as have others (54). Furthermore, each of the six encoded SENPs (SENP1-3 and SENP5-7) has been visualized at the nucleus with sub-structure localization determined by their six unique N- termini (55). Thus, consistent with reports by others that the N-terminus of SENP2 mediates association with the endoplasmic reticulum and inner nuclear membranes and its deletion leads to enzyme in the cytosol (56); Fig 5 shows that in our hands WT SENP2 tagged with an mCherry fluorescent protein (SENP2-mCherry) labels the nuclear membrane and a truncation lacking just the N-terminal 18 residues (SENP2-del 18-mCherry) is seen throughout the cytosol (Fig 5), similar to a SENP2 splice variant lacking the first 50 residues (57). This demonstrates an approach we propose to apply to study the six encoded SENPs. Furthermore, we observe the N-terminal 18 residues of SENP2 carries Cysl6 within a motif for S-palmitoylation, a post-translational modification by a palmitic acid via a thioester linkage that mediates protein association with membranes, compart-mentalization in membrane subdomains, trafficking, and stability. Hypoxia-induces excess ILATEand AP prolongation in hiPS-CMs in a SUMO- dependent manner. Validating the utility of the hiPS-CMs for these studies. Fig 6 shows that hypoxia rapidly induces native ILATEin human iPS-CMs in a SUMO- dependent manner. At 21% O2, INa activated rapidly to a mean peak of 40 ± 4 pA / pF at -30 mV in whole-cell mode and then inactivated to produce a normal residual ILATEthat was 0.46% ± 0.1% of peak current. When 02 was lowered to 1.5%, mean peak INa was unchanged (42 ± 3 pA / pF), but ILATEincreased approximately 10-fold to 4.4% (1.85 pA / pF) of peak after 100 s. The magnitude of the changes in ILATEwere like those observed in tissue culture cells (Figs 3 and 4), isolated rat ventricular myocytes6 and human cardiac muscle subjected to hypoxia, approximately 4-5% over 5 -15 min (2, 58). As expected from our studies of cloned NaV1.5, SUMO1 in the pipette induced changes in native INa like those produced by hypoxia: ILATEincreased to 4.2% of peak INa without a change in the peak magnitude. With SUMO in the pipette, changing the bath from 21% to 1.5% O2did not further alter INa peak nor ILATE, supporting the conclusion that hypoxic regulation of INa proceeds via SUMOylation. Further, delivering 1 nM SENP1 into the hiPS-CMs by the pipette fully suppressed the increase in ILATEinduced by hypoxia. Of note, we deliver 1 nM SUMO 1 into cells because that level evokes maximal effects on NaV 1.2 channels in cerebellar granule neurons (CGNs) and three different K+ channels in CGNs, hippocampal neurons, and rat cardiac myocytes (26-28, 34).

[0796] By recording spontaneous APs in current-clamp mode we could show SUMO- dependent, hypoxia-induced increases ILATEled to APD prolongation to levels observed in patients with VA and sudden death (Fig 7). The changes are suppressed by ranolazine, a drug that inhibits NaV1.5 ILATE. Including half of the hypoxia-induced increase in ILATEin hiPS-CMs into the O’Hara- Rudy model for human cardiac action potentials (59) reproduced the observed pro-arrhythmic increase in APD.

[0797] We have shown that hypoxia produces pathologic levels of ILATEdue to SUMOylation of NaV1.5 channels on K4421 and demonstrate here for the first time that SUMO pathway inhibitors of E1 and E2, TAK- 981 and 2-D08, respectively, suppress SUMOylation and ILATEin vitro and VA and LV dysfunction in WT mice subjected to LAD ligation. Here, we assess the kinetics and duration of protection and the time course for recovery with two proven agents and two alternative inhibitors.

[0798] We can use whole-cell patch clamp withNaV1.5 (with NaVβ1) expressed in HEK293T cells to study (a) the dose-dependent kinetics of ILATEonset with hypoxia in absence and presence of TAK-981 and 2-D08 at various concentrations, (b) the dose- dependent kinetics of ILATEresolution with normoxic reperfusion in the absence and presence of TAK-981 and 2-D08 at various concentrations, (c) The dose- dependent kinetics of ILATEonset with hypoxia with two less studied E1 inhibitors, ML-792 and COH000, and further analyses if indicated. Despite their use only in preclinical studies, ML-792 is of interest because it is the most potent El inhibitor identified thus far and inhibits SUMOylation rapidly in vitro (60) and COH000 is reported to be selective with IC50 of 0.2 μM and a delineated molecular mechanism of action (61, 62). These studies are performed as in Figure 3.

[0799] We can first to express NaV1.5 (with NaVβ1) in HEK293T cells to study (a) the kinetics of SUMO forward transport to the PM and colocalization at sites of NaV1.5 in response to hypoxia without and with TAK-981 and 2-D08 using TIRFm, FRET, and Western Blot, (b) To determine the rate of loss of NaV1.5 from the membrane at baseline and after 20 min hypoxia without and with TAK-981 and 2-D08 via surface biotinylation and membrane protein purification with avidin at various times, followed by SDS-PAGE and NaV1.5 visualization with epitopes to polyHisl or 1D4 and HA23; whole-cell INa is measured at the time of purification to estimate the rate of restoration of new, non-biotinylated channels on the surface; (c) the rate of deSUMOylation of PM channels when delivery of new channels to the surface is suppression by cycloheximide (68). (d) Comparative studies of channel turnover can be performed and cardiomyocytes from WT and SCN5AK442Q mice using surface biotinylation / avidin purification and antibodies directed to mNaV1.5 (69, 70).

[0800] We posit that the enzy mes required to activate, mature and conjugate SUMO are present in the PM of Xenopus oocytes, tissue culture cells and neurons (1,23-28) because (1) covalent attachment is required to regulate the channels; (2) pro-SUMO can modify channels in off-cell, inside-out membrane patches and this requires maturation of the pro-protein by a SENP, as well as an El- and E2-like ligase and conjugase activities; (3) although SENPs are synthesized in the cytoplasm, the cytosol diffuses away from inside-out off-cell patches; (4) we used confocal microscopy to visualize GFP-E2 at the PM of Xenopus oocytes (23) and SAE1 , E2 and SUMO1 at the PM of CHO-K1 cells;24 and others have reported that E2 can reach the PM and co-localizes with the dopamine transporter in HEK293T cells (72).

[0801] Using on and off-cell patch recording, FRET, and TIRFm (Figures 3-6 and 1,23-28) we can confirm and expand the findings to quantify surface expression of (a) FP-tagged E1, E2, SUMO1, SUMO2 / 3, and the six encoded SENPs in HEK293T cells using TIRFm and seek evidence for co-localization withNaV1.5. (b) Validate use of commercial antibodies for SUMO pathway proteins for study of native proteins in iPS-CMs and mouse cells, (c) Study the role of the first 18 residues in the PM binding of SENP2 in HEK293T cells using the full-length and truncated SENP2 constructs for TIRF and FRET, (d) Study SENP2 C16S mutation to test our hypothesis that palmitoylation mediates PM binding, (e) Apply palmitoyl-protein thioesterase in the pipette and in inside-out patch membranes to test the hypothesis that SENP2 matures pro-SUMO and leads to the SUMOylation ofNavl.5 in HEK293T cells to uncover mechanism and for potential later use with iPS-CMs Human cardiac myocytes derived from pluripotent stem cells (iPS-CMs) are now widely used to screen for drug-induced alterations in cardiac cellular contractility', electrophysiology, and viability because they recapitulate many of the attributes of cardiac physiology. Thus, we showed hypoxia induces a rapid increase in ILATEand APD in iPS-CMs (Figure 6) to levels associated with human pathology and studying the parameters of electrical function and employing the O’Hara Rudy model for simulations, as described.1 Here, we seek to expand our prior studies to assess the effect of the SUMO inhibitors, SUMO enhancers (overexpression of SUMO + / - E2) and, if indicated palmitoyl-protein thioesterase. Current-clamp can be used to record action potentials in iPS-CMs under normoxia and hypoxia without and with TAK-981 and 2-D08 and the alternative inhibitors, ML-792 and COH000, if indicated. Second, to study intact cells, iPS-CMs genetically encoded with a fluorescent voltage indicator (as described by Chiamvimonvat (73)) can be imaged under the same experimental conditions.

[0802] After covalent attachment of SUMO, regulatory- actions are via non-covalent binding of a conserved hydrophobic groove in SUMO and hydrophobic target residues called the “SUMO interacting motif (SIM)” stabilized by charge-charge interactions (74, 75). SUMO-SIM interactions alter function directly (76, 77) and can also recruit non-SUMOylated proteins bearing SIMs into complexes with SUMOylated targets (78).

[0803] We can (a) test the 6 high-likelihood SIMs in NaV1.5 predicted by two validated tools79,80 by site-directed mutagenesis and functional assessment of ILATE; (b) identify SIM and non-SIM interactions of Navi.5 with SUMO by cross-linking mass spectrometry (XL-MS) using novel agents like Alkyne- A-DSBSO, a strategy method that allow s purification and secondary cleavage to improve MS resolution (81, 82); and (c) use molecular dynamics (MD) simulations to predict energetically important residue-residue interactions (e.g.. mutations alter AAG by > 2 kcal / mol). and confirm them by thermodynamic mutant-cycle analysis to support the structural model of the complex produced with know n structures for SUMO175 and NaV1.5 (83, 84) as we have before for complexes of the Hvl channel with Alb, and Hvl with a designed protein blocker (85, 86).

[0804] Inhibition of Nav1.5 SUMOylation and ILATEcan result in a decrease in AP prolongation, reduction in intracellular Na+ loading, prevention of the subsequent increase in intracellular Ca2+ from Na+ / Ca2+ exchanger activities, leading to a decrease in early and delayed afterdepolarization (EAD and DAD) and adverse LV remodeling post I / R. Therefore, inhibition of NaV1.5 SUMOylation represents a viable strategy to reduce VA and LV dysfunction in acute and post-acute phases of MI. We demonstrated that hypoxia produces pathologic levels of ILATEdue to SUMOylation of NaV1.5 channels on a single residue, lysine 442 (K442), and that ILATEis eliminated by mutation of the site, or by application of a deSUMOylating enzyme (1). We can take advantage of a knock-in (KI) mouse model. NaV1.5- K442Q, that we have generated. Eata demonstrate that ablation of Nav1.5 SUMOylation and hypoxia-induced ILATEresult in a significant decrease of VA and LV dysfunction compared to the WT mice. Moreover, SUMO pathway inhibitors suppress Nav1.5 SUMOylation, ILATE, ventricular arrhythmia (VA), and ameliorate LV dysfunction in WT mice subjected to I / R injury.

[0805] Figs 9-18 and (33, 91-98) support the feasibility of the techniques to quantify the arrhythmia burdens, infarct size, and systolic and diastolic function during the acute, sub-acute (2 weeks) and chronic phase (3 months) after I / R injury. Multidisciplinary techniques include ECG recordings during I / R surgeries, ambulatory ECG, ECHO, in vivo hemodynamic monitoring, histological analyses, single-cell analyses including quantification of local and global Ca2+ concentrations, and patch-clamp analyses. Here, we show the beneficial effects of ablation of SUMOylation site in Nav1.5 as well as SUMO pathway inhibitors, offering preliminary demonstration that VA and pump failure are suppressed.

[0806] NaV1.5-K442Q mice show normal systolic and diastolic function as well as heart rate compared to WT animals as assessed by echocardiography. To create the KI mice, replacing the only SUMO- conjugation site in Navl.5, we used CRISPR / Cas9 and FVB mice, a strain commonly used to study cardiac function (99-102). Briefly, Scn5AK442Q / WT heterozygous mice were generated by Biocytogen (Wakefield, MA) as follows. Two sgRNAs in introns 7 and 10 were designed with a tool from Wellcome Sanger Institute and screened for on-target activity using a Universal CRISPR Activity Assay (UCATM, Biocytogen Pharmaceuticals Co., Ltd). AT7 promoter sequence was added to the Cas9 or sgRNA template by PCR amplification. Different concentrations of donor vector and purified in vitro transcribed Cas9 mRNA and sgRNA were mixed and co-injected into the cytoplasm of one- cell stage FVB fertilized eggs. Injected zygotes were transferred into oviducts of Kunming pseudo- pregnant females to generate FO mice. PCR and sequencing verification of founder pups with the intended mutation were then crossed with wild-type (WT) mice for germline transmission, which was further confirmed by PCR, sequencing, and Southern blot analysis. Each liter is genotyped and cared for according to 1ACUC protocols. We reported an analogous strategy to create SCN2AK38Q mice, demonstrating that single SUMOylation site in NaV1.2 regulates AP backpropagation in cortical neurons playing a prominent role in synaptic integration and plasticity the CNS (46).

[0807] Echocardiography was used to determine the systolic and diastolic function in the WT and NaV1.5-K442Q mice as we have described (33, 93, 95-97). Diastolic function is assessed using the ratio of the blood flow velocities through the mitral valve during early filling (E) and late filling (A), E / A ratio, as well as deceleration time. Fig. 9 shows normal systolic and diastolic function in the KI mice compared to WT littermates. Mice is implanted with ECG transmitters. Longitudinal ambulatory ECG telemetry and echocardiographic recordings is performed every 2 weeks for up to 3 months. There were no significant differences in RR, PR intervals, QRS duration or QTc intervals at baseline between WT and KI mice.

[0808] NaVl .5-K442Q mice show protection from VA and improvement in LV dysfunction compared to WT in response to LAD ligation. To support the central hypothesis, we performed 45-minute ischemia followed by reperfusion (I / R) surgeries in WT and KI mice. Fig IOC and E show a significant decrease in VA burdens in the KI compared to the WT littermates. Moreover, at 2 weeks after I / R surgeries, WT mice show a significant decrease in left ventricular ejection fraction (LVEF) and fractional shortening (FS) compared to sham animals (Fig 11, D-E). In contrast, even though KI mice show a significant decrease in LVEF and FS compared to sham, the left ventricular function is better preserved compared to WT animals post 1 / R. To further confirm the findings, we perform hemodynamic monitoring as we have previously described (33, 98) at 2 weeks to assess changes in LV pressure, volume, developed pressure (dP / dt), end- systolic and end-diastolic volumes (Fig 12). Our exciting preliminary data show a significant preservation in cardiac function in KI mice compared to WT animals (Fig. 11).

[0809] SUMO pathway inhibitors suppress VA and improve pump function in WT mice in response to LAD ligation. Two different SUMO pathway inhibitors were used (TAK-981, an El enzyme inhibitor and 2- D08, an E2 enzyme inhibitor). WT mice were injected (IP) with either vehicle (DMSO) or TAK-981 (7.5 mg / kg, resulting in the blood level of 160 pM acutely), or 2-D08 (10 mg / kg, resulting in the blood level of 460 μM acutely). The dosage were chosen based on prior published studies (48. 49, 103-108). I / R surgeries were performed after the injection. Fig 13 shows a significant decrease in VA burdens in the mice that received SUMO pathway inhibitors during ischemia. There were VAs during reperfusion in all 3 groups of mice, but the numbers of VAs were significantly reduced in the treated mice compared to the vehicle controls (Fig. 13C and E), both during ischemia and reperfusion phases Moreover, the left ventricular function is better preserved compared to WT animals treated with SUMO pathway inhibitors post I / R (Fig. 14).

[0810] To determine VA burdens and LV dysfunction in WT and KI mice as well as SUMO pathway inhibitors after I / R injury, we can directly test the arrhythmia burdens and adverse cardiac remodeling in male and female WT and NaV1.5-K442Q KI mice at 12-16 w eeks during the acute phase, sub-acute (2 weeks), and chronic (3 months) after I / R injury (96, 98, 109) SUMO pathway inhibitors is tested in the WT animals. Multidisciplinary techniques can be used including echocardiography, hemodynamic monitoring, ambulatory ECG recordings, in vivo electrophysiologic studies (EPS), assessment of infarct size and adverse cardiac remodeling. Mice can then be sacrificed for in vitro mechanistic studies.

[0811] Echocardiography can be performed to determine the LVEF, FS, LV wall thickness, dimension and volume. Diastolic function is assessed using mitral inflow and tissue Doppler to quantify E / A ratios and isovolumic relaxation time (IVRT).

[0812] Pressure-volume (PV) analyses. Intact heart hemodynamic analysis can be performed (Millar Instruments) to record chamber volume by impedance and pressure by micromanometry (33, 110). Different parameters is assessed including LV end diastolic pressure (LVEDP). PV loops can be constructed before and during transient reduction of preload to generate specific systolic and diastolic function indexes, LV afterload (indexed by arterial elastance), ejection fraction, contractile function as assessed through load-independent parameters (maximal power index and preload recruitable stroke work), and diastolic function (LV stiffness constant, tau and peak rate of pressure decline) (Fig. 12) as we have described (33).

[0813] Quantification of arrhythmia burdens in homozygous KI mice compared to WT animals. Two complementary techniques are used. Ambulatory ECG Recordings. Implanted ETA-F20™ telemetric transmitters (DSI, St. Paul, MN) is used to obtain continuous lead II ECG recordings on awake mice. Animals are allowed to recover from the telemetry implantation for 7 days before the I / R surgeries. Telemetry recordings are performed over a total of 24-hour period with 30 min of recording for each hour over 2 consecutive days (Fig. 14). Recordings are obtained at exact time of the day and night for all animals to control for diurnal variations. Data is analyzed in a blinded manner using PONEMAH SOFTWARE SYSTEM™ (DSI). Atrial and VA is quantified. VA can include premature ventricular contractions (PVCs), couplets, triplets and non-sustained ventricular tachycardia (NSVT). ECG are analyzed for RR, PR and QTc intervals. The rate-corrected QT interval (QTc) is calculated using modified Bazett's formula (QTc = QT interval (in ms) / (RR / 100)l / 2) (111).

[0814] In vivo electrophysiologic studies (EPS). In vivo EPS are performed as we have described previously to test for arrhythmia inducibility (91, 92). Briefly, standard pacing protocols is used to determine the electrophysiologic parameters, including sinus node recovery time, atrial, AV nodal, and ventricular refractory periods and AV nodal conduction properties. Each animal can undergo an identical pacing and programmed stimulation protocol. To induce atrial and ventricular tachycardia and fibrillation, programmed extra-stimulation techniques and burst pacing is utilized as we have previously described (91, 92) (Fig. 15). Programmed right atrial and right ventricular double and triple extra-stimulation techniques is performed at 100-ms drive cycle length, down to a minimum coupling interval of 10 ms. Right atrial and right ventricular burst pacing is performed as eight 50-ms and four 30-ms cycle length trains repeated several times, up to a maximum 1-min time limit of total stimulation. Reproducibility is defined as greater than one episode of induced atrial or ventricular tachycardia. The exact same programmed stimulation protocols are used in all animals. Infarct size and adverse cardiac remodeling. Infarct size is quantified in different groups of animals as we have described using TTC stain.109 For sub-acute and chronic animals, cardiac sections is stained using antibodies against collagen la, Illa, and V. and fibronectin. Picrosirius Red and wheat germ agglutinin (WGA) stains is used to quantify the amount of collagen (96-98). Flow cytometric analyses is performed as we have described to quantify the degrees of fibrosis and cardiac fibroblast activation (96-98).

[0815] To determine the mechanisms underlying the reduction in VA and LV dysfunction by Navi.5 SUMOylation ablation or SUMO inhibitors, experiments are performed using isolated ventricular myocytes from WT and KI mice during acute, sub-acute and chronic phase. We can use two SUMO- specific inhibitors (TAK-981 and 2-D08) to directly test the effects of the inhibitors on ILATE, AP, global and local Ca2+, EAD, and DAD in WT ventricular myocytes exposed to hypoxia.

[0816] To investigate the mechanistic effects of SUMO inhibitors and ablation of Nav1.5 SUMOylation on cellular arrhythmogenic mechanisms using AP recordings and AP-clamp, we can use AP-clamp as described by (112-115) followed by sequential dissection, using internal and external solutions resembling the physiological ionic milieu. The AP waveform is applied as the command voltage onto the same cell. A specific channel blocker is added to block the current of interest, and the compensation current is recorded. GS-458967, ORM-10962, and nifedipine is used to record ILATE, Na+ / Ca2+ exchanger current (INCX), and E-type Ca2+ current (ICaL). respectively, as we have described. The current of interest is obtained by subtracting the compensation current from the baseline (112-115) AP clamp (Fig. 16) can determine if late Na+ currents (ILate) are altered in 1 / R and if NaV1.5-K442Q KI or SUMO inhibitors rescue those alterations. Complementary experiments are performed using cell-attached single channel recordings as described (116). We can quantify the open probability (Po), open and closed time of the channels (116, 117). APs are recorded using perforated patch-clamp techniques to maintain intracellular milieu as we have described to quantify EAD and DAD (116, 118-122).

[0817] To determine the mechanistic effects of Nav1.5 SUMOylation ablation or SUMO inhibitors on intracellular Ca2+. Ca2+ transients (CaT). Ca2+ sparks, and SR Ca2+ load, two complementary techniques are used to investigate the critical roles of SUMO inhibitor treatment and NaV1.5-K442Q KI on local and global Ca2+ release. IONOPTIX™ sarcomere detection (lonOptix Co) and fast Fourier transform (FFT) method (33, 95, 123) rather than Fluo-4 confocal imaging edge detection are used to measure single cardiomyocyte contraction because the latter may lose precision if the cell's ends move out of the focal plane during contraction. Contraction is measured using a high-speed camera. The sarcomere pattern is used to calculate the sarcomere length using an FFT algorithm. The fractional shortening is calculated as the percentage change in sarcomere length during contraction (Fig. 17). Simultaneous Ca2+ transients (CaT) are studied using Fura-2 dual- wavelength ratiometric method, 123 which yields absolute [Ca2+], Additional analyses of excitation- contraction (EC) coupling are performed including assessment of sarcoplasmic reticulum (SR) Ca2+ load ([Ca]SRT), Ca2+ current (ICa), and Ca2+ sparks using confocal line scan (Fig. 17).

[0818] Multimodal Second Harmonic Generation (SHG) - Two-Photon Fluorescence (TPF) microscopy. We employed SHG microscopy, a non-linear label-free technique, to directly image the sarcomeres based on the unique intrinsic property of the SUMO rod domains to generate a signal at twice the frequency and half the wavelength of the laser beam used to excite the sarcomeres (124). We utilized multimodal SHG-TPF imaging techniques to simultaneously image Ca2+ levels and sarcomere contraction at high spatial and temporal resolutions (Fig. 18) (32, 33). The method has multiple advantages: 1) it images the A-band and is independent of T- tubule morphology which varies in diseased hearts and in different cell types, and 2) it enables simultaneous measurements of microdomain contraction and local Ca2+ levels in live cardiomyocytes. Simultaneous SHG to non-invasively image the sarcomere and TPF to image the Ca2+signal is obtained in ventricular myocytes isolated from KI models (Fig. 18B). The experiments allow us to directly quantify the single sarcomere contraction and relaxation of cardiac myocytes. We can determine if alterations in local and global Ca2+ signaling, sarcomere contraction, and relaxation can be rescued with SUMO inhibitors and in the KI mouse model.

[0819] Statistical analysis: All data is tested for normality by the Shapiro-Wilk test and homogeneity of variances are assessed using Levene’s test (125). Statistical significance is determined using appropriate unpaired two-tailed Student’s t-test, nonparametric tests, one-way analysis of variance (ANOVA) or two-way ANOVA for multiple comparisons with appropriate post hoc analyses. p<0.05 are considered significant.

[0820] Methods

[0821] IV drug administration. Mice are anesthetized with inhaled isoflurane, and 40 pl of drug or drug vehicle is instilled retro-orbitally (IV) using a 27-gauge needle.

[0822] Adult mouse cardiomyocyte isolation. Before the procedure, we record the mouse body weight, strain, sex and date of birth. Heparin (200 pl) are injected intraperitoneal (i.p.) 10 min prior to anesthesia to prevent coagulation of blood in the coronary arteries. The mouse is anesthetized with ketamine and xylazine by i.p. injection (80- 120 / 5-10 mg / kg), secured in the supine position by gently fixing the forepaws and hind paws to a work surface on a surgery tray near the perfusion system. A midline skin incision from mid abdomen to the diaphragm is made and cuts bilaterally to retroflect the thoracic cage to expose the heart. The heart is lifted slightly using fine curved serrated and atraumatic forceps and dissected out of the thoracic cavity. The heart is transferred to a 100 mm dish containing cold perfusion buffer and then connected to the cannula. The heart is perfused with calcium-free perfusion buffer at flow rate of 4 ml / min for 4-5 min until the effluent becomes clear, then we switch to digestion buffer for 3.5-20 min depending on perfusion pressure and collagenase activity. Digestion is stopped when the heart becomes slightly pale and flaccid. Cells are dissociated and calcium reintroduced. Cells are collected and suspended in 5 ml plating medium. We count the total number of myocytes and percentage of rod-shaped myocytes and adjust the concentration of rod-shaped myocytes to 25.000 / ml in a 50 ml tube. Gently suspension with a 10 ml pipette is followed by aspiration of the coating solution, and plating to dishes. Dishes are placed in a CO2 incubator at 37°C for 1-3 h to allow myocyte attachment to about 80%. We gently remove unattached myocytes and cell debris and add new culture medium.

[0823] Ischemia-Reperfusion Model (I / R model): The procedure is performed as described in detail in previous publication (1-3). Ischemia-reperfusion is induced in 10-week-old male Slc26a6- / - (KO) mice and wild-ty pe (WT) littermates. The procedure is performed using aseptic techniques. The fur is removed from the surgical incision sites (Nair) and the skin cleaned with Betadine followed by 70% alcohol 3 times. Surgery is performed on a clean and disinfected lab bench, with sterilized instruments (autoclaved). Mice is placed on a sterilized cloth on a circulating water blanket to maintain body temperature. Surgeons scrub with Betadine, mask and wear sterile gloves. Animals is anesthetized with intraperitoneal ketamine 100 mg / kg and xylazine 5 mg / kg. Intubation is done perorally and mechanical ventilation is started. The procedure is performed as described in detail in previous publication.1 An oblique 8-mm incision is made 2 mm away from the left sternal border in the 4th intercostal space. The chest cavity is then opened. The chest retractor is inserted and opened gently to spread the wound 8-10 mm in width. The heart partially covered by the lung is then visualized. The pericardium is gently picked up with curved and straight forceps, pulled apart, and placed behind the arms of the retractor. The left anterior descending (LAD) coronary artery is then visualized and ligated 1-2 mm below the tip of the left auricle in its normal position, which induces roughly 40-50% ischemia of the LV. Occlusion is confirmed by the change of color of the anterior wall of the LV and is left for a period of 45 minutes after which the occlusion is removed. The retractor is removed, and the lungs are reinflated by shutting oft" the ventilator outflow as previously described. The chest cavity is then closed. The sham-operated mice undergo the same procedure without tying the suture but moving it behind the LAD artery. The chest is closed with 3-0 DEXON™ rib sutures, 5-0 DEXON II™ muscle sutures and buried skin sutures (buried continuous intradermal suture pattern with 4-0 absorbable suture). Negative plural pressure is re-established via a temporary chest tube until spontaneous breathing occurs. A bolus injection of 0.1 mg / kg of buprenorphine SC is given. Recovery (until the animal is ambulatory) includes visual monitoring of the animal for a period of 2-4 hours. Additional postoperative treatment includes supplemental buprenorphine (0.1 mg / kg) every 12 hours as an analgesic (through at least the second day) and a warm environment.

[0824] Electrocardiographic recordings: ECG recordings is performed using BIOAMPLIFIER™ (BMA 831, CWE, Incorporated, Ardmore, PA) as we have previously described4. The intraperitoneal pentobarbital 40 mg / kg is used to maintain anesthesia during recording. The animals are placed on a temperature-controlled warming blanket at 37°C. Four consecutive two-minute epochs of ECG data is obtained from each animal. Signals were low-pass filtered at 0.2 kHz and digitized using DIG1DATA 1200™ (Axon Instrument, CA). A total of 100 beats is analyzed from each animal in a blinded fashion. The Q-T interval is determined manually by placing cursors on the beginning of the QRS and the end of the T wave. The rate- corrected QT interval (QTc) is calculated using modified Bazett's formula as reported by Mitchell et al for mouse models, whereby the RR interval is expressed as a unitless ratio (RR in ms / 100 ms). QTc interval is defined as (QT interval (in ms) / (RR / 100)l / 2) (5).

[0825] Radiotelemetry ECG monitors: Mice are anesthetized with intraperitoneal ketamine 100 mg / kg and xylazine 5 mg / kg, intubated and mechanically ventilated with supplemental oxygen at a respiratory rate of 100 breaths / min and tidal volume of 0.2 ml. The fur is removed from the surgical incision sites (Nair) and the skin cleaned with Betadine followed by 70% alcohol 3 times. Surgery is performed on a clean and disinfected lab bench, with sterilized instruments (autoclaved). Mice is on a sterilized cloth on a circulating w ater blanket to maintain body temperature. Surgeons scrub with Betadine, mask and w ear sterile gloves. A sterile ECG transmitter (Data Sciences International. volume=2.1 cc and weight=3.4 g) is implanted to monitor heart rate. A 10-mm incision is made in the abdomen and the sterile transmitter placed within the peritoneal cavity and is secured to the inside wall of the abdomen with non- absorbable sutures. The two ECG leads is tunneled subcutaneously and tied to right shoulder and xyphoid space caudal to the ribcage. The abdominal muscle is closed with 6-0 Nylon sterile sutures and the skin is closed with 4-0 absorbable sutures (Ethilon). The whole procedure should take about 15-20 minutes. A bolus injection of 0.1 mg / kg of buprenorphine SC is given. Recovery (until the animal is ambulatory) includes visual monitoring of the animal for a period of 2-4 hours. Additional postoperative treatment includes supplemental buprenorphine every 12 hours as an analgesic (through at least the second day) and a warm environment. Post-operative records is kept in the housing room, noting the condition and date of PI inspection. The mice is allowed to recover for at least 7 days, at which point, 24 hour of non- invasive telemetry is recorded. Ischemia-reperfusion or sham surgery is performed and monitoring is continued for up to 6 weeks.

[0826] In vivo Electrophysiologic Studies: In vivo electrophysiologic studies is performed as we have previously described (4). All surgical procedures follow the UC Davis Guidelines for Aseptic Surgical Technique. Intraperitoneal pentobarbital 40 mg / kg is used to maintain anesthesia during surgery which is expected to last -30-45 minutes. A small skin incision of 3-5 mm. is made on the right side of the neck. Right jugular vein is dissected under direct visualization under dissecting microscope and catheter inserted. A special 1.7 French octapolar catheter with an interelectrode spacing of 0.5 mm (CIBER MOUSE EP™, NuMed, Hopkinton, NY) is used via right jugular vein into the right atrium and right ventricle.

[0827] Pacing protocols is performed via the catheter under anesthesia. Standard pacing protocols is used to determine the electrophysiologic parameters, including sinus node recovery time, atrial, AV nodal, and ventricular refractory periods and AV nodal conduction properties. Pacing is performed through the catheter using an external stimulator (Bloom stimulator). Sinus node recovery time is measured between the last paced atrial depolarization and the first sinus return cycle after 15 second of atrial pacing at several pacing cycle lengths (CL).

[0828] AV nodal conduction properties is obtained by atrial and ventricular incremental pacing methods until Wenckebach Periods are defined such as 2: 1 AV block or retrograde ventriculoatrial block. Atrial, AV, and ventricular effective refractor) period (ERP) is analyzed by the extra-stimulus method. Each animal undergoes an identical pacing and programmed stimulation protocol.

[0829] To induce atrial and ventricular tachycardia and fibrillation, programmed extra-stimulation techniques and burst pacing is utilized. Programmed right atrial and right ventricular double and triple extra-stimulus techniques is performed at 150-ms drive cycle length, down to a minimum coupling interval of 10 ms. Right atrial and right ventricular burst pacing is performed as eight 50-ms and four 30-ms cycle length trains episodes repeated several times, up to a maximum 1-min time limit of total stimulation. Carbachol (a muscarinic receptor agonist, 50-100 ng / g i.p.) is administered during the pacing protocol to further assess atrial arrhythmias inducibility as previously described above. For comparison of the inducibility in each animal, programmed extra- stimulus techniques and stimulation duration of atrial and ventricular burst pacing is kept uniformly in all animals. Reproducibility is defined as greater than one episode of induced atrial or ventricular tachycardia. All the animals remain under anesthesia for the catheter placement and the pacing procedures. Noninvasive echocardiographic imaging: Cardiac function in the I / R animals is assessed using noninvasive imaging with echocardiogram in conscious state as we have previously described4. The chest is shaved. The animal is placed in a supine position on a warming blanket (37). The mice are restrained in a 50 ml conical tube with an opening for the nose and chest, this is a standard procedure for imaging mice for this kind of study. The imaging generally lasts for 5 minutes. We can determine the fractional shortening, left ventricular wall thickness, dimension and volume. Diastolic function is assessed using mitral inflow and tissue Doppler imaging (TDI).6 Altered patterns of valve annulus motion can be early indicators of diastolic dysfunction. Hemodynamic Monitoring: Hemodynamic monitoring is performed as previously described using AD instrument. The animals is injected with pentobarbital i.p. (40-80 mg / kg) to achieve surgical anesthesia. Intact heart hemodynamic analysis is performed using a four-electrode PV catheter (Millar Instruments) to record chamber volume by impedance and pressure by micromanometry: 7 Different parameters is assessed including left ventricular (LV) end diastolic pressure (LVEDP). PV loops is constructed before and during transient reduction of preload to generate specific systolic and diastolic function indexes, LV afterload (indexed by arterial elastance), ejection fraction, contractile function as assessed through load-independent parameters (maximal power index and preload recruitable stroke work), and diastolic function (LV stiffness constant, tau and peak rate of pressure decline (dP / dtmin)) (8). All animals are euthanized 3 weeks after the surgery. Intraperitoneal pentobarbital 40- 80 mg / kg is used to achieve surgical anesthesia. The chest is opened via a midline thoracotomy. Exsanguination can occur upon removal of the heart. The animals are expected to develop compensated cardiac hypertrophy at 3 weeks with no clinical detectable sign or symptoms and can develop early decompensated heart failure at 6 w eeks at which time the animals can develop sign and symptoms of heart failure including decreasing appetite, and weight gain.

[0830] References:

[0831] 1. Tamavski O, et al. Mouse cardiac surgery: comprehensive techniques for the generation of mouse models of human diseases and their application for genomic studies. Physiol Genomics. 2004;16:349-60.

[0832] 2. Li N, et al. Beneficial effects of soluble epoxide hydrolase inhibitors in myocardial infarction model: Insight gained using metabolomic approaches. J Mol Cell Cardiol. 2009:47:835-45. 3. Sirish P, et al. Unique mechanistic insights into the beneficial effects of soluble epoxide hydrolase inhibitors in the prevention of cardiac fibrosis. Proc Natl Acad Sci U S A. 2013;110:5618-23. 3619365

[0833] 4. Zhang XD, et al. Critical roles of a small conductance Ca2+-activated K+ channel (SK3) in the repolarization process of atrial myocytes. Cardiovasc Res. 2014;101:317-25. 3896251

[0834] 5. Mitchell GF, et al. Measurement of heart rate and Q-T interval in the conscious mouse. Am J Physiol. 1998;274:H747-51.

[0835] 6. Daneshvar D. et al. Diastolic dysfunction: improved understanding using emerging imaging techniques. Am Heart J. 2010;160:394-404.

[0836] 7. Takimoto E, et al. Chronic inhibition of cyclic GMP phosphodiesterase 5A prevents and reverses cardiac hypertrophy. Nat Med. 2005;11:214-22.

[0837] 8. Zhang XD. et al. Prestin amplifies cardiac motor functions. Cell Rep. 2021;35:109097.

[0838] References Example 1 :

[0839] 1. Plant LD, et al. Hypoxia Produces Pro-arrhythmic Late Sodium Current in Cardiac Myocytes by SUMOylation ofNaV1.5 Channels. Cell Rep. 2020;30:2225- 2236 e2224. doi: 10.1016 / j.celrep.2020.01.025

[0840] 2. Makielski JC. Late sodium current: A mechanism for angina, heart failure, and arrhythmia. Trends Cardiovasc Med. 2016;26:115-122. doi:

[0841] 10.1016 / j.tcm.2015.05.006

[0842] 3. Bennett PB, et al. Molecular mechanism for an inherited cardiac arrhythmia. Nature. 1995;376:683-685.

[0843] 4. Plant LD, et al. A common cardiac sodium channel variant associated with sudden infant death in African Americans, SCN5 A S 1103 Y. J Clin Invest.

[0844] 2006:116:430-435. doi: 10.1172 / JCI25618

[0845] 5. Belardinelli L, Giles WR, Rajamani S, Karagueuzian HS, Shryock JC. Cardiac late Na(+) current: proarrhythmic effects, roles in long QT syndromes, and pathological relationship to CaMKII and oxidative stress. Heart Rhythm.

[0846] 2015:12:440-448. doi: 10.1016 / j.hrthm.2014.11.009

[0847] 6. Ju YK, Saint DA. Gage PW. Hypoxia increases persistent sodium current in rat ventricular myocytes. J Physiol. 1996;497 ( Pt 2):337-347. 7. Carmeliet E. Cardiac ionic currents and acute ischemia: from channels to arrhythmias. Physiol Rev. 1999;79:917-1017.

[0848] 8. Belardinelli L, et al. Inhibition of the late sodium current as a potential cardioprotective principle: effects of the late sodium current inhibitor ranolazine. Heart. 2006;92 Suppl 4:iv6-ivl4. doi: 10.1136 / hrt.2005.078790

[0849] 9. Gaur N, Rudy Y, Hool L. Contributions of ion channel currents to ventricular action potential changes and induction of early afterdepolarizations during acute hypoxia. Circ Res. 2009;105:1196-1203. doi: 10.1161 / CIRCRESAHA.109.202267

[0850] 10. Shryock JC, et al. The arrhythmogenic consequences of increasing late INa in the cardiomyocyte. Cardiovasc Res. 2013;99:600-611. doi: 10.1093 / cvr / cvtl45

[0851] 11. Chadda KR, et al. Sodium channel biophysics, late sodium current and genetic arrhythmic syndromes. Pflugers Arch. 2017:469:629-641. doi: 10.1007 / s00424-017- 1959-1

[0852] 12. Bui AH, Waks JW. Risk Stratification of Sudden Cardiac Death After Acute Myocardial Infarction. J Innov Card Rhythm Manag. 2018;9:3035-3049. doi: 10.19102 / icrm.2018.090201

[0853] 13. Yeh RW, Sidney S, Chandra M, Sorel M, Selby JV, Go AS. Population trends in the incidence and outcomes of acute myocardial infarction. N Engl J Med.

[0854] 2010;362:2155-2165. doi: 10.1056 / NEJMoa0908610

[0855] 14. Anderson JL, Morrow DA. Acute Myocardial Infarction. N Engl J Med.

[0856] 2017:376:2053-2064. doi: 10.1056 / NEJMral606915

[0857] 15. Antzelevitch C, Nesterenko V, Shryock JC, Rajamani S. Song Y, Belardinelli L. The role of late I Na in development of cardiac arrhythmias. Handb Exp Pharmacol. 2014;221:137-168. doi: 10.1007 / 978-3-642- 41588-3_7

[0858] 16. Komyeyev D, et al. Contribution of the late sodium current to intracellular sodium and calcium overload in rabbit ventricular myocytes treated by anemone toxin. Am J Physiol Heart Circ Physiol. 2016;310:H426-435. doi:

[0859] 10.1152 / ajpheart.00520.2015

[0860] 17. Horvath B, et al. Late Sodium Current of the Heart: Where Do We Stand and Where Are We Going? Pharmaceuticals (Basel). 2022;15. doi: 10.3390 / phl5020231

[0861] 18. West JB. Physiological Effects of Chronic Hypoxia. N Engl J Med.

[0862] 2017;376:1965-1971. doi: 10.1056 / NEJMral612008 19. Song Y, et al. Blocking late sodium current reduces hydrogen peroxideinduced arrhythmogenic activity and contractile dysfunction. J Pharmacol Exp Ther. 2006;318:214-222. doi: 10.1124 / jpet.106.101832

[0863] 20. Mahajan R. et al. A Small Ubiquitin-Related Polypeptide Involved in Targeting RanGAPl to Nuclear Pore Complex Protein RanBP2. Cell. 1997:88:97.

[0864] 21. Matunis MJ, et al. A novel ubiquitin-like modification modulates the partitioning of the Ran- GTPase-activating protein RanGAPl between the cytosol and the nuclear pore complex. J Cell Biol. 1996;135:1457-1470. doi: 10.1083 / jcb.l35.6.1457

[0865] 22. Melchior F. SUMO— nonclassical ubiquitin. Annu Rev Cell Dev Biol. 2000;16:591-626.

[0866] 23. Rajan S, et al. Sumoylation silences the plasma membrane leak K+ channel K2P1. Cell. 2005;121:37-47. doi: 10.1016 / j.cell.2005.01.019

[0867] 24. Plant LD, et al. One SUMO is sufficient to silence the dimeric potassium channel K2P1. Proceedings of the National Academy of Sciences of the United States of America. 2010;107:10743-10748. doi: 10.1073 / pnas.1004712107

[0868] 25. Plant LD. et al. K2P1 assembles with K2P3 or K2P9 to form SUMO- regulated TASK background channels. Biophys J. 2010;98:710a. doi:

[0869] 10.1016 / j.bpj.2009.12.3894

[0870] 26. Plant LD, et al. SUMOylation silences heterodimeric TASK potassium channels containing K2P1 subunits in cerebellar granule neurons. Science signaling. 2012;5:ra84. doi: 10.1126 / scisignal.2003431

[0871] 27. Xiong D, et al. SUMOylation determines the voltage required to activate cardiac IKs channels. Proceedings of the National Academy of Sciences of the United States of America. 2017;114:E6686-E6694. doi: 10.1073 / pnas.1706267114

[0872] 28. Plant LD. Marks JD, Goldstein SA. SUMOylation ofNaV1.2 channels mediates the early response to acute hypoxia in central neurons. eLife. 2016;5. doi: 10.7554 / eLife.20054

[0873] 29. Lee A, et al. Post-translational Modifications in Heart Failure: Small Changes, Big Impact. Heart Lung Circ. 2016;25:319-324. doi: 10.1016 / j.hlc.2015.11.008

[0874] 30. Zhao W, Zhang X, Rong J. SUMOylation as a Therapeutic Target for Myocardial Infarction. Front Cardiovasc Med. 2021;8:701583. doi: 10.3389 / fcvm.2021.701583 31. Kotler O, et al. SUMOylation of NaV1.2 channels regulates the velocity of backpropagating action potentials in cortical pyramidal neurons. bioRxiv.

[0875] 2022:2022.2008.2002.502500. doi: 10.1101 / 2022.08.02.502500

[0876] 32. Awasthi S, et al. Multimodal SHG-2PF Imaging of Microdomain Ca2+- Contraction Coupling in Live Cardiac Myocytes. Circ Res. 2016;118:el9-28. doi: 10.1161 / CIRCRESAHA.115.307919

[0877] 33. Zhang XD, et al. Prestin amplifies cardiac motor functions. Cell Rep. 2021:35:109097. doi: 10.1016 / j.celrep.2021.109097

[0878] 34. Plant LD. et al. SUMO modification of cell surface Kv2.1 potassium channels regulates the activity of rat hippocampal neurons. J Gen Physiol. 2011 ; 137 :441 -454. doi: 10.1085 / jgp.201110604

[0879] 35. Johnson ES, Blobel G. Cell cycle-regulated attachment of the ubiquitin-related protein sumo to the yeast septins. J Cell Biol. 1999;147:981-994. doi:

[0880] 10.1083 / jcb.147.5.981

[0881] 36. Johnson ES, Gupta AA. An E3-like factor that promotes SUMO conjugation to the yeast septins. Cell. 2001;106:735-744.

[0882] 37. Henley JM. Craig TJ, Wilkinson KA. Neuronal SUMOylation: mechanisms, physiology, and roles in neuronal dysfunction. Physiol Rev. 2014;94:1249-1285. doi: 10.1152 / physrev.00008.2014

[0883] 38. Hotz PW, Muller S, Mendler L. SUMO-specific Isopeptidases Tuning Cardiac SUMOylation in Health and Disease. Front Mol Biosci. 2021;8:786136. doi: 10.3389 / fmolb.2021.786136

[0884] 39. Wilkinson KA, et al. Commentary: Analysis of SUMO1 -conjugation at synapses. Front Cell Neuros ci. 2017;ll:345. doi: 10.3389 / fncel.2017.00345

[0885] 40. Choi JH, et al. Regulation of mGluR7 trafficking by SUMOylation in neurons. Neuropharmacology. 2016;102:229-235. doi: 10.1016 / j.neuropharm.2015.11.021

[0886] 41. Dai XQ, Kobe J, Marchi P, Sipione S, Macdonald PE. SUMOylation regulates Kv2.1 and modulates pancreatic beta-cell excitability. J Cell Sci. 2009;122:775-779. doi: 10.1242 / jcs.036632

[0887] 42. Lee YJ. et al. Protein SUMOylation is massively increased in hibernation torpor and is critical for the cytoprotection provided by ischemic preconditioning and hypothermia in SHSY5Y cells. J Cereb Blood Flow Metab. 2007;27:950-962. doi: 10.1038 / sj.jcbfm.9600395 43. Boening JA, et al. The effect of blocking sodium influx on anoxic damage in the rat hippocampal slice. Neuroscience. 1989;33:263-268.

[0888] 44. Stys PK, et al. Ionic mechanisms of anoxic injury in mammalian CNS white matter: role ofNa+ channels and Na(+)-Ca2+ exchanger. JNeurosci. 1992:12:430- 439.

[0889] 45. Sundt TM, Jr., et al. Correlation of cerebral blood flow and electroencephalographic changes during carotid endarterectomy: with results of surgery and hemodynamics of cerebral ischemia. Mayo Clin Proc. 1981;56:533-543.

[0890] 46. Kotler O. et al. SUMOylation of Na(V)1.2 channels regulates the velocity of backpropagating action potentials in cortical pyramidal neurons. eLife. 2023;12. doi: 10.7554 / eLife.81463

[0891] 47. Seel er JS, Dejean A. SUMO and the robustness of cancer. Nat Rev Cancer. 2017:17: 184-197. doi: 10.1038 / nrc.2016.143

[0892] 48. Langston SP, et al. Discovery' of TAK-981, a First-in-Class Inhibitor of SUMO-Activating Enzyme for the Treatment of Cancer. J Med Chem. 2021;64:2501- 2520. doi: 10.1021 / acs.jmedchem.0c01491

[0893] 49. Kumar S. et al. Targeting pancreatic cancer by TAK-981 : a SUMOylation inhibitor that activates the immune system and blocks cancer cell cycle progression in apreclinical model. Gut. 2022;71:2266-2283. doi: 10.1136 / gutjnl-2021-324834

[0894] 50. Jones SP, et al. The NHLBLsponsored Consortium for preclinicAl assESsment of cARdioprotective therapies (CAESAR): a new paradigm for rigorous, accurate, and reproducible evaluation of putative infarct-sparing interventions in mice, rabbits, and pigs. Circulation research. 2015;116:572-586. doi:

[0895] 10.1161 / CIRCRESAHA.116.305462

[0896] 51. Peters RW, Gold MR. The influence of gender on arrhythmias. Cardiol Rev.

[0897] 2004:12:97-105. doi: 10.1097 / 01. crd.0000096416.94304. bd

[0898] 52. Doris U, et al. A sexy approach to pacemaking: differences in function and molecular make up of the sinoatrial node. Histol Histopathol. 2019;34:1255-1268. doi: 10.14670 / HH-18-115

[0899] 53. Jeevaratnam K. et al. Differences in sino- atrial and atrio-ventricular function with age and sex attributable to the Scn5a+ / - mutation in a murine cardiac model. Acta Physiol (Oxf). 2010;200:23-33. doi: 10.1111 / j .1748-1716.2010.02110.x 54. Qi Y, et al. Hyper- SUMOylation of the Kv7 potassium channel diminishes the M-current leading to seizures and sudden death. Neuron. 2014;83:1159-1171. doi:

[0900] 10.1016 / j.neuron.2014.07.042

[0901] 55. Hickey CM, Wilson NR, Hochstrasser M. Function and regulation of SUMO proteases. Nat Rev Mol Cell Biol. 2012;13:755-766. doi: 10.1038 / nrm3478

[0902] 56. Odeh HM, et al. The SUMO-specific isopeptidase SENP2 is targeted to intracellular membranes via a predicted N-terminal amphipathic alpha-helix. Mol Biol Cell. 2018;29:1878- 1890. doi: 10.1091 / mbc.E17-07-0445

[0903] 57. Nishida T, et al. Characterization of a novel mammalian SUMO-1 / Smt3- specific isopeptidase, a homologue of rat axam, which is an axin-binding protein promoting beta-catenin degradation. J Biol Chem. 2001;276:39060-39066. doi: 10.1074 / jbc.M103955200

[0904] 58. Wei X, et al. Pre- and Delayed Treatments With Ranolazine Ameliorate Ventricular Arrhythmias and Navi.5 Downregulation in Ischemic / Reperfused Rat Hearts. J Cardio vase Pharmacol. 2016;68:269-279. doi:

[0905] 10.1097 / FJC.0000000000000412

[0906] 59. O'Hara T. et al. Simulation of the undiseased human cardiac ventricular action potential: model formulation and experimental validation. PLoS Comput Biol.

[0907] 2011;7:el002061. doi: 10.1371 / joumal.pcbi.1002061

[0908] 60. He Xet al. Probing the roles of SUMOylation in cancer cell biology by using a selective SAE inhibitor. Nat Chem Biol. 2017;13:1164-1171. doi:

[0909] 10.1038 / nchembio.2463

[0910] 61. Li YJ, et al. Allosteric Inhibition of Ubiquitin-like Modifications by a Class of Inhibitor of SUMO-Activating Enzy me. Cell Chem Biol. 2019;26:278-288 e276. doi: 10.1016 / j.chembiol.2018.10.026

[0911] 62. Lv Z. et al. Molecular mechanism of a covalent allosteric inhibitor of SUMO El activating enzyme. Nat Commun. 2018;9:5145. doi: 10.1038 / s41467-018-07015-l

[0912] 63. Maltsev VA, Kyle JW, Mishra S, Undrovinas A. Molecular identity of the late sodium current in adult dog cardiomyocytes identified by Navi.5 antisense inhibition. Am J Physiol Heart Circ Physiol. 2008;295:H667- 676. doi:

[0913] 10.1152 / ajpheart.00111.2008 64. O'Kelly I, Butler MH, Zilberberg N, Goldstein SA. Forward transport. 14-3-3 binding overcomes retention in endoplasmic reticulum by dibasic signals. Cell.

[0914] 2002;111:577-588. doi: 10.1016 / S0092-8674(02)01040-l

[0915] 65. O'Kelly I, Goldstein SAN. Forward transport of K2P3.1: mediation by 14-3-3 and COPI, modulation by pl 1.

[0916] Traffic. 2008;9:72-78.

[0917] 66. Levy DI, et al. The membrane protein MiRP3 regulates Kv4.2 channels in a KChlP-dependent manner. J Physiol. 2010;588:2657-2668. doi:

[0918] 10.1113 / jphysiol.2010.191395

[0919] 67. Rafizadeh S, et al. Functional interaction with filamin A and intracellular Ca2+ enhance the surface membrane expression of a small-conductance Ca2+-activated K+ (SK2) channel. Proceedings of the National Academy of Sciences of the United States ofAmerica. 2014;111:9989-9994. doi: 10.1073 / pnas.1323541111

[0920] 68. Belle A, et al. Quantification of protein half-lives in the budding yeast proteome. Proceedings of the National Academy of Sciences of the United States of America. 2006;103:13004-13009. doi: 10.1073 / pnas.0605420103

[0921] 69. Ziane R, et al. Cell membrane expression of cardiac sodium channel Na(v) 1.5 is modulated by alpha-actinin-2 interaction. Biochemistry. 2010;49:166- 178. doi: 10.1021 / bi901086v

[0922] 70. Lopes CMB, et al. Single-Cell Electrophysiology and Ion Channel opathies. In: Berul CI, Towbin JA, eds. Molecular Genetics of Cardiac Electrophysiology. Boston, MA: Springer US; 2000:3- 21.

[0923] 71. Kalogeris T, et al. Cell biology of ischemia / reperfusion injury. Int Rev Cell Mol Biol. 2012;298:229-317. doi: 10.1016 / B978-0-12-394309-5.00006-7

[0924] 72. Cartier E. et al. The SUMO-Conjugase Ubc9 Prevents the Degradation of the Dopamine Transporter. Enhancing Its Cell Surface Level and Dopamine Uptake.

[0925] Front Cell Neurosci. 2019;13:35. doi: 10.3389 / fncel.2019.00035

[0926] 73. Sun YH, et al. Human induced pluripotent stem cell line with genetically encoded fluorescent voltage indicator generated via CRISPR for action potential assessment post-cardiogenesis. Stem Cells. 2020;38:90-101. doi: 10.1002 / stem.3085

[0927] 74. Yau TY, Sander W. Eidson C, Courey AJ. SUMO Interacting Motifs: Structure and Function. Cells. 2021;10. doi: 10.3390 / cellslOl 12825 75. Lussier-Price M, et al. Characterization of a C-Terminal SUMO-Interacting Motif Present in Select PIAS-Family Proteins. Structure. 2020;28:573-585 e575. doi: 10.1016 / j.str.2020.04.002

[0928] 76. Reverter D. Lima CD. Insights into E3 ligase activity revealed by a SUMO- RanGAPl-Ubc9-Nup358 complex. Nature. 2005;435:687-692. doi:

[0929] 10.1038 / nature03588

[0930] 77. Baba D, et al. Crystal structure of thymine DNA glycosylase conjugated to SUMO-1. Nature. 2005;435:979-982.

[0931] 78. Ryu HY, Hochstrasser M. Histone sumoylation and chromatin dynamics. Nucleic Acids Res. 2021;49:6043- 6052. doi: 10.1093 / nar / gkab280

[0932] 79. Zhao Q, et al, GPS-SUMO: a tool for the prediction of sumoylation sites and SUMO-interaction motifs. Nucleic Acids Res. 2014;42:W325-330. doi:

[0933] 10.1093 / nar / gku383

[0934] 80. Beauclair G, et al. JASSA: a comprehensive tool for prediction of SUMOylation sites and SIMs. Bioinformatics. 2015;31:3483-3491. doi: 10.1093 / bioinformatics / btv403

[0935] 81. Wheat A, et al. Protein interaction landscapes revealed by advanced in vivo cross-linking-mass spectrometry. Proceedings of the National Academy of Sciences of the United States of America. 2021;118. doi: 10.1073 / pnas.2023360118

[0936] 82. Gutierrez C, et al. Enabling Photoactivated Cross-Linking Mass Spectrometric Analysis of Protein Complexes by Novel MS-Cleavable Cross-Linkers. Mol Cell Proteomics. 2021;20: 100084. doi: 10.1016 / j.mcpro.2021.100084

[0937] 83. Jiang D, et al. Structure of the Cardiac Sodium Channel. Cell. 2020:180:122- 134 e110. doi: 10.1016 / j.cell.2019.11.041

[0938] 84. Li J, et al. Fibroblast Growth Factor 21 Ameliorates Na(V)1.5 and Kir2.1 Channel Dysregulation in Human AC 16 Cardiomyocytes. Front Pharmacol.

[0939] 2021; 12:715466. doi: 10.3389 / fphar.202L 715466

[0940] 85. Zhao R, et al. Direct activation of the proton channel by albumin leads to human sperm capacitation and sustained release of inflammatory mediators by neutrophils. Nat Commun. 2021;12:3855. doi: 10.1038 / s41467-021- 24145-1

[0941] 86. Zhao R, et al. Molecular determinants of inhibition of the human proton channel hHvl by the designer peptide C6 and a bivalent derivative. Proceedings of the National Academy of Sciences of the United States of America. 2022;119:e2120750119. doi: 10.1073 / pnas.2120750119

[0942] 87. Hay RT. SUMO: AHistory of Modification. Molecular Cell. 2005;18:1-12.

[0943] 88. Jakobs A. Koehnke J, Himstedt F, Funk M, Korn B, Gaestel M, Niedenthal R. Ubc9 fusion-directed SUMOylation (UFDS): a method to analyze function of protein SUMOylation. Nat Meth. 2007;4:245-250.

[0944] 89. Takahashi H, et al. Noncovalent SUMO-1 binding activity of thymine DNA glycosylase (TDG) is required for its SUMO-1 modification and colocalization with the promyelocytic leukemia protein. J Biol Chem. 2005;280:5611-5621. doi: 10.1074 / jbc.M408130200

[0945] 90. Bekes M, Prudden J, Srikumar T, Raught B, Boddy MN, Salvesen GS. The dynamics and mechanism of SUMO chain deconjugation by SUMO-specific proteases. J Biol Chem. 2011;286:10238-10247. doi: 10.1074 / jbc.Ml 10.205153

[0946] 91. Zhang XD, et al. Critical roles of a small conductance Ca(2)(+)-activated K(+) channel (SK3) in the repolarization process of atrial myocytes. Cardiovasc Res. 2014:101:317-325. doi: 10.1093 / cvr / cvt262

[0947] 92. Zhang Z, et al. Functional roles of Cavl.3 (alphalD) calcium channels in atria: insights gained from gene-targeted null mutant mice. Circulation. 2005;112:1936- 1944. doi: CIRCULATIONAHA.105.540070 [pii] 10.1161 / CIRCULATIONAHA. 105.540070

[0948] 93. Ren L, et al. Disruption of mitochondria-sarcoplasmic reticulum microdomain connectomics contributes to sinus node dysfunction in heart failure. Proc Natl Acad Sci U S A. 2022;119:e2206708119. doi: 10.1073 / pnas.2206708119

[0949] 94. Ren L, et al. Adenylyl cyclase isoform 1 contributes to sinoatrial node automaticity via functional microdomains. JCI Insight. 2022;7. doi: 10.1172 / jci.insight.l62602

[0950] 95. Sirish P, et al. Action Potential Shortening and Impairment of Cardiac Function by Ablation of Slc26a6. Circ Arrhythm Electrophysiol. 2017;10. doi: 10.1161 / CIRCEP.117.005267

[0951] 96. Sirish P, et al. Unique mechanistic insights into the beneficial effects of soluble epoxide hydrolase inhibitors in the prevention of cardiac fibrosis. Proc Natl Acad Sci U S A. 2013;110:5618-5623. doi: 1221972110

[0952] [pii] 10.1073 / pnas.1221972110 97. Sirish P, et al. Molecular Mechanisms and New Treatment Paradigm for Atrial Fibrillation. Circ Arrhythm Electrophysiol. 2016;9:e003721. doi:

[0953] 10.1161 / CIRCEP.115.003721

[0954] 98. Sirish P, et al. Suppression of inflammation and fibrosis using soluble epoxide hydrolase inhibitors enhances cardiac stem cell-based therapy. Stem Cells Transl Med. 2020;9:1570-1584. doi: 10.1002 / sctm.20-0143

[0955] 99. Remme CA, et al. Genetically determined differences in sodium current characteristics modulate conduction disease severity in mice with cardiac sodium channelopathy. Circ Res. 2009;104:1283-1292. doi:

[0956] 10.1161 / CIRCRESAHA.109.194423

[0957] 100. Wan E, et al. Aberrant sodium influx causes cardiomyopathy and atrial fibrillation in mice. J Clin Invest. 2016;126:112-122. doi: 10.1172 / JCI84669

[0958] 101. Portero V, et al. K(V)4.3 Expression Modulates Na(V)1.5 Sodium Current. Front Physiol. 2018;9: 178. doi: 10.3389 / fphys.2018.00178

[0959] 102. Abrams J, et al. Fibroblast grow th factor homologous factors tune arrhythmogenic late NaV1.5 current in calmodulin binding-deficient channels. JCI Insight. 2020;5. doi: 10.1172 / j ci. insight.141736

[0960] 103. Nakamura A, et al. The SUMOylation inhibitor subasumstat potentiates rituximab activity by IFNI -dependent macrophage and NK cell stimulation. Blood. 2022;139:2770-2781. doi: 10.1182 / blood.2021014267

[0961] 104. Du L. et al. SUMOylation inhibition enhances dexamethasone sensitivity in multiple myeloma. J Exp Clin Cancer Res. 2022;41:8. doi: 10.1186 / sl 3046-021 - 02226-9

[0962] 105. Hanel W, et al. A sumoylation program is essential for maintaining the mitotic fidelity in proliferating mantle cell lymphoma cells. Exp Hematol Oncol. 2022:11 :40. doi: 10.1186 / S40164-022-00293 -y

[0963] 106. Xiao J, et al. UBC9 deficiency enhances immunostimulatory macrophage activation and subsequent antitumor T cell response in prostate cancer. J Clin Invest.

[0964] 2023;133. doi: 10.1172 / JCI158352

[0965] 107. Huang J, et al. Inhibition of Drpl SUMOylation by ALR protects the liver from ischemia- reperfusion injury. Cell Death Differ. 2021;28:1174-1192. doi:

[0966] 10.1038 / s41418-020-00641 -7 108. Baik H, et al. Targeting the SUMO Pathway Primes All-trans Retinoic Acid- Induced Differentiation of Nonpromyelocytic Acute Myeloid Leukemias. Cancer Res.

[0967] 2018;78:2601-2613. doi: 10.1158 / 0008- 5472.CAN-17-3361

[0968] 109. Dehghani T, et al. Selectin-targeting glycosaminoglycan-peptide conjugate limits neutrophil-mediated cardiac reperfusion injury. Cardiovasc Res. 2022;118:267- 281. doi: 10.1093 / cvr / cvaa312

[0969] 110. Takimoto E, et al. Chronic inhibition of cyclic GMP phosphodiesterase 5 A prevents and reverses cardiac hypertrophy. Nat Med. 2005;11:214-222.

[0970] 111. Mitchell GF, et al. Measurement of heart rate and Q-T interval in the conscious mouse. Am J Physiol. 1998;274:H747-751.

[0971] 112. Banyasz T, et al. Sequential dissection of multiple ionic currents in single cardiac myocytes under action potential-clamp. J Mol Cell Cardiol. 2011;50:578-581. doi : 10.1016 / j .yj mcc.2010.12.020

[0972] 113. Banyasz T, et al . Profile of L-type Ca2+ current and Na+ / Ca2+ exchange current during cardiac action potential in ventricular myocytes. Heart Rhythm. 2012:9:134-142. doi: 10.1016 / j.hrthm.2011.08.029

[0973] 114. Banyasz T, Jian Z, Horvath B, Khabbaz S, Izu LT, Chen-Izu Y. Beta- adrenergic stimulation reverses the IKr-IKs dominant pattern during cardiac action potential. Pflugers Arch. 2014;466:2067-2076. doi: 10.1007 / s00424-014-1465-7

[0974] 115. Hegyi B, et al. Complex electrophysiological remodeling in postinfarction ischemic heart failure. Proc Natl Acad Sci U S A. 2018;115:E3036-E3044. doi: 10.1073 / pnas.1718211115

[0975] 116. Xu Y, et al. Molecular identification and functional roles of a Ca2+-activated K+ channel in human and mouse hearts. J Biol Chem. 2003;278:49085-49094.

[0976] 117. Chiamvimonvat N, et al. Functional consequences of sulfhydryl modification in the pore-forming subunits of cardiovascular Ca2+ and Na+ channels. Circ Res. 1995;76:325-334. doi: 10.1161 / 01.res.76.3.325

[0977] 118. Tuteja D, et al. Differential expression of small-conductance Ca2+-activated K+ channels SK1, SK2, and SK3 in mouse atrial and ventricular myocytes. Am J Physiol Heart Circ Physiol. 2005;289:H2714-2723. doi: 00534.2005 [pn]10.1152 / ajpheart.00534.2005

[0978] 119. Tuteja D, et al. Cardiac small conductance Ca2+-activated K+ channel subunits form heteromultimers via the coiled-coil domains in the C termini of the channels. Circ Res. 2010;107:851-859. doi: CIRCRESAHA.109.215269

[0979] [pii] 10.1161 / CIRCRES AH A.109.215269

[0980] 120. Lu L, et al. Molecular coupling of a Ca2+-activated K+ channel to L-type Ca2+ channels via alpha-actinin2. Circ Res. 2007:100:112- 120. doi:

[0981] 01. RES.0000253095.44186.72 [ph]10.1161 / 01.RES.0000253095.44186.72

[0982] 121. Lu L, et al. Alpha-actinin2 cytoskeletal protein is required for the functional membrane localization of a Ca2+-activated K+ channel (SK2 channel). Proc Natl Acad Sci U S A. 2009;106:18402-18407. doi: 0908207106

[0983] [pii] 10.1073 / pnas.0908207106

[0984] 122. Li N, et al. Ablation of a Ca2+-activated K+ channel (SK2 channel) results in action potential prolongation in atrial myocytes and atrial fibrillation. J Physiol.

[0985] 2009;587:1087-1100. doi: jphysiol.2008.167718 [pii] 10.1113 / jphysiol.2008.167718

[0986] 123. Jian Z, et al. Mechanochemotransduction during cardiomyocyte contraction is mediated by localized nitric oxide signaling. Sci Signal. 2014;7:ra27. doi:

[0987] 10.1126 / scisignal.2005046

[0988] 124. Awasthi S, et al. Label-free identification and characterization of human pluripotent stem cell-derived cardiomyocytes using second harmonic generation (SHG) microscopy. J Biophotonics. 2012;5:57-66. doi: 10.1002 / jbio.201100077

[0989] 125. Levene H. Contributions to Probability and Statistics. In: Olkin I, ed. Essays in Honor of Harold Hotelling. Stanford University Press; 1960:278-292.

[0990] A number of embodiments of the invention have been described.

[0991] Nevertheless, it can be understood that various modifications may be made without departing from the spirit and scope of the invention. Accordingly, other embodiments are within the scope of the following claims.

Claims

WHAT IS CLAIMED IS:

1. A method for treating, ameliorating, lessening the symptoms of, or preventing: angina, myocardial infarction, reperfusion injury, a ventricular arrhythmia (VA), cardiac pump failure due to myocardial infarction (MI) (optionally cardiac pump failure due to myocardial infarction (Ml) in an early ischerma / reperfusion (I / R) period), congestive heart failure (CHF), hypovolemia, acidosis, hyperkalemia, hypokalemia, a cardiac poisoning or cardiopathology due to a toxin or poisoning, cardiac tamponade, tension pneumothorax, coronary thrombosis, sepsis, shock, pulmonary thrombosis, pulmonary embolism, respiratory decompensation, arrythmia a cardiac event caused by hypoperfusion and / or cardiac arrest, comprising administering to an individual in need thereof a drug or composition capable of suppressing hypoxia-induced SUMOylation of a cardiac sodium channel NaV1.5, thereby preventing or inhibiting or lessening the amount or severity of hypoxia-induced post-translational modification (PTM) of cardiac sodium channel NaV1.5, and preventing, inhibiting or lessening pathologic elevations in intracellular Na+ current (ILATE), or treating, ameliorating, lessening the symptoms of. or preventing: angina, myocardial infarction, reperfusion injury, ventricular arrhythmias (VA), pump failure due to myocardial infarction (MI) in an early ischemia / reperfusion (ER) period, and / or congestive heart failure (CHF), hypovolemia, acidosis, hyperkalemia, hypokalemia, cardiopathology due to a toxin, cardiac tamponade, tension pneumothorax, coronary thrombosis, sepsis, shock, pulmonary thrombosis, pulmonary embolism, respiratory decompensation, arrythmia and other causes of cardiac hypoperfusions and / or cardiac arrest.

2. The method of claim 1, wherein the drug or composition capable of suppressing hypoxia-induced SUMOylation of a cardiac sodium channel NaV1.5 comprises:(a) subasumstat, or CAS No. 1858276-04-6, or a compound having the formula (also called TAK-981):(b) a compound having the formula:(c) a compound having the formula,(also called 2-D08);(d) ginkgolic acid or ginkgolic acid derivatives thereof, or a compound having the formula:(e) anacardic acid or anacardic acid derivatives thereof, or 2-Hydroxy-6- pentadecylbenzoic acid;(f) kerriamycin B or kerriamycin B derivatives thereof, or 9-[4-[5-(4.5-dihydroxy-6- methyloxan-2-yl)oxy-6-methyloxan-2-yl]oxy-5-hydroxy-6-methyloxan-2-yl] -3 ,4a, 8-trihydroxy-12b-(5-hydroxy-6-methyloxan-2-yl)oxy-3-methyl-2.4-dihydrobenzo[a]anthracene- 1,7,12-trione;(g) davidiin or davidiin derivatives thereof, or |(1S,19R,2OR,21S,22R)-6,7,8,11,12,13-hexahydroxy-3,16-dioxo-21,22-bis[(3,4,5-trihydroxybenzoyl)oxy]-2,17,23- trioxatetracyclo[17.3.1.04,9.010,15]tricosa-4,6,8.10,12.14-hexaen-20-yl] 3,4,5- trihydroxybenzoate;(h) tannic, or gallotannic acid, or 1.2.3.4.6-penta-O-{3,4-dihydroxy-5-[(3.4,5- trihydroxybenzoyl)oxy]benzoyl}-D-glucopyranose, or a compound having the structure:(i) a SUMO adenylate (AMSN) or a tetrahedral SUMO intermediate (AVSN) analogues;(j) MLN4924, or pevonedistat (CAS no. 905579-51-3), or a compound having the structure:(k) Dimethyl 1-(1-anilinobut-3-enyl)-7-oxabicyclo[2.2.1]hepta-2,5-diene-2,3- dicarboxylate (or 1-(1-Phenylamino-but-3-enyl)-7-oxa-bicyclo[2.2.1]hepta-2,5-diene- 2, 3-dicarboxylic acid dimethyl ester), or PubChem CID no. 9549553;(1) compound COH000, or CAS No. 1534358-79-6, or a compound having the structure:(m) ML-792, or CAS No. 1644342-14-2, or a compound having the structure:(n) ML-93, or a compound having the structure or formula or formula:(o) McM025044, or a compound having the structure or formula or formula:(p) glycyrrhizin (or glycyrrhizic acid or glycyrrhizinic acid) I, or a compound having the structure:(q) spectomycin B1, or a compound having the structure:(r) chaetochromin A, or PubChem CID no. 6712966, or (2R,3R)-5,6,8- trihydroxy-2,3-dimethyl-9-[(2R,3R)-5,6,8-trihydroxy-2,3-dimethyl-4-oxo-2,3- dihydrobenzo[g]chromen-9-yl]-2,3-dihydrobenzo[g]chromen-4-one, or a compound having the structure:(s) viomellein, or PubChem CID no. 3033108. or (3R)-8-[(3R)-9,10- dihydroxy-7-methoxy-3-methyl-1-oxo-3,4-dihydrobenzo[g]isochromen-8-yl]-10- hydroxy-7-methoxy-3-methyl-3,4-dihydrobenzo[g]isochromene-1,6,9-trione, or a compound having the structure:(t) a compound having the structure or formula:(u) GSK145A, or CAS No. 1609945-27-8, or a compound having the structure:(v) topotecan, or HYC AMTIN™ or POTACTASOL™, or a compound having the structure:(w) nocardione A, or a compound having the structure:(x) 33-DINOR-dunnione, or a compound having the structure or formula:33-DINOR-dehydrodunnione, or a compound having the structure or formula:(y) [3-lapachone, or 3,4-Dihydro-2.2-dimethyl-2H-naphtho[1,2-b]pyran-5.6- dione, or CAS No. 4707-32-8, , or a compound having the structure or formula:(z) macrophilone A and its methoxy derivative, or a compound having the structure or formula:(aa) a Ubc9 inhibitor having the structure or formula:(bb) an SENP inhibitor (or an aza-epoxide SENP inhibitor) having the structure or formula:(also called momordin Ic),(also called Streptonigrin),(also called NSC45384),(also called vialinin A),(also called atromentin).(cc) a compound as set forth in U.S. patent application publication no. US20170002032A9, and described in WO2016004136, as set forth in further detail, below; and / or(dd) an isomer, deuterated isoform, optical isomer or stereoisomer, a racemate or racemic mixture, an enantiomer, an individual diastereomer or a diastereomeric mixture, an analog, a crystalline product or a crystalline intermediate, a pharmaceutically acceptable salt thereof, a prodrug or a bioisostere of any of (a) to (cc); and / or(ee) a SUMO protease or a SENP protein, or a catalytic domain of a SUMO protease or a SENP protein, or a nucleic acid encoding the SUMO protease or SENP protein, or the catalytic domain of the SUMO protease or SENP protein, wherein optionally the SUMO protease or a SENP protein, or a catalytic domain of a SUMO protease or a SENP protein, comprises a:(i) SENP1, sentrin-specific protease 1 Homo sapiens, optionally a sentrin- specific protease 1 having an amino acid (aa) sequence SEQ ID NO:3, orNCBI Reference Sequence NP_OO 1254524.1, or a SENP1 catalytic domain comprising aa residues from 419 to 644 of SEQ ID NO:3;(ii) SENP2, sentrin-specific protease 2 Homo sapiens, optionally a SENP2, sentrin-specific protease 2 having an amino acid (aa) sequence SEQ ID NO:4, or NCBI Reference Sequence NP_067640.2, or a SENP2 catalytic domain compnsing aa residues 336 to 584 of SEQ ID NO:4:(iii) SENP3, sentrin-specific protease 3 Homo sapiens, optionally a sentrin-specific protease 3 having an amino acid (aa) sequence SEQ ID NO: 5, or NCBI Reference Sequence NP_056485.2, or a SENP3 catalytic domain comprising aa residues 400 to 572 of SEQ ID NO:5;(iv) SENP5, isoform 1, sentrin-specific protease 5 isoform 1 Homo sapiens, optionally a sentrin-specific protease 5 isoform 1 having an amino acid (aa) sequence SEQ ID NO:6, or NCBI Reference Sequence NP_689912.2, or a sentrin-specific protease 5 isoform 1 catalytic domain comprising aa residues 581 to 753 of SEQ ID NO:6;(v) SENP5, isoform 2, sentrin-specific protease 5 isoform 2 Homo sapiens, optionally an isoform 2, sentrin-specific protease 5 isoform 1 having an amino acid (aa) sequence SEQ ID NO:7, or NCBI Reference Sequence NP_001294974.1, or a sentrin-specific protease 5 isoform 2 catalytic domain comprising aa residues 581 to 707 of SEQ ID NO:7;(vi) SENP6, sentrin-specific protease 6 isoform 1 Homo sapiens, optionally a sentrin-specific protease 6 isoform 1 having an amino acid (aa) sequence SEQ ID NO:8, or protein Accession no. NP_056386.2, or a sentrin- specific protease 6 isoform 1 catalytic domain comprising aa residues 973 to 1067 of SEQ ID NO:8;(vii) Sentrin-specific protease 6 isoform 2 Homo sapiens, optionally a Sentrin-specific protease 6 isoform 2 having an ammo acid (aa) sequence SEQ ID NO:9, or protein Accession no. NP 001093879.1, or a Sentrin-specific protease 6 isoform 2 catalytic domain comprising aa residues 966 to 1060 of SEQ ID NO:9;(viii) Sentrin-specific protease 6 isoform 3 Homo sapiens, optionally a Sentrin-specific protease 6 isoform 3 having an amino acid (aa) sequence protein Accession no. NP_001291721.1, or a Sentrin-specific protease 6 isoform 3 catalytic domain comprising aa residues 966 to 1060 of protein Accession no. NP_001291721.1;(ix) SENP7, sentrin-specific protease 7 isoform 1 Homo sapiens, optionally a sentrin-specific protease 7 isoform 1 having an amino acid (aa) sequence SEQ ID NO: 10, or protein Accession no. NP_065705.3, or a sentrin- specific protease 7 isoform 1 catalytic domain comprising aa residues 774 to 1011 of SEQ ID NO: 10;(x) Sentrin-specific protease 7 isoform 2 Homo sapiens, optionally a Sentrin-specific protease 7 isoform 2 having an amino acid (aa) sequence SEQ ID NO:11, orprotein Accession no. NP 001070671.1, or a C -terminal catalytic domain thereof;(xi) sentrin-specific protease 7 isoform 3 Homo sapiens, optionally a sentrin-specific protease 7 isoform 3 protein Accession no. NP 001269730.1, or a C-terminal catalytic domain thereof;(xii) sentrin-specific protease 7 isoform 4 Homo sapiens, optionally a sentrin-specific protease 7 isoform 4 protein Accession no. NP 001269731.1, or a C-terminal catalytic domain thereof;(xiii) sentrin-specific protease 7 isoform 6 Homo sapiens, optionally a sentrin-specific protease 7 isoform 6 protein Accession no. NP 001269733.1, or a C-terminal catalytic domain thereof; and / or(xiv) sentrin-specific protease 7 isoform 5 Homo sapiens, optionally a sentrin-specific protease 7 isoform 5 protein Accession no. NP_001269732.1, or a C-terminal catalytic domain thereof.

3. The method of claim 1 or 2, wherein the drug or composition capable of suppressing hypoxia-induced SUMOylation of a cardiac sodium channel NaV1.5 is formulated:(a) in a liquid, a gel, a hydrogel, a vesicle, a liposome, a nanoparticle, ananolipid particle, a tablet, a gel, a powder or an aqueous or a saline formulation, or for administration in vitro or in vivo.,(b) for enteral or parenteral administration;(c) in or as a liposome, an exosome, a nanoparticle, or a nanoliposome, wherein optionally the liposome, nanoparticle, or nanoliposome is fabricated to comprise or express moieties or molecules that target the liposome, nanoparticle, or nanoliposome to a particular tissue or organ, wherein optionally the liposome, nanoparticle, or nanoliposome is fabricated to target heart tissue, myocardial tissue, and / or a coronary artery;(d) in or as a dendrimer, a tablet, a pill, a capsule, a gel, a hydrogel, a geltab, a liquid, a powder, an emulsion, a lotion, an aerosol, a spray, a lozenge, an aqueous or a sterile or an injectable solution, an eye drop, or an implant; or(e) for intravenous injection, subcutaneous injection, intramuscular injection, inhalation, or intravitreal injection.

4. The method of any of claims 1 to 3, wherein the drug or composition capable of suppressing hypoxia-induced SUMOylation of a cardiac sodium channel NaV1.5 is administered after, or immediately after, or between about 5 minutes and 5 hours of, or between about 15 minutes and 15 hours of, an individual in need thereof presenting with symptoms of angina, myocardial infarction, reperfusion injury, ventricular arrhythmia (VA). pump failure due to myocardial infarction (MI) in an early ischemia / reperfusion (I / R) period, congestive heart failure (CHF), hypovolemia, acidosis, hyperkalemia, hypokalemia, cardiopathology due to a toxin, cardiac tamponade, tension pneumothorax, coronary thrombosis, sepsis, shock, pulmonary thrombosis, pulmonary' embolism, respiratory' decompensation, arrythmia and / or cardiac arrest.

5. The method of any of claims 1 to 3, wherein the drug or composition capable of suppressing hypoxia-induced SUMOylation of a cardiac sodium channel NaV1.5 is administered to the individual in need thereof intravenously (IV), and optionally is administered IV via catheter to the heart.

6. The method of any of claims 1 to 3, wherein the drug or composition capable of suppressing hypoxia-induced SUMOylation of a cardiac sodium channel NaV1.5 is administered to the individual in need intramuscularly (IM), or by injection directly into or approximate to heart tissue, or cardiac myocytes, or heart vasculature.

7. The method of any of claims 1 to 5, wherein the drug or composition capable of suppressing hypoxia-induced SUMOylation of a cardiac sodium channel NaV1.5 is administered to the individual in need thereof by implantation of an implant in the individual in need thereof, wherein the implant contains therein or comprises the drug or composition, and the drug or composition is released to tissue (optionally a heart) surrounding or approximate to the implant by elution or effusion of the drug or composition, or by controlled release of the drug or composition from the implant into the surrounding tissue, wherein optionally the implant is fabricated as a stent.

8. The method of any of claims 1 to 5, wherein the drug or composition capable of suppressing hypoxia-induced SUMOylation of a cardiac sodium channel NaV1.5 is administered to the individual in need thereof by oral, rectal, topical routes, or by inhalation, or by vaginal, topical, nasal or pulmonary administration, or by parenteral (optionally subcutaneous, intramuscular, intravenous and intradermal) infusion.

9. Use of a drug or composition capable of suppressing hypoxia-induced SUMOylation of a cardiac sodium channel NaV1.5 for treating, ameliorating, lessening the symptoms of, or preventing: angina, myocardial infarction, reperfusion injury, ventricular arrhythmias (VA), pump failure due to myocardial infarction (MI) in an early ischemia / reperfusion (I / R) period, and / or congestive heart failure (CHF), hypovolemia, acidosis, hyperkalemia, hypokalemia, cardiopathology due to a toxin, cardiac tamponade, tension pneumothorax, coronary thrombosis, sepsis, shock, pulmonary thrombosis, pulmonary embolism, respiratory decompensation, arrythmia and other causes of cardiac hypoperfusions and / or cardiac arrest.

10. A drug or composition capable of suppressing hypoxia-induced SUMOylation of a cardiac sodium channel NaV1.5 for use in treating, ameliorating, lessening the symptoms of, or preventing: angina, myocardial infarction, reperfusion injury, ventricular arrhythmias (VA), pump failure due to myocardial infarction (MI) in an early ischemia / reperfusion (I / R) period, and / or congestive heart failure (CHF), hypovolemia, acidosis, hyperkalemia, hypokalemia, cardiopathology due to a toxin, cardiac tamponade, tension pneumothorax, coronary thrombosis, sepsis, shock, pulmonary thrombosis, pulmonary embolism, respiratory decompensation, arrythmia and other causes of cardiac hypoperfusions and / or cardiac arrest.

11. A method for treating, ameliorating, lessening the symptoms of, or preventing: angina, myocardial infarction, reperfusion injury, ventricular arrhythmias (VA), pump failure due to myocardial infarction (MI) in an early ischemia / reperfusion (I / R) period, and / or congestive heart failure (CHF), hypovolemia, acidosis, hyperkalemia, hypokalemia, cardiopathology due to a toxin, cardiac tamponade, tension pneumothorax, coronary thrombosis, sepsis, shock, pulmonary thrombosis, pulmonary embolism, respiratory decompensation, arrythmia and other causes of cardiac hypoperfusions and / or cardiac arrest, comprising administering to an individual in need thereof a compound, a composition or a nucleic acid capable of modifying a NaV1.5 K442 residue to another residue that cannot be SUMOylated.

12. The method of claim 11, wherein the nucleic acid capable of modifying a NaV1.5 K.442 residue to another residue that cannot be SUMOylated: is delivered using CRISPR technology ; or, is or comprises: an RNAi inhibitory nucleic acid molecule, a double-stranded RNA (dsRNA) molecule, a microRNA (mRNA), a small interfering RNA (siRNA), an antisense RNA, a short hairpin RNA (shRNA), an inhibitory ribozyme, or a CRISPR system such as CRISPR-Cas9.

13. The method of claim 11 or 12, wherein the nucleic acid capable of modifying a NaV1.5 K442 residue to another residue that cannot be SUMOylated is contained in an expression construct, plasmid, expression vehicle, virus or vector, orthe nucleic acid capable of modifying a NaV1.5 K442 residue to another residue that cannot be SUMOylated is formulated:(a) in a liquid, a gel, a hydrogel, a vesicle, a liposome, a nanoparticle, a nanolipid particle, a powder or an aqueous or a saline formulation, or for administration In vitro or in vivo;(b) for enteral or parenteral administration;(c) in or as a liposome, an exosome, a nanoparticle, or a nanoliposome, wherein optionally the liposome, nanoparticle, or nanoliposome is fabricated to comprise or express moieties or molecules that target the liposome, nanoparticle, or nanoliposome to a particular tissue or organ, wherein optionally the liposome, nanoparticle, or nanoliposome is fabricated to target heart tissue, myocardial tissue, and / or a coronary artery;(d) in or as a tablet, a pill, a capsule, a gel, a hydrogel, a geltab, a liquid, a powder, an emulsion, a lotion, an aerosol, a spray, a lozenge, an aqueous or a sterile or an injectable solution, an eye drop, or an implant; or(e) for intravenous injection, subcutaneous injection, intramuscular injection, inhalation, or intravitreal injection.

14. The method of any of claims 11 to 13: wherein the nucleic acid capable of modifying a NaV 1.5 K442 residue to another residue that cannot be SUMOylated is contained in an expression construct, a plasmid, an expression vehicle, a virus or a vector, and the expression construct, plasmid, expression vehicle, virus or vector is delivered or administered to the individual in need thereof, wherein optionally the expression vehicle or vector is selected from the group consisting of a herpes simplex virus, a human immunodeficiency virus (HIV), a synthetic vector, an adeno-associated virus (AAV), a lentivirus. an adenovirus and a plasmid, and optionally the AAV is selected from the group consisting of AAV1. AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10 AAV11, AAV12, pseudotyped AAV, a rhesus-derived AAV, AAVrh8, AAVrh10 and AAV- DJan AAV capsid mutant, an AAV hybrid serotype, an organ-tropic AAV, a cardiotropic AAV, and a cardiotropic AAVM41 mutant.

Images