Alpha-Vbeta-6 (alphavbeta6) integrin ligands for extrahepatic delivery

Abstract

The present invention provides: a double-stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of a target gene, comprising an antisense strand complementary to the target gene; a sense strand complementary to the antisense strand and forming a double strand region with the antisense strand; and at least one alpha-v-beta-6 (alphavbeta6) integrin targeting ligand bound to the at least one strand that mediates delivery to extrahepatic tissue, such as muscular tissue, such as skeletal muscle tissue and / or cardiac muscle tissue, or lung tissue; compositions comprising such dsRNA agents; and methods of using the same to treat an individual having a condition that would benefit from reduced expression of the gene of interest.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to U.S. Provisional Application No. 63 / 432,448, filed on December 14, 2022, the entire contents of which are incorporated herein by reference. Background Art

[0003] Effective delivery of RNAi agents to cells in vivo requires specific targeting and substantial protection from the extracellular environment, particularly serum proteins. RNAi-based therapeutics have shown promising clinical data for the treatment of liver-related disorders. However, obstacles to RNAi delivery to extrahepatic tissues remain, limiting the use of RNAi-based therapies.

[0004] One of the limiting factors is the ability to effectively deliver intact RNAi to extrahepatic tissues, such as muscle tissue, eg, skeletal muscle tissue and / or cardiac muscle tissue, or lung tissue.

[0005] Previous studies have used delivery agents such as liposomes, cationic lipids, and nanoparticles to form complexes to help RNAi agents be internalized into extrahepatic cells. However, only limited success has been reported for delivering RNAi agents to extrahepatic tissues, such as muscle tissue, after systemic administration. For example, although cholesterol is delivered to muscle in conjunction with RNAi agents after intravenous injection, high doses (50 mg / kg) are required to achieve sustainable gene silencing. In addition, cholesterol conjugates are highly toxic at high concentrations, limiting their potential for clinical application.

[0006] Thus, systemic delivery of oligonucleotides to muscle tissue remains a challenge, and thus, there is a continuing need for new and improved compositions and methods for the in vivo delivery of RNAi agents.

[0007] Sequence Listing

[0008] This application contains a sequence listing, which has been submitted electronically in .XML format and is incorporated herein by reference in its entirety. This .XML copy was created on December 12, 2023, is named "121301_21120_SL.xml", and is 7,359,147 bytes in size. The sequence listing contained in this .XML file is part of this specification and is hereby incorporated by reference in its entirety. Summary of the Invention

[0009] The present invention is based, at least in part, on the discovery of alpha-v-beta-6 (αvβ6) integrin binding compounds suitable for binding to cargo molecules (e.g., single-stranded and double-stranded oligonucleotides) to be delivered to cells, and the unexpected discovery that conjugating at least one such alpha-v-beta-6 (αvβ6) integrin binding compound to at least one strand, e.g., the sense strand, of a dsRNA agent provides unexpectedly effective in vivo delivery to extrahepatic tissues expressing αvβ6, i.e., muscle tissue, such that the dsRNA agent effectively enters and is internalized into the extrahepatic tissue, e.g., muscle tissue, e.g., skeletal muscle tissue and / or cardiac muscle tissue, or lung tissue, and unexpectedly well inhibits target gene expression in the extrahepatic tissue, e.g., muscle tissue, e.g., skeletal muscle tissue and / or cardiac muscle tissue, or lung tissue.

[0010] Thus, in one aspect, the present invention provides a double-stranded ribonucleic acid (dsRNA) agent for inhibiting target gene expression, comprising: an antisense strand that is complementary to the target gene; a sense strand that is complementary to the antisense strand and forms a double-stranded region with the antisense strand; and at least one α-v-β-6 (αvp6) integrin targeting ligand bound to at least one strand, which mediates delivery to muscle tissue, such as skeletal muscle tissue and / or cardiac muscle tissue.

[0011] In another aspect, the present invention provides a double-stranded ribonucleic acid (dsRNA) agent for inhibiting target gene expression, comprising: an antisense strand that is complementary to the target gene; a sense strand that is complementary to the antisense strand and forms a double-stranded region with the antisense strand; and at least one alpha-v-beta-6 (αvβ6) integrin targeting ligand bound to at least one strand, which mediates delivery to lung tissue.

[0012] Exemplary ligands can be found in U.S. Patent Nos. 10,023,568, 10,450,312, 10,144,733, 10,487,080, 105,13,517, and 10,000,489, the entire contents of each of which are incorporated herein by reference.

[0013] In one embodiment, the ligand is conjugated to the dsRNA agent via a linker comprising a compound of the following structure:

[0014]

[0015] In one embodiment, the ligand comprises a compound having the structure:

[0016]

[0017] In one embodiment, the ligand comprises a compound having the structure:

[0018]

[0019] In one embodiment, the ligand comprises a compound having the structure:

[0020]

[0021] In one embodiment, the ligand comprises a compound having the structure:

[0022]

[0023] In some embodiments, the ligand comprises a compound of formula (I):

[0024]

[0025] in:

[0026] Q1 is selected from the group consisting of H, halogen, OR1, NR1R2, optionally substituted sulfonyl, optionally substituted C1-C6 alkyl, optionally substituted C3-C7 cycloalkyl, optionally substituted heterocycle, optionally substituted aryl, and optionally substituted heteroaryl;

[0027] Q2 and Q4 are each independently selected from the group consisting of optionally substituted C1-C6 alkyl, optionally substituted C3-C7 cycloalkyl, optionally substituted heterocycle, optionally substituted aryl, and optionally substituted heteroaryl;

[0028] Q3 is the connector;

[0029] Q5 is optionally substituted C1-C6 alkyl, optionally substituted C3-C7 cycloalkyl, O or NR1; and

[0030] Q6 is optionally substituted C4-C7 cycloalkyl, optionally substituted heterocycle, optionally substituted aryl or optionally substituted heteroaryl; and

[0031] R1 and R2 are each independently selected from the group consisting of H, optionally substituted carbonyl, optionally substituted alkyl, optionally substituted aryl, and optionally substituted heterocyclic group.

[0032] In some embodiments, the ligand comprises a compound of formula (II):

[0033]

[0034] in:

[0035] Q1, Q7, Q8, Q9 and Q10 are each independently selected from the group consisting of H, halogen, OR1, NR1R2, optionally substituted sulfonyl, guanidino, optionally substituted C1-C6 alkyl, optionally substituted C3-C7 cycloalkyl, optionally substituted heterocycle, optionally substituted aryl and optionally substituted heteroaryl;

[0036] Q2 and Q4 are independently selected from the group consisting of optionally substituted C1-C6 alkyl, optionally substituted C3-C7 cycloalkyl, optionally substituted heterocycle, optionally substituted aryl, and optionally substituted heteroaryl;

[0037] Q3 is the connector;

[0038] Q5 is optionally substituted C1-C6 alkyl, optionally substituted C3-C7 cycloalkyl, O or NR1; and

[0039] R1 and R2 are each independently selected from the group consisting of H, optionally substituted carbonyl, optionally substituted alkyl, optionally substituted aryl, and optionally substituted heterocyclic group.

[0040] Alternatively, Q9 and Q10, together with the atoms to which they are attached, can combine to form an optionally substituted carbocyclic or heterocyclic ring.

[0041] In some embodiments, the ligand comprises a compound of formula (III):

[0042]

[0043] in:

[0044] Q1 is selected from the group consisting of H, halogen, NR1R2, OR3, optionally substituted sulfonyl, optionally substituted C1-C6 alkyl, optionally substituted C3-C7 cycloalkyl, optionally substituted heterocycle, optionally substituted aryl, and optionally substituted heteroaryl;

[0045] Q2 and Q4 are each independently selected from the group consisting of optionally substituted C1-C6 alkyl, optionally substituted C3-C7 cycloalkyl, optionally substituted heterocycle, optionally substituted aryl, and optionally substituted heteroaryl;

[0046] Q3 is the connector;

[0047] Q5 is optionally substituted C1-C6 alkyl, optionally substituted C3-C7 cycloalkyl, O or NR3;

[0048] Q6 is optionally substituted C4-C7 cycloalkyl, optionally substituted heterocycle, optionally substituted aryl or optionally substituted heteroaryl.

[0049] L1 and L2 are each independently selected from the group consisting of optionally substituted alkyl, carbonyl, sulfonyl and NR3;

[0050] R1 and R2 are each independently selected from the group consisting of H, optionally substituted carbonyl, optionally substituted alkyl, optionally substituted aryl, and optionally substituted heterocyclic group; and

[0051] R3 is selected from the group consisting of H, optionally substituted alkyl, optionally substituted aryl, and optionally substituted heterocyclic group.

[0052] In one embodiment, the ligand comprises the structure:

[0053] where * indicates linkage to the oligonucleotide; for example, via a phosphodiester or phosphorothioate linkage to the 3' end of the oligonucleotide.

[0054] In one embodiment, the ligand comprises the structure:

[0055] where * indicates linkage to the oligonucleotide; for example, via a phosphodiester or phosphorothioate linkage to the 3' end of the oligonucleotide.

[0056] In one embodiment, the ligand comprises the structure:

[0057] where * indicates linkage to the oligonucleotide; for example, via a phosphodiester or phosphorothioate linkage to the 3' end of the oligonucleotide.

[0058] In one embodiment, the ligand comprises the structure:

[0059] or a salt thereof, wherein * represents a bond to the oligonucleotide; for example, to the 5' end of the oligonucleotide.

[0060] In one embodiment, the ligand comprises the structure:

[0061] or a salt thereof, wherein * represents a bond to the oligonucleotide; for example, to the 5' end of the oligonucleotide.

[0062] In some embodiments, suitable ligands and / or ligands and linkers for use in the present invention are selected from any one of the compounds shown in the following table:

[0063]

[0064] wherein Z1 is a linker group connected to the oligonucleotide.

[0065] In one embodiment, the ligand is bound to the sense strand, eg, the 3' end of the sense strand; the 5' end of the sense strand; or both the 5' end and the 3' end of the sense strand.

[0066] In one embodiment, the ligand binds to an internal position on the sense strand, such as the 2' position on a nucleotide or a modified internucleotide linkage. In one embodiment, the ligand binds to the 2' position on a nucleotide of the sense strand.

[0067] In another embodiment, the ligand is bound to the antisense strand, eg, the 3' end of the antisense strand; the 5' end of the antisense strand; or both the 5' end and the 3' end of the antisense strand.

[0068] In one embodiment, the ligand binds to an internal position on the antisense strand, such as the 2' position on a nucleotide or a modified internucleotide linkage. In one embodiment, the ligand binds to the 2' position on a nucleotide of the antisense strand.

[0069] In one embodiment, the target gene is selected from the group consisting of myostatin (MSTN); choline receptor nicotinic alpha 1 subunit (CHRNA1); choline receptor nicotinic beta 1 subunit (CHRNB1); choline receptor nicotinic delta subunit (CHRND); choline receptor nicotinic epsilon subunit (CHRNE); choline receptor nicotinic gamma subunit (CHRNG); collagen type XIII alpha 1 chain (COL13A1); docking protein 7 (DOK7); LDL receptor-related protein 4 (LRP4); muscle-associated receptor tyrosine kinase (MUSK); synaptic receptor-associated protein (RAPSN); sodium voltage-gated channel alpha subunit 4 (SCN4A); and double homeobox 4 (DUX4), dystrophic myotonic protein kinase (DMPK), glycogen synthase 1 (GYS1), motor neuron survival factor 1 (SMN1), alpha glucosidase (GAA); adrenergic receptor beta 1 (ADRB1); calcium voltage-gated channel subunit alpha 1C (CACNA1C); calcium voltage-gated channel subunit alpha 1G (CACNA1G) (T-type calcium C channel); angiotensin II receptor type 1 (AGTR1); sodium voltage-gated channel alpha subunit 2 (SCN2A); hyperpolarization-activated cyclic nucleotide-gated potassium channel 1 (HCN1); hyperpolarization-activated cyclic nucleotide-gated potassium channel 4 (HCN4); hyperpolarization-activated cyclic nucleotide-gated potassium channel 3 (HCN3); potassium voltage-gated channel subfamily A, member 5 (KCNA5); potassium inwardly rectifier channel subfamily J, member 3 (KCNJ3); potassium inwardly rectifier channel subfamily J, member 4 (KCNJ4); phospholamban (PLN); calcium / calmodulin-dependent protein kinase II delta (CAMK2D); or phosphodiesterase 1 (PDE1).

[0070] In one embodiment, the target gene is selected from the group consisting of: myostatin (MSTN); choline receptor nicotinic alpha 1 subunit (CHRNA1); choline receptor nicotinic beta 1 subunit (CHRNB1); choline receptor nicotinic delta subunit (CHRND); choline receptor nicotinic epsilon subunit (CHRNE); choline receptor nicotinic gamma subunit (CHRNG); collagen type XIII alpha 1 chain (COL13A1); docking protein 7 (DOK7); LDL receptor-related protein 4 (LRP4); muscle-associated receptor tyrosine kinase (MUSK); synaptic receptor-associated protein (RAPSN); sodium voltage-gated channel alpha subunit 4 (SCN4A); and double homeobox 4 (DUX4), dystrophic myotonic protein kinase (DMPK), glycogen synthase 1 (GYS1), motor neuron survival factor 1 (SMN1) and alpha-glucosidase (GAA).

[0071] In one embodiment, the target gene is selected from the group consisting of: adrenergic receptor beta 1 (ADRB1); calcium voltage-gated channel subunit alpha 1C (CACNA1C); calcium voltage-gated channel subunit alpha 1G (CACNA1G) (T-type calcium c channel); angiotensin II receptor type 1 (AGTR1); sodium voltage-gated channel alpha subunit 2 (SCN2A); hyperpolarization-activated cyclic nucleotide-gated potassium channel 1 (HCN1); hyperpolarization-activated cyclic nucleotide-gated potassium channel 4 (HCN4); hyperpolarization-activated cyclic nucleotide-gated potassium channel 3 (HCN3); potassium voltage-gated channel subfamily A member 5 (KCNA5); potassium inward rectifier channel subfamily J member 3 (KCNJ3); potassium inward rectifier channel subfamily J member 4 (KCNJ4); phospholamban (PLN); calcium / calmodulin-dependent protein kinase II delta (CAMK2D); or phosphodiesterase 1 (PDE1).

[0072] In one embodiment, the target gene is selected from the group consisting of: MUC5B, TSLP, IL33, ALOX15, AGER (RAGE), MUC5AC, and STAT6.

[0073] In one aspect, the invention provides cells containing any of the dsRNA agents of the invention.

[0074] In another aspect, the present invention provides a pharmaceutical composition for inhibiting target gene expression, comprising any one of the dsRNA agents of the present invention.

[0075] In one embodiment, the dsRNA agent is present in a buffered solution, such as a buffered solution comprising acetate, citrate, prolamin, carbonate, or phosphate, or any combination thereof. In one embodiment, the buffered solution is phosphate buffered saline (PBS). In other embodiments, the dsRNA agent is present in an unbuffered solution, such as water or normal saline.

[0076] In one aspect, the present invention provides a method for inhibiting the expression of a target gene in a skeletal muscle cell and / or a cardiomyocyte. The method comprises contacting a cell with, for example, any one of the dsRNA agents of the present invention or any one of the pharmaceutical compositions of the present invention, and maintaining the cell produced in step (a) for a time sufficient to achieve degradation of the mRNA transcript of the target gene in the skeletal muscle cell and / or the cardiomyocyte, thereby inhibiting the expression of the target gene in the skeletal muscle cell and / or the cardiomyocyte.

[0077] In one aspect, the present invention provides a method for inhibiting the expression of a target gene in a lung cell. The method comprises contacting the cell with, for example, any one of the dsRNA agents of the present invention or any one of the pharmaceutical compositions of the present invention, and maintaining the cell produced in step (a) for a time sufficient to achieve degradation of the mRNA transcript of the target gene in the lung cell, thereby inhibiting the expression of the target gene in the lung cell.

[0078] In one embodiment, the cell is within a subject, eg, a human subject.

[0079] In certain embodiments, contacting a cell with a dsRNA agent or pharmaceutical composition inhibits target gene expression by at least about 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 95%.

[0080] In one embodiment, inhibiting the expression of the target gene reduces the target gene protein level in the serum of the individual by at least 30%, 40%, 50%, 60%, 70%, 80%, 90% or 95%.

[0081] In one aspect, the present invention provides a method for treating an individual suffering from a muscle disorder, such as a skeletal muscle disorder and / or a cardiac muscle disorder. The method comprises administering to the individual a therapeutically effective amount of any one of the dsRNA agents of the present invention or any one of the pharmaceutical compositions of the present invention, thereby treating the individual.

[0082] In one embodiment, the muscle disorder is selected from the group consisting of myostatin-associated muscle hypertrophy, congenital myasthenic syndrome, facioscapulohumeral muscular dystrophy (FSHD), spinal muscular atrophy (SMA), myotonic dystrophy type 1 (DM1), Pompe disease, PLN cardiomyopathy, spasticity, obstructive hypertrophic cardiomyopathy (HOCM); familial hypertrophic cardiomyopathy (FHC); heart failure with preserved ejection fraction (HFPEF); atrial fibrillation (AFIB); ventricular fibrillation (VFIB); angina; myocardial infarction (MI); heart failure or heart failure with reduced ejection fraction (HFREF); supraventricular tachycardia (SVT); hypertrophic cardiomyopathy (HCM); and PLN cardiomyopathy.

[0083] In one embodiment, the skeletal muscle disorder is selected from the group consisting of myostatin-associated muscle hypertrophy, congenital myasthenic syndrome, facioscapulohumeral muscular dystrophy (FSHD), spinal muscular atrophy (SMA), myotonic dystrophy type 1 (DMI), Pompe disease, PLN cardiomyopathy, and spasticity.

[0084] In one embodiment, the cardiomyopathy is selected from the group consisting of obstructive hypertrophic cardiomyopathy (HOCM); familial hypertrophic cardiomyopathy (FHC); heart failure with preserved ejection fraction (HFPEF); atrial fibrillation (AFIB); ventricular fibrillation (VFIB); angina; myocardial infarction (MI); heart failure or heart failure with reduced ejection fraction (HFREF); supraventricular tachycardia (SVT); hypertrophic cardiomyopathy (HCM); and PLN cardiomyopathy.

[0085] In one aspect, the present invention provides a method of treating a subject suffering from a pulmonary disorder. The method comprises administering to the subject a therapeutically effective amount of any of the dsRNA agents of the present invention or any of the pharmaceutical compositions of the present invention, thereby treating the subject.

[0086] In one embodiment, the pulmonary disorder is selected from the group consisting of: pulmonary fibrosis (e.g., idiopathic pulmonary fibrosis), nonspecific interstitial pneumonia (NSIP), usual interstitial pneumonia (UIP), Hermansky-Pudlak syndrome, progressive massive fibrosis (complication of coal workers' pneumoconiosis), connective tissue disease-associated pulmonary fibrosis, airway fibrosis in asthma and COPD, fibrosis associated with acute respiratory distress syndrome (ARDS), acute lung injury; radiation-induced fibrosis; familial pulmonary fibrosis; pulmonary hypertension, asthma, asthma and chronic sinusitis; and nasal polyps and chronic sinusitis.

[0087] The dsRNA agent or pharmaceutical composition can be administered to a subject subcutaneously, intramuscularly, intravenously, or via inhalation.

[0088] In one embodiment, the treatment methods of the present invention further comprise administering to the individual an additional agent or therapy useful for treating or preventing an extrahepatic disorder. BRIEF DESCRIPTION OF THE DRAWINGS

[0090] Figure 1 Graph depicting inhibition of Sod1 mRNA expression in quadriceps muscle, heart, liver, and lung of mice 21 days after administration of a single 2 mg / kg intravenous dose of AD-1427062, AD-1534458, AD-1534460, AD-1481903, AD-1481901, or AD-1481902, or PBS control.

[0091] Figure 2 Graph depicting the inhibition of Sod1 mRNA expression in the gastrocnemius and quadriceps muscles of mice 21 days after administration of a single 2 mg / kg intravenous dose of AD-2032892 or PBS control.

[0092] Figure 3Graph depicting inhibition of Sod1 mRNA expression in heart, quadriceps, liver, gastrocnemius, and kidney of cynomolgus monkeys 30 days after administration of a single 10 mg / kg intravenous dose of AD-2032892 or PBS control.

[0093] Figure 4 Graph depicting inhibition of Sod1 mRNA expression in mouse lungs 21 days after administration of a single 0.5 mg / kg intratracheal dose of AD-1481901, AD-2032892, or PBS control.

[0094] Figure 5 Graph depicting inhibition of Sod1 mRNA expression in the lungs of mice 21 days after administration of a single 0.5 mg / kg intranasal dose of AD-1481901, AD-2032892, or PBS control.

[0095] Figure 6 Graph depicting the inhibition of Sod1 mRNA expression in the lungs of mice 21 days after administration of a single 2 mg / kg oropharyngeal inhalation dose of AD-2032892 or PBS control.

[0096] Figure 7 Graph depicting the inhibition of Sod1 mRNA expression in the lungs of cynomolgus monkeys 30 days after administration of a single 5 mg / kg or 10 mg / kg intravenous dose of AD-2032892 or PBS control.

[0097] Figure 8 Graph depicting inhibition of Sod1 mRNA expression in the lungs of mice 10 days after administration of a single 1 mg / kg or 10 mg / kg intranasal dose of the indicated agents or PBS control.

[0098] Figure 9 Formulas (IV), (V), (X), (XII), and (XV) herein are illustrated comparatively; as depicted by the columns bounded by vertical dashed lines, embodiments of the variables in one formula also apply to the variables in the other formula; in each case, except for L′ in formula (X) and T in formulas (V) and (XV), the variables in each formula have the same names.

[0099] Figure 10 A process for preparing the oligonucleotide conjugates of the present invention is illustrated wherein the first members of the reactive pairs (Z and Z, respectively) are each 0 ]) is contacted with an oligonucleotide comprising a second member (Z′) of a reactive pair to provide a bound oligonucleotide having a ZZ covalent construct generated by the reactive pair; the oligonucleotide may be modified at the 5′ end, the 3′ end, or at an internal position (e.g., 2′-O or at an internucleotide linkage of a nucleoside) by the second member of the reactive pair.

[0100] Figure 11 Representative embodiments of formula (X) are illustrated, wherein R T1 is any of the following: a phosphorus coupling group (provided, for example, by a phosphoramidite); a divalent linking group (L L ) connected oligonucleotide; or a solid supported ligand suitable for solid phase oligonucleotide synthesis, wherein the ligand of formula (X) is connected via a divalent support linking group (L K ) is connected to the surface functional group of the solid support; a divalent linking group (L L ) can be attached to the oligonucleotide at the 5' terminus (eg, 5'-O), the 3' terminus (eg, 3'-O), or at an internal position (eg, 2'-O or at an internucleotide linkage of a nucleoside).

[0101] Figure 12 Representative embodiments of formula (XV) are illustrated, wherein R T1 is any of the following: a phosphorus coupling group (provided, for example, by a phosphoramidite); a divalent linking group (L L ) connected oligonucleotide; or a solid supported ligand suitable for solid phase oligonucleotide synthesis, wherein the ligand of formula (XV) is connected to the surface functional group of the solid support via a divalent supporting linking group (LK); the divalent linking group (L L ) can be attached to the oligonucleotide at the 5' terminus (eg, 5'-O), the 3' terminus (eg, 3'-O), or at an internal position (eg, 2'-O or at an internucleoside linkage).

[0102] Detailed description

[0103] The present invention is based, at least in part, on the discovery of alpha-v-beta-6 (αvβ6) integrin compounds; and the unexpected discovery that conjugating at least one such alpha-v-beta-6 (αvβ6) integrin compound to at least one strand, e.g., the sense strand, of a dsRNA agent provides unexpectedly effective in vivo delivery to extrahepatic tissues, i.e., muscle tissue, such that the dsRNA agent effectively enters and is internalized into extrahepatic tissues, e.g., muscle tissue, e.g., skeletal muscle tissue and / or cardiac muscle tissue, or lung tissue, and unexpectedly well inhibits target gene expression in extrahepatic tissues, e.g., muscle tissue, e.g., skeletal muscle tissue and / or cardiac muscle tissue, or lung tissue.

[0104] The following embodiments disclose how to make and use alpha-v-beta-6 (αvβ6) integrin compounds; compositions for inhibiting target gene expression containing dsRNA agents comprising such compounds that mediate delivery to extrahepatic tissue, such as muscle tissue, such as skeletal muscle tissue and / or cardiac tissue, or lung tissue, that bind to at least one chain; and compositions, uses, and methods for treating individuals who would benefit from inhibition and / or reduction of target gene expression.

[0105] I. Definition

[0106] In order to make the present invention easier to understand, some terms are first defined. In addition, it should be noted that each time a value or value range of a parameter is described, the intermediate values ​​and ranges of the values ​​are also intended to be part of the present invention.

[0107] In order to make the present invention more easily understood, certain terms are first defined. In addition, it should be noted that whenever a value or range of values ​​of a parameter is recited, the values ​​and ranges between the recited values ​​are also intended to be part of the present invention.

[0108] "A" and "an" are used herein to refer to one or more than one (ie, at least one) of the grammatical object of the article. By way of example, "an element" means one element or more than one element, eg, a plurality of elements.

[0109] The term "including" is used herein to mean the phrase "including but not limited to", and they are used interchangeably.

[0110] Unless the context clearly indicates otherwise, the term "or" is used herein to mean the term "and / or" and can be used interchangeably with each other. For example, "sense strand or antisense strand" should be understood as "sense strand or antisense strand or sense strand and antisense strand".

[0111] The term "about" is used herein to mean within the typical tolerance range in the art. For example, "about" can be understood as about 2 standard deviations from the mean. In certain embodiments, about means +10%. In certain embodiments, about means +5%. When about appears before a series of numbers or a range, it should be understood that "about" can modify each number in the series or range.

[0112] The terms "at least," "not less than," or "or more" preceding a number or a range of numbers should be understood to include the number adjacent to the term "at least" and all subsequent numbers or integers that can be logically included, as the context dictates. For example, the number of nucleotides in a nucleic acid molecule must be an integer. For example, "at least 19 nucleotides of a 21-nucleotide nucleic acid molecule" means that 19, 20, or 21 nucleotides have the specified property. When at least precedes a range of numbers or a range, it should be understood that "at least" can modify each of the numbers in the range or series.

[0113] As used herein, "no more than" or "or less" should be understood to refer to the value adjacent to the phrase, and a reasonable lower value or integer to zero as indicated by the logic of the context. For example, a duplex having an overhang of "no more than 2 nucleotides" has an overhang of 2, 1, or 0 nucleotides. When "no more than" is present before a series of numbers or a range, it should be understood that "no more than" can modify each of the numbers in the series or range. As used herein, a range includes both an upper limit and a lower limit.

[0114] As used herein, a detection method can include determining that an analyte is present in an amount below the level of detection of the method.

[0115] In the event of a conflict between the indicated target site and the nucleotide sequence of either the sense or antisense strand, the indicated sequence takes precedence.

[0116] In the event of a conflict between a sequence and its indicated site in a transcript or other sequence, the nucleotide sequence described in this specification takes precedence.

[0117] The terms used herein may be preceded and / or followed by a single dash, or a double dash "=" to indicate the order of the bonds between the named substituent and its parent moiety; a single dash indicates a single bond, and a double dash indicates a double bond or, in the case of a spiro-substituent, a pair of single bonds. In the absence of a single or double dash, it is understood that a single bond is formed between the substituent and its parent moiety; furthermore, unless a dash indicates otherwise, substituents are intended to be read "left to right." For example, C 1-6 Alkoxycarbonyloxy and -OC(O)C 1-6 6-alkyl refers to the same functionality; similarly, arylalkyl, arylalkyl-, and -alkylaryl refer to the same functionality.

[0118] In addition, certain terms herein can function as monovalent and divalent linking groups, as will be familiar to those skilled in the art, and through their presence provide a link between two other moieties. For example, an alkyl group can be a monovalent group or a divalent group; in the latter case, it will be clear to those skilled in the art that removal of additional hydrogen atoms from a monovalent alkyl group provides a suitable divalent moiety.

[0119] Throughout this invention, Used to refer to oligonucleotides; such oligonucleotides can be RNA, DNA, single-stranded RNA such as antisense oligonucleotides (ASOs), double-stranded RNA such as siRNA, and oligonucleotide derivatives such as phosphorodiamidate morpholino oligomers (PMOs).

[0120] As used herein, the term "alkenyl" means a straight or branched chain hydrocarbon containing 2 to 10 carbon atoms and, unless otherwise specified, at least one carbon-carbon double bond. Representative examples of alkenyl include, but are not limited to, ethenyl, 2-propenyl, 2-methyl-2-propenyl, 3-butenyl, 4-pentenyl, 5-hexenyl, 2-heptenyl, 2-methyl-1-heptenyl, 3-decenyl, and 3,7-dimethylocta-2,6-dienyl.

[0121] As used herein, the term "alkynyl" means a straight or branched hydrocarbon chain containing 2 to 10 carbon atoms and, unless otherwise specified, at least one carbon-carbon triple bond. Representative examples of alkynyl include, but are not limited to, 1-butynyl, 2-butynyl, 1-propynyl, and the like.

[0122] As used herein, the term "alkoxy" means an alkyl group, as defined herein, attached to the parent molecular moiety through an oxygen atom. Representative examples of alkoxy include, but are not limited to, methoxy, ethoxy, n-propoxy, 2-propoxy, n-butoxy, tert-butoxy, n-pentyloxy, and n-hexyloxy.

[0123] Unless otherwise specified, as used herein, the term "alkyl" means a straight or branched chain hydrocarbon containing 1 to 10 carbon atoms. Representative examples of alkyl include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, 3-methylhexyl, 2,2-dimethylpentyl, 2,3-dimethylpentyl, n-heptyl, n-octyl, n-nonyl, and n-decyl. When "alkyl" is a divalent linking group between two other moieties, it can also be straight or branched; examples include, but are not limited to, -CH2-, -CH2CH2-, -CH2CH2CHC(CH3)-, and -CH2CH(CH2CH3)CH2-.

[0124] As used herein, the term "aryl" means phenyl (ie, a monocyclic aryl); naphthyl or azulenyl; a bicyclic ring system containing a phenyl ring fused to a cycloalkyl, cycloalkenyl, or heterocyclyl ring. Representative examples of bicyclic aryl groups include, but are not limited to, azulenyl, naphthyl, 2,3-dihydroindan-1-yl, 2,3-dihydroindan-2-yl, 2,3-dihydroindan-3-yl, 2,3-dihydroindan-4-yl, 2,3-dihydroindan-5-yl, 2,3-dihydroindole-1-yl, indolin-2-yl, indolin-3-yl, indolin-4-yl, indolin-5-yl, indolin-6-yl, indolin-7-yl, inden-1-yl, inden-2-yl, inden-3-yl, inden-4-yl, dihydronaphthalen-2-yl, dihydronaphthalen-3-yl, yl, dihydronaphthalen-4-yl, dihydronaphthalen-1-yl, 5,6,7,8-tetrahydronaphthalen-1-yl, 5,6,7,8-tetrahydronaphthalen-2-yl, 2,3-dihydrobenzofuran-2-yl, 2,3-dihydrobenzofuran-3-yl, 2,3-dihydrobenzofuran-4-yl, 2,3-dihydrobenzofuran-5-yl, 2,3-dihydrobenzofuran-6-yl, 2,3-dihydrobenzofuran-7-yl, 2,3-dihydrobenzothiophen-2-yl, 2,3-dihydrobenzothiophen-3-yl, 2,3-dihydrobenzothiophen- 4-yl, 2,3-dihydrobenzothiophene-5-yl, 2,3-dihydrobenzothiophene-6-yl, 2,3-dihydrobenzothiophene-7-yl, 2,3-dihydrobenzothiophene-8-yl, benzo[d][1,3]dioxol-4-yl, benzo[d][1,3]dioxol-5-yl, 2H-chromen-2-on-3-yl, 2H-chromen-2-on-4-yl, 2H-chromen-2-on-5-yl, 2H-chromen-2-on-6-yl, 2H-chromen-2 -one-7-yl, 2H-benzopyran-2-one-8-yl, isoindoline-1,3-dione-2-yl, isoindoline-1,3-dione-4-yl, isoindoline-1,3-dione-5-yl, inden-1-one-2-yl, inden-1-one-3-yl, inden-1-one-4-yl, inden-1-one-5-yl, inden-1-one-6-yl, inden-1-one-7-yl, 2,3-dihydrobenzo[b][1,4]dioxan-2-yl, 2,3-dihydrobenzo[b][1,4]dioxan-5-yl, 2,3-dihydrobenzo[b][1,4]dioxan-6-yl, 2H-benzo[b][1,4]oxazin-3(4H)-on-5-yl, 2H-benzo[b][1,4]oxazin-3(4H)-on-6-yl, 2H-benzo[b][1,4]oxazin-3(4H)-on-7-yl, 2H-benzo[b][1,4]oxazin-3(4H)-on-8-yl, quinazolin-4(3H)-on-5-yl, quinazolin-4(3H)-on-6-yl, quinazolin-4(3H)-on-7-yl, Quinazolin-4(3H)-on-8-yl, quinoxalin-2(1H)-on-5-yl, quinoxalin-2(1H)-on-6-yl, quinoxalin-2(1H)-on-7-yl, quinoxalin-2(1H)-on-8-yl, benzo[d]thiazol-2(3H)-on-3-yl, benzo[d]thiazol-2(3H)-on-4-yl, benzo[d]thiazol-2(3H)-on-5-yl, benzo[d]thiazol-2(3H)-on-6-yl, and benzo[d]thiazol-2(3H)-on-7-yl.

[0125] In certain embodiments, the bicyclic aryl is (i) naphthyl or (ii) a benzene ring fused to a 5- or 6-membered monocyclic cycloalkyl, a 5- or 6-membered monocyclic cycloalkenyl, or a 5- or 6-membered monocyclic heterocyclyl, wherein the fused cycloalkyl, cycloalkenyl, and heterocyclyl are optionally substituted with one or two groups which are independently oxo or thia.

[0126] As used herein, the terms "arylalkyl," "aralkyl," "-alkylaryl," and "arylalkyl-" refer to an aryl group, as defined herein, appended to the parent molecular moiety through an alkyl group, as defined herein. Representative examples of arylalkyl include, but are not limited to, benzyl, 2-phenylethyl, 3-phenylpropyl, and 2-naphth-2-ylethyl.

[0127] The term "azido" refers to a -N3 group.

[0128] The term "carboxy" means a -COOH group.

[0129] As used herein, the terms "cyano" and "nitrile" refer to a -CN group.

[0130] As used herein, the term "cycloalkyl" means a monocyclic or bicyclic cycloalkyl ring system.

[0131] Monocyclic ring systems are cyclic hydrocarbon groups containing 3 to 10 carbon atoms, wherein such groups are saturated. Examples of monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Bicyclic cycloalkyl ring systems are bridged monocyclic or fused bicyclic rings. Bridged monocyclic rings contain monocyclic cycloalkyl rings in which two non-adjacent carbon atoms of the monocyclic ring are separated by an alkylene bridge (i.e., of the form -(CH2)) having one to three additional carbon atoms.w -bridge group, wherein w is 1, 2 or 3). Representative examples of bridged bicyclic ring systems include, but are not limited to, bicyclo[3.1.1]heptane, bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane, bicyclo[3.2.2]nonane, bicyclo[3.3.1]nonane and bicyclo[4.2.1]nonane. Fused bicyclic cycloalkyl ring systems contain a monocyclic cycloalkyl ring fused to a monocyclic cycloalkyl. Representative examples of fused bicyclic ring systems include, but are not limited to, decalinyl. The cycloalkyl is optionally substituted with one or two groups that are independently oxy or thia. In certain embodiments, the fused bicyclic cycloalkyl is a 5- or 6-membered monocyclic cycloalkyl ring fused to a 5- or 6-membered monocyclic cycloalkyl, wherein the fused bicyclic cycloalkyl is optionally substituted with one or two groups that are independently oxy or thia.

[0132] As used herein, "cycloalkenyl" refers to a monocyclic or bicyclic cycloalkenyl ring system. A monocyclic ring system is a cyclic hydrocarbon group containing 3 to 8 carbon atoms, wherein such group is unsaturated (i.e., contains at least one cyclic carbon-carbon double bond), but is not aromatic. Examples of monocyclic ring systems include cyclopentenyl and cyclohexenyl. Bicyclic cycloalkenyl rings are bridged monocyclic or fused bicyclic rings. Bridged monocyclic rings contain monocyclic cycloalkenyl rings in which two non-adjacent carbon atoms of the monocyclic ring are separated by an alkylene bridge (i.e., of the form -(CH2)) having one to three additional carbon atoms. w -bridge group, wherein w is 1, 2, or 3). Representative examples of bicyclic cycloalkenyl groups include, but are not limited to, norbornenyl and bicyclo[2.2.2]oct-2-enyl. Fused bicyclic cycloalkenyl ring systems contain a monocyclic cycloalkenyl ring fused to a monocyclic cycloalkyl or monocyclic cycloalkenyl group. The cycloalkenyl group is optionally substituted with one or two groups that are independently oxy or thia.

[0133] As used herein, the term "monocyclic" includes monocyclic aryl, monocyclic cycloalkyl, monocyclic cycloalkenyl, and monocyclic heterocyclyl.

[0134] As used herein, the term "halo" or "halogen" means -Cl, -Br, -I, or -F.

[0135] The term "H" means hydrogen.

[0136] As used herein, the term "haloalkyl" means at least one halogen, as defined herein, appended to the parent molecular moiety through an alkyl, as defined herein. Representative examples of haloalkyl include, but are not limited to, chloromethyl, 2-fluoroethyl, trifluoromethyl, pentafluoroethyl, and 2-chloro-3-fluoropentyl.

[0137] As used herein, the term "heteroaryl" means a monocyclic heteroaryl or a bicyclic ring system containing at least one heteroaromatic ring (i.e., a monocyclic or bicyclic aromatic ring system containing at least one heteroatom within the aromatic system). A monocyclic heteroaryl can be a 5- or 6-membered ring. A 5-membered ring consists of two double bonds and one, two, three, or four nitrogen atoms and, optionally, one oxygen or sulfur atom. A 6-membered ring consists of three double bonds and one, two, three, or four nitrogen atoms. A 5- or 6-membered heteroaryl is attached to the parent molecular moiety through any carbon atom or any nitrogen atom contained within the heteroaryl. Representative examples of monocyclic heteroaryls include, but are not limited to, furanyl, imidazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, oxazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, pyrazolyl, pyrrolyl, tetrazolyl, thiadiazolyl, thiazolyl, thienyl, triazolyl, and triazinyl. Bicyclic heteroaryl is composed of a monocyclic heteroaryl fused to a phenyl, monocyclic cycloalkyl, monocyclic cycloalkenyl, monocyclic heterocyclic or monocyclic heteroaryl. The fused cycloalkyl or heterocyclic moieties of the bicyclic heteroaryl are optionally substituted by one or two groups independently being oxo or thia. Representative examples of bicyclic heteroaryl include, but are not limited to, benzimidazolyl, benzofuranyl, benzothienyl, benzoxadiazolyl, benzoxathiadiazolyl, benzothiazolyl, cinnolinyl, 5,6-dihydroquinolin-2-yl, 5,6-dihydroquinolin-8-yl, 5,6-dihydroisoquinolin-1-yl, furopyridinyl, indazolyl, indolyl, isoquinolinyl, naphthyridinyl, quinolyl, purinyl, 5,6,7,8-tetrahydroquinolin-2-yl, 5,6,7,8-tetrahydroquinolin-3-yl, 5,6,7,8-tetrahydroquinolin-3-yl, 5,6,7,8-tetrahydroquinolin-4-yl, 5,6,7,8-tetrahydroquinolin-5-yl, 5,6,7,8-tetrahydroquinolin-6-yl, 5,6,7,8-tetrahydroquinolin-7-yl, 5,6,7,8-tetrahydroquinolin-8-yl, 5,6,7,8-tetrahydroquinolin-9-yl, 5,6,7,8-tetrahydroquinolin-10-yl, 5,6,7,8-tetrahydroquinolin-20-yl, 5,6,7,8-tetrahydroquinolin-3-yl, 5,6,7,8-tetrahydroquinolin-10-yl, 5,6,7,8-tetrahydroquinolin-20-yl, 5,6,7 6,7,8-tetrahydroquinolin-4-yl, 5,6,7,8-tetrahydroquinolin-5-yl, 5,6,7,8-tetrahydroquinolin-6-yl, 5,6,7,8-tetrahydroquinolin-7-yl, 5,6,7,8-tetrahydroquinolin-8-yl, 5,6,7,8-tetrahydroisoquinolin-1-yl, thienopyridinyl, 4,5,6,7-tetrahydrobenzo[c][1,2,5]oxadiazol-4-yl and 6,7-dihydrobenzo[c][1,2,5]oxadiazol-4(5H)-onyl. In certain embodiments, the fused bicyclic heteroaryl is a 5- or 6-membered monocyclic heteroaryl ring fused to a phenyl ring, a 5- or 6-membered monocyclic cycloalkyl, a 5- or 6-membered monocyclic cycloalkenyl, a 5- or 6-membered monocyclic heterocyclyl, or a 5- or 6-membered monocyclic heteroaryl, wherein the fused cycloalkyl, cycloalkenyl, and heterocyclyl are optionally substituted with one or two groups that are independently oxo or thia.

[0138] As used herein, the terms "heteroarylalkyl" and "-alkylheteroaryl" refer to a heteroaryl group, as defined herein, appended to the parent molecular moiety through an alkyl group, as defined herein. Representative examples of heteroarylalkyl include, but are not limited to, furan-3-ylmethyl, 1H-imidazol-2-ylmethyl, 1H-imidazol-4-ylmethyl, 1-(pyridin-4-yl)ethyl, pyridin-3-ylmethyl, pyridin-4-ylmethyl, pyrimidin-5-ylmethyl, 2-(pyrimidin-2-yl)propyl, thien-2-ylmethyl, and thien-3-ylmethyl.

[0139] As used herein, the term "heterocyclyl" refers to a monocyclic heterocycle or a bicyclic heterocycle. A monocyclic heterocycle is a 3-, 4-, 5-, 6-, or 7-membered ring containing at least one heteroatom independently selected from the group consisting of O, N, and S, wherein the ring is saturated or unsaturated but not aromatic. A 3- or 4-membered ring contains one heteroatom selected from the group consisting of O, N, and S. A 5-membered ring may contain zero or one double bond and one, two, or three heteroatoms selected from the group consisting of O, N, and S. A 6- or 7-membered ring contains zero, one, or two double bonds and one, two, or three heteroatoms selected from the group consisting of O, N, and S. Representative examples of monocyclic heterocycles include, but are not limited to, azetidinyl, azepanyl, aziridinyl, diazepanyl, 1,3-dioxolan-2-yl, 1,3-dithiolan-2-yl, 1,2-dithiolan-3-yl, 1,2-dithiolan-4-yl, 1,3-dithian-2 ...2-yl, 1,3-dithian-3-yl, 1,2-dithian-4-yl, 1,3-dithian-2-yl, 1,3-dithian-2-yl, 1,3-dithian-2-yl, 1,3-dithian-2-yl, 1,3-dithian-2-yl, 1,3-dithian-3-yl, 1,2-dithian-4-yl, 1,3-dithian-2-yl, 1,3-dithian-2-yl, 1,3-dithian-2-yl, 1,3-dithian-2-yl, 1,3-dithian-2-yl, 1,3-dithian-2-yl, 1,3-dithian-2-yl, 1,3-dithian-2- 1,2-dithian-3-yl, 1,2-dithian-4-yl, imidazolinyl, imidazolidinyl, isothiazolinyl, isothiazolidinyl, isoxazolinyl, isoxazolidinyl, morpholinyl, oxadiazolinyl, oxadiazolidinyl, oxazolinyl, oxazolidinyl, piperazinyl, piperidinyl, pyranyl, pyrazolinyl, pyrazolidinyl, pyrrolinyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydrothiophenyl, thiadiazolinyl, thiadiazolidinyl, thiazolinyl, thiazolidinyl, thiamorpholinyl, 1,1-dioxothiomorpholinyl (thiomorpholine sulfone), thiopyranyl, and trithianyl. A bicyclic heterocycle is a monocyclic heterocycle fused to a monocyclic cycloalkyl, monocyclic cycloalkenyl, or monocyclic heterocycle. Representative examples of bicyclic heterocyclyls include, but are not limited to, decahydroquinolinyl, decahydroisoquinolinyl, octahydro-1H-indolyl, and octahydrobenzofuranyl. The heterocyclyl is optionally substituted with one or two groups that are independently oxy or thia. In certain embodiments, the bicyclic heterocyclyl is a 5- or 6-membered monocyclic heterocyclyl ring fused to a 5- or 6-membered monocyclic cycloalkyl, a 5- or 6-membered monocyclic cycloalkenyl, or a 5- or 6-membered monocyclic heterocyclyl, wherein the bicyclic heterocyclyl is optionally substituted with one or two groups that are independently oxy or thia.

[0140] As used herein, the term "hydroxy" or "hydroxyl" refers to an -OH group. As used herein, the term "thiol" refers to an -SH group.

[0141] As used herein, the term "nitro" refers to a -NO2 group.

[0142] As used herein, the term "oxy" refers to a =0 group.

[0143] As used herein, the term "saturated" means that the referenced chemical structure does not contain any multiple carbon-carbon bonds. For example, saturated cycloalkyl groups as defined herein include cyclohexyl, cyclopropyl, and the like.

[0144] As used herein, the term "thia" refers to a =S group.

[0145] The term "amine" or "amino" encompasses compounds wherein the nitrogen atom is covalently bonded to at least one carbon or heteroatom. The term "alkylamino" includes groups and compounds wherein the nitrogen atom is bonded to at least one additional alkyl group. The term "dialkylamino" includes groups wherein the nitrogen atom is bonded to at least two additional alkyl groups.

[0146] As used herein, the term "unsaturated" means that the referenced chemical structure contains at least one multiple carbon-carbon bond (i.e., double or triple bonds or both), but is not aromatic. For example, unsaturated cycloalkyl groups as defined herein include cyclohexenyl, cyclopentenyl, cyclohexadienyl, and the like.

[0147] As used herein, the term "leaving group" means an atom or group (charged or uncharged) that leaves an atom in a substance considered to be the remainder or major portion of a substrate in a given reaction. For example, unless otherwise indicated, reactions are given herein as S as understood by those skilled in the art. N 1 or S N 2 reaction. In certain embodiments, the reaction is designated as S N 2 reactions.

[0148] As used herein, the term "substituted", whether or not preceded by the term "optionally", means that at least one hydrogen present on a group (e.g., a carbon or nitrogen atom) is replaced with a permissible substituent, e.g., a substituent that upon substitution results in a stable compound, e.g., a compound that does not spontaneously undergo transformation, such as by rearrangement, cyclization, elimination, or other reaction. Unless otherwise indicated, a "substituted" group has substituents at one or more substitutable positions of the group, and when more than one position in any given structure is substituted, the substituents may be the same or different at each position. Suitable substituents include, but are not limited to, halogen, hydroxy, thiol, nitro, alkoxy, azido, carboxyl, cyano, amino, C 1-6 Alkyl, C 2-6 Alkenyl, C 1-6 Alkoxy, C 1-6 As used herein, the term "support linker" means a divalent chemical moiety that covalently links a surface-binding functional group of a solid support (eg, an amino group of an amino-modified solid support) to another chemical moiety.

[0149] As used herein, the term "reactive pair" means two functional groups known to those skilled in the art that are capable of reacting, alone or in the presence of other reagents, to form a covalent bond between two chemical entities, each of which comprises one member of the reactive pair; the latter is referred to herein as the "linker formed by the reactive pair." In some embodiments, the reactive pair is a click pair, i.e., two functional groups capable of reacting in a click reaction to form a covalent linkage.

[0150] As used herein, the term "Michael acceptor" means an α,β-unsaturated compound that is capable of reacting with a nucleophile at the β-carbon of the electrophilic olefin of the α,β-unsaturated compound. Examples of α,β-unsaturated compounds include, but are not limited to, groups such as α,β-unsaturated aldehydes, esters, amides, sulfonyls, ketones, nitrile, or nitro groups. "α,β-unsaturated" refers to a carbon-carbon multiple bond that connects the carbon atom immediately adjacent to the mentioned aldehyde, ester, amide, sulfonyl, ketone, nitrile, or nitro group to its adjacent carbon atom. Examples of Michael acceptor groups include, but are not limited to, N-maleimide, acrolein, acrylonitrile, acrylic acid, acrylamides (e.g., N-isopropyl (isopropryl) acrylamide), acrylates (e.g., methyl acrylate), vinyl sulfones, vinyl sulfonates, and vinyl sulfonamides.

[0151] The term "hydroxy protecting group" or "hydroxy protecting group" as used herein means those functional groups well known in the art, and include those described in detail, for example, in Protecting Groups in Organic Synthesis, TW Greene and PGM Wuts, 3rd Edition, John Wiley & Sons, 1999, the entire contents of which are incorporated herein by reference. Suitable hydroxy protecting groups include, but are not limited to, acetyl, trifluoroacetyl, trichloroacetyl, pivaloyl, tert-butyl, allyl, optionally substituted benzyl (such as benzyl, 2-nitrobenzyl, 4-nitrobenzyl, 2,6-dichlorobenzyl, 4-chlorobenzyl, 4-fluorobenzyl, 4-bromobenzyl, 4-methoxybenzyl, 3,4-dimethoxybenzyl, 2-cyanobenzyl, 4-cyanobenzyl, 4-phenylbenzyl), 2-picolyl, 4-picolyl, methoxymethyl (MOM), methylthiomethyl (MTM), ethoxymethyl, 2-methoxyethoxymethyl (MEM), 2,2,2-trichloroethoxymethyl, tert-butoxymethyl, benzyloxymethyl (BOM), 4-methoxybenzyloxymethyl (Mbom), (phenyldimethylsilyl)methoxymethyl (SMOM), 2-(trimethylsilyl)ethoxymethyl (SEM), tert-butylthiomethyl, 2-tetramethylsilyl Tetrahydropyranyl (THP), 4-methoxytetrahydropyran-2-yl (MTHP), 4-methoxytetrahydrothiopyran-2-yl, 3-bromotetrahydropyran-2-yl, 2-tetrahydrothiopyranyl, trimethylsilyl (TMS), triethylsilyl (TES), triisopropylsilyl (TIPS), tert-butyldimethylsilyl (TBDMS), tert-butyldiphenylsilyl (TBDPS), isopropyldimethylsilyl (IPDMS), diethylisopropylsilyl (DEIPS), diphenylmethyl, 9-phenylxanthin-9-yl (Pixyl), 9-(p-methoxyphenyl)xanthin-9-yl (MOX), and optionally substituted trityl groups such as trityl (Trt), 2-chlorotrityl (Clt), 4-methoxytrityl (Mmt), 4-methyltrityl (Mtt), 4,4′-dimethoxytrityl (DMT), and 4,4′,4″-trimethoxytrityl.

[0152] As used herein, the term "nitrogen protecting group" means those functional groups well known in the art and includes those described in detail, for example, in Protecting Groups in Organic Synthesis, TW Greene and PGM Wuts, 3rd edition, John Wiley & Sons, 1999, the entire contents of which are incorporated herein by reference. Suitable nitrogen protecting groups include, but are not limited to, allyl (Alloc), acetyl (Ac), Adpoc (1-(1-adamantyl)-1-methylethoxycarbonyl), Boc (tert-butyloxycarbonyl), Dde ivDde Dnp (2,4-dinitrophenyl), Mmt (4-methoxytrityl), Mtt (4-methyltrityl), Teoc (2-trimethylsilylethoxycarbonyl), Tfa (trifluoroacetyl), optionally substituted trityl (e.g., trityl (Trt), 2-chlorotrityl (Clt), 4-methoxytrityl (Mmt), 4-methyltrityl (Mtt), 4,4′-dimethoxytrityl (DMT)), and 4,4′,4″-trimethoxytrityl), optionally substituted benzyloxycarbonyl (e.g., benzyloxycarbonyl (Z) or 2-chlorobenzyloxycarbonyl (2ClZ)), fluorenylmethoxycarbonyl (Fmoc), methoxyacetyl (mac), phenoxyacetyl (pac), 2-chlorophenoxyacetyl, 3-chlorophenoxyacetyl, phenoxyacetyl, 2-(tert-butyl)phenoxyacetyl, 3-(tert-butyl)phenoxyacetyl, 4-(tert-butyl)phenoxyacetyl, 2-fluorophenoxyacetyl, 3-fluorophenoxyacetyl, 4-fluorophenoxyacetyl, 2,4-difluorophenoxyacetyl, 4-(trifluoromethoxy)phenoxyacetyl, 2-phenoxypropanoyl, 2-(4-chloro-2-methylphenoxy)propanoyl, and 2-(4-chlorophenoxy)propanoyl.

[0153] As used herein, the term "phosphorus coupling group" means an H-phosphonate or phosphoroamidite that is reactive with a hydroxyl group and can form a phosphite triester or phosphorothioate triester when used within a process to make an internucleotide linkage, such as a phosphodiester or phosphorothioate linkage.

[0154] As used herein, the term "solid support" refers to any form of polymer or composite material that is not completely soluble in a solvent. By way of example only, solid supports include colloids (isolated or suspended), gels, resins, membranes, and any other form of polymer or composite material that retains properties different from those of the solvent. The term is not intended to limit in any way the size, shape, form, or chemical structure of the polymer or composite material. Such polymers or composite materials are well known in the art and include, by way of example only, cellulose, microporous glass, silica, polystyrene, polystyrene cross-linked with divinylbenzene, polyacrylamide, latex, dimethylacrylamide, dimethylacrylamide cross-linked with N,N'-bis-acryloylethylenediamine, glass, glass coated with a hydrophobic polymer, composite materials, or any other material conventionally used for solid-phase organic synthesis. In addition, the term solid support is not limited by the presence and nature of cross-linking groups and the nature of exposed functional groups. The exposed functional groups are moieties on the solid support that can react with guest molecules to form guest molecules bound to the support; preferred exposed functional groups include -OH, -SH, -NH2, silanoloxy, alkylamino, NH2NH, COOH, ester, aldehyde, -Br, -I, halomethyl (e.g., bromomethyl), and alkenyl. In addition, the exposed functional groups can be located on the surface of the solid support or dispersed throughout the solid support. In one embodiment, the solid support has a rigid or semi-rigid surface. Specific examples of solid supports include amino-terminated controlled pore glass (CPG), such as long chain alkylamine CPG (LCAA-CPG), and amino-terminated or hydroxyl-terminated cross-linked polystyrene, such as NittoPhase TM Solid support (about 420 μmol / g hydroxyl), HL solid support (about 550 μmol / g hydroxyl) and UnyLinker TM Solid supports, each available from Kinovate Life Sciences (Oceanside, CA).

[0155] As used herein, the term "activated ester" refers to a derivative of a carboxyl group that is more readily displaced by nucleophilic addition and elimination than an ethyl ester group (e.g., NHS ester, sulfonyl-NHS ester, PAM ester, or halophenyl ester). Representative carbonyl substituents of activated esters include succinimidyloxy, sulfonylsuccinimidyloxy, -1-oxybenzotriazolyl; 4-sulfonyl-2,3,5,6-tetrafluorophenyl; or an aryloxy group optionally substituted one or more times with an electron-withdrawing substituent such as nitro, fluorine, chlorine, cyano, trifluoromethyl, or a combination thereof (e.g., pentafluorophenyloxy). In certain embodiments, activated esters include succinimidyloxy and sulfonylsuccinimidyloxy esters.

[0156] As used herein, "target sequence" or "target nucleic acid" refers to a contiguous portion of the nucleotide sequence of an mRNA molecule formed during transcription of a target gene, including mRNA that is an RNA processing product of the primary transcription product. In one embodiment, the target portion of the sequence will be at least long enough to serve as a substrate for RNAi-guided cleavage at or near that portion of the nucleotide sequence of the mRNA molecule formed during transcription of the target gene. In one embodiment, the target sequence is within the protein coding region of the target gene. In another embodiment, the target sequence is within the 3'UTR of the target gene. The target nucleic acid may be a cellular gene (or mRNA transcribed from a gene) whose expression is associated with a particular condition or disease state.

[0157] The target sequence may be about 9 to 36 nucleotides in length, such as about 15 to 30 nucleotides in length. For example, the target sequence may be about 15-30 nucleotides, 15-29, 15-28, 15-27, 15-26, 15-25, 15-24, 15-23, 15-22, 15-21, 15-20, 15-19, 15-18, 15-17, 18-30, 18-29, 18-28, 18-27, 18-26, 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-30, 19 In some embodiments, the target sequence has a length of about 19 to about 30 nucleotides. In other embodiments, the target sequence has a length of about 19 to about 25 nucleotides. In yet other embodiments, the target sequence has a length of about 19 to about 23 nucleotides. In some embodiments, the target sequence has a length of about 21 to about 23 nucleotides. Intermediate ranges and lengths between the above recited ranges and lengths are also contemplated as part of the present invention.

[0158] As used herein, the term "strand comprising a sequence" refers to an oligonucleotide comprising a chain of nucleotides described by a sequence referred to using standard nucleotide nomenclature.

[0159] "G", "C", "A", "T" and "U" each generally represent a nucleotide containing guanine, cytosine, adenine, thymidine and uracil as a base, respectively. However, it should be understood that the term "ribonucleotide" or "nucleotide" can also refer to a modified nucleotide, as further described below, or an alternative replacement moiety (see, for example, Table 1). It is well known to those skilled in the art that guanine, cytosine, adenine and uracil can be replaced by other moieties without substantially changing the base pairing properties of the oligonucleotide comprising the nucleotide carrying such replacement moiety. It should be understood that when a cDNA sequence is provided, the corresponding mRNA or RNAi agent will include U in place of T. By way of example, but not limitation, a nucleotide comprising inosine as its base can base pair with a nucleotide containing adenine, cytosine or uracil. Thus, a nucleotide containing uracil, guanine or adenine can be replaced with a nucleotide containing, for example, inosine in the dsRNA nucleotide sequence characterized in the present invention. In another example, an adenine and cytosine residue at any position in the oligonucleotide can be replaced with a guanine residue and a uracil residue, respectively, to form a GU wobble base pairing with the target mRNA. Sequences containing such replacement moieties are suitable for use in the compositions and methods characterized herein. In addition, those skilled in the art will appreciate that in the RNAi agents of the present invention, T residues in the target gene sequence or its reverse complement sequence will typically be replaced with U residues.

[0160] The terms "iRNA," "RNAi agent," "iRNA agent," and "RNA interfering agent," as used interchangeably herein, refer to agents that contain RNA, as such terms are defined herein, and that mediate targeted cleavage of RNA transcripts via the RNA-induced silencing complex (RISC) pathway. RNA interference (RNAi) is a process that directs the sequence-specific degradation of mRNA. RNAi modulates, for example, inhibits, the expression of a target gene in a cell, for example, a cell within a subject, such as a mammalian subject.

[0161] In one embodiment, the RNAi agents of the present invention comprise single-stranded RNAi that interacts with a target RNA sequence (e.g., a target mRNA sequence) to direct the cleavage of the target RNA. Without wishing to be bound by theory, it is believed that long double-stranded RNA introduced into cells is broken down by a type III endonuclease called Dicer into double-stranded short interfering RNAs (siRNAs) comprising a sense strand and an antisense strand (Sharp et al., (2001) Genes Dev. 15:485). Dicer, an enzyme similar to ribozyme III, processes these dsRNAs into 19-23 base pair short interfering RNAs with a characteristic two-base 3′ overhang (Bernstein et al., (2001) Nature 409:363). These siRNAs are then incorporated into the RNA-induced silencing complex (RISC), where one or more helicases unwind the siRNA duplex, enabling the complementary antisense strand to direct target recognition (Nykanen et al., (2001) Cell 107:309). Upon binding to the appropriate target mRNA, one or more endonucleases within RISC cleave the target to induce silencing (Elbashir et al., (2001) Genes Dev. 15:188). Thus, in one aspect, the present invention relates to single-stranded RNA (ssRNA) (the antisense strand of the siRNA duplex) produced within a cell and which promotes the formation of the RISC complex to achieve silencing of the target gene. Thus, the term "siRNA" is also used herein to refer to RNAi as described above.

[0162] In another embodiment, the RNAi agent may be a single-stranded RNA introduced into a cell or organism to inhibit a target mRNA. The single-stranded RNAi agent binds to the RISC endonuclease Argonaute 2, which then cleaves the target mRNA. Single-stranded siRNAs are generally 15 to 30 nucleotides and are chemically modified. The design and testing of single-stranded RNAs are described in U.S. Patent No. 8,101,348 and Lima et al. (2012) Cell 150: 883-894, the entire contents of each of which are hereby incorporated herein by reference. Any of the antisense nucleotide sequences described herein can be used as a single-stranded siRNA as described herein, or chemically modified as described by the methods described in Lima et al. (2012) Cell 150: 883-894.

[0163] In another embodiment, the "RNAi agent" used in the compositions and methods of the present invention is double-stranded RNA, and is referred to herein as a "double-stranded RNAi agent," "double-stranded RNA (dsRNA) molecule," "dsRNA agent," or "dsRNA." The term "dsRNA" refers to a complex of ribonucleic acid molecules having a double-helical structure comprising two antiparallel and substantially complementary nucleic acid strands, referred to as having "sense" and "antisense" orientations relative to a target RNA, i.e., a target mRNA sequence. In some embodiments of the present invention, double-stranded RNA (dsRNA) triggers degradation of a target RNA (e.g., mRNA) through a post-transcriptional gene silencing mechanism, referred to herein as RNA interference or RNAi.

[0164] Generally, dsRNA molecules may include ribonucleotides, but as described in detail herein, each strand or both strands may also include one or more non-ribonucleotides, such as deoxyribonucleotides or modified nucleotides. Furthermore, as used herein, "RNAi agents" may include ribonucleotides with chemical modifications; RNAi agents may include substantial modifications at multiple nucleotides.

[0165] As used herein, the term "modified nucleotide" refers to a nucleotide having an independently modified sugar moiety, a modified internucleotide linkage, or a modified nucleobase. Thus, the term modified nucleotide encompasses substitutions, additions, or removals of, for example, functional groups or atoms, of internucleoside linkages, sugar moieties, or nucleobases. Modifications suitable for use in the agents of the present invention include all types of modifications disclosed herein or known in the art. Any such modification, such as that used in siRNA-type molecules, is encompassed within an "RNAi agent" for purposes of this specification and claims.

[0166] In certain embodiments of the invention, deoxy-nucleotides, including naturally occurring forms of nucleotides identified as nucleotides, may be considered to constitute modified nucleotides if present in an RNAi agent.

[0167] The duplex region can be of any length that allows for specific degradation of the desired target RNA by the RISC pathway, and can be in the range of about 9 to 36 base pairs in length, for example, about 15-30 base pairs in length, for example: about 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, or 36 base pairs in length, such as about 15-30, 15-29, 15-28, 15-27, 15-26, 15-25, 15-24, 15-23, 15-22, 15-21, 15-20, 15-19, 15-18, 15-19, 15-21 8-30, 18-29, 18-28, 18-27, 18-26, 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-30, 19-29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-21, 1 Intermediate ranges and lengths between the above-recited ranges and lengths are also considered to be part of the present invention.

[0168] The two strands forming a double helix structure may be different parts of a larger RNA molecule, or they may be separate RNA molecules. Where the two strands are part of a larger molecule and are therefore connected without interruption by nucleotides between the 3' end of one strand and the 5' end of the corresponding other strand forming the double helix structure, the connected RNA strands are referred to as "hairpin loops". A hairpin loop may comprise at least one unpaired nucleotide. In some embodiments, the hairpin loop may comprise at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 20, at least 23 or more unpaired nucleotides or nucleotides that are not directed to the target site of the dsRNA. In some embodiments, the hairpin loop may be 10 or fewer nucleotides. In some embodiments, the hairpin loop may be 8 or fewer unpaired nucleotides. In some embodiments, the hairpin loop may be 4-10 unpaired nucleotides. In some embodiments, the hairpin loop may be 4-8 nucleotides.

[0169] In certain embodiments, the two chains of the double-stranded oligomeric compound may be connected together. The two chains may be connected to each other at both ends, or only at one end. Connecting at one end means that the 5' end of the first chain is connected to the 3' end of the second chain or the 3' end of the first chain is connected to the 5' end of the second chain. When the two chains are connected to each other at both ends, the 5' end of the first chain is connected to the 3' end of the second chain and the 3' end of the first chain is connected to the 5' end of the second chain. The two chains may be connected together by an oligonucleotide linker, which includes but is not limited to (N)n; wherein N is independently a modified or unmodified nucleotide and n is 3-23. In some embodiments, n is 3-10, for example 3, 4, 5, 6, 7, 8, 9 or 10. In some embodiments, the oligonucleotide linker is selected from the group consisting of: GNRA, (G)4, (U)4 and (dT)4, wherein N is a modified or unmodified nucleotide and R is a modified or unmodified purine nucleotide. Some nucleotides in the linker may be involved in base pair interactions with other nucleotides in the linker. The two chains can also be linked together by a non-nucleoside linker, such as the linkers described herein. It will be appreciated by those skilled in the art that any oligonucleotide chemical modification or variation described herein can be used in an oligonucleotide linker.

[0170] Hairpin and dumbbell oligomeric compounds will have a duplex region of equal to or at least 14, 15, 15, 16, 17, 18, 19, 29, 21, 22, 23, 24, or 25 nucleotide pairs. The length of the duplex region may be equal to or less than 200, 100, or 50. In some embodiments, the length of the duplex region ranges from 15-30, 17 to 23, 19 to 23, and 19 to 21 nucleotide pairs.

[0171] The hairpin oligomeric compound may have a single-stranded overhang or terminal unpaired region, in some embodiments at the 3′, and in some embodiments at the antisense side of the hairpin. In some embodiments, the overhang is 1-4, more typically 2-3 nucleotides in length. Hairpin oligomeric compounds that can induce RNA interference are also referred to herein as "shRNAs."

[0172] Where the two substantially complementary strands of a dsRNA are composed of separate RNA molecules, those molecules need not be but may be covalently linked. The connection between the two strands is by means of covalent linkage, rather than by nucleotides on one strand extending from the 3' end of one strand to the 5' end of the other strand forming the double helix structure without interrupting the strands. This connection structure is referred to as a "linker." RNA strands may have the same or different numbers of nucleotides. The maximum number of base pairs is the number of nucleotides in the shortest strand of the dsRNA minus any overhangs present in the double helix. In addition to the double helix structure, RNAi may comprise one or more nucleotide overhangs.

[0173] In one embodiment, the RNAi agent of the present invention is a dsRNA, wherein each strand is 24-30 nucleotides in length, which interacts with a target RNA sequence, such as a target mRNA sequence, to guide cleavage of the target RNA. Without wishing to be bound by theory, long double-stranded RNA introduced into cells is broken down into siRNAs by a type III endonuclease called Dicer (Sharp et al., (2001) Genes Dev. 15:485). Dicer, a ribozyme III-like enzyme, processes dsRNA into 19-23 base pair short interfering RNAs with a characteristic two-base 3′ overhang (Bemstein et al., (2001) Nature 409:363). The siRNA is then incorporated into the RNA-induced silencing complex (RISC), where one or more helicases unwind the siRNA duplex, allowing the complementary antisense strand to guide target recognition (Nykanen et al., (2001) Cell 107:309). Upon binding to the appropriate target mRNA, one or more endonucleases within RISC cleave the target, inducing silencing (Elbashir et al. (2001) Genes Dev. 15:188).

[0174] In one embodiment, the RNAi agent of the present invention is a dsRNA agent, wherein each strand comprises 19-23 nucleotides that interact with a target mRNA sequence to guide cleavage of the target RNA. Without wishing to be bound by theory, long double-stranded RNA introduced into cells is broken down into siRNAs by a type III endonuclease called Dicer (Sharp et al., (2001) Genes Dev. 15:485). Dicer, a ribozyme III-like enzyme, processes dsRNA into 19-23 base pair short interfering RNAs with a characteristic two-base 3′ overhang (Bernstein et al., (2001) Nature 409:363). The siRNA is then incorporated into the RNA-induced silencing complex (RISC), where one or more helicases unwind the siRNA duplex, allowing the complementary antisense strand to guide target recognition (Nykanen et al., (2001) Cell 107:309). Upon binding to the appropriate target mRNA, one or more endonucleases within RISC cleave the target to induce silencing (Elbashir et al., (2001) Genes Dev. 15:188). In one embodiment, the RNAi agent of the present invention is a 24-30 nucleotide dsRNA that interacts with the target mRNA sequence to guide the cleavage of the target RNA.

[0175] As used herein, the term "nucleotide overhang" refers to at least one unpaired nucleotide that protrudes from the double helical structure of an RNAi agent, such as a dsRNA. For example, a nucleotide overhang exists when the 3' end of one strand of a dsRNA extends beyond the 5' end of the other strand, or vice versa. A dsRNA may comprise an overhang of at least one nucleotide; alternatively, the overhang may comprise at least two nucleotides, at least three nucleotides, at least four nucleotides, at least five nucleotides, or more. The nucleotide overhang may comprise or consist of nucleotide / nucleoside analogs, including deoxynucleotides / nucleosides. The one or more overhangs may be on the sense strand, the antisense strand, or any combination thereof. Furthermore, the one or more nucleotides of the overhang may be present on the 5' end, the 3' end, or both ends of the antisense strand or the sense strand of the dsRNA.

[0176] In one embodiment of a dsRNA, at least one strand comprises a 3' overhang of at least 1 nucleotide. In another embodiment, at least one strand comprises a 3' overhang of at least 2 nucleotides, such as 2, 3, 4, 5, 6, 7, 9, 10, 11, 12, 13, 14, or 15 nucleotides. In other embodiments, at least one strand of the RNAi agent comprises a 5' overhang of at least 1 nucleotide. In certain embodiments, at least one strand comprises a 5' overhang of at least 2 nucleotides, such as 2, 3, 4, 5, 6, 7, 9, 10, 11, 12, 13, 14, or 15 nucleotides. In yet other embodiments, both the 3' and 5' ends of one strand of the RNAi agent comprise an overhang of at least 1 nucleotide.

[0177] In one embodiment, the antisense strand of the dsRNA has an overhang of 1-10 nucleotides, e.g., 0-3, 1-3, 2-4, 2-5, 4-10, 5-10, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides at the 3' or 5' end. In one embodiment, the sense strand of the dsRNA has an overhang of 1-10 nucleotides, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides at the 3' or 5' end. In another embodiment, one or more of the nucleotides in the overhang are substituted with phosphorothioate nucleosides.

[0178] In certain embodiments, the overhang on the sense strand or the antisense strand, or both, may include an extension length greater than 10 nucleotides, such as 1-30 nucleotides, 2-30 nucleotides, 10-30 nucleotides, or 10-15 nucleotides in length. In certain embodiments, the extension overhang is located on the sense strand of the duplex. In certain embodiments, the extension overhang is present at the 3′ end of the sense strand of the duplex. In certain embodiments, the extension overhang is present at the 5′ end of the sense strand of the duplex. In certain embodiments, the extension overhang is on the antisense strand of the duplex. In certain embodiments, the extension overhang is present at the 3′ end of the antisense strand of the duplex. In certain embodiments, the extension overhang is present at the 5′ end of the antisense strand of the duplex. In certain embodiments, one or more of the nucleotides in the overhang is replaced by a phosphorothioate nucleoside. In certain embodiments, the overhang includes a self-complementary portion, such that the overhang is capable of forming a hairpin structure that is stable under physiological conditions.

[0179] As used herein with respect to dsRNA, the term "blunt" or "blunt ended" refers to the absence of unpaired nucleotides or nucleotide analogs at a given end of the dsRNA, i.e., the absence of nucleotide overhangs. A dsRNA may be blunt at one or both ends. Where both ends of a dsRNA are blunt, the dsRNA is referred to as blunt-ended. For clarity, a "blunt-ended" dsRNA is a dsRNA that is blunt at both ends, i.e., there are no nucleotide overhangs at either end of the molecule. Typically, such molecules will be double-stranded throughout their entire length.

[0180] The term "antisense strand" or "guide strand" refers to the strand of an iRNA, such as a dsRNA, that includes a region that is substantially complementary to a target sequence, such as a target mRNA sequence.

[0181] As used herein, the term "complementarity region" refers to a region on the antisense strand that is substantially complementary to a sequence as defined herein (e.g., a target sequence, e.g., a target nucleotide sequence). When the complementary region is not completely complementary to the target sequence, the mismatch may be in an internal or terminal region of the molecule. Generally, the most tolerated mismatch is in the terminal region, e.g., within 5, 4, 3, or 2 nucleotides of the 5' or 3' end of the RNAi agent.

[0182] In some embodiments, the double-stranded RNA agent of the present invention includes a nucleotide mismatch in the antisense strand. In some embodiments, the antisense strand of the double-stranded RNA agent of the present invention includes no more than 4 mismatches with the target mRNA, for example, the antisense strand includes 4, 3, 2, 1 or 0 mismatches with the target mRNA. In some embodiments, the antisense strand of the double-stranded RNA agent of the present invention includes no more than 4 mismatches with the sense strand, for example, the antisense strand includes 4, 3, 2, 1 or 0 mismatches with the sense strand. In some embodiments, the double-stranded RNA agent of the present invention includes a nucleotide mismatch in the sense strand. In some embodiments, the sense strand of the double-stranded RNA agent of the present invention includes no more than 4 mismatches with the antisense strand, for example, the sense strand includes 4, 3, 2, 1 or 0 mismatches with the antisense strand. In some embodiments, the nucleotide mismatch is, for example, within 5, 4, or 3 nucleotides from the 3' end of the iRNA. In another embodiment, the nucleotide mismatch is, for example, in the 3' end nucleotide of the iRNA agent. In some embodiments, one or more mismatches are not in the seed region.

[0183] Therefore, RNAi agents as described herein may contain one or more mismatches with the target sequence. In one embodiment, RNAi agents as described herein contain no more than 3 mismatches (i.e., 3, 2, 1, or 0 mismatches). In one embodiment, RNAi agents as described herein contain no more than 2 mismatches. In one embodiment, RNAi agents as described herein contain no more than 1 mismatch. In one embodiment, RNAi agents as described herein contain 0 mismatches. In certain embodiments, if the antisense strand of the RNAi agent contains a mismatch with the target sequence, the mismatch may optionally be limited to the last 5 nucleotides from the 5' or 3' end of the complementary region. For example, in such embodiments, for RNAi agents of 23 nucleotides, the strand complementary to the region of the target gene generally does not contain any mismatch within the central 13 nucleotides. Methods described herein or methods known in the art can be used to determine whether RNAi agents containing mismatches with the target sequence can effectively inhibit the expression of the target gene. It is important to consider the efficacy of RNAi agents with mismatches in inhibiting target gene expression, especially when the specific complementary region in the target gene is known to change.

[0184] As used herein, the term "sense strand" or "transfer strand" refers to the strand of the RNAi agent that includes a region that is substantially complementary to a region of the antisense strand as defined herein.

[0185] As used herein, "substantially all nucleotides are modified" means that most, but not all, nucleotides are modified, and may include no more than 5, 4, 3, 2, or 1 unmodified nucleotides.

[0186] As used herein, the term "cleavage zone" refers to a region located adjacent to a cleavage site. A cleavage site is a site on a target where cleavage occurs. In some embodiments, the cleavage zone comprises three bases located on either end of the cleavage site and adjacent to the cleavage site. In some embodiments, the cleavage zone comprises two bases located at either end of the cleavage site and adjacent to the cleavage site. In some embodiments, the cleavage site specifically occurs at the site where nucleotides 10 and 11 of the antisense strand bind, and the cleavage zone comprises nucleotides 11, 12, and 13.

[0187] As used herein, unless otherwise indicated, the term "complementary" when used to describe a first nucleotide sequence relative to a second nucleotide sequence, as will be understood by the skilled artisan, refers to the ability of an oligonucleotide or polynucleotide comprising the first nucleotide sequence to hybridize and form a duplex structure with an oligonucleotide or polynucleotide comprising the second nucleotide sequence under specific conditions. Such conditions may be, for example, "stringent conditions," wherein stringent conditions may include: 400 mM NaCl, 40 mM PIPES pH 6.4, 1 mM EDTA, 50° C. or 70° C. for 12-16 hours, followed by washing (see, for example, "Molecular Cloning: A Laboratory Manual, Sambrook et al. (1989) Cold Spring Harbor Laboratory Press). Other conditions may be applicable, such as physiologically relevant conditions that may be encountered within an organism. A skilled artisan will be able to determine the most appropriate set of conditions for testing the complementarity of two sequences based on the ultimate application of the hybridizing nucleotides.

[0188] Complementary sequences within an RNAi agent, such as within a dsRNA as described herein, include base pairing of an oligonucleotide or polynucleotide comprising a first nucleotide sequence to an oligonucleotide or polynucleotide comprising a second nucleotide sequence over the full length of one or both nucleotide sequences. Such sequences may be referred to herein as "fully complementary" to one another. However, when a first sequence is referred to herein as "substantially complementary" to a second sequence, the two sequences may be fully complementary, or they may form one or more, but generally no more than 5, 4, 3, or 2, mismatched base pairs when a duplex of up to 30 base pairs hybridizes, while retaining the ability to hybridize under conditions most relevant to their ultimate application, such as inhibition of gene expression in vitro or in vivo. However, when two oligonucleotides are designed to form one or more single-stranded overhangs upon hybridization, such overhangs should not be considered mismatches with respect to complementarity determinations. For example, for the purposes described herein, a dsRNA comprising an oligonucleotide of 21 nucleotides in length and a dsRNA comprising another oligonucleotide of 23 nucleotides in length, wherein the longer oligonucleotide comprises a 21 nucleotide sequence that is fully complementary to the shorter oligonucleotide, may still be referred to as "fully complementary."

[0189] As used herein, "complementary" sequences may also include or consist entirely of non-Watson-Crick base pairs or base pairs formed by non-natural and modified nucleotides, as long as the above requirements regarding their hybridization ability are met. Such non-Watson-Crick base pairs include, but are not limited to, G:U wobble or Hoogsteen base pairs.

[0190] As will be understood from the context of their use, the terms "complementary," "fully complementary," and "substantially complementary" herein can be used with respect to base matching between the sense and antisense strands of a dsRNA, or between two oligonucleotides or polynucleotides, such as the antisense strand of an RNAi agent and a target sequence.

[0191] As used herein, a polynucleotide that is "substantially complementary to at least a portion of a messenger RNA (mRNA) or a target sequence refers to a polynucleotide that is substantially complementary to a contiguous portion of the mRNA or target sequence of interest (e.g., an mRNA encoding a target gene). For example, a polynucleotide is complementary to at least a portion of a target RNA if the sequence is substantially complementary to a non-interrupted portion of an mRNA encoding a target gene.

[0192] Thus, in some embodiments, the antisense polynucleotides disclosed herein are fully complementary to the target gene sequence. In other embodiments, the antisense polynucleotides disclosed herein are substantially complementary to the target sequence and comprise a contiguous nucleotide sequence that is at least 80% complementary to the equivalent region of the nucleotide sequence of the target sequence over its entire length, such as about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% complementary.

[0193] Exemplary target genes include, for example, adrenergic receptor β1 (ADRB1); calcium voltage-gated channel subunit α1C (CACNA1C); calcium voltage-gated channel subunit α1G (CACNA1G) (T-type calcium c channel); angiotensin II receptor type I (AGTR1); sodium voltage-gated channel alpha subunit 2 (SCN2A); hyperpolarization-activated cyclic nucleotide-gated potassium channel 1 (HCN1); hyperpolarization-activated cyclic nucleotide-gated potassium channel 4 (HCN4); hyperpolarization-activated cyclic nucleotide-gated potassium channel 3 (HCN3); potassium voltage-gated channel subfamily A, member 5 (KCNA5); potassium inwardly rectifier channel subfamily J, member 3 (KCNJ3); potassium inwardly rectifier channel subfamily J, member 4 (KCNJ4); phospholamban (PLN); calcium / calmodulin-dependent protein kinase II delta (CAMK2D); or phosphodiesterase 1 (PDE1).

[0194] Additional exemplary target genes also include, for example, myostatin (MSTN); choline receptor nicotinic alpha 1 subunit (CHRNA1); choline receptor nicotinic beta 1 subunit (CHRNB1); choline receptor nicotinic delta subunit (CHRND); choline receptor nicotinic epsilon subunit (CHRNE); choline receptor nicotinic gamma subunit (CHRNG); collagen type XIII alpha 1 chain (COL13A1); docking protein 7 (DOK7); LDL receptor-related protein 4 (LRP 4); muscle-associated receptor tyrosine kinase (MUSK); receptor-associated synaptic protein (RAPSN); sodium voltage-gated channel alpha subunit 4 (SCN4A); dual homeobox 4 (DUX4); dystrophic myotonic protein kinase (DMPK); glycogen synthase 1 (GYS1); survival motor neuron 1 (SMN1); alpha-glucosidase (GAA); receptor specific for advanced glycation end products (AGER); oligomeric mucus / gel-forming mucin 5AC (MUC5AC); and signal transducer and activator of transcription 6 (STAT6).

[0195] As used herein, "adrenergic receptor β1" is used interchangeably with the term "ADRB1" to refer to a member of the adrenergic receptor family. Adrenergic receptors are the prototypical family of guanine nucleotide-binding regulatory protein-coupled receptors that mediate the physiological actions of the hormone epinephrine and the neurotransmitter norepinephrine. β-1 adrenergic receptors are primarily located in the heart. Specific polymorphisms in this gene have been shown to affect resting heart rate and may be involved in heart failure. ADRB1 is also known as ADRB1R, β-1 adrenergic receptor, B1AR, β1AR, FNSS2, or RHR.

[0196] Exemplary sequences of human ADRB1 mRNA transcripts can be found, for example, in GenBank Accession No. GI:1653960731 (NM_000684.3; SEQ ID NO:1; reverse complement, SEQ ID NO:5). The sequence of mouse ADRB1 mRNA can be found, for example, in GenBank Accession No. GI:1693744501 (NM_007419.3; SEQ ID NO:2; reverse complement, SEQ ID NO:6). The sequence of rat ADRB1 mRNA can be found, for example, in GenBank Accession No. GI:6978458 (NM_012701.1; SEQ ID NO:3; reverse complement, SEQ ID NO:7). The sequence of rhesus macaque ADRB1 mRNA can be found, for example, in GenBank Accession No. GI:577861029 (NM_001289866.1; SEQ ID NO:4; reverse complement, SEQ ID NO:8). The sequence of cynomolgus macaque ADRB1 mRNA can be found, for example, in GenBank Accession No. GI:985482105 (NM_001319353.1; SEQ ID NO:9; reverse complement sequence, SEQ ID NO:10).

[0197] Additional examples of ADRB1 mRNA sequences are readily available through publicly available databases such as GenBank, UniProt, OMIM, the UCSC Genome Browser, and the Macaque Genome Project website.

[0198] Additional information about ADRB1 can be found, for example, at www.ncbi.nlm.nih.gov / gene / ?term=ADRB1.

[0199] The entire contents of each of the aforementioned GenBank accession numbers and gene database numbers are incorporated herein by reference as of the filing date of the present application.

[0200] As used herein, the term ADRB1 also refers to variations of the ADRB1 gene, including variants provided in SNP databases. Many sequence variations within the ADRB1 gene have been identified and can be found, for example, in NCBI dbSNP and UniProt (see, for example, www.ncbi.nlm.nih.gov / snp / ?term=ADRB1, the entire contents of which are incorporated herein by reference as of the filing date of this application).

[0201] In one embodiment, the target gene is calcium voltage-gated channel subunit α1C (CACNA1C).

[0202] As used herein, "calcium voltage-gated channel subunit α1C" is used interchangeably with the term "CACNA1C" and refers to the α-1 subunit of the voltage-dependent calcium channel. The calcium channel mediates the influx of calcium ions into the cell under membrane polarization. The α-1 subunit is composed of 24 transmembrane segments and forms a pore through which ions enter the cell. The calcium channel is composed of a complex of α-1, α-2 / δ, β, and γ subunits in a 1:1:1:1 ratio. Each of these proteins exists in multiple isoforms, encoded by different genes or as a result of alternative splicing of transcripts. The proteins encoded by these genes bind to and are inhibited by dihydropyridines. CACNA1C is also known as calcium channel voltage-dependent L-type α1C subunit; voltage-dependent L-type calcium channel subunit α-1C; voltage-gated L-type calcium channel Cav1.2 α1 subunit splice variant 10; cardiac calcium channel L-type α-1 polypeptide isoform 1; calcium channel cardiac dihydropyridine-sensitive α-1 subunit; voltage-dependent L-type Ca2+ channel α1 subunit; voltage-gated calcium channel subunit αCaV1.2; DHPR, α-1 subunit; CACH2, CACN2, CACNL1A1, CCHL1A1, CaV1.2, LQT8, TS, or TS.LQT8

[0203] Exemplary sequences of human CACNA1C mRNA transcripts can be found, for example, in GenBank Accession No. GI:1890333913 (NM_199460.4; SEQ ID NO:11; reverse complement, SEQ ID NO:12). The sequence of mouse CACNA1C mRNA can be found, for example, in GenBank Accession No. GI:594140631 (NM_009781.4; SEQ ID NO:13; reverse complement, SEQ ID NO:14). The sequence of rat CACNA1C mRNA can be found, for example, in GenBank Accession No. GI:158186632 (NM_012517.2; SEQ ID NO:15; reverse complement, SEQ ID NO:16). The sequence of the rhesus macaque CACNA1C mRNA can be found, for example, in GenBank Accession No. GI: ​​1622843324 (XM_028829106.1; SEQ ID NO: 17; reverse complement sequence, SEQ ID NO: 18).

[0204] Additional examples of CACNA1C mRNA sequences are readily available through publicly available databases such as GenBank, UniProt, OMIM, the UCSC Genome Browser, and the Macaque Genome Project website.

[0205] Additional information about CACNA1C can be found, for example, at www.ncbi.nlm.nih.gov / gene / ?term=CACNA1C.

[0206] The entire contents of each of the aforementioned GenBank accession numbers and gene database numbers are incorporated herein by reference as of the filing date of the present application.

[0207] As used herein, the term CACNA1C also refers to variations of the CACNA1C gene, including variants provided in SNP databases. Many sequence variations within the CACNA1C gene have been identified and can be found, for example, in NCBI dbSNP and UniProt (see, for example, www.ncbi.nlm.nih.gov / gene / ?term=CACNA1C, the entire contents of which are incorporated herein by reference as of the filing date of this application).

[0208] As used herein, "calcium voltage-gated channel subunit α1G" is used interchangeably with the term "CACNA1G" and refers to the T-type low-voltage-activated calcium channel. Voltage-sensitive calcium channels mediate the entry of calcium ions into excitable cells and are also involved in a variety of calcium-dependent processes, including muscle contraction, hormone or neurotransmitter release, gene expression, cell motility, cell division, and cell death. T-type channels generate currents that are short-lived due to rapid inactivation and small due to low conductance. T-type channels are believed to be involved in pacemaker activity, low-threshold calcium spikes, neuronal oscillations and resonance, and rebound burst firing. CACNA1G is also known as calcium channel voltage-dependent T-type α1G subunit; voltage-dependent T-type calcium channel subunit α-1G; voltage-gated calcium channel subunit α Cav3.1; NBR13; Cav3.1c; Ca(V)T.1; KIAA1123; SCA42ND; or SCA42.

[0209] Exemplary sequences of human CACNA1G mRNA transcripts can be found, for example, in GenBank Accession No. GI:1519244109 (NM_018896.5; SEQ ID NO:21; reverse complement, SEQ ID NO:22). The sequence of mouse CACNA1G mRNA can be found, for example, in GenBank Accession No. GI:295444826 (NM_009783.3; SEQ ID NO:23; reverse complement, SEQ ID NO:24). The sequence of rat CACNA1G mRNA can be found, for example, in GenBank Accession No. GI:1995160279 (NM_001308302.2; SEQ ID NO:25; reverse complement, SEQ ID NO:26). The sequence of rhesus macaque CACNA1G mRNA can be found, for example, in GenBank Accession No. GI: ​​1622879013 (XM_015119270.2; SEQ ID NO: 27; reverse complement sequence, SEQ ID NO: 28). The sequence of cynomolgus macaque CACNA1G mRNA can be found, for example, in GenBank Accession No. GI: ​​982305044 (XM_005583707.2; SEQ ID NO: 29; reverse complement sequence, SEQ ID NO: 30).

[0210] Additional illustrative examples of CACNA1G mRNA sequences are readily available through publicly available databases such as GenBank, UniProt, OMIM, the UCSC Genome Browser, and the Macaque Genome Project website.

[0211] Additional information about CACNA1G can be found, for example, at www.ncbi.nlm.nih.gov / gene / ?term=CACNA1G.

[0212] The entire contents of each of the aforementioned GenBank accession numbers and gene database numbers are incorporated herein by reference as of the filing date of the present application.

[0213] As used herein, the term CACNA1G also refers to variants of the CACNA1G gene, including variants provided in SNP databases. Many sequence variations within the CACNA1G gene have been identified and can be found, for example, in NCBI dbSNP and UniProt (see, for example, www.ncbi.nlm.nih.gov / gene / ?term=CACNA1G, the entire contents of which are incorporated herein by reference as of the filing date of this application).

[0214] As used herein, "angiotensin II receptor type 1" is used interchangeably with the term "AGTR1" and refers to the receptor for the vasoconstrictor peptide angiotensin II. Angiotensin II is a potent vasopressor hormone and the primary regulator of aldosterone secretion. AGTR1 is activated by angiotensin II. Activated receptors, in turn, couple to G proteins, thereby activating phospholipase C and increasing cytosolic Ca2+ concentrations, triggering cellular responses such as stimulation of protein kinase C. AGTR1 plays an important role in blood pressure control and is implicated in the pathogenesis of hypertension. AGTR1 is also known as angiotensin receptor 1B, AT1, AT2R1, AGTR1A, AT2R1B, AGTR1B, HAT1R, AG2S, AT1B, AT2R1A, AT1AR, AT1BR, or AT1R.

[0215] Exemplary sequences of human AGTR1 mRNA transcripts can be found, for example, in GenBank Accession No. GI:1820101583NM_000685.5 (SEQ ID NO:31; reverse complement, SEQ ID NO:32). The sequence of mouse AGTR1 mRNA can be found, for example, in GenBank Accession No. GI:158937294 (NM_177322.3; SEQ ID NO:33; reverse complement, SEQ ID NO:34). The sequence of rat AGTR1 mRNA can be found, for example, in GenBank Accession No. GI:140969764 (NM_030985.4; SEQ ID NO:35; reverse complement, SEQ ID NO:36). The sequence of rhesus macaque AGTR1 mRNA can be found, for example, in GenBank Accession No. GI: ​​1622904093 (XM_028843763.1; SEQ ID NO: 37; reverse complement, SEQ ID NO: 38). The sequence of cynomolgus macaque AGTR1 mRNA can be found, for example, in GenBank Accession No. GI: ​​5444411901 (XM_005546040.1; SEQ ID NO: 39; reverse complement, SEQ ID NO: 40).

[0216] Additional examples of AGTR1 mRNA sequences are readily available through publicly available databases such as GenBank, UniProt, OMIM, the UCSC Genome Browser, and the Macaque Genome Project website.

[0217] Additional information about AGTR1 can be found, for example, at www.ncbi.nlm.nih.gov / gene / ?term=AGTR1.

[0218] The entire contents of each of the aforementioned GenBank accession numbers and gene database numbers are incorporated herein by reference as of the filing date of the present application.

[0219] As used herein, the term AGTR1 also refers to variations of the AGTR1 gene, including variants provided in SNP databases. Many sequence variations within the AGTR1 gene have been identified and can be found, for example, in NCBI dbSNP and UniProt (see, for example, www.ncbi.nlm.nih.gov / gene / ?term=AGTR1, the entire contents of which are incorporated herein by reference as of the filing date of this application).

[0220] As used herein, "sodium voltage-gated channel alpha subunit 2" is used interchangeably with the term "SCN2A" to refer to a member of the voltage-gated sodium channel family. Voltage-gated sodium channels are transmembrane glycoprotein complexes composed of a large alpha subunit with four repeating domains and one or more regulatory beta subunits, each of which consists of six transmembrane segments. Voltage-gated sodium channels play a role in the generation and propagation of action potentials in neurons and muscles. Specifically, SCN2A allows sodium to flow from the extracellular space into the cytosol after depolarization of the neural membrane. Allele variants of SCN2A are associated with epilepsy and autism spectrum disorder. SCN2A is also known as Nav1.2, HBSCII, SCN2A1, SCN2A2, HBSCI, EIEE11, BFIC3, BFIS3, BFNIS, DEE11, EA9, or HBA.

[0221] Exemplary sequences of human SCN2A mRNA transcripts can be found, for example, in GenBank Accession No. GI: ​​1697699196 (NM_021007.3; SEQ ID NO: 41; reverse complement, SEQ ID NO: 42). The sequence of mouse SCN2A mRNA can be found, for example, in GenBank Accession No. GI: ​​1114439824 (NM_001099298.3; SEQ ID NO: 43; reverse complement, SEQ ID NO: 44). The sequence of rat SCN2A mRNA can be found, for example, in GenBank Accession No. GI: ​​1937915892 (NM_012647.2; SEQ ID NO: 45; reverse complement, SEQ ID NO: 46). The sequence of rhesus macaque SCN2A mRNA can be found, for example, in GenBank Accession No. GI: ​​1622850108 (XM_001100368.4; SEQ ID NO: 47; reverse complement sequence, SEQ ID NO: 48). The sequence of cynomolgus macaque SCN2A mRNA can be found, for example, in GenBank Accession No. GI: ​​544475515 (XM_005573351.1; SEQ ID NO: 49; reverse complement sequence, SEQ ID NO: 50).

[0222] Additional examples of SCN2A mRNA sequences are readily available through publicly available databases such as GenBank, UniProt, OMIM, the UCSC Genome Browser, and the Macaque Genome Project website.

[0223] Additional information about SCN2A can be found, for example, at www.ncbi.nlm.nih.gov / gene / ?term=SCN2A.

[0224] The entire contents of each of the aforementioned GenBank accession numbers and gene database numbers are incorporated herein by reference as of the filing date of the present application.

[0225] As used herein, the term SCN2A also refers to variations of the SCN2A gene, including variants provided in SNP databases. Many sequence variations within the SCN2A gene have been identified and can be found, for example, in NCBI dbSNP and UniProt (see, for example, www.ncbi.nlm.nih.gov / gene / ?term=SCN2A, the entire contents of which are incorporated herein by reference as of the filing date of this application).

[0226] As used herein, "hyperpolarization-activated cyclic nucleotide-gated potassium channel 1" is used interchangeably with the term "HCN1" to refer to members of the hyperpolarization-activated cyclic nucleotide-gated (HCN) channel family. These channels are primarily expressed in the heart and the central and peripheral nervous systems. HCN channels mediate the rhythmic electrical activity of cardiac pacemaker cells and, in neurons, play an important role in setting the resting membrane potential, dendritic integration, neuronal pacing, and establishing the action potential threshold. The HCN1 protein can homo- or heterodimerize with other pore-forming subunits to form potassium channels. HCN1 is also known as potassium channel 1, BCNG-1, HAC-2, BCNG1, potassium / sodium hyperpolarization-activated cyclic nucleotide-gated channel 1; brain cyclic nucleotide-gated channel 1; hyperpolarization-activated cyclic nucleotide-gated potassium channel 1; GEFSP10, EIEE24, or DEE24.

[0227] Exemplary sequences of human HCN1 mRNA transcripts can be found, for example, in GenBank Accession No. GI:1519313076 (NM_021072.4; SEQ ID NO:51; reverse complement, SEQ ID NO:52). The sequence of mouse HCN1 mRNA can be found, for example, in GenBank Accession No. GI:283837798 (NM_010408.3; SEQ ID NO:53; reverse complement, SEQ ID NO:54). The sequence of rat HCN1 mRNA can be found, for example, in GenBank Accession No. GI:2000186052 (NM_053375.2; SEQ ID NO:55; reverse complement, SEQ ID NO:56). The sequence of rhesus macaque HCN1 mRNA can be found, for example, in GenBank Accession No. GI:1622944535 (XM_015140004.2; SEQ ID NO:57; reverse complement, SEQ ID NO:58). The sequence of cynomolgus macaque HCN1 mRNA can be found, for example, in GenBank Accession No. GI:982252681 (XM_005556858.2; SEQ ID NO:59; reverse complement, SEQ ID NO:60).

[0228] Additional examples of HCN1 mRNA sequences are readily available through publicly available databases such as GenBank, UniProt, OMIM, the UCSC Genome Browser, and the Macaque Genome Project website.

[0229] Additional information about HCN1 can be found, for example, at www.ncbi.nlm.nih.gov / gene / ?term=HCN1.

[0230] The entire contents of each of the aforementioned GenBank accession numbers and gene database numbers are incorporated herein by reference as of the filing date of the present application.

[0231] As used herein, the term HCN1 also refers to variations of the HCN1 gene, including variants provided in SNP databases. Many sequence variations within the HCN1 gene have been identified and can be found, for example, in NCBI dbSNP and UniProt (see, for example, www.ncbi.nlm.nih.gov / gene / ?term=HCN1, the entire contents of which are incorporated herein by reference as of the filing date of this application).

[0232] As used herein, "hyperpolarization-activated cyclic nucleotide-gated potassium channel 4" is used interchangeably with the term "HCN4" to refer to a member of the hyperpolarization-activated cyclic nucleotide-gated (HCN) channel family. The HCN4 channel transports positively charged ions into cardiac myocytes. This channel is primarily located in the sinoatrial (SA) node, an area of ​​specialized cells in the heart that acts as a natural pacemaker. The HCN4 channel allows potassium and sodium ions to flow into cells of the SA node. This ion flow is commonly referred to as the "pacemaker current" because it generates the electrical impulse that starts each heartbeat and is involved in maintaining a regular heart rhythm. HCN4 is also known as potassium / sodium hyperpolarization-activated cyclic nucleotide-gated channel 4, hyperpolarization-activated cyclic nucleotide-gated potassium channel 4, hyperpolarization-activated cyclic nucleotide-gated cation channel 4, or SSS2.

[0233] Exemplary sequences of human HCN4 mRNA transcripts can be found, for example, in GenBank Accession No. GI:1519312820 (NM_005477.3; SEQ ID NO:61; reverse complement, SEQ ID NO:62). The sequence of mouse HCN4 mRNA can be found, for example, in GenBank Accession No. GI:1686254400 (NM_001081192.3; SEQ ID NO:63; reverse complement, SEQ ID NO:64). The sequence of rat HCN4 mRNA can be found, for example, in GenBank Accession No. GI:1937893976 (NM_021658.2; SEQ ID NO:65; reverse complement, SEQ ID NO:66). The sequence of rhesus macaque HCN4 mRNA can be found, for example, in GenBank Accession No. GI: ​​1622953870 (XM_002804859.3; SEQ ID NO: 67; reverse complement, SEQ ID NO: 68). The sequence of cynomolgus macaque HCN4 mRNA can be found, for example, in GenBank Accession No. GI: ​​982258526 (XM_005559993.2; SEQ ID NO: 69; reverse complement, SEQ ID NO: 70).

[0234] Additional examples of HCN4 mRNA sequences are readily available through publicly available databases such as GenBank, UniProt, OMIM, the UCSC Genome Browser, and the Macaque Genome Project website.

[0235] Additional information about HCN4 can be found, for example, at www.ncbi.nlm.nih.gov / gene / ?term=HCN4.

[0236] The entire contents of each of the aforementioned GenBank accession numbers and gene database numbers are incorporated herein by reference as of the filing date of the present application.

[0237] As used herein, the term HCN4 also refers to variations of the HCN4 gene, including variants provided in the SNP database. Many sequence variations within the HCN4 gene have been identified and can be found, for example, in NCBI dbSNP and UniProt (see, for example, www.ncbi.nlm.nih.gov / gene / ?term=HCN4, the entire contents of which are incorporated herein by reference as of the filing date of this application).

[0238] As used herein, "hyperpolarization-activated cyclic nucleotide-gated potassium channel 3" is used interchangeably with the term "HCN3" to refer to members of the hyperpolarization-activated cyclic nucleotide-gated (HCN) channel family. Studies in mice suggest that HCN3 channels may be involved in regulating the circadian rhythm system. HCN3 channels have also been reported to be present in the intergeniculate leaflet of the hypothalamus. HCN3 is also known as potassium / sodium hyperpolarization-activated cyclic nucleotide-gated channel 3, hyperpolarization-activated cyclic nucleotide-gated potassium channel 3, or KIAA1535.

[0239] Exemplary sequences of human HCN3 mRNA transcripts can be found, for example, in GenBank Accession No. GI:1519312303 (NM_020897.3; SEQ ID NO:71; reverse complement, SEQ ID NO:72). The sequence of mouse HCN3 mRNA can be found, for example, in GenBank Accession No. GI:6680190 (NM_008227.1; SEQ ID NO:73; reverse complement, SEQ ID NO:74). The sequence of rat HCN3 mRNA can be found, for example, in GenBank Accession No. GI:16758501 (NM_053685.1; SEQ ID NO:75; reverse complement, SEQ ID NO:76). The sequence of rhesus macaque HCN3 mRNA can be found, for example, in GenBank Accession No. GI:1622829938 (XM_001115891.4; SEQ ID NO:77; reverse complement, SEQ ID NO:78). The sequence of cynomolgus monkey HCN3 mRNA can be found, for example, in GenBank Accession No. GI:982225310 (XM_005541549.2; SEQ ID NO:79; reverse complement sequence, SEQ ID NO:80).

[0240] Additional examples of HCN3 mRNA sequences are readily available through publicly available databases such as GenBank, UniProt, OMIM, the UCSC Genome Browser, and the Macaque Genome Project website.

[0241] Additional information about HCN3 can be found, for example, at www.ncbi.nlm.nih.gov / gene / ?term=HCN3.

[0242] The entire contents of each of the aforementioned GenBank accession numbers and gene database numbers are incorporated herein by reference as of the filing date of the present application.

[0243] As used herein, the term HCN3 also refers to variations of the HCN3 gene, including variants provided in the SNP database. Many sequence variations within the HCN3 gene have been identified and can be found, for example, in NCBI dbSNP and UniProt (see, for example, www.ncbi.nlm.nih.gov / gene / ?term=HCN3, the entire contents of which are incorporated herein by reference as of the filing date of this application).

[0244] As used herein, "potassium voltage-gated channel subfamily A member 5" is used interchangeably with the term "KCNA5" to refer to a member of the voltage-gated potassium channel family. Voltage-gated potassium channels mediate transmembrane potassium transport in excitable membranes. These channels form tetrameric potassium-selective channels through which potassium ions pass according to their electrochemical gradient and alternate between open and closed configurations in response to a voltage difference across the membrane. KCNA5 contains six transmembrane domains, including a shaker-type repeat in the fourth segment. It belongs to the class of delayed rectifiers, whose function is to restore the resting membrane potential of beta cells after depolarization and thereby contribute to the regulation of insulin secretion. KCNA5 is also known as HPCN1, HK2, potassium voltage-gated channel oscillator-related subfamily member 5; voltage-gated potassium channel subunit Kv1.5; voltage-gated potassium channel HK2; Kv1.5; insulinoma and pancreatic islet potassium channel; cardiac potassium channel; potassium channel 1; ATFB7, HCK1, or PCN1.

[0245] Exemplary sequences of human KCNA5 mRNA transcripts can be found, for example, in GenBank Accession No. GI:1653961222 (NM_002234.4; SEQ ID NO:81; reverse complement, SEQ ID NO:82). The sequence of mouse KCNA5 mRNA can be found, for example, in GenBank Accession No. GI:158937280 (NM_145983.2; SEQ ID NO:83; reverse complement, SEQ ID NO:84). The sequence of rat KCNA5 mRNA can be found, for example, in GenBank Accession No. GI:6981117 (NM_012972.1; SEQ ID NO:85; reverse complement, SEQ ID NO:86). The sequence of rhesus macaque KCNA5 mRNA can be found, for example, in GenBank Accession No. GI: ​​1622843572 (XM_001102294.4; SEQ ID NO: 87; reverse complement sequence, SEQ ID NO: 88). The sequence of cynomolgus macaque KCNA5 mRNA can be found, for example, in GenBank Accession No. GI: ​​982279162 (XM_005569870.2; SEQ ID NO: 89; reverse complement sequence, SEQ ID NO: 90).

[0246] Additional examples of KCNA5 mRNA sequences are readily available through publicly available databases such as GenBank, UniProt, OMIM, the UCSC Genome Browser, and the Macaque Genome Project website.

[0247] Additional information about KCNA5 can be found, for example, at www.ncbi.nlm.nih.gov / gene / ?term=KCNA5.

[0248] The entire contents of each of the aforementioned GenBank accession numbers and gene database numbers are incorporated herein by reference as of the filing date of the present application.

[0249] As used herein, the term KCNA5 also refers to variants of the KCNA5 gene, including variants provided in SNP databases. Many sequence variations within the KCNA5 gene have been identified and can be found, for example, in NCBI dbSNP and UniProt (see, for example, www.ncbi.nlm.nih.gov / gene / ?term=KCNA5, the entire contents of which are incorporated herein by reference as of the filing date of this application).

[0250] As used herein, "potassium inward rectifier channel subfamily J member 3" can be used interchangeably with the term "KCNJ3" to refer to integral membrane proteins and inward rectifier potassium channels. Inward rectifier potassium channels have a greater tendency to allow potassium to flow into cells rather than out of cells. This asymmetry in potassium ion conduction plays a key role in the excitability of muscle cells and neurons. KCNJ3 is controlled by G proteins and plays an important role in regulating heartbeat. It binds to three other G protein-activated potassium channels to form a heteromultimeric pore-forming complex, which is also coupled to neurotransmitter receptors in the brain. These multimeric G protein-gated inward rectifier potassium (GIRK) channels have a wide range of physiological effects, including regulating heartbeat, reward mechanisms, learning and memory function, platelet aggregation, insulin secretion and lipid metabolism. KCNJ3 is also known as GIRK1, G protein-activated inward rectifier potassium channel 1, KGA; potassium channel inward rectifier family J member 3; inward rectifier K(+) channel Kir3.1; or potassium inward rectifier family J member 3 splice variant 1e.

[0251] Exemplary sequences of human KCNJ3 mRNA transcripts can be found, for example, in GenBank Accession No. GI: ​​1519246021 (NM_002239.4; SEQ ID NO: 91; reverse complement, SEQ ID NO: 92). The sequence of mouse KCNJ3 mRNA can be found, for example, in GenBank Accession No. GI: ​​756398330 (NM_008426.2; SEQ ID NO: 93; reverse complement, SEQ ID NO: 94). The sequence of rat KCNJ3 mRNA can be found, for example, in GenBank Accession No. GI: ​​148747456 (NM_031610.3; SEQ ID NO: 95; reverse complement, SEQ ID NO: 96). The sequence of rhesus macaque KCNJ3 mRNA can be found, for example, in GenBank Accession No. GI:387849010 (NM_001261696.1; SEQ ID NO:97; reverse complement sequence, SEQ ID NO:98). The sequence of cynomolgus macaque KCNJ3 mRNA can be found, for example, in GenBank Accession No. GI:982285759 (XM_005573205.2; SEQ ID NO:99; reverse complement sequence, SEQ ID NO:100).

[0252] Additional examples of KCNJ3 mRNA sequences are readily available through publicly available databases such as GenBank, UniProt, OMIM, the UCSC Genome Browser, and the Macaque Genome Project website.

[0253] Additional information about KCNJ3 can be found, for example, at www.ncbi.nlm.nih.gov / gene / ?term=KCNJ3.

[0254] The entire contents of each of the aforementioned GenBank accession numbers and gene database numbers are incorporated herein by reference as of the filing date of the present application.

[0255] As used herein, the term KCNJ3 also refers to variations of the KCNJ3 gene, including variants provided in SNP databases. Many sequence variations within the KCNJ3 gene have been identified and can be found, for example, in NCBI dbSNP and UniProt (see, for example, www.ncbi.nlm.nih.gov / gene / ?term=KCNAJ3, the entire contents of which are incorporated herein by reference as of the filing date of this application).

[0256] As used herein, "potassium inward rectifier channel subfamily J member 4" can be used interchangeably with the term "KCNJ4" to refer to integral membrane proteins and inward rectifier type potassium channels. Inward rectifier type potassium channels have a greater tendency to allow potassium to flow into cells rather than out of cells. This asymmetry of potassium ion conduction plays a key role in the excitability of muscle cells and neurons. KCNJ4 can tetramerize to form a functional inward rectifier channel, wherein each monomer contains two transmembrane helical domains, an ion-selective P loop and a cytoplasmic N-terminal domain and a C-terminal domain. The distribution of KCNJ4 is mainly concentrated in both the heart and the brain, especially in cardiomyocytes and forebrain regions. KCNJ4 can play an important role in regulating the resting membrane potential, cell excitability and potassium homeostasis in the nervous system and various peripheral tissues. KCNJ4 is also known as HIRK2, HRK1, IRK3, HIR, Kir2.3, inward rectifier potassium channel 4; inward rectifier K(+) channel Kir2.3; potassium voltage-gated channel subfamily J member 4; hippocampal inward rectifier potassium channel; or hippocampal inward rectifier.

[0257] Exemplary sequences of human KCNJ4 mRNA transcripts can be found, for example, in GenBank Accession No. GI: ​​1732746379 (NM_152868.3; SEQ ID NO: 101; reverse complement, SEQ ID NO: 102). The sequence of mouse KCNJ4 mRNA can be found, for example, in GenBank Accession No. GI: ​​1720383422 (XM_006520486.4; SEQ ID NO: 103; reverse complement, SEQ ID NO: 104). The sequence of rat KCNJ4 mRNA can be found, for example, in GenBank Accession No. GI: ​​1937901561 (NM_053870.3; SEQ ID NO: 105; reverse complement, SEQ ID NO: 106). The sequence of rhesus macaque KCNJ4 mRNA can be found, for example, in GenBank Accession No. GI: ​​1622838042 (XM_015150354.2; SEQ ID NO: 107; reverse complement sequence, SEQ ID NO: 108). The sequence of cynomolgus macaque KCNJ4 mRNA can be found, for example, in GenBank Accession No. GI: ​​544461851 (XM_005567299.1; SEQ ID NO: 109; reverse complement sequence, SEQ ID NO: 110).

[0258] Additional examples of KCNJ4 mRNA sequences are readily available through publicly available databases such as GenBank, UniProt, OMIM, the UCSC Genome Browser, and the Macaque Genome Project website.

[0259] Additional information about KCNJ4 can be found, for example, at www.ncbi.nlm.nih.gov / gene / ?term=KCNJ4.

[0260] The entire contents of each of the aforementioned GenBank accession numbers and gene database numbers are incorporated herein by reference as of the filing date of the present application.

[0261] As used herein, the term KCNJ4 also refers to variations of the KCNJ4 gene, including variants provided in SNP databases. Many sequence variations within the KCNJ4 gene have been identified and can be found, for example, in NCBI dbSNP and UniProt (see, for example, www.ncbi.nlm.nih.gov / gene / ?term=KCNAJ4, the entire contents of which are incorporated herein by reference as of the filing date of this application. As used herein, "phosphodiesterase 1" is used interchangeably with the term "PDE1" to refer to members of the cyclic nucleotide phosphodiesterase family. Cyclic nucleotide phosphodiesterases (PDEs) are a superfamily of enzymes that regulate the spatial and temporal relationships of second messenger signaling in cellular systems. Among the 11 different families of PDEs, the phosphodiesterase 1 (PDE1) subfamily of enzymes hydrolyzes 3',5'-cyclic adenosine monophosphate (cAMP) and 3',5'-cyclic guanosine monophosphate (cGMP) in a competitive manner. The catalytic activity of PDE1 is stimulated by its binding to Ca2+ / calmodulin (CaM), leading to the integration of Ca2+- and cyclic nucleotide-mediated signaling in various diseases. The PDE1 family includes three isoforms, PDE1A, PDE1B, and PDE1C, which differ in their relative affinities for cAMP and cGMP. These isoforms are differentially expressed throughout the body, including in the cardiovascular, central nervous system, and other organs. Therefore, the PDE1 enzyme plays a key role in the pathophysiology of disease through its fundamental regulation of cAMP and cGMP signaling. PDE1 is also known as calcium / calmodulin-dependent 3′,5′-cyclic nucleotide phosphodiesterase 1; calcium / calmodulin-stimulated cyclic nucleotide phosphodiesterase; CAM-PDE 1, HSPDE1, HCAM1, or EC 3.1.4.

[0262] Exemplary sequences of human PDEl mRNA transcripts can be found, for example, in GenBank Accession No. GI:2062580163 (NM_005019.7; SEQ ID NO:111; reverse complement, SEQ ID NO:112). The sequence of mouse PDEl mRNA can be found, for example, in GenBank Accession No. GI:227330628 (NM_001159582.1; SEQ ID NO:113; reverse complement, SEQ ID NO:114). The sequence of rat PDEl mRNA can be found, for example, in GenBank Accession No. GI:13540702 (NM_030871.1; SEQ ID NO:115; reverse complement, SEQ ID NO:116). The sequence of rhesus macaque PDEl mRNA can be found, for example, in GenBank Accession No. GI:383872283 (NM_001257584.1; SEQ ID NO:117; reverse complement, SEQ ID NO:118). The sequence of cynomolgus macaque PDEl mRNA can be found, for example, in GenBank Accession No. GI:982286500 (XR_001483985.1; SEQ ID NO:119; reverse complement, SEQ ID NO:120).

[0263] Additional examples of PDE1 mRNA sequences are readily available through publicly available databases such as GenBank, UniProt, OMIM, the UCSC Genome Browser, and the Macaque Genome Project website.

[0264] Additional information about PDEl can be found, for example, at www.ncbi.nlm.nih.gov / gene / ?term=PDEl.

[0265] The entire contents of each of the aforementioned GenBank accession numbers and gene database numbers are incorporated herein by reference as of the filing date of the present application.

[0266] As used herein, the term PDE1 also refers to variations of the PDE1 gene, including variants provided in SNP databases. Many sequence variations within the PDE1 gene have been identified and can be found, for example, in NCBI dbSNP and UniProt (see, for example, www.ncbi.nlm.nih.gov / gene / ?term=PDE1, the entire contents of which are incorporated herein by reference as of the filing date of this application).

[0267] As used herein, "myostatin" is used interchangeably with the term "MSTN" and refers to a secreted ligand of the transforming growth factor-β (TGF-β) superfamily of proteins. Ligands from this family bind to various TGF-β receptors, leading to the recruitment and activation of SMAD family transcription factors that regulate gene expression. The protein encodes a preproprotein that undergoes proteolytic processing to produce disulfide-linked homodimers. This protein negatively regulates skeletal muscle cell proliferation and differentiation. Mutations in this gene are associated with increased skeletal muscle mass in humans and other mammals. Myostatin is also known as GDF8, growth / differentiation factor 8, or MSLHP.

[0268] The sequence of human myostatin mRNA transcripts can be found, for example, in GenBank Accession No. GI:1653961810 (NM_005259.3; SEQ ID NO:221; reverse complement, SEQ ID NO:222). The sequence of mouse myostatin mRNA can be found, for example, in GenBank Accession No. GI:922959927 (NM_010834.3; SEQ ID NO:223; reverse complement, SEQ ID NO:224). The sequence of rat myostatin mRNA can be found, for example, in GenBank Accession No. GI:9506906 (NM_019151.1; SEQ ID NO:225; reverse complement, SEQ ID NO:226). The sequence of cynomolgus macaque myostatin mRNA can be found, for example, in GenBank Accession No. NM_001287623.1; SEQ ID NO:227; reverse complement, SEQ ID NO:228. The sequence of rhesus macaque myostatin mRNA can be found, for example, in GenBank Accession No. GI: ​​121583757 (NM_001080119.1; SEQ ID NO: 229; reverse complement sequence, SEQ ID NO: 230).

[0269] Additional examples of myostatin mRNA sequences are readily available through publicly available databases such as GenBank, UniProt, OMIM, and the Macaque Genome Project website. Further information on myostatin can be found, for example, at www.ncbi.nlm.nih.gov / gene / ?term=myostatin.

[0270] The entire contents of each of the aforementioned GenBank accession numbers and gene database numbers are incorporated herein by reference as of the filing date of the present application.

[0271] As used herein, the term myostatin also refers to variants of the myostatin gene, including variants provided in SNP databases. Many sequence variations within the myostatin gene have been identified and can be found, for example, in NCBI dbSNP and UniProt (see, for example, www.ncbi.nlm.nih.gov / snp / ?term=myostatin, the entire contents of which are incorporated herein by reference as of the filing date of this application).

[0272] As used herein, "cholinergic receptor nicotinic alpha 1 subunit" is used interchangeably with the term "CHRNA1" and refers to the alpha subunit of the muscle acetylcholine receptor (AChR). The muscle acetylcholine receptor is composed of five subunits of four different types: two alpha subunits and one each of beta, gamma, and delta subunits. This protein plays a role in acetylcholine binding / channel gating. Upon binding acetylcholine, the AChR responds with a large-scale conformational change that affects all subunits and opens an ion-conducting channel across the plasma membrane. CHRNA1 is associated with diseases such as myasthenic syndrome. CHRNA1 is also known as cholinergic receptor nicotinic alpha polypeptide 1; acetylcholine receptor nicotinic alpha 1 (muscle); ACHRA; CHRNA; muscle nicotinic acetylcholine receptor; CMS1A, CMS1B, CMS2A, FCCMS, SCCMS, or ACHRD.

[0273] The sequence of the human CHRNA1 mRNA transcript can be found, for example, in GenBank Accession No. GI:1676317412 (NM_001039523.3; SEQ ID NO:231; reverse complement, SEQ ID NO:232). The sequence of the mouse CHRNA1 mRNA can be found, for example, in GenBank Accession No. GI:425905338 (NM_007389.5; SEQ ID NO:233; reverse complement, SEQ ID NO:234). The sequence of the rat CHRNA1 mRNA can be found, for example, in GenBank Accession No. GI:1937369362 (NM_024485.2; SEQ ID NO:235; reverse complement, SEQ ID NO:236). The sequence of cynomolgus monkey CHRNA1 mRNA can be found, for example, in GenBank Accession No. GI:982286285 (XM_015432377.1; SEQ ID NO:237; reverse complement sequence, SEQ ID NO:238). The sequence of rhesus monkey CHRNA1 mRNA can be found, for example, in GenBank Accession No. GI:1622850381 (XM_001091711.4; SEQ ID NO:239; reverse complement sequence, SEQ ID NO:240).

[0274] Additional examples of CHRNA1 mRNA sequences are readily available through publicly available databases such as GenBank, UniProt, OMIM, and the Macaque Genome Project website. Further information about CHRNA1 can be found, for example, at www.ncbi.nlm.nih.gov / gene / ?term=CHRNA1.

[0275] The entire contents of each of the aforementioned GenBank accession numbers and gene database numbers are incorporated herein by reference as of the filing date of this application.

[0276] As used herein, the term CHRNA1 also refers to variations of the CHRNA1 gene, including variants provided in the SNP database. Many sequence variations within the CHRNA1 gene have been identified and can be found, for example, in NCBI dbSNP and UniProt (see, for example, www.ncbi.nlm.nih.gov / snp / ?term=CHRNA1, the entire contents of which are incorporated herein by reference as of the filing date of this application).

[0277] As used herein, "cholinergic receptor nicotinic beta 1 subunit" is used interchangeably with the term "CHRNB1" and refers to the beta subunit of the muscle acetylcholine receptor (AChR). The muscle AChR is composed of five subunits of four different types: two alpha subunits and one each of beta, gamma, and delta subunits. This protein plays a role in acetylcholine binding / channel gating. Upon binding acetylcholine, the AChR responds with a large-scale conformational change that affects all subunits and opens an ion-conducting channel across the plasma membrane. CHRNB1 is associated with diseases such as myasthenic syndrome. CHRNB1 is also known as cholinergic receptor nicotinic beta polypeptide 1; acetylcholine receptor nicotinic beta 1 (muscle); ACHRB; CHRNB; CMS1D, CMS2C, CMS2A, or SCCMS.

[0278] The sequence of the human CHRNB1 mRNA transcript can be found, for example, in GenBank Accession No. GI:1519313560 (NM_000747.3; SEQ ID NO:241; reverse complement, SEQ ID NO:242). The sequence of the mouse CHRNB1 mRNA can be found, for example, in GenBank Accession No. GI:160358781 (NM_009601.4; SEQ ID NO:243; reverse complement, SEQ ID NO:244). The sequence of the rat CHRNB1 mRNA can be found, for example, in GenBank Accession No. GI:2048631755 (NM_001395118.1; SEQ ID NO:245; reverse complement, SEQ ID NO:246). The sequence of cynomolgus monkey CHRNB1 mRNA can be found, for example, in GenBank Accession No. GI:982302904 (XM_005582753.2; SEQ ID NO:247; reverse complement, SEQ ID NO:248). The sequence of rhesus monkey CHRNB1 mRNA can be found, for example, in GenBank Accession No. GI:1622877217 (XM_015118481.2; SEQ ID NO:249; reverse complement, SEQ ID NO:250).

[0279] Additional examples of CHRNB1 mRNA sequences are readily available through publicly available databases such as GenBank, UniProt, OMIM, and the Macaque Genome Project website. Further information about CHRNB1 can be found, for example, at www.ncbi.nlm.nih.gov / gene / ?term=CHRNB1.

[0280] The entire contents of each of the aforementioned GenBank accession numbers and gene database numbers are incorporated herein by reference as of the filing date of the present application.

[0281] As used herein, the term CHRNB1 also refers to variants of the CHRNB1 gene, including variants provided in the SNP database. Many sequence variations within the CHRNB1 gene have been identified and can be found, for example, in NCBI dbSNP and UniProt (see, for example, www.ncbi.nlm.nih.gov / snp / ?term=CHRNB1, the entire contents of which are incorporated herein by reference as of the filing date of this application).

[0282] As used herein, "cholinergic receptor nicotinic delta subunit" is used interchangeably with the term "CHRND" and refers to the delta subunit of the muscle acetylcholine receptor (AChR). The muscle acetylcholine receptor is composed of five subunits of four different types: two alpha subunits and one each of beta, gamma, and delta subunits. Upon binding acetylcholine, the AChR responds by undergoing a large-scale conformational change that affects all subunits and opens ion-conducting channels across the plasma membrane. CHRND is associated with diseases such as myasthenic syndrome. CHRND is also known as ACHRD, cholinergic receptor nicotinic delta polypeptide; acetylcholine receptor nicotinic delta (muscle); CMS2A; CMS3A, CMS3B, CMS3C, FCCMS, or SCCMS.

[0283] The sequence of the human CHRND mRNA transcript can be found, for example, in GenBank Accession No. GI:1519243557 (NM_000751.3; SEQ ID NO:251; reverse complement, SEQ ID NO:252). The sequence of the mouse CHRND mRNA can be found, for example, in GenBank Accession No. GI:426214082 (NM_021600.3; SEQ ID NO:253; reverse complement, SEQ ID NO:254). The sequence of the rat CHRND mRNA can be found, for example, in GenBank Accession No. GI:9506486 (NM_019298.1; SEQ ID NO:255; reverse complement, SEQ ID NO:256). The sequence of cynomolgus macaque CHRND mRNA can be found, for example, in GenBank Accession No. GI:982288086 (XM_005574618.2; SEQ ID NO:257; reverse complement, SEQ ID NO:258). The sequence of rhesus macaque CHRND mRNA can be found, for example, in GenBank Accession No. GI:1622852529 (XM_028831231.1; SEQ ID NO:259; reverse complement, SEQ ID NO:260).

[0284] Additional examples of CHRND mRNA sequences are readily available through publicly available databases such as GenBank, UniProt, OMIM, and the Macaque Genome Project website. Further information on CHRND can be found, for example, at www.ncbi.nlm.nih.gov / gene / ?term=CHRND.

[0285] The entire contents of each of the aforementioned GenBank accession numbers and gene database numbers are incorporated herein by reference as of the filing date of the present application.

[0286] As used herein, the term CHRND also refers to variants of the CHRND gene, including variants provided in SNP databases. Many sequence variations within the CHRND gene have been identified and can be found, for example, in NCBI dbSNP and UniProt (see, for example, www.ncbi.nlm.nih.gov / snp / ?term=CHRND, the entire contents of which are incorporated herein by reference as of the filing date of this application.

[0287] As used herein, "cholinergic receptor nicotinic epsilon subunit" is used interchangeably with the term "CHRNE" to refer to a subunit of the acetylcholine receptor. The acetylcholine receptor at the neuromuscular junction of mature mammals is composed of a pentameric protein complex composed of four subunits, with a ratio of two alpha subunits to one beta, one epsilon, and one delta subunit. The acetylcholine receptor changes subunit composition shortly after birth, when the epsilon subunit replaces the gamma subunit found in the embryonic receptor. Mutations in the epsilon subunit are associated with congenital myasthenic syndrome. CHRNE is also known as cholinergic receptor nicotinic epsilon; acetylcholine receptor nicotinic epsilon; ACHRE; CMS1D, CMS1E, CMS2A, CMS4A, CMS4B, CMS4C, FCCMS, or SCCMS.

[0288] The sequence of the human CHRNE mRNA transcript can be found, for example, in GenBank Accession No. GI:1433531118 (NM_000080.4; SEQ ID NO:261; reverse complement, SEQ ID NO:262). The sequence of the mouse CHRNE mRNA can be found, for example, in GenBank Accession No. GI:6752949 (NM_009603.1; SEQ ID NO:263; reverse complement, SEQ ID NO:264). The sequence of the rat CHRNE mRNA can be found, for example, in GenBank Accession No. GI:8393128 (NM_017194.1; SEQ ID NO:265; reverse complement, SEQ ID NO:266). The sequence of cynomolgus macaque CHRNE mRNA can be found, for example, in GenBank Accession No. GI:982302635 (XM_015437499.1; SEQ ID NO:267; reverse complement, SEQ ID NO:268). The sequence of rhesus macaque CHRNE mRNA can be found, for example, in GenBank Accession No. GI:1622876897 (XM_015118354.2; SEQ ID NO:269; reverse complement, SEQ ID NO:270).

[0289] Additional examples of CHRNE mRNA sequences are readily available through publicly available databases such as GenBank, UniProt, OMIM, and the Macaque Genome Project website. Further information about CHRNE can be found, for example, at www.ncbi.nlm.nih.gov / gene / ?term=CHRNE.

[0290] The entire contents of each of the aforementioned GenBank accession numbers and gene database numbers are incorporated herein by reference as of the filing date of the present application.

[0291] As used herein, the term CHRNE also refers to variations of the CHRNE gene, including variants provided in SNP databases. Many sequence variations within the CHRNE gene have been identified and can be found, for example, in NCBI dbSNP and UniProt (see, for example, www.ncbi.nlm.nih.gov / snp / ?term=CHRNE, the entire contents of which are incorporated herein by reference as of the filing date of this application).

[0292] As used herein, "choline receptor nicotinic gamma subunit" is used interchangeably with the term "CHRNG" to refer to the subunit of the acetylcholine receptor. The mammalian muscle acetylcholine receptor is a transmembrane pentameric glycoprotein with two alpha subunits, one beta, one delta, and one epsilon (in adult skeletal muscle) or gamma (in embryonic and denervated muscle) subunit. This gene encoding the gamma subunit is expressed in humans before the 33rd week of gestation. The gamma subunit of the acetylcholine receptor plays a role in neuromuscular organogenesis and ligand binding, and disruption of gamma subunit expression prevents the correct positioning of the receptor in the cell membrane. Mutations in the subunit are associated with congenital myasthenic syndrome. CHRNG is also known as choline receptor nicotinic gamma; acetylcholine receptor nicotinic gamma; or ACHRG.

[0293] The sequence of the human CHRNG mRNA transcript can be found, for example, in GenBank Accession No. GI:1441481359 (NM_005199.5; SEQ ID NO:271; reverse complement, SEQ ID NO:272). The sequence of the mouse CHRNG mRNA can be found, for example, in GenBank Accession No. GI:119964695 (NM_009604.3; SEQ ID NO:273; reverse complement, SEQ ID NO:274). The sequence of the rat CHRNG mRNA can be found, for example, in GenBank Accession No. GI:9506488 (NM_019145.1; SEQ ID NO:275; reverse complement, SEQ ID NO:276). The sequence of cynomolgus monkey CHRNG mRNA can be found, for example, in GenBank Accession No. GI:982288092 (XM_005574625.3; SEQ ID NO:277; reverse complement sequence, SEQ ID NO:278). The sequence of rhesus monkey CHRNG mRNA can be found, for example, in GenBank Accession No. GI:1622852538 (XM_028831233.1; SEQ ID NO:279; reverse complement sequence, SEQ ID NO:280).

[0294] Additional examples of CHRNG mRNA sequences are readily available through publicly available databases such as GenBank, UniProt, OMIM, and the Macaque Genome Project website. Further information on CHRNG can be found, for example, at www.ncbi.nlm.nih.gov / gene / ?term=CHRNG.

[0295] The entire contents of each of the aforementioned GenBank accession numbers and gene database numbers are incorporated herein by reference as of the filing date of the present application.

[0296] As used herein, the term CHRNG also refers to variants of the CHRNG gene, including variants provided in SNP databases. Many sequence variations within the CHRNG gene have been identified and can be found, for example, in NCBI dbSNP and UniProt (see, for example, www.ncbi.nlm.nih.gov / snp / ?term=CHRNG, the entire contents of which are incorporated herein by reference as of the filing date of this application).

[0297] As used herein, "collagen type XIII alpha 1 chain" is used interchangeably with the term "COL13A1" and refers to a synaptic extracellular matrix protein involved in the formation and maintenance of neuromuscular synapses. COL13A1 encodes collagen type XIII alpha 1 chain (COL13A1), a single-pass type II transmembrane protein composed of a short intracellular domain, a single transmembrane domain, and a triple-helical collagen extracellular domain. Studies have shown that patients with COL13A1 mutations are susceptible to a myasthenic syndrome characterized by early onset of weakness, significant eating and breathing difficulties, and the need for ventilation and artificial feeding. COL13A1 is also known as COLXIIIA1, collagen alpha-1 (XIII) chain, or CMS19.

[0298] The sequence of human COL13A1 mRNA transcript can be found, for example, in GenBank Accession No. GI:1677498641 (NM_001130103.2; SEQ ID NO:281: reverse complement, SEQ ID NO:282). The sequence of mouse COL13A1 mRNA can be found, for example, in GenBank Accession No. GI:755571593 (NM_007731.3; SEQ ID NO:283; reverse complement, SEQ ID NO:284). The sequence of rat COL13A1 mRNA can be found, for example, in GenBank Accession No. GI:157821424 (NM_001109172.1; SEQ ID NO:285; reverse complement, SEQ ID NO:286). The sequence of cynomolgus macaque COL13A1 mRNA can be found, for example, in GenBank Accession No. GI:982269148 (XM_015456252.1; SEQ ID NO:287; reverse complement, SEQ ID NO:288). The sequence of rhesus macaque COL13A1 mRNA can be found, for example, in GenBank Accession No. GI:1622966101 (XM_015147482.2; SEQ ID NO:289; reverse complement, SEQ ID NO:290).

[0299] Additional examples of COL13A1 mRNA sequences are readily available through publicly available databases such as GenBank, UniProt, OMIM, and the Macaque Genome Project website. Additional information about COL13A1 can be found, for example, at www.ncbi.nlm.nih.gov / gene / ?term=COL13A1.

[0300] The entire contents of each of the aforementioned GenBank accession numbers and gene database numbers are incorporated herein by reference as of the filing date of the present application.

[0301] As used herein, the term COL13A1 also refers to variants of the COL13A1 gene, including variants provided in SNP databases. Many sequence variations within the COL13A1 gene have been identified and can be found, for example, in NCBI dbSNP and UniProt (see, for example, www.ncbi.nlm.nih.gov / snp / ?term=COL13A1, the entire contents of which are incorporated herein by reference as of the filing date of this application).

[0302] As used herein, "Docking protein 7" is used interchangeably with the term "DOK7" and refers to a protein essential for neuromuscular synaptogenesis. The protein functions through aneural activation of muscle-specific receptor kinase, which is required for postsynaptic differentiation and subsequent clustering of acetylcholine receptors in myotubes. This protein also induces autophosphorylation of muscle-specific receptor kinase. Mutations in this gene are the cause of myasthenic congenital syndrome. DOK7 is also known as C4orf25, downstream tyrosine kinase 7, FLJ33718, FLJ39137, chromosome 4 open reading frame 25, CMS10, CMS1B, or FADS3.

[0303] The sequence of human DOK7 mRNA transcripts can be found, for example, in GenBank Accession No. GI:1519242777 (NM_173660.5; SEQ ID NO:291; reverse complement, SEQ ID NO:292). The sequence of mouse DOK7 mRNA can be found, for example, in GenBank Accession No. GI:1143077055 (NM_001348478.1; SEQ ID NO:293; reverse complement, SEQ ID NO:294). The sequence of rat DOK7 mRNA can be found, for example, in GenBank Accession No. GI:194240570 (NM_001130062.1; SEQ ID NO:295; reverse complement, SEQ ID NO:296). The sequence of cynomolgus macaque DOK7 mRNA can be found, for example, in GenBank Accession No. GI:982247946 (XM_015450057.1; SEQ ID NO:297; reverse complement sequence, SEQ ID NO:298). The sequence of rhesus macaque DOK7 mRNA can be found, for example, in GenBank Accession No. GI:1622938489 (XM_015137905.2; SEQ ID NO:299; reverse complement sequence, SEQ ID NO:300).

[0304] Additional examples of DOK7 mRNA sequences are readily available through publicly available databases such as GenBank, UniProt, OMIM, and the Macaque Genome Project website. Additional information about DOK7 can be found, for example, at www.ncbi.nlm.nih.gov / gene / ?term=DOK7.

[0305] The entire contents of each of the aforementioned GenBank accession numbers and gene database numbers are incorporated herein by reference as of the filing date of the present application.

[0306] As used herein, the term DOK7 also refers to variants of the DOK7 gene, including variants provided in SNP databases. Many sequence variations within the DOK7 gene have been identified and can be found, for example, in NCBI dbSNP and UniProt (see, for example, www.ncbi.nlm.nih.gov / snp / ?term=DOK7, the entire contents of which are incorporated herein by reference as of the filing date of this application).

[0307] As used herein, "LDL receptor-related protein 4" is used interchangeably with the term "LRP4" to refer to a member of the low-density lipoprotein receptor-related protein family. LRP4 is a single transmembrane protein with a large extracellular domain containing multiple LDLR repeats, EGF-like and β-propeller repeats; a transmembrane domain; and a short C-terminal region without an identifiable catalytic motif. Mice lacking LRP4 die at birth and do not form NMJs, indicating a key role in the formation of the neuromuscular junction (NMJ). LPR4 mutations or dysfunction are implicated in conditions including congenital myasthenic syndrome, myasthenia gravis, and skeletal or renal diseases. LRP4 is also known as MEGF7, LRP-4, SOST2, CLSS, low-density lipoprotein receptor-related protein 4, multiple epidermal growth factor-like domain 7, LRP10, KIAA0816, or CMS17.

[0308] The sequence of the human LRP4 mRNA transcript can be found, for example, in GenBank Accession No. GI:1519312025 (NM_002334.4; SEQ ID NO:301; reverse complement, SEQ ID NO:302). The sequence of the mouse LRP4 mRNA can be found, for example, in GenBank Accession No. GI:224994222 (NM_172668.3; SEQ ID NO:303; reverse complement, SEQ ID NO:304). The sequence of the rat LRP4 mRNA can be found, for example, in GenBank Accession No. GI:329112575 (NM_031322.3; SEQ ID NO:305; reverse complement, SEQ ID NO:306). The sequence of cynomolgus monkey LRP4 mRNA can be found, for example, in GenBank Accession No. GI:982294148 (XM_005578015.2; SEQ ID NO:307; reverse complement, SEQ ID NO:308). The sequence of rhesus monkey LRP4 mRNA can be found, for example, in GenBank Accession No. GI:1622863351 (XM_015114355.2; SEQ ID NO:309; reverse complement, SEQ ID NO:310).

[0309] Additional examples of LRP4 mRNA sequences are readily available through publicly available databases such as GenBank, UniProt, OMIM, and the Macaque Genome Project website. Additional information about LRP4 can be found, for example, at www.ncbi.nlm.nih.gov / gene / ?term=LRP4.

[0310] The entire contents of each of the aforementioned GenBank accession numbers and gene database numbers are incorporated herein by reference as of the filing date of the present application.

[0311] As used herein, the term LRP4 also refers to variants of the LRP4 gene, including variants provided in SNP databases. Many sequence variations within the LRP4 gene have been identified and can be found, for example, in NCBI dbSNP and UniProt (see, for example, www.ncbi.nlm.nih.gov / snp / ?term=LRP4, the entire contents of which are incorporated herein by reference as of the filing date of this application).

[0312] As used herein, "muscle-associated receptor tyrosine kinase" is used interchangeably with the term "MUSK" and refers to a muscle-specific tyrosine kinase receptor that plays a central role in the formation and maintenance of the synapse between motor neurons and skeletal muscle at the neuromuscular junction (NMJ). AGRIN is recruited to the MUSK signaling complex via LRP4 to induce phosphorylation and activation of MUSK, i.e., the complex's kinase. Activation of MUSK in myotubes regulates NMJ formation by regulating various processes, including specific gene expression in the subsynaptic nucleus, reorganization of the actin cytoskeleton, and clustering of acetylcholine receptors in the postsynaptic membrane. Mutations in this gene are associated with congenital myasthenic syndrome. MUSK is also known as EC 2.7.10.1, FADS1, CMS9, FADS, musculoskeletal receptor tyrosine kinase, or muscle-specific kinase receptor.

[0313] The sequence of human MUSK mRNA transcripts can be found, for example, in GenBank Accession No. GI:1609044119 (NM_005592.4; SEQ ID NO:311; reverse complement, SEQ ID NO:312). The sequence of mouse MUSK mRNA can be found, for example, in GenBank Accession No. GI:260267047 (NM_001037127.2; SEQ ID NO:313; reverse complement, SEQ ID NO:314). The sequence of rat MUSK mRNA can be found, for example, in GenBank Accession No. GI:1937920431 (NM_031061.2; SEQ ID NO:315; reverse complement, SEQ ID NO:316). The sequence of cynomolgus macaque MUSK mRNA can be found, for example, in GenBank Accession No. GI:982300549 (XM_005581093.2; SEQ ID NO:317; reverse complement, SEQ ID NO:318). The sequence of rhesus macaque MUSK mRNA can be found, for example, in GenBank Accession No. GI:1622871800 (XM_015117113.2; SEQ ID NO:319; reverse complement, SEQ ID NO:320).

[0314] Additional examples of MUSK mRNA sequences are readily available through publicly available databases such as GenBank, UniProt, OMIM, and the Macaque Genome Project website. Further information about MUSK can be found, for example, at www.ncbi.nlm.nih.gov / gene / ?term=MUSK.

[0315] The entire contents of each of the aforementioned GenBank accession numbers and gene database numbers are incorporated herein by reference as of the filing date of the present application.

[0316] As used herein, the term MUSK also refers to variants of the MUSK gene, including variants provided in SNP databases. Many sequence variations within the MUSK gene have been identified and can be found, for example, in NCBI dbSNP and UniProt (see, for example, www.ncbi.nlm.nih.gov / snp / ?term=MUSK, the entire contents of which are incorporated herein by reference as of the filing date of this application).

[0317] As used herein, "synaptic receptor-associated protein" is used interchangeably with the term "RAPSN" to refer to a member of the family of proteins that are synaptic receptor-associated proteins. The encoded protein contains a conserved cAMP-dependent protein kinase phosphorylation site and plays a key role in clustering and anchoring nicotinic acetylcholine receptors at synaptic sites by linking the receptor to the underlying postsynaptic cytoskeleton, possibly through direct binding to actin or spectrin. Mutations in this gene may play a role in postsynaptic congenital myasthenic syndrome. RAPSN is also known as RNF205, synaptic 43 kDa receptor-associated protein, RING finger protein 205, CMS1D, CMS1E, acetylcholine receptor-associated 43 kDa protein, RAPSYN, CMS11, CMS4C, FADS2, or FADS.

[0318] The sequence of the human RAPSN mRNA transcript can be found, for example, in GenBank Accession No. GI: ​​1519241818 (NM_005055.5; SEQ ID NO: 321; reverse complement, SEQ ID NO: 322). The sequence of the mouse RAPSN mRNA can be found, for example, in GenBank Accession No. GI: ​​224967080 (NM_009023.3; SEQ ID NO: 323; reverse complement, SEQ ID NO: 324). The sequence of the rat RAPSN mRNA can be found, for example, in GenBank Accession No. GI: ​​157819696 (NM_001108584.1; SEQ ID NO: 325; reverse complement, SEQ ID NO: 326). The sequence of cynomolgus macaque RAPSN mRNA can be found, for example, in GenBank Accession No. GI:982294016 (XM_015434747.1; SEQ ID NO:327; reverse complement, SEQ ID NO:328). The sequence of rhesus macaque RAPSN mRNA can be found, for example, in GenBank Accession No. GI:1622863236 (XM_015114296.2; SEQ ID NO:329; reverse complement, SEQ ID NO:330).

[0319] Additional examples of RAPSN mRNA sequences are readily available through publicly available databases such as GenBank, UniProt, OMIM, and the Macaque Genome Project website. Further information on RAPSN can be found, for example, at www.ncbi.nlm.nih.gov / gene / ?term=RAPSN.

[0320] The entire contents of each of the aforementioned GenBank accession numbers and gene database numbers are incorporated herein by reference as of the filing date of the present application.

[0321] As used herein, the term RAPSN also refers to variants of the RAPSN gene, including variants provided in the SNP database. Many sequence variations within the RAPSN gene have been identified and can be found, for example, in NCBI dbSNP and UniProt (see, for example, www.ncbi.nlm.nih.gov / snp / ?term=RAPSN, the entire contents of which are incorporated herein by reference as of the filing date of this application.

[0322] As used herein, "sodium voltage-gated channel alpha subunit 4" is used interchangeably with the term "SCN4A" to refer to a member of the voltage-gated sodium channel family. Voltage-gated sodium channels are transmembrane glycoprotein complexes composed of a large alpha subunit with 24 transmembrane domains and one or more regulatory beta subunits. They are responsible for the generation and propagation of action potentials in neurons and muscles. This gene encodes a member of the sodium channel alpha subunit gene family. It is expressed in skeletal muscle, and mutations in this gene have been associated with congenital myasthenic syndromes and several myotonic and periodic paralysis disorders. SCN4A is also known as SkM1, Nav1.4, HYPP, sodium channel protein skeletal muscle subunit alpha, voltage-gated sodium channel subunit alpha Nav1.4, HYKPP, skeletal muscle voltage-dependent sodium channel type IV alpha subunit, CTC-264K15.6, Na(V)1.4, HOKPP2, CMS16, or NAC1A.

[0323] The sequence of the human SCN4A mRNA transcript can be found, for example, in GenBank Accession No. GI:93587341 (NM_000334.4; SEQ ID NO:331; reverse complement, SEQ ID NO:332). The sequence of the mouse SCN4A mRNA can be found, for example, in GenBank Accession No. GI:134948031 (NM_133199.2; SEQ ID NO:333; reverse complement, SEQ ID NO:334). The sequence of the rat SCN4A mRNA can be found, for example, in GenBank Accession No. GI:1937369400 (NM_013178.2; SEQ ID NO:335; reverse complement, SEQ ID NO:336). The sequence of cynomolgus macaque SCN4A mRNA can be found, for example, in GenBank Accession No. GI:982306407 (XM_015438708.1; SEQ ID NO:337; reverse complement, SEQ ID NO:338). The sequence of rhesus macaque SCN4A mRNA can be found, for example, in GenBank Accession No. GI:1622880585XM_015120096.2; SEQ ID NO:339; reverse complement, SEQ ID NO:340).

[0324] Additional examples of SCN4A mRNA sequences are readily available through publicly available databases such as GenBank, UniProt, OMIM, and the Macaque Genome Project website. Additional information about SCN4A can be found, for example, at www.ncbi.nlm.nih.gov / gene / ?term=SCN4A.

[0325] The entire contents of each of the aforementioned GenBank accession numbers and gene database numbers are incorporated herein by reference as of the filing date of the present application.

[0326] As used herein, the term SCN4A also refers to variations of the SCN4A gene, including variants provided in SNP databases. Many sequence variations within the SCN4A gene have been identified and can be found, for example, in NCBI dbSNP and UniProt (see, for example, www.ncbi.nlm.nih.gov / snp / ?term=SCN4A, the entire contents of which are incorporated herein by reference as of the filing date of this application).

[0327] As used herein, "double homeobox 4" is used interchangeably with the term "DUX4" to refer to a transcriptional activator of many genes. DUX4 is normally expressed during early embryonic development and is subsequently silenced in virtually all tissues except the testis and thymus. DUX4 has been implicated in cell death, oxidative stress, muscle differentiation and growth, epigenetic regulation, and many other signaling pathways in skeletal muscle. Inappropriate expression of DUX4 in muscle cells is the cause of facioscapulohumeral muscular dystrophy (FSHD), a disease characterized by muscle weakness and atrophy (dystrophy) that slowly worsens over time. DUX4 is also known as double homeobox protein 10, double homeobox protein 4, double homeobox protein 4 / 10, DUX4L, and DUX10.

[0328] The sequence of the human DUX4 mRNA transcript can be found, for example, in GenBank Accession No. GI: ​​1774753171 (NM_001306068.3; SEQ ID NO: 341; reverse complement, SEQ ID NO: 342). The sequence of the mouse DUX4 mRNA can be found, for example, in GenBank Accession No. GI: ​​126432555 (NM_001081954.1; SEQ ID NO: 343; reverse complement, SEQ ID NO: 344). The sequence of the rat DUX4 mRNA can be found, for example, in GenBank Accession No. GI: ​​1958689769 (XM_008771031.3; SEQ ID NO: 345; reverse complement, SEQ ID NO: 346). The sequence of rhesus macaque DUX4 mRNA can be found, for example, in GenBank Accession No. GI: ​​1622942424 (XM_028848991.1; SEQ ID NO: 347; reverse complement sequence, SEQ ID NO: 348).

[0329] Additional examples of DUX4 mRNA sequences are readily available through publicly available databases such as GenBank, UniProt, OMIM, and the Macaque Genome Project website. Additional information about DUX4 can be found, for example, at www.ncbi.nlm.nih.gov / gene / ?term=DUX4.

[0330] The entire contents of each of the aforementioned GenBank accession numbers and gene database numbers are incorporated herein by reference as of the filing date of the present application.

[0331] As used herein, the term DUX4 also refers to variants of the DUX4 gene, including variants provided in the SNP database. Many sequence variations within the DUX4 gene have been identified and can be found, for example, in NCBI dbSNP and UniProt (see, for example, www.ncbi.nlm.nih.gov / snp / ?term=DUX4, the entire contents of which are incorporated herein by reference as of the filing date of this application).

[0332] As used herein, "phospholamban" is used interchangeably with the term "PLN" and refers to a key regulator of cardiac contractility. PLN is the primary substrate of cAMP-dependent protein kinase in the myocardium. The encoded protein is an inhibitor of the cardiac sarcoplasmic reticulum Ca(2+)-ATPase in its unphosphorylated state, but inhibition is reduced upon protein phosphorylation. Subsequent activation of the Ca(2+) pump leads to an enhanced rate of muscle relaxation, thereby promoting the muscle contraction response induced by β-agonists in the heart. The encoded protein is a key regulator of cardiac diastolic function. Mutations in this gene are the cause of hereditary human dilated cardiomyopathy with refractory congestive heart failure, as well as familial hypertrophic cardiomyopathy. PLN is also known as CMD1P, PLB, cardiophospholamban, or CMH.

[0333] Exemplary sequences of human PLN mRNA transcripts can be found, for example, in GenBank Accession No. GI:1519242997 (NM_002667.5; SEQ ID NO:349; reverse complement, SEQ ID NO:350). The sequence of mouse PLN mRNA can be found, for example, in GenBank Accession No. GI:213512815 (NM_001141927.1; SEQ ID NO:351; reverse complement, SEQ ID NO:352). The sequence of rat PLN mRNA can be found, for example, in GenBank Accession No. GI:399124783 (NM_022707.2; SEQ ID NO:353; reverse complement, SEQ ID NO:354). The sequence of rhesus macaque PLN mRNA can be found, for example, in GenBank Accession No. GI: ​​1863319929 (NM_001190894.2; SEQ ID NO: 355; reverse complement sequence, SEQ ID NO: 356).

[0334] Additional examples of PLN mRNA sequences are readily available through publicly available databases such as GenBank, UniProt, OMIM, the UCSC Genome Browser, and the Macaque Genome Project website. Additional information about PLN can be found, for example, at www.ncbi.nlm.nih.gov / gene / ?term=PLN.

[0335] The entire contents of each of the aforementioned GenBank accession numbers and gene database numbers are incorporated herein by reference as of the filing date of the present application.

[0336] As used herein, the term PLN also refers to variations of the PLN gene, including variants provided in the SNP database. Many sequence variations within the PLN gene have been identified and can be found, for example, in NCBI dbSNP and UniProt (see, for example, www.ncbi.nlm.nih.gov / snp / ?term=PLN, the entire contents of which are incorporated herein by reference as of the filing date of this application.

[0337] As used herein, "calcium / calmodulin-dependent protein kinase IIδ" can be used interchangeably with the term "CAMK2D" to refer to a member of the serine / threonine protein kinase family and the Ca(2+) / calmodulin-dependent protein kinase subfamily. CAMK2D is involved in regulating Ca(2+) homeostasis and excitation-contraction coupling in the heart by targeting ion channels, transport proteins, and auxiliary proteins involved in Ca(2+) influx into myocytes, Ca(2+) release from the sarcoplasmic reticulum (SR), SR Ca(2+) absorption, and Na(+) and K(+) channel transport. CAMK2D also targets transcription factors and signaling molecules to regulate cardiac function. In its activated form, CAMK2D is involved in the pathogenesis of dilated cardiomyopathy and heart failure. CAMK2D contributes to cardiac decompensation and heart failure by regulating SR Ca(2+) release through direct phosphorylation of the RYR2 Ca(2+) channel. In the nucleus, CAMK2D phosphorylates the MEF2 inhibitor HDAC4, promoting its nuclear export and binding to 14-3-3 proteins, as well as the expression of MEF2 and genes involved in the hypertrophic process. CAMK2D is crucial for the left ventricular remodeling response to myocardial infarction. In pathological myocardial remodeling, CAMK2D acts downstream of the β-adrenergic receptor signaling cascade to regulate key proteins involved in excitation-contraction coordination. CAMK2D regulates Ca(2+) influx into myocytes by binding to and phosphorylating the L-type Ca(2+) channel subunit β-2CACNB2. In addition to Ca(2+) channels, CAMK2D can also target and regulate the cardiac sarcolemma Na(+) channel Nav1.5 / SCN5A and the K+ channel Kv4.3 / KCND3, which contribute to the development of arrhythmias in heart failure. CAMK2D phosphorylates phospholamban (PLN), an endogenous inhibitor of SERCA2A / ATP2A2, leading to enhanced SR Ca(2+) uptake during acidosis, which may be important in frequency-dependent acceleration of relaxation and maintenance of contractile function. CAMK2D may participate in regulating skeletal muscle function in response to exercise by modulating SR Ca(2+) transport through phosphorylation of PLN and trinexin, a ryanodine receptor coupling factor. CAMK2D is also known as calcium / calmodulin-dependent protein kinase type II delta chain, CaM kinase II delta subunit, CaM kinase II subunit delta, CAMKD, EC 2.7.11.17, or EC 2.7.11.

[0338] Exemplary sequences of human CAMK2D mRNA transcripts can be found, for example, in GenBank Accession No. GI: ​​1519243899 (NM_001321571.2; SEQ ID NO: 357; reverse complement, SEQ ID NO: 358). The sequence of mouse CAMK2D mRNA can be found, for example, in GenBank Accession No. GI: ​​654824235 (NM_001025439.2; SEQ ID NO: 359; reverse complement, SEQ ID NO: 360). The sequence of rat CAMK2D mRNA can be found, for example, in GenBank Accession No. GI: ​​144922682 (NM_012519.2; SEQ ID NO: 361; reverse complement, SEQ ID NO: 362). The sequence of rhesus macaque CAMK2D mRNA can be found, for example, in GenBank Accession No. GI: ​​1622941163 (XM_015139100.2; SEQ ID NO: 363; reverse complement sequence, SEQ ID NO: 364).

[0339] Additional examples of CAMK2D mRNA sequences are readily available through publicly available databases such as GenBank, UniProt, OMIM, the UCSC genome browser, and the Macaque Genome Project website. Additional information about CAMK2D can be found, for example, at www.ncbi.nlm.nih.gov / gene / ?term=CAMK2D.

[0340] The entire contents of each of the aforementioned GenBank accession numbers and gene database numbers are incorporated herein by reference as of the filing date of the present application.

[0341] As used herein, the term CAMK2D also refers to variations of the CAMK2D gene, including variants provided in SNP databases. Many sequence variations within the CAMK2D gene have been identified and can be found, for example, in NCBI dbSNP and UniProt (see, for example, www.ncbi.nlm.nih.gov / snp / ?term=CAMK2D, the entire contents of which are incorporated herein by reference as of the filing date of this application).

[0342] As used herein, "dystrophin-myotonic protein kinase" is used interchangeably with the term "DMPK" and refers to a non-receptor serine / threonine protein kinase that is essential for maintaining skeletal muscle structure and function. DMPK plays a role in myocyte differentiation and survival by regulating the integrity of the nuclear envelope and the expression of muscle-specific genes. DMPK also phosphorylates PPP1R12A and inhibits myosin phosphatase activity to regulate myosin phosphorylation. DMPK is also critical for regulating myocardial contractility and maintaining proper cardiac conduction activity, possibly through regulation of cellular calcium homeostasis. DMPK phosphorylates PLN, a regulator of the calcium pump, and can regulate sarcoplasmic reticulum calcium uptake in myocytes. DMPK also phosphorylates FXYD1 / PLM, which can induce chloride currents, and can play a role in synaptic plasticity. DMPK is also known as DM1 protein kinase, DM1PK, DM1, MT-PK, MDPK, DMK, myotonic kinase, myotonic dystrophy-related protein kinase, dystrophic myotonic protein kinase, myotonic protein kinase A, thymopoietin homolog, or EC 2.7.11.1.

[0343] Exemplary sequences of human DMPK mRNA transcripts can be found, for example, in GenBank Accession No. GI:571026697 (NM_001081563.2; SEQ ID NO:365; reverse complement, SEQ ID NO:366). The sequence of mouse DMPK mRNA can be found, for example, in GenBank Accession No. GI:1824718155 (NM_032418.3; SEQ ID NO:367; reverse complement, SEQ ID NO:368). The sequence of rat DMPK mRNA can be found, for example, in GenBank Accession No. GI:1719749725 (NM_001372064.1; SEQ ID NO:369; reverse complement, SEQ ID NO:370). The sequence of cynomolgus macaque DMPK mRNA can be found, for example, in GenBank Accession No. GI:2161880869 (XM_045381179.1; SEQ ID NO:371; reverse complement sequence, SEQ ID NO:372).

[0344] Additional examples of DMPK mRNA sequences are readily available through publicly available databases such as GenBank, UniProt, OMIM, the UCSC genome browser, and the Macaque Genome Project website. Additional information about DMPK can be found, for example, at www.ncbi.nlm.nih.gov / gene / ?term=DMPK.

[0345] The entire contents of each of the aforementioned GenBank accession numbers and gene database numbers are incorporated herein by reference as of the filing date of the present application.

[0346] As used herein, the term DMPK also refers to variants of the DMPK gene, including variants provided in the SNP database. Many sequence variations within the DMPK gene have been identified and can be found, for example, in NCBI-dbSNP and UniProt (see, for example, www.ncbi.nlm.nih.gov / snp / ?term=DMPK, the entire contents of which are incorporated herein by reference as of the filing date of this application.

[0347] As used herein, "glycogen synthase 1" is used interchangeably with the term "GYS1" and refers to an enzyme that catalyzes the addition of glucose monomers to growing glycogen molecules. Glycogen is the body's primary source of stored energy. Most of the glucose obtained from food is stored in muscle cells as glycogen. During cardiac contraction or rapid or sustained action of skeletal muscle, glycogen stored in muscle cells is broken down to supply energy to the cells. GYS1 is produced in most cells but is most abundant in cardiac (heart) muscle and muscles used for movement (skeletal muscle). Mutations in the GYS1 gene have been found to cause a form of glycogen storage disease type 0 (GSDO) that affects cardiac and skeletal muscle. Most GYS1 gene mutations that cause this condition result in a lack of functional muscle glycogen synthase, resulting in a complete absence of glycogen in muscle cells. Normally, glycogen is formed from excess glucose that is not immediately used by cells after glucose is ingested during a meal. In people with GSDO who are unable to form glycogen, the extra sugar is released by the body. As a result, people with muscle GSD0 lack any stored energy, which leads to muscle pain, weakness, or fainting episodes after moderate physical activity. Due to the absence of glycogen in the myocardium, affected individuals are also at increased risk of cardiac arrest and sudden death, especially after physical activity. GYS1 is also known as muscle glycogen synthase, GSY, GYS, or EC 2.4.1.11.

[0348] Exemplary sequences of human GYS1 mRNA transcripts can be found, for example, in GenBank Accession No. GI:1519246122NM_002103.5 (SEQ ID NO:373; reverse complement, SEQ ID NO:374). The sequence of mouse GYS1 mRNA can be found, for example, in GenBank Accession No. GI:119672917 (NM_030678.3; SEQ ID NO:375; reverse complement, SEQ ID NO:376). The sequence of rat GYS1 mRNA can be found, for example, in GenBank Accession No. GI:157823921 (NM_001109615.1; SEQ ID NO:377; reverse complement, SEQ ID NO:378). The sequence of cynomolgus macaque GYS1 mRNA can be found, for example, in GenBank Accession No. GI:2161874347 (XM_005589837.3; SEQ ID NO:379; reverse complement sequence, SEQ ID NO:380).

[0349] Additional examples of GYS1 mRNA sequences are readily available through publicly available databases such as GenBank, UniProt, OMIM, the UCSC Genome Browser, and the Macaque Genome Project website. Additional information about GYS1 can be found, for example, at www.ncbi.nlm.nih.gov / gene / ?term=GYS1.

[0350] The entire contents of each of the aforementioned GenBank accession numbers and gene database numbers are incorporated herein by reference as of the filing date of the present application.

[0351] As used herein, the term GYS1 also refers to variants of the GYS1 gene, including variants provided in SNP databases. Many sequence variations within the GYS1 gene have been identified and can be found, for example, in NCBI dbSNP and UniProt (see, for example, www.ncbi.nlm.nih.gov / snp / ?term=GYS1, the entire contents of which are incorporated herein by reference as of the filing date of this application).

[0352] As used herein, "survival motor neuron 1" is used interchangeably with the term "SMN1" and refers to one of a group of proteins called the SMN complex, which is important for maintaining specialized nerve cells called motor neurons. These cells are located in the spinal cord and the part of the brain connected to the spinal cord (the brainstem). Motor neurons transmit signals from the brain and spinal cord that tell skeletal muscles to tense (contract), which enables body movement. Within the cell, the SMN complex plays an important role in processing mRNA. The SMN complex helps assemble the cellular machinery needed to process pre-mRNA. The SMN complex is also important for the development of specialized outgrowths called dendrites and axons from nerve cells. Dendrites and axons are required for transmitting impulses between neurons and from neurons to muscles. Many mutations in the SMN1 gene have been found to cause spinal muscular atrophy. This condition is characterized by the loss of motor neurons, leading to weakness and atrophy (dystrophy) of muscles used for movement (skeletal muscles) that worsens with age. SMN1 is also known as SMNT, TDRD16A, Gemin-1, BCD541, GEMIN1, SMA1, SMA2, SMA3, SMA4, SMN, SMNT, TUDOE domain-containing 16A, gems complementation sequence 1, or SMNC.

[0353] Exemplary sequences of human SMN1 mRNA transcripts can be found, for example, in GenBank Accession No. GI:663070993 (NM_001297715.1; SEQ ID NO:381; reverse complement, SEQ ID NO:382). The sequence of mouse SMN1 mRNA can be found, for example, in GenBank Accession No. GI:145386573 (NM_011420.2; SEQ ID NO:383; reverse complement, SEQ ID NO:384). The sequence of rat SMN1 mRNA can be found, for example, in GenBank Accession No. GI:1939402010 (NM_022509.2; SEQ ID NO:385; reverse complement, SEQ ID NO:386). The sequence of rhesus macaque SMN1 mRNA can be found, for example, in GenBank Accession No. GI:386781228 (NM_001260664.1; SEQ ID NO:387; reverse complement sequence, SEQ ID NO:388).

[0354] Additional examples of SMN1 mRNA sequences are readily available through publicly available databases such as GenBank, UniProt, OMIM, the UCSC Genome Browser, and the Macaque Genome Project website. Additional information about SMN1 can be found, for example, at www.ncbi.nlm.nih.gov / gene / ?term=SMN1.

[0355] The entire contents of each of the aforementioned GenBank accession numbers and gene database numbers are incorporated herein by reference as of the filing date of the present application.

[0356] As used herein, the term SMN1 also refers to variants of the SMN1 gene, including variants provided in SNP databases. Many sequence variations within the SMN1 gene have been identified and can be found, for example, in NCBI dbSNP and UniProt (see, for example, www.ncbi.nlm.nih.gov / snp / ?term=SMN1, the entire contents of which are incorporated herein by reference as of the filing date of this application).

[0357] As used herein, "α-glucosidase" is used interchangeably with the term "GAA" and refers to the enzyme in the lysosome that is responsible for the degradation of glycogen into glucose. More than 200 mutations in the GAA gene have been identified in people with Pompe disease. Many of these mutations change one of the protein building blocks (amino acids) used to make acid α-glucosidase. Other mutations insert or delete genetic material in the GAA gene. Mutations in this gene significantly reduce the activity of acid α-glucosidase, effectively preventing the enzyme from breaking down glycogen. As a result, this complex sugar can accumulate to toxic levels in the lysosomes. Abnormal accumulation of glycogen damages organs and tissues throughout the body, especially muscles, causing progressive muscle weakness, heart problems, and other characteristics of Pompe disease. GAA is also known as lysosomal α-glucosidase, acid maltase, EC 3.2.1.20, type II glycogen storage disease, or LYAG.

[0358] Exemplary sequences of human GAA mRNA transcripts can be found, for example, in GenBank Accession No. GI:1519245858 (NM_000152.5; SEQ ID NO:389; reverse complement, SEQ ID NO:390). The sequence of mouse GAA mRNA can be found, for example, in GenBank Accession No. GI:957579368 (NM_008064.4; SEQ ID NO:391; reverse complement, SEQ ID NO:392). The sequence of rat GAA mRNA can be found, for example, in GenBank Accession No. GI:40018605 (NM_199118.1; SEQ ID NO:393; reverse complement, SEQ ID NO:394). The sequence of rhesus macaque GAA mRNA can be found, for example, in GenBank Accession No. GI: ​​1622881859 (XM_015120499.2; SEQ ID NO: 395; reverse complement sequence, SEQ ID NO: 396).

[0359] Additional examples of GAA mRNA sequences are readily available through publicly available databases such as GenBank, UniProt, OMIM, the UCSC genome browser, and the Macaque Genome Project website. Further information on GAA can be found, for example, at www.ncbi.nlm.nih.gov / gene / ?term=GAA.

[0360] The entire contents of each of the aforementioned GenBank accession numbers and gene database numbers are incorporated herein by reference as of the filing date of the present application.

[0361] As used herein, the term GAA also refers to variants of the GAA gene, including variants provided in the SNP database. Many sequence variations within the GAA gene have been identified and can be found, for example, in NCBI dbSNP and UniProt (see, for example, www.ncbi.nlm.nih.gov / snp / ?term=GAA, the entire contents of which are incorporated herein by reference as of the filing date of this application.

[0362] As used herein, "oligomeric mucus / gel-forming mucin 5B" is used interchangeably with the term "MUC5B" to refer to a member of the mucin family of proteins that are highly glycosylated macromolecular components of mucus secretions. MUB5B is the major gel-forming mucin in mucus. It is a major contributor to the lubricity and viscoelastic properties of whole saliva, normal lung mucus, and cervical mucus. This gene has been found to be upregulated in several human diseases, including the sinus mucosa of chronic rhinosinusitis (CRS), CRS with nasal polyps, chronic obstructive pulmonary disease (COPD), and Helicobacter pylori (H. pylori)-associated gastropathy. MUC5B is also known as mucin-5B, MUC5, MG1, high molecular weight salivary mucin MG1; tracheobronchial mucin 5 subtype B; sublingual gland mucin; cervical mucin, or MUC9.

[0363] Exemplary sequences of human MUC5B mRNA transcripts can be found, for example, in GenBank Accession No. GI:1519244536 (NM_002458.3; SEQ ID NO:397; reverse complement, SEQ ID NO:398). The sequence of mouse MUC5B mRNA can be found, for example, in GenBank Accession No. GI:147905739 (NM_028801.2; SEQ ID NO:399; reverse complement, SEQ ID NO:400). The sequence of rat MUC5B mRNA can be found, for example, in GenBank Accession No. GI:1958654562 (XM_039101271.1; SEQ ID NO:401; reverse complement, SEQ ID NO:402). The sequence of rhesus macaque MUC5B mRNA can be found, for example, in GenBank Accession No. GI: ​​1622861542 (XM_028833012.1; SEQ ID NO: 403; reverse complement sequence, SEQ ID NO: 404).

[0364] Additional examples of MUC5B mRNA sequences are readily available through publicly available databases such as GenBank, UniProt, OMIM, the UCSC Genome Browser, and the Macaque Genome Project website.

[0365] Additional information about MUC5B can be found, for example, at www.ncbi.nlm.nih.gov / gene / ?term=MUC5B.

[0366] The entire contents of each of the aforementioned GenBank accession numbers and gene database numbers are incorporated herein by reference as of the filing date of the present application.

[0367] As used herein, the term MUC5B also refers to variants of the MUC5B gene, including variants provided in SNP databases. Many sequence variations within the MUC5B gene have been identified and can be found, for example, in NCBI dbSNP and UniProt (see, for example, www.ncbi.nlm.nih.gov / gene / ?term=MUC5B, the entire contents of which are incorporated herein by reference as of the filing date of this application).

[0368] As used herein, "thymic stromal lymphopoietin" is used interchangeably with the term "TSLP" and refers to a hematopoietic cytokine that signals through a heterodimeric receptor complex composed of the TSLP receptor and the IL-7Rα chain. TSLP primarily affects myeloid cells and induces the release of T cell-attracting chemokines from monocytes and enhances the maturation of CD11c(+) dendritic cells. TSLP promotes T helper type 2 (TH2) cell responses associated with immunity in various inflammatory diseases, including asthma, allergic inflammation, and chronic obstructive pulmonary disease.

[0369] Exemplary sequences of human TSLP mRNA transcripts can be found, for example, in GenBank Accession No. GI: ​​1519241510 (NM_033035.5; SEQ ID NO: 405; reverse complement, SEQ ID NO: 406). The sequence of mouse TSLP mRNA can be found, for example, in GenBank Accession No. GI: ​​283945612 (NM_021367.2; SEQ ID NO: 407; reverse complement, SEQ ID NO: 408). The sequence of rat TSLP mRNA can be found, for example, in GenBank Accession No. GI: ​​1958745494 (XM_039097381.1; SEQ ID NO: 409; reverse complement, SEQ ID NO: 410). The sequence of rhesus macaque TSLP mRNA can be found, for example, in GenBank Accession No. GI: ​​1622946249 (XM_001100503.4; SEQ ID NO: 411; reverse complement sequence, SEQ ID NO: 412).

[0370] Additional examples of TSLP mRNA sequences are readily available through publicly available databases such as GenBank, UniProt, OMIM, the UCSC Genome Browser, and the Macaque Genome Project website.

[0371] Additional information about TSLP can be found, for example, at www.ncbi.nlm.nih.gov / gene / ?term=TSLP.

[0372] The entire contents of each of the aforementioned GenBank accession numbers and gene database numbers are incorporated herein by reference as of the filing date of the present application.

[0373] As used herein, the term TSLP also refers to variations of the TSLP gene, including variants provided in SNP databases. Many sequence variations within the TSLP gene have been identified and can be found, for example, in NCBI dbSNP and UniProt (see, for example, www.ncbi.nlm.nih.gov / gene / ?term=TSLP, the entire contents of which are incorporated herein by reference as of the filing date of this application).

[0374] As used herein, "interleukin 33" is used interchangeably with the term "IL33" and refers to an alarmin cytokine from the II-1 family. IL33 binds to the IL1RLI / ST2 receptor and transmits signals through the receptor, which in turn activates the NF-κ-B and MAPK signaling pathways in target cells. IL33 is involved in the maturation of Th2 cells that induce the secretion of T helper type 2-related cytokines. IL33 is also involved in the activation of mast cells, basophils, eosinophils, and natural killer cells. IL33 acts as a chemoattractant for Th2 cells and acts as an alarmin that enhances the immune response during tissue damage. IL33 is also known as NF-HEV, IL1F11, C9orf26, DVS27, nuclear factor from high endothelial venules, interleukin-1 family member 11, chromosome 9 open reading frame 26 (NF-HEV), or DKFZp586H0523.

[0375] Exemplary sequences of human IL33 mRNA transcripts can be found, for example, in GenBank Accession No. GI:1677537223 (NM_033439.4; SEQ ID NO:413; reverse complement, SEQ ID NO:414). The sequence of mouse IL33 mRNA can be found, for example, in GenBank Accession No. GI:1341395582 (NM_001164724.2; SEQ ID NO:415; reverse complement, SEQ ID NO:416). The sequence of rat IL33 mRNA can be found, for example, in GenBank Accession No. GI:62079056 (NM_001014166.1; SEQ ID NO:417; reverse complement, SEQ ID NO:418). The sequence of rhesus macaque IL33 mRNA can be found, for example, in GenBank Accession No. GI: ​​1622872669 (XM_015117709.2; SEQ ID NO: 419; reverse complement sequence, SEQ ID NO: 420).

[0376] Additional examples of IL33 mRNA sequences are readily available through publicly available databases such as GenBank, UniProt, OMIM, the UCSC Genome Browser, and the Macaque Genome Project website.

[0377] Additional information about IL33 can be found, for example, at www.ncbi.nlm.nih.gov / gene / ?term=IL33.

[0378] The entire contents of each of the aforementioned GenBank accession numbers and gene database numbers are incorporated herein by reference as of the filing date of the present application.

[0379] As used herein, the term IL33 also refers to variants of the IL33 gene, including variants provided in SNP databases. Many sequence variations within the IL33 gene have been identified and can be found, for example, in NCBI dbSNP and UniProt (see, for example, www.ncbi.nlm.nih.gov / gene / ?term=IL33, the entire contents of which are incorporated herein by reference as of the filing date of this application).

[0380] As used herein, "arachidonic acid 15-lipoxygenase" is used interchangeably with the term "ALOX15" to refer to members of the lipoxygenase family of proteins. ALOX15 acts on various polyunsaturated fatty acid substrates to produce various bioactive lipid mediators, such as eicosanoids, hepoxilins, lipoxins, and other molecules. The encoded enzyme and its reaction products have been shown to regulate inflammation and immunity. ALOX15 plays an important role in maintaining self-tolerance by peroxidase-bound phosphatidylethanolamine, which can then signal the sorting process to eliminate apoptotic cells during inflammation and prevent autoimmune reactions. In addition to its role in immune and inflammatory responses, ALOX15 may play a role in epithelial wound healing in the cornea by producing lipoxin A4 (LXA(4)) and docosahexaenoic acid-derived neuroprotectin D1, both of which exhibit anti-inflammatory and neuroprotective properties. Furthermore, ALOX15 regulates actin polymerization, which is crucial for several biological processes, such as phagocytosis of apoptotic cells. ALOX15 is also implicated in the production of endogenous ligands for peroxisome proliferator-activated receptor γ (PPAR-γ), thereby regulating macrophage development and function. ALOX15 may also negatively impact skeletal development by regulating bone mass through this pathway. Finally, ALOX15 is involved in the cellular response to IL-13. ALOX15 is also known as 15-LOX-1, polyunsaturated fatty acid lipoxygenase ALOX15, leukocyte-type arachidonic acid 12-lipoxygenase, arachidonic acid ω-6 lipoxygenase, hepatic arachidonic acid synthase Alox15, linoleic acid 13S-lipoxygenase, 12 / 15-lipoxygenase, 12-LOX, LOG15, 15-lipoxygenase type I, EC 1.13.11.31, EC 1.13.11.33, EC 1.13.11.12, EC 1.13.11, or EC 1.13.11.

[0381] Exemplary sequences of human ALOX15 mRNA transcripts can be found, for example, in GenBank Accession No. GI:1698254589 (NM_001140.5; SEQ ID NO:421; reverse complement, SEQ ID NO:422). The sequence of mouse ALOX15 mRNA can be found, for example, in GenBank Accession No. GI:134948632 (NM_009660.3; SEQ ID NO:423; reverse complement, SEQ ID NO:424). The sequence of rat ALOX15 mRNA can be found, for example, in GenBank Accession No. GI:31542124 (NM_031010.2; SEQ ID NO:425; reverse complement, SEQ ID NO:426). The sequence of rhesus macaque ALOX15 mRNA can be found, for example, in GenBank Accession No. GI: ​​1622876822 (XM_028835918.1; SEQ ID NO: 427; reverse complement sequence, SEQ ID NO: 428).

[0382] Additional examples of ALOX15 mRNA sequences are readily available through publicly available databases such as GenBank, UniProt, OMIM, the UCSC Genome Browser, and the Macaque Genome Project website.

[0383] Additional information about ALOX15 can be found, for example, at www.ncbi.nlm.nih.gov / gene / ?term=ALOX15.

[0384] The entire contents of each of the aforementioned GenBank accession numbers and gene database numbers are incorporated herein by reference as of the filing date of the present application.

[0385] As used herein, the term ALOX15 also refers to variants of the ALOX15 gene, including variants provided in SNP databases. Many sequence variations within the ALOX15 gene have been identified and can be found, for example, in NCBI dbSNP and UniProt (see, for example, www.ncbi.nlm.nih.gov / gene / ?term=ALOX15, the entire contents of which are incorporated herein by reference as of the filing date of this application).

[0386] As used herein, "receptor for advanced glycation end products," which is used interchangeably with the terms "AGER" and "RAGE," refers to a member of the immunoglobulin superfamily of cell-surface receptors. It is a multiligand receptor and, in addition to AGEs, interacts with other molecules involved in homeostasis, development, and inflammation, as well as certain diseases such as diabetes and Alzheimer's disease.

[0387] Exemplary sequences of human AGER mRNA transcripts can be found, for example, in GenBank Accession No. NM_001136.5 (SEQ ID NO: 429; reverse complement, SEQ ID NO: 430). The sequence of mouse AGER mRNA can be found, for example, in GenBank Accession No. NM_007425.3 (SEQ ID NO: 431; reverse complement, SEQ ID NO: 432). The sequence of rat AGER mRNA can be found, for example, in GenBank Accession No. NM_053336.2 (SEQ ID NO: 433; reverse complement, SEQ ID NO: 434). The sequence of rhesus macaque AGER mRNA can be found, for example, in GenBank Accession No. NM_001205117.1 (SEQ ID NO: 435; reverse complement, SEQ ID NO: 436).

[0388] Additional examples of AGER mRNA sequences are readily available through publicly available databases such as GenBank (e.g., NM_001136.5; NM_001206929.2; NM_001206932.2; NM_001206934.2; NM_001206936.; NM_001206940.2; NM_001206954.2; NM_001206966.2; NM_172197.3), UniProt, OMIM, the UCSC Genome Browser, and the Macaque Genome Project website.

[0389] Additional information about AGER can be found, for example, at www.ncbi.nlm.nih.gov / gene / ?term=AGER.

[0390] The entire contents of each of the aforementioned GenBank accession numbers and gene database numbers are incorporated herein by reference as of the filing date of the present application.

[0391] As used herein, the term AGER also refers to variants of the AGER gene, including variants provided in SNP databases. Many sequence variations within the AGER gene have been identified and can be found, for example, in NCBI dbSNP and UniProt (see, for example, www.ncbi.nlm.nih.gov / snp / ?term=AGER, the entire contents of which are incorporated herein by reference as of the filing date of this application).

[0392] As used herein, "oligomeric mucus / gel-forming mucin 5AC" is used interchangeably with the terms "MUC5AC," "MUC5," and "TBM," and refers to a member of the extracellular matrix structural components involved in phosphatidylcyclohexane hexol-mediated signaling. Located in the cytoplasm; the extracellular space; and the mucus layer. A biomarker for several diseases, including Sjögren's syndrome; biliary tract disease (multiple); cystic fibrosis; eye disease (multiple); and pancreatic cancer (multiple). MUC5AC is known to contribute to severe muco-obstructive lung disease, worsening the pathogenesis of chronic obstructive pulmonary disease (COPD). Genome-wide association studies have highlighted the pathogenic role of increased MUC5AC expression in the pathogenesis of moderate and severe asthma. Overexpression of respiratory mucins, such as MUC5AC, has been described in idiopathic pulmonary fibrosis (IPF) lungs.

[0393] Exemplary sequences of human MUC5AC mRNA transcripts can be found, for example, in GenBank Accession No. NM_001304359.2 (SEQ ID NO: 437; reverse complement, SEQ ID NO: 438). The sequence of mouse MUC5AC mRNA can be found, for example, in GenBank Accession No. NM_010844.3 (SEQ ID NO: 439 reverse complement, SEQ ID NO: 440). The sequence of rat MUC5AC mRNA can be found, for example, in GenBank Accession No. NM_001419868 (SEQ ID NO: 441; reverse complement, SEQ ID NO: 442). The sequence of rhesus macaque MUC5AC mRNA can be found, for example, in GenBank Accession No. XM_028832999.1 (SEQ ID NO: 443; reverse complement, SEQ ID NO: 444).

[0394] Additional examples of MUC5AC mRNA sequences are readily available through publicly available databases such as GenBank, UniProt, OMIM, the UCSC Genome Browser, and the Macaque Genome Project website.

[0395] Additional information about MUC5AC can be found, for example, at www.ncbi.nlm.nih.gov / gene / ?term=MUC5AC.

[0396] The entire contents of each of the aforementioned GenBank accession numbers and gene database numbers are incorporated herein by reference as of the filing date of the present application.

[0397] As used herein, the term MUC5AC also refers to variants of the MUC5AC gene, including variants provided in SNP databases. Many sequence variations within the MUC5AC gene have been identified and can be found, for example, in NCBI dbSNP and UniProt (see, for example, www.ncbi.nlm.nih.gov / snp / ?term=MUC5AC, the entire contents of which are incorporated herein by reference as of the filing date of this application).

[0398] As used herein, "signal transducer and activator of transcription 6" is used interchangeably with the term "STAT6" to refer to members of the STAT transcription factor family. In response to cytokines and growth factors, STAT family members are phosphorylated by receptor-associated kinases and subsequently form homodimers or heterodimers that translocate to the nucleus, where they act as transcriptional activators. STAT6 has been shown to regulate many pathological features of lung inflammatory responses in animal models, including respiratory eosinophilia, epithelial mucus production, smooth muscle changes, Th2 cell differentiation, and IgE production by B cells (Wurster AL et al., Oncogene 2000; 19: 2577-84). STAT6 is also known as interleukin-4 inducer, IL-4-STAT, D12S1644, STAT6B, or STAT6C.

[0399] The sequence of human STAT6 mRNA transcripts can be found, for example, in GenBank Accession No. GI: ​​1519313969 (NM_003153.5; SEQ ID NO: 445; reverse complement, SEQ ID NO: 446). The sequence of mouse STAT6 mRNA can be found, for example, in GenBank Accession No. GI: ​​128485773 (NM_009284.2; SEQ ID NO: 447; reverse complement, SEQ ID NO: 448). The sequence of rat STAT6 mRNA can be found, for example, in GenBank Accession No. GI: ​​113205499 (NM_001044250.1; SEQ ID NO: 449; reverse complement, SEQ ID NO: 450). The sequence of cynomolgus macaque STAT6 mRNA can be found, for example, in GenBank Accession No. GI:982282006 (XM_005571286.2; SEQ ID NO:451; reverse complement sequence, SEQ ID NO:452). The sequence of rhesus macaque STAT6 mRNA can be found, for example, in GenBank Accession No. GI:1622842915 (XM_015152044.2; SEQ ID NO:453; reverse complement sequence, SEQ ID NO:454).

[0400] Additional examples of STAT6 mRNA sequences are readily available through publicly available databases such as GenBank, UniProt, OMIM, and the Macaque Genome Project website. Additional information about STAT6 can be found, for example, at www.ncbi.nlm.nih.gov / gene / ?term=STAT6.

[0401] The entire contents of each of the aforementioned GenBank accession numbers and gene database numbers are incorporated herein by reference as of the filing date of the present application.

[0402] As used herein, the term STAT6 also refers to variations of the STAT6 gene, including variants provided in SNP databases. Many sequence variations within the STAT6 gene have been identified and can be found, for example, in NCBI dbSNP and UniProt (see, for example, www.ncbi.nlm.nih.gov / snp / ?term=STAT6, the entire contents of which are incorporated herein by reference as of the filing date of this application.

[0403] In some embodiments, the double-stranded region of the double-stranded iRNA agent is equal to or at least 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 23, 24, 25, 26, 27, 28, 29, 30 or more nucleotide pairs in length.

[0404] In some embodiments, the antisense strand of a double-stranded iRNA agent is equal to or at least 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides in length.

[0405] In some embodiments, the sense strand of the double-stranded iRNA agent is equal to or at least 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides in length.

[0406] In one embodiment, the sense and antisense strands of the double-stranded iRNA agent are each independently 15 to 30 nucleotides in length.

[0407] In one embodiment, the sense and antisense strands of the double-stranded iRNA agent are each independently 19 to 25 nucleotides in length.

[0408] In one embodiment, the sense and antisense strands of the double-stranded iRNA agent are each independently 21 to 23 nucleotides in length.

[0409] In one embodiment, the sense strand of the iRNA agent is 21 nucleotides in length and the antisense strand is 23 nucleotides in length, wherein the strands form a double-stranded region of 21 consecutive base pairs and have a 2 nucleotide long single-stranded overhang at the 3' end.

[0410] In one aspect of the invention, the agent used in the methods and compositions of the present invention is a single-stranded antisense nucleic acid molecule that inhibits a target mRNA via an antisense inhibition mechanism. The single-stranded antisense RNA molecule is complementary to a sequence within the target mRNA. Single-stranded antisense oligonucleotides can inhibit translation stoichiometrically by base-pairing with the mRNA and physically hindering the translation machinery, see Dias, N. et al., (2002) Mol Cancer Ther 1: 347-355. The single-stranded antisense RNA molecule can be about 15 to about 30 nucleotides in length and have a sequence complementary to the target sequence. For example, the single-stranded antisense RNA molecule can comprise a sequence of at least about 15, 16, 17, 18, 19, 20 or more consecutive nucleotides from any one of the antisense sequences described herein.

[0411] In one embodiment, at least partial inhibition of target gene expression is assessed by a decrease in the amount of target mRNA that can be isolated or detected in the first cell or cell group in which the target gene is transcribed or has been treated to inhibit target gene expression, compared to a second cell or cell group that is substantially identical to the first cell or cell group or has not been subjected to the treatment (control cells). The degree of inhibition can be represented by the following formula:

[0412]

[0413] In one embodiment, inhibition of expression is determined by a dual luciferase method, wherein the RNAi agent is present at 10 nM.

[0414] As used herein, the phrase "contacting a cell with an RNAi agent," such as dsRNA, includes contacting the cell by any possible means. Contacting the cell with the RNAi agent includes contacting the cell with the RNAi agent in vitro or contacting the cell with the RNAi agent in vivo. Contacting can be performed directly or indirectly. Thus, for example, the RNAi agent can be physically contacted with the cell by the individual performing the method, or the RNAi agent can be placed in a situation that allows or enables subsequent contact with the cell.

[0415] Contacting cells in vitro can be performed, for example, by incubating the cells with an RNAi agent. Contacting cells in vivo can be performed, for example, by injecting the RNAi agent into or near the tissue where the cells are located, or by injecting the RNAi agent into another area, or into the bloodstream or subcutaneous space, so that the agent will then reach the tissue where the cells are located. In some embodiments, the RNAi agent may contain or be coupled to a ligand, for example, one or more α-v-β-6 (ɑvβ6) integrin targeting ligands.

[0416] As used herein, "αvβ6 (αvβ6) integrin targeting ligands" include any moiety (e.g., peptides and small molecules) that binds to αvβ6 integrin and is capable of mediating delivery of its linked dsRNA agent to skeletal muscle (e.g., skeletal muscle cells or skeletal muscle tissue) and / or cardiac muscle (e.g., cardiac muscle cells or cardiac muscle tissue). αvβ6 integrin targeting ligands bind to αvβ6 integrin or αvβ6 integrin receptors on skeletal muscle / cardiac muscle cells (cells). Exemplary αvβ6 integrin targeting ligands are described in Section II below.

[0417] In one embodiment, contacting a cell with an RNAi agent includes "introducing" or "delivering" the RNAi agent into the cell by promoting or enabling uptake or absorption into the cell. Uptake or absorption of the RNAi agent can occur by unassisted diffusion or active cellular processes, or by an adjuvant or device. The RNAi agent can be introduced into the cell in vitro or in vivo. For example, for in vivo introduction, the RNAi agent can be injected into a tissue site or administered systemically. In vitro introduction into cells includes methods known in the art, such as electroporation and lipofection. Other methods are described below or are known in the art.

[0418] As used herein, "subject" is an animal that expresses a target gene endogenously or heterologously, such as a mammal, including primates (such as humans, non-human primates, e.g., monkeys and chimpanzees) or non-primates (such as cattle, pigs, horses, goats, rabbits, sheep, hamsters, guinea pigs, cats, dogs, rats or mice) or birds. In one embodiment, as described herein, the subject is a human, such as a human being who is being treated or evaluated for a disease, disorder or condition that will benefit from reduced target gene expression; a human being at risk of a disease, disorder or condition that will benefit from reduced target gene expression; a human being suffering from a disorder or condition that will benefit from reduced target gene expression; or a human being being treated for a disease, disorder or condition that will benefit from reduced target gene expression. In some embodiments, the subject is a female human. In other embodiments, the subject is a male human. In one embodiment, the subject is an adult subject. In another embodiment, the subject is a pediatric subject.

[0419] As used herein, the term "treating" or "treatment" refers to beneficial or desired results, including but not limited to alleviating or improving one or more signs or symptoms associated with target gene expression or target gene protein production, such as target gene-related diseases, such as muscle disorders, such as skeletal muscle disorders and / or cardiac muscle disorders, or symptoms associated with undesirable target gene expression; reducing the extent of undesirable target activation or stabilization; alleviating or alleviating undesirable target activation or stabilization. "Treatment" can also mean prolonging survival compared to expected survival in the absence of treatment.

[0420] In the case of target gene or disease marker or symptom levels in an individual, the term "reduce" refers to a statistically significant reduction in such levels. The reduction can be, for example, at least 10%, 15%, 20%, 25%, 30%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or more. In certain embodiments, the reduction is at least 20%. In certain embodiments, disease markers, such as protein or gene expression, are reduced by at least 50%. In the case of target gene levels in an individual, "reduction" refers to a reduction to a level within the normal range for individuals generally recognized as having no such conditions. In certain embodiments, the expression of the target is normalized, that is, a reduction towards or towards a level within the normal range for individuals generally recognized as having no such conditions, such as blood glucose levels, blood uric acid levels, blood lipid levels, blood oxygen levels, white blood cell counts, renal function, spleen function, liver function. For example, chronic hyperuricemia is defined as a serum urate level greater than 6.8 mg / dl (greater than 360 mmol / ), above which the level exceeds the physiological saturation threshold (Mandell, Cleve. Clin. Med. 75: S5-S8, 2008). As used herein, "reduction" in an individual may refer to a reduction in gene expression or protein production in cells in the individual, without necessarily reducing expression in all cells or tissues of the individual. For example, as used herein, reduction in an individual may include reducing gene expression or protein production in the individual.

[0421] The term "reduce" can also be used in conjunction with normalizing the symptoms of a disease or condition, i.e., reducing the difference between the levels of individuals suffering from the target gene-associated disease and the levels of normal individuals who do not suffer from the target gene-associated disease. As used herein, if the disease is associated with elevated symptom values, "normal" should be considered the upper limit of normal values. If the disease is associated with decreased symptom values, "normal" should be considered the lower limit of normal values.

[0422] As used herein, "prevention" or "preventing" when used with respect to a disease, disorder, or condition thereof that would benefit from reduced expression of a target gene or reduced production of a target protein refers to a reduction in the likelihood that an individual will develop symptoms associated with such a disease, disorder, or condition, such as symptoms of a target gene-associated disease. Failure to develop a disease, disorder, or condition, or a reduction in the development of symptoms associated with such a disease, disorder, or condition (e.g., at least about 10% on a clinically recognized scale for the disease or condition), or a delay in the onset of symptoms (e.g., days, weeks, months, or years) is considered effective prevention.

[0423] As used herein, the term "target gene-associated disease" is a disease or condition that would benefit from reduced expression or activity of a target gene. The term "target gene-associated disease" is a disease or condition caused by or associated with the expression or protein production of a target gene. The term "target gene-associated disease" includes diseases, disorders, or conditions that would benefit from reduced expression or protein activity of a target gene. Additional information about specific target genes and diseases that would benefit from reduced target gene expression is described below.

[0424] In one embodiment, the target gene-associated disease is a muscle disorder.

[0425] Exemplary muscle disorders include myostatin-associated muscle hypertrophy, congenital myasthenic syndrome, facioscapulohumeral muscular dystrophy (FSHD), spinal muscular atrophy (SMA), myotonic dystrophy type 1 (DM1), Pompe disease, PLN cardiomyopathy, spasticity, obstructive hypertrophic cardiomyopathy (HOCM); familial hypertrophic cardiomyopathy (FHC); heart failure with preserved ejection fraction (HFPEF); atrial fibrillation (AFIB); ventricular fibrillation (VFIB); angina; myocardial infarction (MI); heart failure or heart failure with reduced ejection fraction (HFREF); supraventricular tachycardia (SVT); hypertrophic cardiomyopathy (HCM); and PLN cardiomyopathy.

[0426] In one embodiment, the target gene-associated disease is a skeletal muscle disease or disorder.

[0427] In one embodiment, the target gene-associated disease is a myocardial disease or disorder.

[0428] Exemplary myocardial disorders include obstructive hypertrophic cardiomyopathy (HOCM); familial hypertrophic cardiomyopathy (FHC); heart failure with preserved ejection fraction (HFPEF); atrial fibrillation (AFIB); ventricular fibrillation (VFIB); angina; myocardial infarction (MI); heart failure or heart failure with reduced ejection fraction (HFREF); supraventricular tachycardia (SVT); hypertrophic cardiomyopathy (HCM); and PLN cardiomyopathy.

[0429] Heart failure ("HF") or "congestive heart failure" ("CHF") is a chronic condition in which the heart cannot pump blood properly. Heart failure occurs when the heart's ability to pump blood is insufficient to keep up with the body's needs. Heart failure can occur if the heart cannot pump (systole) or fill (relax) adequately. As the heart weakens, blood begins to back up and forces fluid through the capillary walls. The term "congestive" refers to the resulting accumulation of fluid in the ankles and feet, arms, lungs, and / or other organs.

[0430] One type of heart failure is "heart failure with preserved left ventricular function" ("HF-pEF"), also known as "heart failure with preserved ejection fraction" ("HF-pEF"), a condition in which the heart contracts and pumps normally, but the ventricles are thicker and stiffer than normal. As a result, the ventricles cannot relax properly and cannot fill completely. With less blood in the ventricles, less blood is pumped to the rest of the body when the heart contracts.

[0431] The most common cause of congestive heart failure is coronary artery disease. Risk factors for coronary artery disease include high cholesterol and / or triglyceride levels, high blood pressure, a poor diet, a sedentary lifestyle, diabetes, smoking, being overweight or obese, and stress. In addition to coronary artery disease, several other conditions can damage the heart muscle, including hereditary and genetic factors, some infections and autoimmune diseases, and some treatments, such as chemotherapy.

[0432] CHF symptoms include shortness of breath, tiredness, leg swelling, and rapid heartbeat.

[0433] Treatment may include eating less salt, restricting fluid intake, and taking prescription medications such as vasodilators, diuretics, aldosterone inhibitors, ACE inhibitors or ARB drugs, digitalis glycosides, anticoagulants or antiplatelet agents, beta-blockers, and sedatives; and surgical procedures including, for example, bypass surgery, heart valve replacement; pacemaker implantation, such as biventricular pacing therapy or an implantable cardioverter-defibrillator; ventricular assist device (VAD therapy) and heart transplantation.

[0434] "Hypertrophic cardiomyopathy" ("HCM") refers to impaired heart function associated with abnormally thick heart muscle in the absence of other heart disease, such as heart valve disease. "Hypertrophic obstructive cardiomyopathy" ("HOCM") is a subtype of HCM in which the wall (septum) between the two bottom chambers of the heart thickens. The walls of the pumping chamber may also become stiff. The thickening of the septum may lead to narrowing, which can block or reduce blood flow from the left ventricle to the aorta, a condition known as "outflow tract obstruction."

[0435] Both HCM and HOCM can be caused by mutations in cardiac myocardial genes and can be inherited. Therefore, multiple family members can be affected by HCM and HOCM. The phenotypic expression of these gene mutations can be variable.

[0436] Both HCM and HOCM can be caused by mutations in cardiac genes and can be inherited. Consequently, multiple family members can be affected by HCM and HOCM. The phenotypic expression of these mutations can be variable. In other words, even with the same mutation, the severity of cardiac impairment can vary between affected patients.

[0437] Symptoms associated with HCM can also vary in severity and characteristics and can include fatigue, chest pain, difficulty breathing, abnormal heart rhythms, heart failure, fainting, and sudden cardiac death.

[0438] Treatment includes pacemakers, defibrillators, alcohol septal ablation, surgical myectomy, advanced heart failure therapy, beta-blockers, calcium channel blockers, and antiarrhythmic agents.

[0439] "Familial hypertrophic cardiomyopathy" is an autosomal dominant disorder characterized primarily by left ventricular enlargement. The thickening typically occurs in the septum between the ventricles. In some cases, the thickening of the septum between the ventricles blocks the flow of oxygen-rich blood from the heart, which can lead to abnormal heart sounds (heart murmurs) during the heartbeat and other signs and symptoms of the condition. Other affected individuals do not have a physical obstruction to blood flow, but instead have less efficient blood pumping, which can also cause symptoms of the condition. Cardiac enlargement typically begins during adolescence or young adulthood, although it can develop at any time in life.

[0440] Symptoms of familial hypertrophic cardiomyopathy vary, even within the same family. Many affected individuals have no symptoms. Others with familial hypertrophic cardiomyopathy may experience chest pain; shortness of breath, especially during exertion; a fluttering or pounding sensation in the chest (palpitations); lightheadedness; dizziness; and fainting.

[0441] Although most people with familial hypertrophic cardiomyopathy have no symptoms or only mild symptoms, the condition can have serious consequences. It can cause abnormal heart rhythms (arrhythmias) that can be life-threatening. People with familial hypertrophic cardiomyopathy have an increased risk of sudden death, even if they have no other symptoms. A small number of affected individuals develop potentially fatal heart failure and may require a heart transplant.

[0442] Mutations in one of several genes can cause familial hypertrophic cardiomyopathy; the genes most commonly involved are MYH7, MYBPC3, TNNT2, and TNNI3. Other genes, including some that have not yet been identified, may also be involved in the condition.

[0443] Treatment includes beta blockers, calcium channel blockers; heart rhythm medications such as amiodarone (Pacerone) or disopyramide (Norpace); and blood viscosity reducers such as warfarin (Coumadin, Jantoven), dabigatran (Pradaxa), rivaroxaban (Xarelto), or apixaban (Eliquis). Surgery or other procedures include apical myectomy, diaphragmatic myectomy, diaphragmatic ablation, and implantable cardioverter-defibrillator (ICD).

[0444] Atrial fibrillation (AFIB) is a disorderly and irregular beating of the atria that is not coordinated with the ventricles. The result is a rapid and irregular heart rhythm. The heart rate in atrial fibrillation can range from 100 to 175 beats per minute. The normal range of heart rate is 60 to 100 beats per minute.

[0445] Atrial fibrillation episodes may come and go, or may disappear, and may require treatment. Although atrial fibrillation itself is not usually life-threatening, it is a serious medical condition that sometimes requires emergency treatment.

[0446] The main problem with atrial fibrillation is the possibility of a blood clot developing in the atria, which can circulate to other organs and cause blood flow blockage (ischemia).

[0447] Causes of AFIB include structural abnormalities or damage to the heart, high blood pressure, heart attack, coronary artery disease, heart valve abnormalities, congenital heart defects, an overactive thyroid or other metabolic imbalances, exposure to stimulants (such as medications, caffeine, tobacco, or alcohol), atrial fibrillation syndrome (in which the heart's natural pacemaker does not function properly), lung disease, previous heart surgery, viral infection, stress from surgery, pneumonia or other medical conditions, and sleep apnea.

[0448] Symptoms include palpitations, which are fast, uncomfortable, irregular heartbeats or a churning feeling in the chest; weakness, decreased ability to exercise, fatigue, lightheadedness, dizziness, shortness of breath, and chest pain.

[0449] Treatment includes cardioversion, antiarrhythmic agents, digoxin, beta-blockers, calcium channel blocker anticoagulants, catheter ablation, the Maze procedure, atrioventricular (AV) node ablation, and left atrial appendage closure.

[0450] "Ventricular fibrillation" ("VFIB") is a type of abnormal heart rhythm (arrhythmia). During VFIB, distorted heart signals cause the ventricles to pump uselessly (quiver). As a result, the heart does not pump blood to the rest of the body.

[0451] Ventricular fibrillation is an emergency that requires immediate medical attention. It is the most common cause of sudden cardiac death.

[0452] Fainting and loss of consciousness are the most common symptoms of ventricular fibrillation. Other symptoms include chest pain, extremely fast heartbeat (tachycardia), dizziness, nausea, and shortness of breath.

[0453] Risk factors include a previous episode of ventricular fibrillation, a previous heart attack, congenital heart defects, heart muscle disease (cardiomyopathy), injury that damages the heart muscle (such as lightning strike, drug abuse, especially cocaine or methamphetamine), and a severe imbalance of potassium or magnesium.

[0454] Treatment includes cardiopulmonary resuscitation (CPR), defibrillation, antiarrhythmic drugs, implantable cardioverter-defibrillator (ICD), cardiac ablation, coronary angioplasty and stent placement, and coronary bypass surgery.

[0455] A "myocardial infarction," or "MI," occurs when blood flow to the heart is blocked. The blockage is most often a buildup of fat, cholesterol, and other substances that form plaque in the arteries that feed the heart (coronary arteries).

[0456] Symptoms include pressure, tightness, pain, or squeezing or aching sensation in the chest or arms, which may spread to the neck, jaw, or back; nausea, indigestion, heartburn or abdominal pain, shortness of breath, cold sweats, fatigue, lightheadedness, or sudden dizziness.

[0457] Heart attack risk factors include age (for example, men aged 45 or older and women aged 55 or older are more likely to have a heart attack than younger men and women), tobacco use, and high blood pressure. Over time, high blood pressure can damage the arteries that reach the heart. High blood pressure that occurs with other conditions such as obesity, high cholesterol, or diabetes increases the risk even more; high cholesterol or triglyceride levels, obesity, diabetes, metabolic syndrome, a family history of heart attacks, physical inactivity, stress, drug use, a history of preeclampsia, and autoimmune conditions.

[0458] Treatment includes aspirin, thrombolytics, antiplatelet agents, other blood thinning drugs, pain relievers, nitroglycerin, beta-blockers, ACE inhibitors, statins, coronary angioplasty and stenting, and coronary artery bypass graft surgery.

[0459] "Supraventricular tachycardia" ("SVT") is an abnormally fast or erratic heartbeat that affects the atria. During an episode of SVT, the heart beats about 150 to 220 times per minute, but it may occasionally beat faster or slower.

[0460] The main symptom of supraventricular tachycardia (SVT) is a very fast heartbeat (100 beats per minute or greater) that can last from a few minutes to a few days. The fast heartbeats can come and go suddenly, with periods of normal heart rate in between.

[0461] Signs and symptoms of supraventricular tachycardia can include very fast (rapid) heartbeats, fluttering or pounding in the chest (palpitations), a pounding sensation in the neck, weakness or feeling very tired (fatigue), chest pain, shortness of breath, lightheadedness or dizziness, sweating, and fainting (syncope) or near-fainting. Some people with SVT have no signs or symptoms at all.

[0462] For some patients, episodes of supraventricular tachycardia are associated with obvious triggers, such as exercise, stress, or lack of sleep. Some people may have no identifiable triggers. Factors that can cause an SVT episode include age, coronary artery disease, previous heart surgery, heart disease, heart failure, other heart problems (such as Wolff-Parkinson-White syndrome), chronic lung disease, consuming too much caffeine, drinking too much alcohol, taking drugs (especially stimulants, such as cocaine and methamphetamine), pregnancy, smoking, thyroid disease, tobacco, sleep apnea, diabetes, and certain medications (including asthma medications and over-the-counter cold and allergy medicines).

[0463] Treatment includes carotid sinus massage, vagus nerve stimulation, cardioversion, beta-blockers, antiarrhythmic drugs, calcium channel blockers, catheter ablation, and pacemakers.

[0464] Hypertrophic cardiomyopathy (HCM) is a condition in which the heart muscle becomes abnormally thickened (enlarged). The thickened heart muscle can make it harder for the heart to pump blood.

[0465] Angina is a type of chest pain caused by reduced blood flow to the heart. Angina is a symptom of coronary artery disease.

[0466] Colic, also known as angina, is often described as squeezing, pressure, heaviness, tightness, or pain in the chest. Some people with colic symptoms say it feels like a vice squeezing their chest or a heavy object pressing against it. Pain may also occur in the arms, neck, jaw, shoulders, or back. Other symptoms that may accompany colic include dizziness, fatigue, nausea, shortness of breath, and sweating.

[0467] Risk factors include tobacco, diabetes, high blood pressure, high cholesterol or triglyceride levels, family history of heart disease, age (for example, men over 45 and women over 55 have a greater risk than younger adults), physical inactivity, obesity, and stress.

[0468] Treatment includes lifestyle changes, nitrates, aspirin, anti-clotting drugs, beta-blockers, statins, calcium channel blockers, blood pressure medications, angiotensin-converting enzyme (ACE) inhibitors or angiotensin II receptor blockers (ARBs), ranolazine (Ranexa), angioplasty and stenting, coronary artery bypass grafting, and external counterpulsation (ECP).

[0469] As used herein, phospholamban (PLN) cardiomyopathy refers to a specific subtype of inherited cardiomyopathy caused by pathogenic variants in PLN p.(Arg14del), a gene encoding PLN, a protein that plays a major role in calcium homeostasis in cardiac tissue. This protein ensures proper contraction and relaxation of the human heart. Carriers of this pathogenic variant are at high risk for developing dilated cardiomyopathy (DCM), arrhythmic cardiomyopathy (ACM), or both. They typically have a high burden of arrhythmias and a positive family history of premature ventricular contractions (PVCs) and ventricular tachycardia, a significantly low-voltage electrocardiogram (ECG), left ventricular dysfunction, and sudden cardiac death. PLN p.(Arg14del) cardiomyopathy has been reported in several European countries, as well as the United States, Canada, and China. Globally, it is a rare disease, but it is particularly common in the Netherlands, where the pathogenic variant is present in 12% of all ACM patients and 15% of all DCM patients.

[0470] Exemplary skeletal muscle disorders include myostatin-associated muscle hypertrophy, congenital myasthenic syndrome, facioscapulohumeral muscular dystrophy (FSHD), spinal muscular atrophy (SMA), myotonic dystrophy type 1 (DM1), Pompe disease, PLN cardiomyopathy, and spasticity.

[0471] Myostatin-associated muscle hypertrophy is a rare condition characterized by decreased body fat and increased muscle size. Affected individuals have up to twice the usual amount of muscle mass in their bodies. They also tend to have increased muscle strength. Myostatin-associated muscle hypertrophy is caused by mutations in the MSTN gene. It follows an incomplete autosomal dominant inheritance pattern.

[0472] Congenital myasthenic syndrome (CMS) is a group of heterogeneous early-onset gene neuromuscular transmission disorders (endplate myopathy) due to proteins involved in the organization, maintenance, function or regulation of motor nerve end plates, such as CHRNA1, CHRNB1, CHRBD, CHRNE, CHRNG, COL13A1, DOX7, LRP4, MUSK, RAPSN or SCN4A mutations. CMS is clinically characterized by abnormal fatigue, or temporary or permanent weakness of the muscles of the eyes, face, medulla oblongata, trunk, respiratory system or limbs. Endplate myopathy attacks are in utero, congenital, in infancy or childhood and rarely in adolescence. The severity ranges from mild staged weakness to disability, permanent muscle weakness, respiratory insufficiency and premature death. All subtypes of CMS have the clinical features of fatigue and muscle weakness, but the age of onset, symptoms and response to treatment vary according to the molecular mechanism produced by the basic gene defect. Since not all CMS are congenital, the term CMS is misleading. See the review by Finsterer (2019) Orphanet J Rare Dis. 14: 57.

[0473] Facioscapulohumeral muscular dystrophy (FSHD) type 1 is an autosomal dominant condition caused by mutations in DUX4. FSHD typically presents before age 20 with weakness of the facial muscles and the scapular stabilizers or foot dorsiflexors. There is extreme clinical variability. In some cases, congenital facial weakness may be present. In FSHD, muscle weakness is slowly progressive, and approximately 20% of affected individuals eventually require a wheelchair. Life expectancy is not shortened. The incidence rate is approximately 4 individuals per 100,000 population.

[0474] As used herein, spinal muscular atrophy refers to a genetic disorder characterized by weakness and atrophy (atrophy) in the muscles used for action (skeletal muscle). It is caused by the loss of specialized nerve cells, called motor neurons, that control muscle action. Compared to muscles far away from the center of the body (remote), weakness tends to be more severe in muscles close to the center of the body (proximal). Muscle weakness usually worsens with age. There are many types of spinal muscular atrophy, which are caused by changes in the same gene. These types differ in the age of onset and severity of muscle weakness; however, there is overlap between these types. Other forms of spinal muscular atrophy and related motor neuron diseases, such as spinal muscular atrophy with progressive myoclonic spastic epilepsy, spinal muscular atrophy dominated by the lower limbs, X-linked infantile spinal muscular atrophy, and spinal muscular atrophy with type 1 respiratory distress are caused by mutations in other genes.

[0475] Mutations in the SMN1 gene cause all of the types of spinal muscular atrophy described above. The number of copies of the SMN2 gene regulates the severity of the condition and helps determine which type develops. Both the SMN1 and SMN2 genes provide instructions for making a protein called survival motor neuron (SMN) protein. Normally, most functional SMN protein is produced by the SMN1 gene, with a smaller amount produced by the SMN2 gene. Several different forms of the SMN protein are produced by the SMN2 gene, but only one form is functional; the others are smaller and break down quickly. The SMN protein is one of a group of proteins called the SMN complex, which is important for maintaining motor neurons. Motor neurons transmit signals from the brain and spinal cord that tell skeletal muscles to tense (contract), which causes the body to move.

[0476] As used herein, myotonic dystrophy refers to part of a group of genetic disorders known as muscular dystrophy. It is the most common form of muscular dystrophy that begins in adulthood. Myotonic dystrophy is characterized by progressive muscle atrophy and weakness. People with this condition typically experience prolonged muscle contractions (myotonia) and are unable to relax certain muscles after use. Other signs and symptoms of myotonic dystrophy include clouding of the lens of the eye (cataracts) and abnormalities in the electrical signals that control the heartbeat (cardiac conduction defects). Some affected individuals develop a condition known as diabetes, in which blood sugar levels can become dangerously high. Although the characteristics of myotonic dystrophy typically develop in people during their twenties or thirties, it can appear at any age. The severity of the symptoms varies widely between affected groups, even between members of the same family. There are two main types of myotonic dystrophy: type 1 and type 2. Their signs and symptoms overlap, but type 2 tends to be milder than type 1. The muscle weakness associated with type 1 particularly affects muscles farthest from the center of the body (remote muscles), such as those in the lower legs, hands, neck, and face. The muscle weakness in type 2 primarily involves muscles closer to the center of the body (proximal muscles), such as those in the neck, shoulders, elbows, and hips. The two types of myotonic dystrophy are caused by mutations in different genes.

[0477] Type 1 myotonic dystrophy is caused by mutations in the DMPK gene, while type 2 results from mutations in the CNBP gene. The protein produced by the DMPK gene may play a role in intracellular communication. It appears to be important for the proper function of cells in the heart, brain, and skeletal muscle (which are used for movement). The protein produced by the CNBP gene is primarily found in the heart and skeletal muscle, where it helps regulate the function of other genes. Similar changes in the structure of the DMPK and CNBP genes lead to type 1 and type 2 myotonic dystrophy. In each case, a segment of DNA is abnormally repeated multiple times, forming unstable regions in the gene. Genes with abnormal segments produce abnormally long messenger RNA, the molecular blueprint for genes that guide protein production. Abnormally long messenger RNA forms clumps inside cells, which interfere with the production of many other proteins. These changes prevent cells in muscle cells and other tissues from functioning normally, leading to the signs and symptoms of myotonic dystrophy. If these changes affect the DMPK gene, the result is type 1 myotonic dystrophy, and if the CNBP gene is affected, the result is type 2 myotonic dystrophy.

[0478] As used herein, Pompe disease refers to a genetic disorder caused by the accumulation of a complex sugar called glycogen in the body's cells. Glycogen accumulation in certain organs and tissues, especially muscles, impairs their ability to function normally. There are three types of Pompe disease, with different severity and age of presentation. These types are called typical infantile-onset, atypical infantile-onset, and delayed-onset. The typical form of infantile-onset Pompe disease begins within a few months of birth. Infants with this disease typically experience muscle weakness (myopathy), poor muscle tone (hypotonia), enlarged liver (hepatomegaly), and heart defects. Affected infants may also be unable to gain weight and grow at the expected rate (stunted growth) and have breathing problems. If left untreated, this form of Pompe disease leads to death from heart failure in the first year of life. The atypical form of infantile-onset Pompe disease typically appears at 1 year old. It is characterized by slowed motor skills (such as turning over and sitting) and progressive muscle weakness. The heart may be abnormally large (cardiomegaly), but affected individuals typically do not experience heart failure. The muscle weakness of this disease causes severe breathing problems, and most children with atypical infantile-onset Pompe disease survive only into early childhood. Late-onset Pompe disease may not become apparent until late childhood, adolescence, or adulthood. Late-onset Pompe disease is usually milder than the infantile-onset form of the disease and is less likely to involve the heart. Most individuals with late-onset Pompe disease experience progressive muscle weakness, especially in the legs and trunk, including the muscles that control breathing. As the disease progresses, breathing problems can lead to respiratory failure.

[0479] Mutations in the GAA gene cause Pompe disease. The GAA gene provides instructions for producing an enzyme called acid α-glucosidase (also known as acid maltase). This enzyme is active in lysosomes, structures that serve as recycling centers within cells. The enzyme normally breaks down glycogen into simpler sugars, namely glucose, which is the main energy source for most cells. Mutations in the GAA gene effectively prevent acid α-glucosidase from breaking down glycogen, which causes this sugar to accumulate to toxic levels in the lysosomes. This accumulation damages organs and tissues throughout the body, especially muscles, leading to the progressive signs and symptoms of Pompe disease.

[0480] As used herein, spasticity refers to a condition in which muscles become stiff or tight, preventing normal fluid movement. Muscles remain contracted and resist stretching, thus affecting movement, speech, and gait. Spasticity is generally caused by damage or destruction to the areas of the brain and spinal cord responsible for controlling muscles and stretch reflexes. These damages can be attributed to an imbalance in inhibitory and excitatory signals sent to muscles, causing them to lock in place. Spasticity can be harmful to growing children because it can affect muscles and joints. People with brain damage, spinal cord injury, cerebral palsy, or multiple sclerosis may suffer from varying degrees of spasticity.

[0481] As used herein, "therapeutically effective amount" is intended to include an amount of an RNAi agent that, when administered to an individual suffering from a disease associated with a target gene, is sufficient to achieve treatment of the disease (e.g., by eliminating, ameliorating, or maintaining an existing disease or one or more disease symptoms). The "therapeutically effective amount" may vary depending on the RNAi agent, how the agent is administered, the disease and its severity, and the medical history, age, weight, family history, genetic makeup, type of previous or concomitant treatment (if any), and other individual characteristics of the individual to be treated.

[0482] As used herein, a "prophylactically effective amount" is intended to include an amount of an RNAi agent that, when administered to an individual suffering from a target gene-related disorder, such as gout or diabetes, is sufficient to prevent or ameliorate the disease or one or more symptoms of the disease. Amelioration of a disease includes slowing the course of the disease or reducing the severity of a disease that subsequently develops. A "prophylactically effective amount" may vary depending on the RNAi agent, how the agent is administered, the degree of disease risk, medical history, age, weight, family medical history, genetic makeup, type of previous or concomitant treatment, if any, and other personal characteristics of the patient to be treated.

[0483] "Therapeutically effective amount" or "prophylactically effective amount" also includes the amount of RNAi agent that produces some desired local or systemic effect at a reasonable benefit / risk ratio applicable to any treatment. The RNAi agent used in the method of the present invention can be administered in an amount sufficient to produce a reasonable benefit / risk ratio applicable to such treatment.

[0484] As used herein, the phrase "pharmaceutically acceptable" refers to those compounds, materials, compositions or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human subjects and animal subjects without excessive toxicity, irritation, allergic response, or other problems or complications, consistent with a reasonable benefit / risk ratio.

[0485] As used herein, the phrase "pharmaceutically acceptable carrier" refers to a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, manufacturing aid (e.g., lubricant, magnesium talc, calcium or zinc stearate or stearic acid), or solvent for an encapsulating material, which participates in carrying the compound of the invention from one organ or part of the body to another. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the subject being treated. Some examples of materials that can serve as pharmaceutically acceptable carriers include: (1) sugars such as lactose, glucose, and sucrose; (2) starches such as corn starch and potato starch; (3) cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) lubricants such as magnesium sulfate, sodium lauryl sulfate, and talc; (8) excipients such as cocoa butter and suppository waxes; (9) oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, and treacle. oil, corn oil, and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerol, sorbitol, mannitol, and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffers, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethanol; (20) pH buffered solutions; (21) polyesters, polycarbonates, or polyanhydrides; (22) bulking agents, such as polypeptides and amino acids; (23) serum components, such as serum albumin, HDL, and LDL; and (22) other nontoxic, compatible substances used in pharmaceutical formulations. Pharmaceutically acceptable carriers for pulmonary delivery are known in the art and will vary depending on the desired site of deposition of the agent, e.g., upper or lower respiratory system, and the type of device to be used for delivery, e.g., nebulizer, atomizer, dry powder inhaler.

[0486] "Pharmaceutically acceptable salts" of each of the RNAi agents include, but are not limited to, sodium salts, calcium salts, lithium salts, potassium salts, ammonium salts, magnesium salts, and mixtures thereof. It will be appreciated by those skilled in the art that when an RNAi agent is provided as a polycationic salt, it has one cation per free acid group of the optionally modified phosphodiester backbone and / or any other acidic modification (e.g., a 5' terminal phosphonate group). For example, an oligonucleotide of "n" nucleotides in length contains n-1 optionally modified phosphodiesters, such that an oligonucleotide of 21 nt in length can be provided in the form of a salt having up to 20 cations (e.g., 20 sodium cations). Similarly, an RNAi agent having a sense strand of 21 nt in length and an antisense strand of 23 nt in length can be provided in the form of a salt having up to 42 cations (e.g., 42 sodium cations). In the aforementioned examples, where the RNAi agent also includes a 5'-terminal phosphate or a 5'-terminal vinylphosphonate group, the RNAi agent can be provided as a salt having up to 44 cations (eg, 44 sodium cations).

[0487] As used herein, the term "sample" includes a group of similar fluids, cells, or tissues isolated from an individual, as well as fluids, cells, or tissues present within an individual. Examples of biological fluids include blood, serum and serosal fluid, plasma, bronchial fluid, sputum, cerebrospinal fluid, ocular fluid, lymph, urine, saliva, sputum, and the like. Tissue samples can include samples from tissues, organs, or localized areas. For example, a sample can be derived from a specific organ, a portion of an organ, or a fluid or cell within such an organ.

[0488] II. Modified α-v-β-6 (ɑvβ6) Integrin Compounds and Ligands

[0489] Integrins are cell surface receptors that activate signal transduction pathways upon ligand binding, including those involved in cytoskeletal organization and cell cycle regulation. Integrins are also involved in cell attachment to the extracellular matrix, and integrin ligands include common extracellular matrix components, including fibronectin, collagen, laminin, fibrinogen, thrombospondin, and glycoproteins such as tenascin C, osteopontin, and nefronectin. In humans, there are at least twenty-four known integrin heterodimers composed of α and β subunits, such as αvβ6. The combination of α and β subunits determines the ligand specificity and function of the integrin. Almost all cells express at least one integrin, and the expression and / or activity of integrins can be affected by other signal-inducing molecules, such as cytokines or steroids.

[0490] The primary function of αvβ6 is the activation of the cytokine transforming growth factor-β1 (TGF-β1). Latent TGF-β1 binds to the extracellular matrix, where it is covered by its propeptide, latency-associated peptide (LAP). αvβ6 binds to LAP and, through cytoskeletal forces, releases TGF-β1. TGF-β1 regulates multiple processes, including cell proliferation, differentiation, angiogenesis, epithelial-mesenchymal transition (EMT), and immunosuppression. These processes combine to heal wounds, but when uncontrolled, they can promote tissue pathology.

[0491] The present invention provides αvβ6 compounds and ligands comprising such αvβ6 compounds that can bind to, for example, dsRNA agents for efficient extrahepatic delivery of dsRNA agents.

[0492] A.ɑ-v-β-6 (αvβ6) integrin compounds

[0493] In one aspect, the present application provides modified integrin modulating compounds in a form suitable for binding to oligonucleotides directly or via a carrier group. In some embodiments, the present invention provides a compound of formula (IV),

[0494]

[0495] or a salt thereof, wherein:

[0496] Y is O, N(H), S or CH2; (e.g., O or CH2)

[0497] R 1 is hydrogen or C 1-6 Alkyl groups (e.g., methyl);

[0498] R Y for in

[0499] m is 0, 1, 2, 3 or 4; and

[0500] Each R 2 R is independently R, or two R on adjacent carbon atoms 2 The group, together with the atoms to which it is bonded, forms a fused 4- to 8-membered ring, which is optionally substituted with 1, 2, 3, or 4 groups independently selected from the group consisting of R and a nitrogen protecting group;

[0501] and

[0502] R L -N(R 3 )(R 4 )、-O(R 5 )、-S(R 5 ) or -R 5 ,in

[0503] R 3 and R 4 Is any of the following:

[0504] (i)R 3 is hydrogen or C 1-6 Alkyl, and R 4 R 5 ;or

[0505] (ii)R 3 and R 4 Together with the nitrogen atom to which it is attached, it forms the 5 substituted 4- to 8-membered monocyclic heterocyclic group;

[0506] and

[0507] R 5 is -LZ, where

[0508] L is -L 1 -[GL 2 ] q -GL 3 -*,in

[0509] * is the bond to Z;

[0510] q is 0 or an integer selected from 1 to 25; (e.g., 1 to 20 or 1 to 15);

[0511] L 1 is a bond or -BA-;

[0512] Each L 2 independently -ABA-;

[0513] L 3 is a bond or -ABA-;

[0514] Each G is independently -DEF-, wherein D, E and F are independently a bond, C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, aryl, or heteroaryl, each of which is optionally substituted with 1, 2, 3, or 4 R groups;

[0515] Each A is independently a bond, -O-, -S- or -N(R N )-;

[0516] Each B is independently a bond, CH2, C(O), C(S), C(NR N ), S(O), S(O)2, P(O)(OH), P(S)(OH) or P(S)(SH);

[0517] Each R N are independently hydrogen or C 1-6 Alkyl, or the two R in the -ABA- group N Together with the atoms to which they are attached, they form a 4- to 8-membered heterocyclic group;

[0518] and

[0519] Z is a member of a reactive pair;

[0520] and

[0521] Each R group is independently selected from the group consisting of: R', C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-8 Cycloalkyl, heterocyclyl, aryl, heteroaryl, C 3-8 Cycloalkyl C 1-6 Alkyl, heterocyclic C 1-6 Alkyl, aryl C 1-6 Alkyl, heteroaryl C 1-6 alkyl, each of which, except R′, is optionally substituted with 1, 2, or 3 R′ groups, wherein

[0522] Each R' is independently halogen, cyano, azido, nitro, -N(R b )2、-O(R a )、-S(R 0 )、-C(O)OR 0 、C(O)R 0 、-C(O)N(R 0 )2、-C(NR 0 )OR 0 、-C(NR 0 )R 0 、-C(NR 0 )N(R 0 )2、-C(S)OR 0 、-C(S)R 0 、-C(S)N(R 0 )2、-S(O)2R 0 、-S(O)2OR 0 、-S(O)2N(R 0 )2、-N(R 0 )C(O)OR 0 、-N(R 0 )C(O)R 0 、-N(R 0 )C(O)N(R 0 )2、-N(R 0)S(O)2R 0 、-N(R 0 )S(O)2OR 0 、-N(R 0 )S(O)2N(R 0 )2、-OC(O)OR 0 、-OC(O)R 0 、-OC(O)N(R 0 )2、-OS(O)2R 0 、-OS(O)2OR 0 、-OS(O)2N(R 0 )2 or -SC(O)R 0 ,in

[0523] Each R 0 are independently hydrogen or C 1-6 Alkyl; each R a are independently hydrogen, C 1-6 an alkyl or hydroxy protecting group; and each R b are independently hydrogen, C 1-6 Alkyl or nitrogen protecting group

[0524] The proviso is that in each -DEF- group, at least one of D, E, and F is not a bond; and R L Not N-morpholinyl.

[0525] In some embodiments, in -N(R 3 )(R 4 ), R 3 and R 4 No N-morpholinyl ring is formed.

[0526] In some embodiments, in each -ABA- group, B is a bond only if one of the A groups is not a bond.

[0527] In some embodiments, each R group is independently selected from the group consisting of: R', C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-8 Cycloalkyl, heterocyclyl, aryl, heteroaryl, C 3-8 Cycloalkyl C 1-6 Alkyl, heterocyclic C 1-6 Alkyl, aryl C 1-6 Alkyl and heteroaryl 1-6 alkyl, each of which, in addition to R′, is optionally substituted with 1, 2, or 3 R′ groups, wherein each R′ is independently halogen, cyano, azido, nitro, —N(R b )2、-O(R a)、-S(R 0 )、-C(O)OR 0 、-C(O)R 0 、-C(O)N(R 0 )2、-N(R 0 )C(O)R 0 、-OC(O)OR 0 or -OC(O)R 0 , where each R 0 are independently hydrogen or C 1-6 Alkyl; each R a are independently hydrogen, C 1-6 an alkyl or hydroxy protecting group; and each R b are independently hydrogen, C 1-6 Alkyl or nitrogen protecting group.

[0528] In some embodiments, each R group is independently selected from the group consisting of: R', C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 cycloalkyl, 5- to 6-membered heterocyclyl, phenyl, 5- to 6-membered heteroaryl, and benzyl, each of which is optionally substituted with 1, 2, or 3 R' groups in addition to R', wherein each R' is independently halogen, cyano, azido, nitro, -N(R b )2、-O(R a )、-S(R 0 )、-C(O)OR 0 、-C(O)R 0 、-C(O)N(R 0 )2、-N(R 0 )C(O)R 0 、-OC(O)OR 0 or -OC(O)R 0 , where each R 0 are independently hydrogen or C 1-6 Alkyl; each R a are independently hydrogen, C 1-6 an alkyl or hydroxy protecting group; and each R b are independently hydrogen, C 1-6 Alkyl or nitrogen protecting group.

[0529] In some embodiments, each R group is independently selected from the group consisting of: R', C 1-6 Alkyl, C 3-6cycloalkyl, 5- to 6-membered heterocyclyl, phenyl, 5- to 6-membered heteroaryl, and benzyl, each of which is optionally substituted with 1, 2, or 3 R' groups in addition to R', wherein each R' is independently halogen, cyano, azido, nitro, -N(R b )2、-O(R a )、-S(R 0 )、-C(O)OR 0 、-C(O)R 0 、-C(O)N(R 0 )2、-N(R 0 )C(O)R 0 、-OC(O)OR 0 or -OC(O)R 0 , where each R 0 are independently hydrogen or C 1-6 Alkyl; each R a are independently hydrogen, C 1-6 an alkyl or hydroxy protecting group; and each R b are independently hydrogen, C 1-6 Alkyl or nitrogen protecting group.

[0530] R Y Example, formula (IV) and (IV-a) to (IV-b)

[0531] In some embodiments of any one of Formula (IV) and Formulas (IV-a) to (IV-b), in R Y In each R 2 R is independently as defined in formula (IV).

[0532] In some embodiments, R Y for (or its tautomers, ).

[0533] In some embodiments, R Y for

[0534] In some embodiments, in R Y In the case of two R 2 The group, together with the atoms to which it is bonded, forms a fused 4- to 8-membered ring, which is optionally substituted with 1, 2, 3, or 4 groups independently selected from the group consisting of R and a nitrogen protecting group, wherein R is as defined in formula (IV).

[0535] In some embodiments, R Y for wherein p is 0, 1, 2, 3 or 4; and each R 21 are independently selected from the group consisting of R and a nitrogen protecting group, wherein R is as defined in formula (IV).

[0536] In some embodiments, R Y for wherein p is 0, 1, 2, 3 or 4; and each R 21 are independently selected from the group consisting of R and a nitrogen protecting group, wherein R is as defined in formula (IV).

[0537] In some embodiments, R Y for

[0538] In some embodiments, R Y for Wherein RP is a nitrogen protecting group.

[0539] In some embodiments, R Y for where R P A nitrogen protecting group.

[0540] In some embodiments, R Y for where R P A nitrogen protecting group.

[0541] In some embodiments, R Y for

[0542] In some embodiments, R Y for where R P A nitrogen protecting group.

[0543] In some embodiments, the compound of formula (IV) is according to formula (IV-a) to (IV-h):

[0544]

[0545] Where p is 0, 1, 2 or 3;

[0546] Each R 21 are independently selected from the group consisting of R and a nitrogen protecting group, and R P is a hydrogen or nitrogen protecting group, and

[0547] R is as defined above.

[0548] L embodiment, formula (IV) and (IV-a) to (IV-r)

[0549] In some embodiments of any one of Formula (IV) and Formulas (IV-a) to (IV-r), L is -L 1 -[GL 2 ]q -GL 3 -*, where * is the bond to Z.

[0550] In another embodiment, wherein L is -L 1 -[GL 2 ] q -GL 3 -*, where q is 0, 1, 2, 3, 4, or 5.

[0551] In another embodiment, L is -L 1 -[GL 2 ] q -GL 3 -*, where q is 0, 1, 2, 3, or 4

[0552] In another embodiment, L is -L 1 -[GL 2 ] q -GL 3 -*, where q is 0, 1, 2, or 3.

[0553] In another embodiment, L is -L 1 -[GL 2 ] q -GL 3 -*, where q is 0, 1, or 2.

[0554] In another embodiment, L is -L 1 -[GL 2 ] q -GL 3 -*, where q is 1, 2, 3, 4, or 5.

[0555] In another embodiment, L is -L 1 -[GL 2 ] q -GL 3 -*, where q is 1, 2, 3, or 4.

[0556] In another embodiment, L is -L 1 -[GL 2 ] q -GL 3 -*, where q is 1, 2, or 3.

[0557] In another embodiment, L is -L 1 -[GL 2 ] q -GL 3 -*, where q is 1 or 2.

[0558] In another embodiment, L is -L1 -[G-L 2 q -G-L 3 -*, where q is 4. In another embodiment, L is -L 1 -[G-L 2 q -G-L 3 -*, where q is 3. In another embodiment, L is -L 1 -[G-L 2 q -G-L 3 -*, where q is 2.

[0559] In another embodiment, L is -L 1 -G-L 2 -G-L 3 -*. In another embodiment nts, L is -L 1 -G-L 3 -*. In another embodiment, L is -G-L 3 -*. In another embodiment, L is -L 1 -G-*. In another embodiment, L is -G-*.

[0560] In some embodiments, each instance of A-B-A- is independently selected from the group consisting of: -C(O)O-, -OC(O)-, -C(O)N(R N )-, -N(R N )C(O)-, -OC(O)O-, -OC(O)N(R N )-, -N(R N )C(O)O-, -N(R N )C(O)N(R N )-, -OP(O)(OH)O-, -OP(S)(OH)O-, -O- and -N(R N )-, where each R N is independently hydrogen or C 1-6 alkyl.

[0561] In some embodiments, each instance of A-B-A- is independently selected from the group consisting of: -C(O)O-, -OC(O)-, -C(O)N(R N )-, -N(R N )C(O)-, -OC(O)N(R N )-, -N(R N )C(O)O-, -N(R N )C(O)N(R N )-, -O- and -N(R N )-, where each R​​​N independently hydrogen or C 1-6 alkyl group.

[0562] In some embodiments, each instance of A-B-A- is independently selected from the group consisting of: -C(O)N(R N ), -N(R N )C(O)-, -OC(O)N(R N ), -N(R N )C(O)O-, -N(R N )C(O)N(R N ), -O- and -N(R N ), where each R N is independently hydrogen or C 1-6 alkyl group.

[0563] In some embodiments, each instance of A-B-A- is independently selected from the group consisting of: -C(O)O-, -OC(O)-, -C(O)N(R N ), -N(R N )C(O)-, -O- and -N(R N ), where each R N is independently hydrogen or C 1-6 alkyl group.

[0564] In some embodiments, each instance of A-B-A- is independently selected from the group consisting of: -C(O)N(R N ), -N(R N )C(O)-, -O- and -N(R N ), where each R N is independently hydrogen or C 1-6 alkyl group.

[0565] In some embodiments, D and F are each independently a bond, C 1-10 alkyl, C 2-10 alkenyl or C 2-10 alkynyl, each of which is optionally substituted with 1, 2, 3 or 4 R groups; and

[0566] E is C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclic group, aryl or heteroaryl, each of which is optionally substituted with 1, 2, 3 or 4 R groups.

[0567] In some embodiments, D and F are each independently a bond or C 1-10 alkyl optionally substituted with 1, 2, 3 or 4 R groups; and

[0568] E is C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, aryl, or heteroaryl, each of which is optionally substituted with 1, 2, 3, or 4 R groups.

[0569] In some embodiments, each G is independently C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, aryl, or heteroaryl, each of which is optionally substituted with 1, 2, 3, or 4 R groups.

[0570] In some embodiments, each G is independently C optionally substituted with 1, 2, or 3 R groups. 1-10 alkyl.

[0571] In some embodiments, each G is independently C optionally substituted with 1 or 2 R groups. 1-10 alkyl.

[0572] In some embodiments, each G is independently C optionally substituted with one R group. 1-10 alkyl

[0573] In some embodiments, L is -L 1 -GL 3 -*, where * is a bond to Z;

[0574] G is -DEF-, wherein D, E and F are independently a bond, C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, aryl, or heteroaryl, each of which is optionally substituted with 1, 2, 3, or 4 R groups;

[0575] L 1 for -BA-;

[0576] L 3 is a bond or -ABA-;

[0577] Each A is independently a bond, -O-, -S- or -N(R N )-, where each R N are independently hydrogen or C 1-6 alkyl.

[0578] each B is independently a bond, CH2, C(O), S(O)2, P(O)(OH) or P(S)(OH); and

[0579] In some embodiments, L is -L 1 -GL 3 -*, where * is a bond to Z;

[0580] G is C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, aryl, or heteroaryl, each of which is optionally substituted with 1, 2, 3, or 4 R groups;

[0581] L 1 for -B-;

[0582] L 3 is a bond or -ABA-;

[0583] Each A is independently a bond, -O-, -S- or -N(R N )-, where each R N are independently hydrogen or C 1-6 alkyl; and

[0584] each B is independently a bond, CH2, C(O), S(O)2, P(O)(OH) or P(S)(OH; and

[0585] In some embodiments, L is -L 1 -G-*, where * is the bond to Z;

[0586] G is C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, aryl, or heteroaryl, each of which is optionally substituted with 1, 2, 3, or 4 R groups;

[0587] L 1 is -BA-, where

[0588] A is a bond, -O-, -S- or -N(R N )-, where each R N are independently hydrogen or C 1-6 alkyl; and

[0589] B is a bond, C(O), C(S), S(O)2, P(O)(OH) or P(S)(OH).

[0590] In some embodiments, L is -L 1 -G-*, where * is the bond to Z;

[0591] G is C 1-10 Alkyl, C2-10 Alkenyl, C 2-10 alkynyl, each of which is optionally substituted with 1 or 2 R groups;

[0592] L 1 is -BA-, where

[0593] A is a bond, -O-, -S- or -N(R N )-, each R N are independently hydrogen or C 1-6 alkyl; and

[0594] B is a bond, C(O), S(O)2, P(O)(OH) or P(S)(OH).

[0595] In some embodiments, L is -L 1 -G-*, where * is the bond to Z;

[0596] G is C 1-10 Alkyl or C 2-10 alkenyl, each of which is optionally substituted with 1 or 2 R groups;

[0597] L 1 is a bond, C(O), S(O)2, P(O)(OH) or P(S)(OH); and

[0598] R N is hydrogen or C 1-6 alkyl.

[0599] In some embodiments, L is Wherein * is a bond to Z; k is an integer from 1 to 10; L 1 for bonds, C(O), C(S), C(NR N ), S(O)2, P(O)(OH) or P(S)(OH); and R N is hydrogen or C 1-6 alkyl.

[0600] In some embodiments, L is Wherein * is a bond to Z; k is an integer from 1 to 10; L 1 is a bond, C(O), P(O)(OH) or P(S)(OH).

[0601] In some embodiments, L is wherein * is a bond to Z; k is an integer from 1 to 10; or an integer from 2 to 10; or an integer from 3 to 10; or an integer from 4 to 10; or an integer from 5 to 10; or an integer from 5 to 9; or an integer from 5 to 8; or an integer from 5 to 7.

[0602] In some embodiments, L is wherein * is a bond to Z; and t is an integer from 0 to 10 (eg, an integer from 1 to 5; or 1; or 2; or 3).

[0603] In some embodiments, L is

[0604] wherein * is a bond to Z, t is an integer from 0 to 10 (e.g., an integer from 1 to 5 or 1; or 2; or 3); a is an integer from 1 to 3; and s and s′ are each independently an integer from 1 to 24 (e.g., an integer from 1 to 16; an integer from 1 to 10; an integer from 3 to 10; an integer from 3 to 7; or an integer from 4 to 6).

[0605] In some embodiments, L is wherein * is a bond to Z; a is 1, 2 or 3; and each s, s′ and s″ is independently an integer from 1 to 24 (e.g., an integer from 1 to 16, an integer from 1 to 10, an integer from 3 to 10, an integer from 3 to 7, or an integer from 4 to 6).

[0606] In some embodiments, L is wherein * is a bond to Z; and s, s′, and s″ are independently an integer from 1 to 24 (eg, an integer from 1 to 16, an integer from 1 to 10, an integer from 3 to 10, an integer from 3 to 7, or an integer from 4 to 6).

[0607] In some embodiments, L is wherein * is a bond to Z and each of s, s′ and s″ is independently an integer from 1 to 24 (e.g., an integer from 1 to 16, an integer from 1 to 10, an integer from 3 to 10, an integer from 3 to 7, or an integer from 4 to 6)

[0608] In some embodiments, L is wherein * is a bond to Z; s and k are independently an integer from 1 to 20 (e.g., an integer from 1 to 16, an integer from 1 to 10, an integer from 3 to 10, an integer from 3 to 7, or an integer from 4 to 6); and w is an integer from 1 to 10 (e.g., an integer from 1 to 16, an integer from 1 to 10, an integer from 3 to 10, an integer from 3 to 7, or an integer from 4 to 6).

[0609] In some embodiments, L is wherein * is a bond to Z and w is an integer from 1 to 20 (eg, an integer from 1 to 16, an integer from 1 to 10, an integer from 3 to 10, an integer from 3 to 7, or an integer from 4 to 6).

[0610] R L Embodiment, formula (IV) and (IV-a) to (IV-h)

[0611] In some embodiments of any one of Formula (IV) and Formulas (IV-a) to (IV-h), RL -N(R 3 )(R 4 ), where R 3 is hydrogen or C 1-6 Alkyl, and R 4 R 5 .

[0612] In some embodiments, R L -N(R 3 )(R 4 ), where R 3 is hydrogen, and R 4 R 5 .

[0613] In some embodiments, R L -N(R 3 )(R 4 ), where R 3 c 1-3 Alkyl, and R 4 R 5 .

[0614] In some embodiments, R L -N(R 3 )(R 4 ), where R 3 is methyl, and R 4 R 5 .

[0615] In some embodiments, R L N(R 3 )(R 4 ), where R 3 and R 4 Together with the nitrogen atom to which it is attached, it forms the 5 a substituted 4- to 8-membered monocyclic heterocyclic group.

[0616] In some embodiments, R L N(R 3 )(R 4 ), wherein R3 and R4 together with the nitrogen atom to which they are attached form 5 Substituted 4- to 8-membered monocyclic heterocyclic group, with the restriction that R 3 and R 4 Together with the nitrogen atom to which it is attached, it does not form an N-morpholinyl group.

[0617] In some embodiments, R L N(R 3 )(R 4 ), where R 3 and R 4Together with the nitrogen atom to which it is attached, it forms the following group: piperidinyl, piperazinyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, oxazolidinyl, thiazolidinyl, azetidinyl, pyrrolinyl, imidazolinyl or pyrazolinyl, each of which is substituted by R 5 replace.

[0618] In some embodiments, R L for

[0619] In some embodiments, R L for

[0620]

[0621] wherein t is an integer from 0 to 10 (e.g., 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10); a is an integer from 1 to 3, and s and s′ are each independently an integer from 1 to 24 (e.g., an integer from 1 to 16, an integer from 1 to 10, an integer from 3 to 10, an integer from 3 to 7, or an integer from 4 to 6).

[0622] In some embodiments, R L for a is an integer from 1 to 3; and s, s′, and s″ are each independently an integer from 1 to 24 (eg, an integer from 1 to 16, an integer from 1 to 10, an integer from 3 to 10, an integer from 3 to 7, or an integer from 4 to 6).

[0623] In some embodiments, R L for and s, s′, and s″ are independently an integer from 1 to 24 (eg, an integer from 1 to 16, an integer from 1 to 10, an integer from 3 to 10, an integer from 3 to 7, or an integer from 4 to 6).

[0624] In some embodiments, R L for wherein s and k are independently an integer from 1 to 20 (e.g., an integer from 1 to 16, an integer from 1 to 10, an integer from 3 to 10, an integer from 3 to 7, or an integer from 4 to 6); and w is an integer from 1 to 10 (e.g., an integer from 1 to 16, an integer from 1 to 10, an integer from 3 to 10, an integer from 3 to 7, or an integer from 4 to 6).

[0625] In some embodiments, R L for

[0626] wherein s, s′, and s″ are independently an integer from 1 to 24 (e.g., an integer from 1 to 16, an integer from 1 to 10, an integer from 3 to 10, an integer from 3 to 7, or an integer from 4 to 6); and t is an integer from 1 to 10 (e.g., an integer from 1 to 8, an integer from 1 to 5, an integer from 1 to 3, or 1, or 2, or 3).

[0627] In some embodiments, R L for

[0628] wherein s is an integer of 1 to 24 (eg, an integer of 1 to 16, an integer of 1 to 10, an integer of 3 to 10, an integer of 3 to 7, or an integer of 4 to 6).

[0629] In some embodiments, R L -O(R 5 ).

[0630] In some embodiments, R L for wherein s is an integer from 1 to 24 (e.g., an integer from 1 to 16, an integer from 1 to 10, an integer from 3 to 10, an integer from 3 to 7, or an integer from 4 to 6); and t is an integer from 0 to 10 (e.g., an integer from 1 to 8, an integer from 1 to 5, an integer from 1 to 3, or 1, or 2, or 3).

[0631] In some embodiments, R L -R 5 .

[0632] In some embodiments, R L for wherein s is 1 to 24 (eg, an integer from 1 to 16, an integer from 1 to 10, an integer from 3 to 10, an integer from 3 to 7, or an integer from 4 to 6).

[0633] In some embodiments, R L for wherein t is 0 to 10 (eg, 1-5; or 1-3; or 1; or 2; or 3).

[0634] In some embodiments, R L for

[0635]

[0636] In some embodiments, R L for

[0637]

[0638] where R Z is hydrogen or C 1-10alkyl, and w is an integer selected from 1 to 10 (e.g., 2-10, 2-8, 4-8). In some embodiments, R Z In some embodiments, R Z In some embodiments, R Z It is a methyl group.

[0639] In some embodiments, R L for

[0640]

[0641]

[0642] wherein Z is as defined in formula (IV) and its examples.

[0643] In some embodiments, R L for

[0644]

[0645] wherein w is an integer selected from 1 to 10 (e.g., 2-10, 2-8, 4-8), and R Z As defined in formula (IV) and its examples.

[0646] In some embodiments, the compound of formula (IV) is according to one of formulas (IV-i) to (IV-1):

[0647]

[0648]

[0649] wherein p is 0, 1, 2 or 3 (e.g., 0); each R 21 is independently selected from the group consisting of R and a nitrogen protecting group, wherein R and the remaining variables are as defined in formula (IV). In one embodiment of formulas (IV-i) to (IV-1), R 1 In another embodiment of formula (IV-i) to (IV-1), R 1 C 1-6 Alkyl (eg, methyl or tert-butyl).

[0650] In some embodiments, the compound of formula (IV) is according to one of formulas (IV-m) to (IV-r):

[0651]

[0652]

[0653] or a salt thereof, wherein p is 0, 1 or 2; each R 21are independently selected from the group consisting of R; and R P is hydrogen or a nitrogen protecting group (e.g., a nitrogen protecting group), wherein R and the remaining variables are as defined in formula (IV). In one embodiment of formulas (IV-m) to (IV-vr), R 1 In another embodiment of formula (IV-m) to (IV-r), R 1 C 1-6 Alkyl (eg, methyl or tert-butyl).

[0654] Z embodiment, formula (IV) and (IV-a) to (IV-r)

[0655] In some embodiments of any one of Formula (IV) and Formulas (IV-a) to (IV-r), Z is azido, hydroxy, amino, -N=C=O, -N=C=S, -SR Z1 、-C(O)H、-c(O)OR Z 、-C(S)OR Z , -CH2-X or Michael acceptor,

[0656] where R Z is hydrogen or C 1-10 alkyl;

[0657] R Z1 is hydrogen, pyridyl or benzotriazolyl;

[0658] X is a leaving group.

[0659] In some embodiments, Z is COOH.

[0660] In some embodiments, Z is NH2.

[0661] In some embodiments, Z is N3.

[0662] In some embodiments, Z is hydroxy.

[0663] In some embodiments, Z is -SH.

[0664] In some embodiments, Z is

[0665] In some embodiments, Z is a Michael acceptor (eg, N-maleimide).

[0666] In some embodiments, Z comprises a terminal alkyne,

[0667] In some embodiments, Z comprises Such as Among them L 53It is a bond, -C(O)-, -C(S)-, -S(O)2-, -C(O)O-, C(O)N(H)-, -S(O)2N(H)-.

[0668] In some embodiments, Z comprises

[0669] In some embodiments, Z comprises For example, Z is Among them L 52 is a bond, -O-, -N(H)-, -S-, -C(O)-, -C(S)-, -S(O)2-, -C(O)O-, -OC(O)-, -C(O)N(H)-, N(H)C(O)-, -OC(O)O-, -OC(O)N(H)-, N(H)C(O)O-, -N(H)C(O)N(H)-, or -S(O)2N(H)- (e.g. ).

[0670] In some embodiments, Z comprises

[0671] In some embodiments, Z 0 Include Such as Among them L 51 is a bond, -O-, -N(H)-, -S-, -C(O)-, -C(S)-, -S(O)2-, -C(O)O-, -OC(O)-, -C(O)N(H)-, N (H)C(O)-, -OC(O)O-, -OC(O)N(H)-, N(H)C(O)O-, -N(H)C(O)N(H)-, -CH2O-, -CH2N (H)-, -CH2S-, -CH2C(O)O-, -CH2OC(O)-, -CH2C(O)N(H)-, -CH2N(H)C(O)-, -CH2OC(O)O-, -CH2OC(O)N(H)-, -CH2N(H)C(O)O-, -CH2N(H)C(O)N(H)-, or -S(O)2N(H)- (e.g. ).

[0672] In some embodiments, Z comprises For example, Z is where R Z10 is hydrogen or C 1-10 alkyl; and L 50is a bond, -O-, -N(H)-, -S-, -C(O)-, -C(S)-, -S(O)2-, -C(O)O-, -OC(O)-, -C(O)N(H)-, N(H)C(O)-, -OC(O)O-, -OC(O)N(H)-, N(H)C(O)O-, -N(H)C(O)N(H)-, -CH2O-, -CH2 N(H)-, -CH2S-, -CH2C(O)O-, -CH2OC(O)-, -CH2C(O)N(H)-, -CH2N(H)C(O)-, --CH2 OC(O)O-, -CH2OC(O)N(H)-, -CH2N(H)C(O)O-, -CH2N(H)C(O)N(H)-, or -S(O)2N(H)-.

[0673] In some embodiments, Z is an activated ester (e.g. ).

[0674] In some embodiments, Z is

[0675] In some embodiments, Z is

[0676] In some embodiments, Z is

[0677] In some embodiments, Z is

[0678] In some embodiments, Z is

[0679] R P Embodiment, formula (IV) and (IV-a) to (IV-r)

[0680] In some embodiments of any one of Formula (IV) and Formulas (IV-a) to (IV-r), R P for Where r is 1, 2 or 3; each R P2 are independently halogen, nitro, cyano, C 1-4 Alkoxy, C 1-4 Alkyl, C 1-4 haloalkyl; and each R P3 are independently hydrogen, methyl or ethyl.

[0681] In some embodiments, when present, R PMethoxyacetyl (MAC), phenoxyacetyl (PAC), 2-chlorophenoxyacetyl, 3-chlorophenoxyacetyl, 4-chlorophenoxyacetyl, 2,4-dichlorophenoxyacetyl, 2-methylphenoxyacetyl, 3-methylphenoxyacetyl, 4-methylphenoxyacetyl, 4-chloro-2-methylphenoxyacetyl, 2-nitrophenoxyacetyl, 3-nitrophenoxyacetyl, 4-nitrophenoxyacetyl, 2-isopropylphenoxyacetyl, 3-isopropylphenoxyacetyl, propylphenoxyacetyl, 4-isopropylphenoxyacetyl, 2-(tert-butyl)phenoxyacetyl, 3-(tert-butyl)phenoxyacetyl, 4-(tert-butyl)phenoxyacetyl, 2-fluorophenoxyacetyl, 3-fluorophenoxyacetyl, 4-fluorophenoxyacetyl, 2,4-difluorophenoxyacetyl, 4-(trifluoromethoxy)phenoxyacetyl, 2-phenoxypropionyl, 2-(4-chloro-2-methylphenoxy)propionyl or 2-(4-chlorophenoxy)propionyl.

[0682] In some embodiments, when present, R P is methoxyacetyl (mac), phenoxyacetyl (pac), 2-chlorophenoxyacetyl, 4-chlorophenoxyacetyl, 2-methylphenoxyacetyl, 4-methylphenoxyacetyl or 4-isopropylphenoxyacetyl.

[0683] In some embodiments, when present, R P is methoxyacetyl (mac).

[0684] In some embodiments, when present, R P It is phenoxyacetyl (pac).

[0685] In some embodiments, when present, R 1 C 1-6 Alkyl, and R P for- Where r is 1, 2 or 3; each R P2 are independently halogen, nitro, cyano, C 1-4 Alkoxy, C 1-4 Alkyl, C 1-4 haloalkyl; and each R P3 are independently hydrogen, methyl or ethyl.

[0686] In some embodiments, when present, R 1 C 1-6 Alkyl, and R PMethoxyacetyl (MAC), phenoxyacetyl (PAC), 2-chlorophenoxyacetyl, 3-chlorophenoxyacetyl, 4-chlorophenoxyacetyl, 2,4-dichlorophenoxyacetyl, 2-methylphenoxyacetyl, 3-methylphenoxyacetyl, 4-methylphenoxyacetyl, 4-chloro-2-methylphenoxyacetyl, 2-nitrophenoxyacetyl, 3-nitrophenoxyacetyl, 4-nitrophenoxyacetyl, 2-isopropylphenoxyacetyl, 3-isopropylphenoxyacetyl, propylphenoxyacetyl, 4-isopropylphenoxyacetyl, 2-(tert-butyl)phenoxyacetyl, 3-(tert-butyl)phenoxyacetyl, 4-(tert-butyl)phenoxyacetyl, 2-fluorophenoxyacetyl, 3-fluorophenoxyacetyl, 4-fluorophenoxyacetyl, 2,4-difluorophenoxyacetyl, 4-(trifluoromethoxy)phenoxyacetyl, 2-phenoxypropionyl, 2-(4-chloro-2-methylphenoxy)propionyl or 2-(4-chlorophenoxy)propionyl.

[0687] In some embodiments, when present, R 1 C 1-6 Alkyl, and R P is methoxyacetyl (mac), phenoxyacetyl (pac), 2-chlorophenoxyacetyl, 4-chlorophenoxyacetyl, 2-methylphenoxyacetyl, 4-methylphenoxyacetyl or 4-isopropylphenoxyacetyl.

[0688] In some embodiments, when present, R 1 C 1-6 Alkyl, and R P is methoxyacetyl (mac).

[0689] In some embodiments, when present, R 1 C 1-6 Alkyl, and R P It is phenoxyacetyl (pac).

[0690] In some embodiments, when present, R 1 is methyl, and RP is Where r is 1, 2 or 3; each R P2 are independently halogen, nitro, cyano, C 1-4 Alkoxy, C 1-4 Alkyl, C 1-4 haloalkyl; and each R P3 are independently hydrogen, methyl or ethyl.

[0691] In some embodiments, when present, R 1 is methyl, and R PMethoxyacetyl (MAC), phenoxyacetyl (PAC), 2-chlorophenoxyacetyl, 3-chlorophenoxyacetyl, 4-chlorophenoxyacetyl, 2,4-dichlorophenoxyacetyl, 2-methylphenoxyacetyl, 3-methylphenoxyacetyl, 4-methylphenoxyacetyl, 4-chloro-2-methylphenoxyacetyl, 2-nitrophenoxyacetyl, 3-nitrophenoxyacetyl, 4-nitrophenoxyacetyl, 2-isopropylphenoxyacetyl, 3-isopropylphenoxyacetyl, propylphenoxyacetyl, 4-isopropylphenoxyacetyl, 2-(tert-butyl)phenoxyacetyl, 3-(tert-butyl)phenoxyacetyl, 4-(tert-butyl)phenoxyacetyl, 2-fluorophenoxyacetyl, 3-fluorophenoxyacetyl, 4-fluorophenoxyacetyl, 2,4-difluorophenoxyacetyl, 4-(trifluoromethoxy)phenoxyacetyl, 2-phenoxypropionyl, 2-(4-chloro-2-methylphenoxy)propionyl or 2-(4-chlorophenoxy)propionyl.

[0692] In some embodiments, when present, R 1 is methyl, and R P is methoxyacetyl (mac), phenoxyacetyl (pac), 2-chlorophenoxyacetyl, 4-chlorophenoxyacetyl, 2-methylphenoxyacetyl, 4-methylphenoxyacetyl or 4-isopropylphenoxyacetyl.

[0693] In some embodiments, when present, R 1 is methyl, and R P is methoxyacetyl (mac).

[0694] In some embodiments, when present, R 1 is methyl, and R P It is phenoxyacetyl (pac).

[0695] In some embodiments, when present, R 1 is hydrogen, and R P for Where r is 1, 2 or 3; each R P2 are independently halogen, nitro, cyano, C 1-4 Alkoxy, C 1-4 Alkyl, C 1-4 haloalkyl; and each R P3 are independently hydrogen, methyl or ethyl.

[0696] In some embodiments, when present, R 1 is hydrogen, and R PMethoxyacetyl (MAC), phenoxyacetyl (PAC), 2-chlorophenoxyacetyl, 3-chlorophenoxyacetyl, 4-chlorophenoxyacetyl, 2,4-dichlorophenoxyacetyl, 2-methylphenoxyacetyl, 3-methylphenoxyacetyl, 4-methylphenoxyacetyl, 4-chloro-2-methylphenoxyacetyl, 2-nitrophenoxyacetyl, 3-nitrophenoxyacetyl, 4-nitrophenoxyacetyl, 2-isopropylphenoxyacetyl, 3-isopropylphenoxyacetyl, propylphenoxyacetyl, 4-isopropylphenoxyacetyl, 2-(tert-butyl)phenoxyacetyl, 3-(tert-butyl)phenoxyacetyl, 4-(tert-butyl)phenoxyacetyl, 2-fluorophenoxyacetyl, 3-fluorophenoxyacetyl, 4-fluorophenoxyacetyl, 2,4-difluorophenoxyacetyl, 4-(trifluoromethoxy)phenoxyacetyl, 2-phenoxypropionyl, 2-(4-chloro-2-methylphenoxy)propionyl or 2-(4-chlorophenoxy)propionyl.

[0697] In some embodiments, when present, R 1 is hydrogen, and R P is methoxyacetyl (mac), phenoxyacetyl (pac), 2-chlorophenoxyacetyl, 4-chlorophenoxyacetyl, 2-methylphenoxyacetyl, 4-methylphenoxyacetyl or 4-isopropylphenoxyacetyl.

[0698] In some embodiments, when present, R 1 is hydrogen, and R P is methoxyacetyl (mac).

[0699] In some embodiments, when present, R 1 is hydrogen, and R P It is phenoxyacetyl (pac).

[0700] Species Example, Formula (IV)

[0701] In some embodiments, the compound of formula (IV) is selected from the group consisting of:

[0702]

[0703]

[0704]

[0705]

[0706]

[0707]

[0708]

[0709]

[0710] In another embodiment, the compound of formula (IV) is selected from the group consisting of:

[0711]

[0712] B.ɑ-v-β-6 (avβ6) integrin ligand

[0713] In another aspect, the present application provides a modified integrin modulating compound conjugated to a carrier group suitable for conjugation to or incorporation into an oligonucleotide. In some embodiments, the present invention provides a compound of formula (X),

[0714]

[0715] or a salt thereof, wherein:

[0716] R 1 、R Y and Y is as defined for formula (IV); and

[0717] R L -N(R 3 )(R 4 )、-O(R 5 )、-S(R 5 ) or -R 5 ,in

[0718] R 3 and R 4 Is any of the following:

[0719] (i)R 3 is hydrogen or C 1-6 Alkyl, and R 4 R 5 ;or

[0720] (ii)R 3 and R 4 Together with the nitrogen atom to which it is attached, it forms the 5 substituted 4- to 8-membered monocyclic heterocyclic group;

[0721] and

[0722] R 5 L-ZZ-L'-R T ,in

[0723] L and L' are independently -L 1 -[GL 2 ] q -GL3 -*,in

[0724] * is the key connected to ZZ;

[0725] q is 0 or an integer selected from 1 to 25; (e.g., 1 to 20 or 1 to 15);

[0726] L 1 is a bond or -BA-;

[0727] Each L 2 independently -ABA-;

[0728] L 3 is a bond or -ABA-;

[0729] Each G is independently -DEF-, wherein D, E and F are independently a bond, C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, aryl, or heteroaryl, each of which is optionally substituted with 1, 2, 3, or 4 R groups;

[0730] Each A is independently a bond, -O-, -S- or -N(R N )-;

[0731] Each B is independently a bond, CH2, C(O), C(S), C(NR N ), S(O), S(O)2, P(O)(OH), P(S)(OH) or P(S)(SH);

[0732] Each R N are independently hydrogen or C 1-6 Alkyl, or the two R in the -ABA- group N Together with the atoms to which they are attached, they form a 4- to 8-membered heterocyclic group;

[0733] and

[0734] ZZ is -A'-B'-A'- or a linker formed by a reactive pair, wherein

[0735] Each A' is independently a bond, -O-, -S- or -N(R N3 )-;

[0736] Each B' is independently a bond, CH2, C(O), C(S), C(NR N3 ), -C=N-, S(O), S(O)2, P(O)(OH), P(S)(OH) or P(S)(SH); and

[0737] Each R N3are independently hydrogen or C 1-6 Alkyl, or the two R in the -A'-B'-A'- group N3 Together with the atoms to which they are attached, they form a 4- to 8-membered heterocyclic group;

[0738] R T R T1 or -G 0 -OR T1 ,in

[0739] G 0 Does not exist or is -D 0 -E 0 -F 0 -, where D 0 、E 0 and F 0 independently for the bond, C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, aryl, or heteroaryl, each of which is optionally substituted with 1, 2, 3, or 4 R groups;

[0740] and

[0741] R T1 is hydrogen, a hydroxyl protecting group, a phosphorus coupling group or -L K -S S or -L L - an oligonucleotide, wherein

[0742] L K is a supporting connecting group;

[0743] L L is an oligonucleotide linking group; and

[0744] S S For solid support, -OR SS or -N(R SS )2 or hydrogen, wherein each R SS are independently hydrogen or C 1-6 alkyl.

[0745] and

[0746] Each R group is independently selected from the group consisting of: R', C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-8 Cycloalkyl, heterocyclyl, aryl, heteroaryl, C 3-8 Cycloalkyl C 1-6 Alkyl, heterocyclic C 1-6Alkyl, aryl C 1-6 Alkyl, heteroaryl C 1-6 alkyl, each of which, except R′, is optionally substituted with 1, 2, or 3 R′ groups, wherein

[0747] Each R' is independently halogen, cyano, azido, nitro, -N(R b )2、-O(R a )、-S(R 0 )、-C(O)OR 0 、C(O)R 0 、-C(O)N(R 0 )2、-C(NR 0 )OR 0 、-C(NR 0 )R 0 、-C(NR 0 )N(R 0 )2、-C(S)OR 0 、-C(S)R 0 、-C(S)N(R 0 )2、-S(O)2R 0 、-S(O)2OR 0 、-S(O)2N(R 0 )2、-N(R 0 )C(O)OR 0 、-N(R 0 )C(O)R 0 、-N(R 0 )C(O)N(R 0 )2、-N(R 0 )S(O)2R 0 、-N(R 0 )S(O)2OR 0 、-N(R 0 )S(O)2N(R 0 )2、-OC(O)OR 0 、-OC(O)R 0 、-OC(O)N(R 0 )2、-OS(O)2R 0 、-OS(O)2OR 0 、-OS(O)2N(R 0 )2 or -SC(O)R 0 ,in

[0748] Each R 0 are independently hydrogen or C 1-6 Alkyl; each R a are independently hydrogen, C 1-6 an alkyl or hydroxy protecting group; and each R bare independently hydrogen, C 1-6 Alkyl or nitrogen protecting group

[0749] The proviso is that in each -DEF- group, at least one of D, E, and F is not a bond; and R L Not N-morpholinyl.

[0750] The same embodiments of the variables of formula (X) as those of formula (IV) are as described above for formula (IV).

[0751] In some embodiments of Formula (X), in -N(R 3 )(R 4 ), R 3 and R 4 No N-morpholinyl ring is formed.

[0752] In some embodiments of Formula (X), in each -ABA- group, B is a bond only if one of the A groups is not a bond.

[0753] R T Example, formula (X)

[0754] In some embodiments of Formula (X), R T R T1 (For example -L L -oligonucleotides).

[0755] In some embodiments of Formula (X), R T -G 0 -OR T1 , where R T1 As defined for formula (X), and G 0 Selected from:

[0756] (a)G 0 Does not exist or is -D 0 -E 0 -F 0 -, where D 0 、E 0 and F 0 independently for the bond, C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, aryl, or heteroaryl, each of which is optionally substituted with 1, 2, 3, or 4 R groups;

[0757] (b)G 0 -D 0 -E 0 -F 0 -,in

[0758] D0 and F 0 is independently a bond or C optionally substituted with 1, 2, 3 or 4 R groups 1-10 alkyl; and

[0759] E 0 C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, aryl, or heteroaryl, each of which is optionally substituted with 1, 2, 3, or 4 R groups;

[0760] (c)G 0 -D 0 -E 0 -F 0 -,in

[0761] D 0 and F 0 is independently a bond or C optionally substituted with 1, 2, 3 or 4 R groups 1-10 alkyl; and

[0762] E 0 is a 3- to 10-membered heterocyclyl group optionally substituted with 1, 2, 3 or 4 R groups; and

[0763] (d)G 0 is a 3- to 10-membered heterocyclic group optionally substituted with 1 or 2 R groups; examples include G 0 is tetrahydrofuranyl, pyrrolidinyl, piperidinyl, piperazinyl or morpholinyl, each of which is optionally substituted with 1 or 2 R groups; examples include:

[0764] and

[0765] (e)G 0 is a 3- to 10-membered heterocyclyl-C optionally substituted with 1, 2, 3 or 4 R groups 1-10 Alkyl; examples include:

[0766] (1) and

[0767] (2)

[0768] (f)G 0 C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 alkynyl, each of which is optionally substituted with 1, 2, 3, or 4 R groups (e.g., 1 or 2 R groups);

[0769] (g)G 0 C 1-10Alkyl or C 2-10 alkenyl, each of which is optionally substituted with 1 or 2 R groups;

[0770] (h)G 0 is C optionally substituted with 1 or 2 R groups 1-10 alkyl;

[0771] (i)G 0 is optionally -O(R a ) substituted C 1-10 Alkyl, where R a are independently hydrogen, C 1-6 Alkyl or hydroxy protecting groups; e.g.

[0772] (j)G 0 C 1-10 alkyl,

[0773] and

[0774] (k)G 0 does not exist (for key);

[0775] Where * indicates the bond to L', and a broken bond indicates the bond to OR T1 The bond, and R is -C 1-6 Alkyl-OR a OR a , where R a are independently hydrogen, C 1-6 alkyl or hydroxy protecting groups;

[0776] -L'-embodiment, formula (X)

[0777] In some embodiments of Formula (X), -L'- is *-GL 1 -, where * is a bond to ZZ; and

[0778] (a)L 1 is a bond or -BA-, where

[0779] A is a bond, -O-, -S- or -N(R N )-;

[0780] B is a bond, CH2, C(O), C(S), C(NR N ), S(O), S(O)2, P(O)(OH), P(S)(OH) or P(S)(SH);

[0781] Each R N are independently hydrogen or C 1-6 alkyl; and

[0782] G is -DEF-, wherein D, E and F are independently a bond, C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, aryl, or heteroaryl, each of which is optionally substituted with 1, 2, 3, or 4 R groups;

[0783] (b)L 1 is a bond or -BA-, where

[0784] A is a bond, -O-, -S- or -N(R N )-;

[0785] B is a bond, CH2, C(O), C(S), C(NR N ), S(O), S(O)2, P(O)(OH), P(S)(OH) or P(S)(SH);

[0786] Each R N are independently hydrogen or C 1-6 alkyl; and

[0787] G is C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, aryl, or heteroaryl, each of which is optionally substituted with 1, 2, 3, or 4 R groups;

[0788] (c)L 1 is a bond, C(O), C(S), S(O)2, P(O)(OH) or P(S)(OH); and

[0789] G is C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 alkynyl, each of which is optionally substituted with 1, 2, 3, or 4 R groups;

[0790] (d)L 1 is a bond, C(O), C(S), S(O)2, P(O)(OH) or P(S)(OH); and

[0791] G is C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 alkynyl, each of which is optionally substituted with 1, 2, 3, or 4 R groups;

[0792] or

[0793] (e)L 1is a bond, C(O), C(S), S(O)2, P(O)(OH) or P(S)(OH); and G is C 1-10 alkyl.

[0794] In some embodiments of Formula (X), -L'- is *-G-[L 2 -G] q -L 1 -, wherein * is a bond to ZZ; q is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 (e.g., an integer from 1 to 8, an integer from 1 to 5, or an integer from 1 to 3); and

[0795] (a)L 1 is a bond or -BA-;

[0796] Each L 2 independently -ABA-;

[0797] Each A is independently a bond, -O-, -S- or -N(R N )-;

[0798] Each B is independently a bond, CH2, C(O), C(S), C(NR N ), S(O), S(O)2, P(O)(OH), P(S)(OH) or P(S)(SH);

[0799] Each R N are independently hydrogen or C 1-6 alkyl;

[0800] Each G is independently -DEF-, wherein D, E and F are independently a bond, C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, aryl, or heteroaryl, each of which is optionally substituted with 1, 2, 3, or 4 R groups;

[0801] (b)L 1 for bonds, CH2, C(O), C(S), C(NR N ), S(O), S(O)2, P(O)(OH), P(S)(OH) or P(S)(SH);

[0802] Each L 2 independently -ABA-;

[0803] Each A is independently a bond, -O-, -S- or -N(R N )-;

[0804] Each B is independently a bond, CH2, C(O), C(S), C(NRN ), S(O), S(O)2, P(O)(OH), P(S)(OH) or P(S)(SH);

[0805] Each R N are independently hydrogen or C 1-6 Alkyl, and

[0806] Each G is independently a bond, a C optionally substituted with 1, 2, 3 or 4 R groups 1-10 alkyl.

[0807] (c)L 1 for bonds, CH2, C(O), C(S), C(NR N ), S(O), S(O)2, P(O)(OH), P(S)(OH) or P(S)(SH);

[0808] Each L 2 independently -ABA-;

[0809] Each A is independently a bond, -O-, -S- or -N(R N )-, where R N is hydrogen or C 1-6 alkyl

[0810] each B is independently a bond, CH2, C(O), C(S), S(O)2, P(O)(OH) or P(S)(OH);

[0811] Each G is independently C optionally substituted with 1, 2, 3 or 4 R groups. 1-10 alkyl

[0812] (d)L 1 is a bond, CH2, C(O), C(S), S(O)2, P(O)(OH) or P(S)(OH)

[0813] Each L 2 independently -ABA-;

[0814] Each A is independently a bond, -O-, -S- or -N(R N )-, where R N is hydrogen or C 1-6 alkyl

[0815] each B is independently a bond, CH2, C(O), C(S), S(O)2, P(O)(OH) or P(S)(OH);

[0816] Each G is independently C optionally substituted with 1, 2, 3 or 4 R groups. 1-10 alkyl;

[0817] or

[0818] (e)L 1 is a bond, CH2, C(O), S(O)2, P(O)(OH) or P(S)(OH);

[0819] Each L 2 independently -ABA-;

[0820] Each A is independently a bond, -O-, -S- or -N(R N )-, where R N is hydrogen or C 1-6 alkyl

[0821] each B is independently a bond, CH2, C(O), C(S), S(O)2, P(O)(OH) or P(S)(OH);

[0822] Each G is independently C 1-10 alkyl;

[0823] In some embodiments of Formula (X), -L'- is -[GL 2 ] q -GL 3 -*, where * is a bond to ZZ; and

[0824] (a) q is 0, 1, 2, 3, 4 or 5;

[0825] Each L 2 independently -ABA-;

[0826] L 3 is a bond or -ABA-;

[0827] Each A is independently a bond, -O-, -S- or -N(R N )-;

[0828] Each B is independently a bond, CH2, C(O), C(S), C(NR N ), S(O), S(O)2, P(O)(OH), P(S)(OH) or P(S)(SH);

[0829] Each R N are independently hydrogen or C 1-6 alkyl;

[0830] Each G is independently C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 alkynyl, each of which is optionally substituted with 1, 2, 3, or 4 R groups;

[0831] (b) q is 0, 1, 2 or 3;

[0832] Each L2 are independently a bond, C(O)O, OC(O), C(O)(NR N )、N(R N )C(O), SO2N(R N )、N(R N )SO2, OP(O)(OH), OP(S)(OH), P(O)(OH)O, P(S)(OH)O, OP(O)(OH)O or OP(S)(OH)O, wherein each R N are independently hydrogen or C 1-6 alkyl;

[0833] Each G is independently C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 alkynyl, each of which is optionally substituted with 1 or 2 R groups;

[0834] (c) q is 0, 1, 2 or 3;

[0835] Each L 2 are independently a bond, C(O)O, OC(O), C(O)(NR N )、N(R N )C(O), OP(O)(OH)O or OP(S)(OH)O, wherein each R N are independently hydrogen or C 1-6 alkyl;

[0836] Each G is independently C 1-10 Alkyl or C 2-10 alkenyl, each of which is optionally substituted with 1 or 2 R groups;

[0837] (d) q is 0, 1, 2 or 3 (e.g., q is 0, 1 or 2; or 0 or 1; or 0; or 1; or 2);

[0838] Each L 2 are independently C(O)O or OC(O);

[0839] Each G is independently C 1-10 alkyl, each of which is optionally substituted with 1 or 2 R groups;

[0840] (e) q is 0, 1, 2 or 3 (e.g., q is 0, 1 or 2; or 0 or 1; or 0; or 1; or 2);

[0841] Each L 2 are independently C(O)(NR N ) or N(R N )C(O), where each R N are independently hydrogen or C 1-6 alkyl;

[0842] Each G is independently C 1-10 alkyl, each of which is optionally substituted with 1 or 2 R groups;

[0843] (f) q is 0, 1, 2 or 3 (e.g., q is 0, 1 or 2; or 0 or 1; or 0; or 1; or 2);

[0844] Each L 2 independently OP(O)(OH)O or OP(S)(OH)O (e.g., each is OP(O)(OH)O);

[0845] Each G is independently C 1-10 alkyl, each of which is optionally substituted with 1 or 2 R groups;

[0846] or

[0847] (g) q is 0, 1, 2 or 3 (e.g., q is 0, 1 or 2; or 0 or 1; or 0; or 1; or 2);

[0848] Each L 2 is the key;

[0849] Each G is independently C 1-10 alkyl, each of which is optionally substituted with 1 or 2 R groups.

[0850] In some embodiments, -L'- is *-L 3 -GL 1 -, where * is a bond to ZZ; and

[0851] (a)L 1 and L 3 independently -ABA-;

[0852] Each G is independently C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, aryl, or heteroaryl, each of which is optionally substituted with 1, 2, or 3 R groups;

[0853] Each A is independently a bond, -O-, -S- or -N(R N )-, where R N is hydrogen or C 1-6 alkyl; and

[0854] each B is independently a bond, C(O), C(S), S(O)2, P(O)(OH) or P(S)(OH);

[0855] (b)L 3 -C(O)O- or C(O)N(R N)-, where R N is hydrogen or C 1-6 alkyl;

[0856] L 1 is -OP(O)(OH)O- or -OP(S)(OH)O-; and

[0857] Each G is independently C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, aryl, or heteroaryl, each of which is optionally substituted with 1, 2, or 3 R groups;

[0858] (c)L 3 -C(O)O- or C(O)N(R N )-, where R N is hydrogen or C 1-6 alkyl;

[0859] L 1 is -OP(O)(OH)O- or -OP(S)(OH)O-; and

[0860] Each G is independently C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, aryl, or heteroaryl, each of which is optionally substituted with one R group;

[0861] (d)L 3 -C(O)O- or C(O)N(R N )-, where R N is hydrogen or C 1-6 alkyl;

[0862] L 1 is -OP(O)(OH)O- or -OP(S)(OH)O-; and

[0863] Each G is independently C 3-10 a cycloalkyl group or a 3- to 10-membered heterocyclic group;

[0864] (e)L 3 -C(O)O- or C(O)N(R N )-, where R N is hydrogen or C 1-6 alkyl;

[0865] L 1 is -OP(O)(OH)O- or -OP(S)(OH)O-; and

[0866] Each G 1 Independently C 3-10 a cycloalkyl group or a 3- to 10-membered heterocyclic group; and

[0867] In some embodiments, -L'- is wherein X is O or S (eg, S).

[0868] In some embodiments, -L'- is *-G-, wherein * is a bond to ZZ; and G is C 1-10 Alkyl, optionally substituted with 1 or 2 R groups.

[0869] In embodiments of formula (X), including embodiments of formula (xa) to (xx), -L'- is -L 1 -[GL 2 ] q -GL 3 -*, where * is the bond to ZZ;

[0870] In some embodiments, -L'- is -L 1 -GL 3 -*, where * is the bond to ZZ;

[0871] In some embodiments, -L'- is -L 1 -[GL 2 ] q -GL 3 -*, where * is the bond to ZZ;

[0872] q is 0, 1, 2, 3, 4, or 5;

[0873] L 1 is a bond or -BA-;

[0874] Each L 2 independently -ABA-;

[0875] L 3 is a bond or -ABA-;

[0876] Each A is independently a bond, -O-, -S- or -N(R N )-;

[0877] Each B is independently a bond, CH2, C(O), C(S), C(NR N ), S(O), S(O)2, P(O)(OH), P(S)(OH) or P(S)(SH);

[0878] Each R N are independently hydrogen or C 1-6 alkyl; and

[0879] Each G is independently C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10alkynyl, each of which is optionally substituted with 1, 2, 3, or 4 R groups.

[0880] In some embodiments, -L'- is -L 1 -[GL 2 ] q -G-*, where * is the bond to ZZ;

[0881] q is 0, 1, 2, or 3;

[0882] L 1 is a bond or -BA-;

[0883] Each L 2 are independently a bond, C(O)O, OC(O), C(O)(NR N )、N(R N )C(O), SO2N(R N )、N(R N )SO2, OP(O)(OH), OP(S)(OH), P(O)(OH)O, P(S)(OH)O, OP(O)(OH)O or OP(S)(OH)O, wherein each R N are independently hydrogen or C 1-6 alkyl; and

[0884] Each G is independently C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 alkynyl, each of which is optionally substituted with 1 or 2 R groups.

[0885] In some embodiments, -L'- is -L 1 -[GL 2 ] q -G-*, where * is the bond to ZZ;

[0886] q is 0, 1, 2, or 3;

[0887] L 1 is a bond or -BA-;

[0888] Each L 2 are independently a bond, C(O)O, OC(O), C(O)(NR N )、N(R N )C(O), OP(O)(OH)O or OP(S)(OH)O, wherein each R N are independently hydrogen or C 1-6 alkyl;

[0889] Each G is independently C 1-10 Alkyl or C 2-10 alkenyl, each of which is optionally substituted with 1 or 2 R groups.

[0890] In some embodiments, -L'- is -[GL 2 ] q -G-*, where * is a bond to ZZ; and

[0891] (a) q is 0, 1, 2 or 3 (e.g., q is 0, 1 or 2; or 0 or 1; or 0; or 1; or 2);

[0892] Each L 2 are independently C(O)O or OC(O);

[0893] Each G is independently C 1-10 alkyl, each of which is optionally substituted with 1 or 2 R groups;

[0894] (b) q is 0, 1, 2 or 3 (e.g., q is 0, 1 or 2; or 0 or 1; or 0; or 1; or 2);

[0895] Each L 2 are independently C(O)(NR N ) or N(R N )C(O), where each R N are independently hydrogen or C 1-6 alkyl; and

[0896] Each G is independently C 1-10 alkyl, each of which is optionally substituted with 1 or 2 R groups;

[0897] (c) q is 0, 1, 2 or 3 (e.g., q is 0, 1 or 2; or 0 or 1; or 0; or 1; or 2);

[0898] Each L 2 is independently OP(O)(OH)O or OP(S)(OH)O (e.g., each is OP(O)(OH)O); and each G is independently C 1-10 alkyl, each of which is optionally substituted with 1 or 2 R groups;

[0899] or

[0900] (d) q is 0, 1, 2 or 3 (e.g., q is 0, 1 or 2; or 0 or 1; or 0; or 1; or 2);

[0901] Each L 2 is a bond; and each G is independently C 1-10 alkyl, each of which is optionally substituted with 1 or 2 R groups.

[0902] In some embodiments, -L'- is -C 2-30 Alkyl-*, where * is a bond to ZZ, such as -C 5-20 Alkyl-* or -C10-20 alkyl-*.

[0903] In some embodiments, -L'- is -C(O)-C 2-30 Alkyl-*, where * is a bond to ZZ, such as -C(O)-C 5-20 Alkyl-* or -C(O)-C 10-20 alkyl-*.

[0904] L′-R T Example

[0905] In some embodiments, -L'-R T for in

[0906] * is the key connected to ZZ;

[0907] L 1 is a bond, C(O), C(S), S(O)2, P(O)(OH) or P(S)(OH); and

[0908] R T1 As defined for formula (X).

[0909] In some embodiments, -L'-R T for

[0910] in

[0911]

[0912] * is the key connected to ZZ;

[0913] L 1 is a bond, C(O), C(S), S(O)2, P(O)(OH) or P(S)(OH); and R T1 As defined for formula (X).

[0914] In some embodiments, -L'-R T for

[0915]

[0916] Where * is the bond to ZZ;

[0917] L 1 is a bond, C(O), C(S), S(O)2, P(O)(OH) or P(S)(OH);

[0918] R is -O(R a ) or -C 1-6 Alkyl-O(R a ), where R a is hydrogen or a hydroxy protecting group; and

[0919] R T1 As defined for formula (X).

[0920] In some embodiments, -L'-R T for

[0921]

[0922] Where * is the bond to ZZ;

[0923] L 1 is a bond, C(O), C(S), S(O)2, P(O)(OH) or P(S)(OH);

[0924] R is -O(R a ) or -C 1-6 Alkyl-O(R a ), where R a is hydrogen or a hydroxy protecting group; and

[0925] R T1 As defined for formula (X).

[0926] In some embodiments, -L'-R T for

[0927]

[0928] Where * is the bond to ZZ;

[0929] L 1 is a bond, C(O), C(S), S(O)2, P(O)(OH) or P(S)(OH);

[0930] R is -O(R a ) or -C 1-6 Alkyl-O(R a ), where R a is hydrogen or a hydroxy protecting group; and

[0931] R T1 As defined for formula (X).

[0932] In some embodiments, -L'-R T for

[0933]

[0934] in

[0935] * is the key connected to ZZ;

[0936] q is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 (e.g., an integer from 1 to 8, an integer from 1 to 5 or an integer from 1 to 3);

[0937] each L 2

[0938] (i) is independently selected from the group consisting of: -C(O)O-, -OC(O)-, -C(O)N(R N )-, -N(R N )C(O)-, -OC(O)O-, -OC(O)N(R N )-, -N(R N )C(O)O-, -N(R N )C(O)N(R N )-, -OP(O)(OH)O-, -OP(S)(OH)O-, -O- and -N(R N )-, where each R <000095​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​)C(O)-, -O- and -N(R N )-, where each R N is independently hydrogen or C 1-6 alkyl; or

[0942] (v) is independently selected from the group consisting of: -C(O)N(R N )-, -N(R N )C(O)-, -O- and -N(R N )-, where each R N is independently hydrogen or C 1-6 alkyl;

[0943] (vi) is independently selected from the group consisting of: -C(O)N(R N )- and -N(R N )C(O), where each R N is independently hydrogen or C 1-6 alkyl;

[0944] L 1 is a bond, C(O), C(S), S(O)2, P(O)(OH) or P(S)(OH); and R T1 is as defined for formula (X).

[0945] In some embodiments, -L'-R T is

[0946] <00​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​Independently hydrogen or C 1-6 alkyl; or

[0952] (ii) independently selected from the group consisting of: -C(O)O-, -OC(O)-, -C(O)N(R N ), -, -N(R N ), -OC(O)N(R N ), -, -N(R N ), -N(R N ), -N(R N ), -, -O- and -N(R N [[ID=1​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​

[0957] L 1 is a key, C(O), C(S), S(O)2, P(O)(OH) or P(S)(OH); and

[0958] R T1 as defined for formula (X). In some embodiments, -L'-R T is

[0959]

[0960] where

[0961] * is a bond connected to ZZ;

[0962] q is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 (for example, an integer from 1 to 8, an integer from 1 to 5 or an integer from 1 to 3);

[0963] each L 2

[0964] (i) is independently selected from the group consisting of: -C(O)O-, -OC(O)-, -C(O)N(R N )-, -N(R N )C(O)-, -OC(O)O-, -OC(O)N(R N )-, -N(R N )C(O)O-, -N(R N )C(O)N(R N )-, -OP(O)(OH)O-, -OP(S)(OH)O-, -O- and -N(R N )-, where each R N is independently hydrogen or C 1-6 alkyl; or

[0965] (ii) is independently selected from the group consisting of: -C(O)O-, -OC(O)-, -C(O)N(R N )-, -N(R N )C(O)-, -OC(O)N(R N )-, -N(R N )C(O)O-, -N(R N )C(O)N(R N )-, -O- and -N(R N )-, where each R N is independently hydrogen or C 1-6 alkyl; or

[0966] (iii) is independently selected from the group consisting of: -C(O)N(R N)-, -N(R N )C(O)-, -OC(O)N(R N )-, -N(R N )C(O)O-, -N(R N )C(O)N(R N )-, -O- and -N(R N )-, where each R N is independently hydrogen or C 1-6 alkyl; or

[0967] (iv) is independently selected from the group consisting of: -C(O)O-, -OC(O)-, -C(O)N(R N )-, -N(R N )C(O)-, -O- and -N(R N )-, where each R N is independently hydrogen or C 1-6 alkyl; or

[0968] (v) is independently selected from the group consisting of: -C(O)N(R N )-, -N(R N )C(O)-, -O- and -N(R N )-, where each R N is independently hydrogen or C 1-6 alkyl;

[0969] (vi) is independently selected from the group consisting of: -C(O)N(R N )- and -N(R N )C(O), where each R N is independently hydrogen or C 1-6 ​​​​​​​​​​​​​​​​​​​​​​​​​​​​​

[0977] (i) independently selected from the group consisting of: -C(O)O-, -OC(O)-, -C(O)N(R N )-, -N(R N )C(O)-, -OC(O)O-, -OC(O)N(R N )-, -N(R N )C(O)O-, -N(R N )C(O)N(R N )-, -OP(O)(OH)O-, -OP(S)(OH)O-, -O- and -N(R N )-, where each R N is independently hydrogen or C[[ID=1​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​

[0981] (v) is independently selected from the group consisting of: -C(O)N(R N )-, -N(R N )C(O)-, -O- and -N(R N ), wherein each R N is independently hydrogen or C 1-6 alkyl;

[0982] (vi) is independently selected from the group consisting of: -C(O)N(R N )- and -N(R N )C(O), wherein each R N is independently hydrogen or C 1-6 alkyl;

[0983] L 1 is a bond, C(O), C(S), S(O)2, P(O)(OH) or P(S)(OH);

[0984] R is -O(R a ) or -C 1-6 alkyl-O(R a ), wherein R a is hydrogen or a hydroxy protecting group; and R T1 is as defined for formula (X).

[0985] In some embodiments, -L'-R T is

[0986]

[0987] where * is a bond connected to ZZ;

[0988] q is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 (for example, an integer from 1 to 8, an integer from 1 to 5 or an integer from 1 to 3);

[0989] each L 2

[0990] (i) is independently selected from the group consisting of: -C(O)O-, -OC(O)-, -C(O)N(R N )-, -N(R N )C(O)-, -OC(O)O-, -OC(O)N(R N )-, -N(R N )C(O)O-, -N(R N )C(O)N(R N )-, -OP(O)(OH)O-, -OP(S)(OH)O-, -O- and -N(R N ), wherein each R NIndependently hydrogen or C 1-6 alkyl; or

[0991] (ii) Independently selected from the group consisting of: -C(O)O-, -OC(O)-, -C(O)N(R N )-, -N(R N )C(O)-, -OC(O)N(R N )-, -N(R N )C(O)O-, -N(R N )C(O)N(R N )-, -O- and -N(R N )-, wherein each R N is independently hydrogen or C 1-6 alkyl; or

[0992] (iii) Independently selected from the group consisting of: -C(O)N(R N ]>)-, -N(R N [[ID=I30]])C(O)-, -OC(O)N(R N )-, -N(R N )C(O)O-, -N(R N )C(O)N(R N )-, -O- and -N(R N )-, wherein each R N is independently hydrogen or C 1-6 alkyl; or

[0993] (iv) Independently selected from the group consisting of: -C(O)O-, -OC(O)-, -C(O)N(R N )-, -N(R N )C(O)-, -O- and -N(R N )-, wherein each R N is independently hydrogen or C 1-6 alkyl; or

[0994] (v) Independently selected from the group consisting of: -C(O)N(R N )-, -N(R N )C(O)-, -O- and -N(R N )-, wherein each R N is independently hydrogen or C 1-6 alkyl;

[0995] (vi) Independently selected from the group consisting of: -C(O)N(R N )- and -N(R N )C(O), wherein each R N is independently hydrogen or C 1-6 alkyl;

[0996] L 1 is a key, C(O), C(S), S(O)2, P(O)(OH) or P(S)(OH);

[0997] R is -O(R a ) or -C 1-6 alkyl-O(R a ), where R a is hydrogen or a hydroxyl protecting group; and

[0998] R T1 is as defined for formula (X).

[0999] In some embodiments, -L'-R T is

[1000] [[ID=3--]]

[1001] where * is the bond connected to ZZ;

[1002] q is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 (e.g., an integer from 1 to 8, an integer from 1 to 5 or an integer from 1 to 3);

[1003] Each L 2

[1004] (i) is independently selected from the group consisting of: -C(O)O-, -OC(O)-, -C(O)N(R N ), -N(R N )C(O)-, -OC(O)O-, -OC(O)N(R N ), -N(R N )C(O)O-, -N(R N )C(O)N(R N ), -OP(O)(OH)O-, -OP(S)(OH)O-, -O- and -N(R N ), where each R N is independently hydrogen or C 1-6 alkyl; or

[1005] (ii) is independently selected from the group consisting of: -C(O)O-, -OC(O)-, -C(O)N(R N ), -N(R N )C(O)-, -OC(O)N(R N ), -N(R N )C(O)O-, -N(R N )C(O)N(R N ), -O- and -N(R<-- N)-, wherein each R N is independently hydrogen or C 1-6 alkyl; or

[1006] (iii) is independently selected from the group consisting of: -C(O)N(R N )-, -N(R N )C(O)-, -OC(O)N(R N )-, -N(R N )C(O)O-, -N(R N )C(O)N(R N )-, -O- and -N(R N )-, wherein each R N [[ID=2)) is independently hydrogen or C 1-6 alkyl; or

[1007] (iv) is independently selected from the group consisting of: -C(O)O-, -OC(O)-, -C(O)N(R N )-, -N(R N )C(O)-, -O- and -N(R N )-, wherein each R N [[ID=3)) is independently hydrogen or C 1-6 alkyl; or

[1008] (v) is independently selected from the group consisting of: -C(O)N(R N )-, -N(R N )C(O)-, -O- and -N(R N )-, wherein each R N is independently hydrogen or C 1-6 alkyl;

[1009] (vi) is independently selected from the group consisting of: -C(O)N(R) N )- and -N(R N )C(O), wherein each R N z is independently hydrogen or C 1-6 alkyl;

[1010] L 1 is a bond, C(O), C(S), S(O)2, P(O)(OH) or P(S)(OH);

[1011] R is -O(R a ) or -C 1-6 q alkyl-O(R a ), wherein R a is hydrogen or a hydroxy protecting group; and

[1012] R T1 is as defined for formula (X). In some embodiments, -L'-RT For

[1013]

[1014] where * is the key connected to ZZ;

[1015] q is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 (for example, an integer from 1 to 8, an integer from 1 to 5, or an integer from 1 to 3);

[1016] each L 2

[1017] (i) is independently selected from the group consisting of: -C(O)O-, -OC(O)-, -C(O)N(R N )-, -N(R N )C(O)-, -OC(O)O-, -OC(O)N(R N )-, -N(R N )C(O)O-, -N(R N )C(O)N(R N )-, -OP(O)(OH)O-, -OP(S)(OH)O-, -O- and -N(R N )-, where each R N is independently hydrogen or C 1-6 alkyl; or

[1018] (ii) is independently selected from the group consisting of: -C(O)O-, -OC(O)-, -C(O)N(R N )-, -N(R N )C(O)-, -OC(O)N(R N )-, -N(R N )C(O)O-, -N(R N )C(O)N(R N )-, -O- and -N(R N )-, where each R N is independently hydrogen or C 1-6 alkyl; or

[1019] (iii) is independently selected from the group consisting of: -C(O)N(R[[ID=六十]] N )-, -N(R N )C(O)-, -OC(O)N(R N )-, -N(R N )C(O)O-, -N(R N )C(O)N(R N )-, -O- and -N(R N )-, where each R N is independently hydrogen or C1-6 alkyl; or

[1020] (iv) independently selected from the group consisting of: -C(O)O-, -OC(O)-, -C(O)N(R N ), -N(R N )C(O)-, -O-, and -N(R N ), where each R N is independently hydrogen or C 1-6 alkyl; or

[1021] (v) independently selected from the group consisting of: -C(O)N(R N ), -N(R N )C(O)-, -O-, and -N(R N ), where each R N is independently hydrogen or C 1-6 alkyl;

[1022] (vi) independently selected from the group consisting of: -C(O)N(R N ), and -N(R N )C(O), where each R N is independently hydrogen or C 1-6 alkyl;

[1023] L 1 is a bond, C(O), C(S), S(O)2, P(O)(OH), or P(S)(OH);

[1024] R is -O(R a ) or -C 1-6 alkyl-O(R a ), where R a is hydrogen or a hydroxyl protecting group; and

[1025] R T1 is as defined for formula (X).

[1026] In some embodiments, -L'-R T is

[1027]

[1028] where * is a bond connected to ZZ;

[1029] L 1 is a bond, C(O), C(S), S(O)2, P(O)(OH), or P(S)(OH);

[1030] R is -O(R a ) or -C 1-6 alkyl-O(R a ), where Ra is hydrogen or a hydroxy protecting group; and

[1031] R T1 As defined for formula (X).

[1032] R L Example, formula (X)

[1033] In one embodiment of Formula (X) or a salt thereof, R L -N(R 3 )(R 4 )、-O(R 5 )、-S(R 5 ) or -R 5 , where R 5 is according to one of formulas (xa) to (xs):

[1034]

[1035]

[1036]

[1037] where R P3 is hydrogen or a hydroxyl protecting group and L, ZZ, L' and R T1 As defined for Formula (X) or any Examples herein.

[1038] In another embodiment, R L is according to one of formulas (xi-a) to (xi-n):

[1039]

[1040]

[1041] where R P3 is a hydrogen or hydroxyl protecting group, and L, ZZ, L' and R T1 As defined for Formula (X) or any Examples herein.

[1042] L embodiment, formula (X)

[1043] In some embodiments of any of Formula (X) and any of its embodiments, L is -L 1 -[GL 2 ] q -GL 3 -*, where * is the bond to ZZ.

[1044] In another embodiment, wherein L is -L 1 -[GL 2 ]q -GL 3 -*, where q is 0, 1, 2, 3, 4, or 5.

[1045] In another embodiment, L is -L 1 -[GL 2 ] q -GL 3 -*, where q is 0, 1, 2, 3, or 4

[1046] In another embodiment, L is -L 1 -[GL 2 ] q -GL 3 -*, where q is 0, 1, 2, or 3.

[1047] In another embodiment, L is -L 1 -[GL 2 ] q -GL 3 -*, where q is 0, 1, or 2.

[1048] In another embodiment, L is -L 1 -[GL 2 ] q -GL 3 -*, where q is 1, 2, 3, 4, or 5.

[1049] In another embodiment, L is -L 1 -[GL 2 ] q -GL 3 -*, where q is 1, 2, 3, or 4.

[1050] In another embodiment, L is -L 1 -[GL 2 ] q -GL 3 -*, where q is 1, 2, or 3.

[1051] In another embodiment, L is -L 1 -[GL 2 ] q -GL 3 -*, where q is 1 or 2.

[1052] In another embodiment, L is -L 1 -[GL 2 ] q -GL 3 -*, wherein q is 4. In another embodiment, L is -L 1 -[GL 2 ]q -G-L 3 -*, where q is 3. In another embodiment, L is -L 1 -[G-L 2 q -G-L 3 -*, where q is 2.

[1053] In another embodiment, L is -L 1 -G-L 2 -G-L 3 -*. In another embodiment, L is -L 1 -G-L 3 -*. In another embodiment, L is -G-L 3 -*. In another embodiment, L is -L 1 -G-*. In another embodiment, L is -G-*.

[1054] In some embodiments, each instance of A-B-A- is independently selected from the group consisting of: -C(O)O-, -OC(O)-, -C(O)N(R N )-, -N(R N )C(O)-, -OC(O)O-, -OC(O)N(R N )-, -N(R N )C(O)O-, -N(R N )C(O)N(R N )-, -OP(O)(OH)O-, -OP(S)(OH)O-, -O- and -N(R N )-, where each R N is independently hydrogen or C 1-6 alkyl.

[1055] In some embodiments, each instance of A-B-A- is independently selected from the group consisting of: -C(O)O-, -OC(O)-, -C(O)N(R N )-, -N(R N )C(O)-, -OC(O)N(R N )-, -N(R N )C(O)O-, -N(R N )C(O)N(R N )-, -O- and -N(R N )-, where each R N is independently hydrogen or C 1-6 alkyl. <​​​​​N ) C(O)-, -OC(O)N(R N ) -, -N(R N ) C(O)O-, -N(R N ) C(O)N(R N ) -, -O- and -N(R N ) -, where each R N is independently hydrogen or C 1-6 alkyl.

[1057] In some embodiments, each instance of A-B-A- is independently selected from the group consisting of: -C(O)O-, -OC(O)-, -C(O)N(R N ) -, -N(R N ) C(O)-, -O- and -N(R N ) -, where each R N is independently hydrogen or C 1-6 alkyl.

[1058] In some embodiments, each instance of A-B-A- is independently selected from the group consisting of: -C(O)N(R N ) -, -N(R N ) C(O)-, -O- and -N(R N ) -, where each R N is independently hydrogen or C 1-6 alkyl.

[1059] In some embodiments,

[1060] D and F are each independently a bond, C 1-10 alkyl, C 2-10 alkenyl or C 2-10 alkynyl, each of which is optionally substituted with 1, 2, 3 or 4 R groups; and

[1061] E is C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl, C 3-10 cycloalkyl, a 3- to 10-membered heterocyclic group, aryl or heteroaryl, each of which is optionally substituted with 1, 2, 3 or 4 R groups.

[1062] In some embodiments,

[1063] D and F are each independently a bond or C 1-10 alkyl optionally substituted with 1, 2, 3 or 4 R groups; and [[ID=7x]]

[1064] E is C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl, C3-10 cycloalkyl, 3- to 10-membered heterocyclyl, aryl, or heteroaryl, each of which is optionally substituted with 1, 2, 3, or 4 R groups.

[1065] In some embodiments, each G is independently C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, aryl, or heteroaryl, each of which is optionally substitut...

Claims

1. A compound of formula (X), or a salt thereof, wherein: Y is O, N(H), S or CH2; R 1 is hydrogen or C 1-6 Alkyl groups (e.g., methyl); R Y for in m is 0, 1, 2, 3 or 4; and Each R 2 R is independently R, or two R on adjacent carbon atoms 2 The group, together with the atoms to which it is bonded, forms a fused 4- to 8-membered ring, which is optionally substituted with 1, 2, 3, or 4 groups independently selected from the group consisting of R and a nitrogen protecting group; and R L -N(R 3 )(R 4 )、-O(R 5 )、-S(R 5 ) or -R 5 ,in R 3 and R 4 Is any of the following: (iii)R 3 is hydrogen or C 1-6 Alkyl, and R 4 R 5 ;or (iv)R 3 and R 4 Together with the nitrogen atom to which it is attached, it forms the 5 substituted 4- to 8-membered monocyclic heterocyclic group; and R 5 -L-ZZ-L′-R T ,in L and L' are independently -L 1 -[GL 2 ] q -GL 3 -*,in * is the key connected to ZZ; q is 0 or an integer selected from 1 to 25; (e.g., 1 to 20 or 1 to 15); L 1 is a bond or -BA-; Each L 2 independently -ABA-; L 3 is a bond or -ABA-; Each G is independently -DEF-, wherein D, E and F are independently a bond, C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, aryl, or heteroaryl, each of which is optionally substituted with 1, 2, 3, or 4 R groups; Each A is independently a bond, -O-, -S- or -N(R N )-; Each B is independently a bond, CH2, C(O), C(S), C(NR N ), S(O), S(O)2, P(O)(OH), P(S)(OH) or P(S)(SH); Each R N are independently hydrogen or C 1-6 Alkyl, or the two R in the -ABA- group N Together with the atoms to which they are attached, they form a 4- to 8-membered heterocyclic group; and ZZ is -A'-B'-A'- or a linking group formed by a reactive pair, wherein Each A' is independently a bond, -O-, -S- or -N(R N3 )-; Each B' is independently a bond, CH2, C(O), C(S), C(NR N3 ), -C=N-, S(O), S(O)2, P(O)(OH), P(S)(OH) or P(S)(SH); and Each R N3 are independently hydrogen or G 1-6 Alkyl, or the two R in the -A'-B'-A'- group N3 Together with the atoms to which they are attached, they form a 4- to 8-membered heterocyclic group; R T -G 0 -OR T1 ,in G 0 Does not exist or is -D 0 -E 0 -F 0 -, where D 0 、E 0 and F 0 independently for the bond, C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, aryl, or heteroaryl, each of which is optionally substituted with 1, 2, 3, or 4 R groups; and R T1 is hydrogen, a hydroxyl protecting group, a phosphorus coupling group or -L K -S S or -L L - an oligonucleotide, wherein L K is a supporting connecting group; L L is an oligonucleotide linking group; and S S For solid support, -OR SS or -N(R SS )2 or hydrogen, wherein each R SS are independently hydrogen or C 1-6 alkyl, and Each R group is independently selected from the group consisting of: R', C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-8 Cycloalkyl, heterocyclyl, aryl, heteroaryl, C 3-8 Cycloalkyl C 1-6 Alkyl, heterocyclic C 1-6 Alkyl, aryl C 1-6 Alkyl, heteroaryl C 1-6 alkyl, each of which, except R′, is optionally substituted with 1, 2, or 3 R′ groups, wherein Each R' is independently halogen, cyano, azido, nitro, -N(R b )2、-O(R a )、-S(R 0 )、-C(O)OR 0 、C(O)R 0 、-C(O)N(R 0 )2、-C(NR 0 )OR 0 、-C(NR 0 )R 0 、-C(NR 0 )N(R 0 )2、-C(S)OR 0 、-C(S)R 0 、-C(S)N(R 0 )2、-S(O)2R 0 、-S(O)2OR 0 、-S(O)2N(R 0 )2、-N(R 0 )C(O)OR 0 、-N(R 0 )C(O)R 0 、-NN(R 0 )C(O)N(R 0 )2、-N(R 0 )S(O)2R 0 、-N(R 0 )S(O)2OR 0 、-N(R 0 )S(O)2N(R 0 )2、-OC(O)OR 0 、-OC(O)R 0 、-OC(O)N(R 0 )2、-OS(O)2R 0 、-OS(O)2OR 0 、-OS(O)2N(R 0 )2 or -SC(O)R 0 ,in Each R 0 are independently hydrogen or C 1-6 Alkyl; each R a are independently hydrogen, C 1-6 an alkyl or hydroxy protecting group; and each R b are independently hydrogen, C 1-6 Alkyl or nitrogen protecting group The proviso is that in each -DEF- group, at least one of D, E, and F is not a bond; and R L Not N-morpholinyl.

2. The compound of claim 1, wherein R Y for 3. The compound of claim 1, wherein R Y for wherein p is 0, 1, 2, 3 or 4; and each R 21 are independently selected from the group consisting of R and a nitrogen protecting group.

4. The compound of claim 1, wherein R Y for where R P A nitrogen protecting group.

5. The compound of claim 1, wherein R Y for where R P A nitrogen protecting group.

6. A compound as claimed in any one of claims 1 to 5, wherein -L'-R T for Where * is the bond to ZZ; and L 1 is a bond, C(O), C(S), S(O)2, P(O)(OH) or P(S)(OH).

7. A compound as claimed in any one of claims 1 to 5, wherein -L'-R T for Where * is the bond to ZZ; L 1 is a bond, C(O), C(S), S(O)2, P(O)(OH) or P(S)(OH); and R is -O(R a ) or -C 1-6 Alkyl-O(R a ), where R a is a hydrogen or hydroxyl protecting group.

8. A compound as claimed in any one of claims 1 to 5, wherein -R L for where R P3 is a hydrogen or hydroxyl protecting group.

9. The compound of claim 8, wherein R L for 10. The compound of claim 9, wherein R P3 is an optionally substituted trityl group.

11. The compound according to any one of claims 1 to 8, wherein -L'- is -L 1 -GL 3 -*, where * is the bond to ZZ.

12. The compound of claim 10, wherein -L'- is -C(O)-C 2-30 alkyl-*.

13. The compound of any one of claims 1 to 12, having the following structure:

14. The compound of any one of claims 1 to 13, wherein L is -L 1 -[GL 2 ] q -GL 3 -*, where q is 0, 1, 2, 3, 4, or 5.

15. The compound of any one of claims 1 to 13, wherein L is -L 1 -G-*, where * is the bond to ZZ; G is C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, aryl, or heteroaryl, each of which is optionally substituted with 1, 2, 3, or 4 R groups; L 1 is -BA-, where A is a bond, -O-, -S- or -N(R N )-, where each R N are independently hydrogen or C 1-6 alkyl; and B is a bond, C(O), C(S), S(O)2, P(O)(OH) or P(S)(OH).

16. A compound as claimed in any one of claims 1 to 13, wherein L is Where * is a bond to ZZ; k is an integer from 1 to 10; L 1 for bonds, C(O), C(S), C(NR N ), S(O)2, P(O)(OH) or P(S)(OH); and R N is hydrogen or C 1-6 alkyl.

17. The compound of any one of claims 1 to 13, wherein L is a group selected from: (a) Wherein * is a bond to ZZ; t is an integer from 0 to 10; (b) L is wherein * is a bond to ZZ, t is an integer from 0 to 10; a is an integer from 1 to 3; and s and s′ are each independently an integer from 1 to 24; (c) wherein * is a bond to ZZ; a is 1, 2 or 3; and each of s, s′ and s″ is independently an integer from 1 to 24; (d) wherein * is a bond to ZZ; and s, s′, and s″ are independently integers from 1 to 24; (e) wherein * is a bond to ZZ, and each of s, s′, and s″ is independently an integer from 1 to 24; and (f) wherein * is a bond to ZZ; s and k are independently integers from 1 to 20; and w is an integer from 1 to 10.

18. A compound as claimed in any one of claims 1 to 13, wherein L is wherein * is a bond to ZZ, and w is an integer from 1 to 20.

19. A compound as claimed in any one of claims 1 to 13, wherein L is wherein * is a bond to ZZ; and k is an integer from 1 to 10.

20. The compound of any one of claims 1 to 19, wherein ZZ comprises a group selected from the group consisting of:

21. The compound of any one of claims 1 to 19, wherein ZZ is -A'-B'-A'-, wherein Each A' is independently a bond, -O-, -S- or -N(R N3 )-, where R N3 are independently hydrogen or C 1-6 Alkyl, and Each B' is independently CH2, C(O), C(S), S(O)2, P(O)(OH) or P(S)(OH).

22. The compound of any one of claims 1 to 19, wherein ZZ is -A'-B'- or -B'-A'-, wherein Each A' is independently -O- or -N(R N3 )-, where R N3 are independently hydrogen or C 1-6 alkyl, each B′ is independently CH 2 , C(O), S(O) 2 , P(O)(OH) or P(S)(OH); and Each R N3 are independently hydrogen or C 1-6 alkyl.

23. The compound of any one of claims 1 to 19, wherein ZZ is -CH2O-, -OCH2-, -SS-, -C=N-, -C=NO-, -C=NN(R N3 )-, -N=C-, -ON=C-, -N(R N3 )-N=C-、-C(O)N(R N3 )-、-N(R N3 )C(O)-, -C(O)O-, -OC(O)-, -OC(O)N(R N3 )-、-N(R N3 )C(O)O-、-N(R N3 )C(O)N(R N3 )-、-S(O)2N(R N3 )-、-N(R N3 )S(O)2-, -OP(O)(OH)O-, -OP(S)(OH)O-, -OP(O)(OH)-, -OP(S)(OH)-, -P(O)(OH)O- or -P(S)(OH)O-, where R N3 are independently hydrogen or C 1-6 alkyl.

24. The compound of any one of claims 1 to 19, wherein ZZ is -C(O)N(R N3 )-or-N(R N3 )C(O)-, where R N3 are independently hydrogen or C 1-6 alkyl.

25. The compound of any one of claims 1 to 19, wherein ZZ is -OP(O)(OH)O-, -OP(S)(OH)O-, -OP(O)(OH)-, -OP(S)(OH)-, -P(O)(OH)O-, or -P(S)(OH)O-.

26. A compound as claimed in any one of claims 1 to 25, wherein R T1 -L L - oligonucleotide, wherein L L is a divalent linker attached to the 3' end of the oligonucleotide, the 5' end of the oligonucleotide, or to an internal 2' or 3' position on an internal nucleotide.

27. The compound of claim 26, wherein L L Attached to the oxygen atom on the nucleoside and is P(O)(OH)-, -P(S)(OH)-, or -P(S)(SH).

28. The compound of claim 27, wherein LL is P(O)(OH)-.

29. The compound of claim 27, wherein LL is P(S)(OH)-.

30. A compound as claimed in any one of claims 1 to 25, wherein R T R T1 , where R T1 -L L - oligonucleotide, wherein L L The oxygen atom attached to the nucleoside of the oligonucleotide, L L is a bond, and the nucleoside has formula (Xf), wherein B is an optionally modified nucleobase and * represents a bond to ZZ.

31. The compound of claim 30, wherein -L'- is -C 4-10 Alkyl-*, where * is a bond to ZZ.

32. The compound of claim 30, wherein the nucleoside has a formula selected from the group consisting of: in B is an optionally modified nucleobase; each n is independently 0 or an integer selected from 1 to 10; and Each m is independently an integer selected from 1 to 20.

33. A compound as described in any one of claims 1 to 25, wherein R T1 -L L -oligonucleotide and binds to the 5' end of the oligonucleotide.

34. The compound of claim 33 having the formula: wherein Y′ is O or S.

35. A compound as described in any one of claims 1 to 25, wherein R T1 -L L -oligonucleotide and binds to the 3' end of the oligonucleotide.

36. The compound of claim 35 having the formula: wherein Y′ is O or S.

37. The compound of claim 35 having the formula: Where Y′ is O or S, R 1 is hydrogen or C 1-6 Alkyl, and R P is a hydrogen or nitrogen protecting group.

38. The compound of claim 35 having the formula: Where Y′ is O or S, R 1 is hydrogen or C 1-6 Alkyl, and R P is a hydrogen or nitrogen protecting group.

39. The compound of claim 35 having the formula: Where Y′ is O or S, R 1 is hydrogen or C 1-6 Alkyl, and R P is a hydrogen or nitrogen protecting group.

40. The compound of claim 35 having the formula: wherein each m is independently an integer selected from 1 to 10; Y′ is O or S, R 1 is hydrogen or C 1-6 Alkyl, and R P is a hydrogen or nitrogen protecting group.

41. The compound of claim 35 having the formula: Where Y′ is O or S, R 1 is hydrogen or C 1-6 Alkyl, and R P is a hydrogen or nitrogen protecting group.

42. The compound of claim 41, wherein R P is hydrogen, and R 1 For hydrogen.

43. The compound of claim 41 or 42, wherein Y' is O.

44. The compound of claim 41 or 42, wherein Y' is S.

45. A compound of formula (XV), (F-ZZ-) x D-TR T (XV) or a salt thereof, wherein x is 2, 3, 4, 5, 6, 7, or 8; T is a divalent linking group; Δ is a branched group; Each ZZ is independently -A'-B'-A'- or a linker formed from a first reactive pair, wherein Each A' is independently a bond, -O-, -S- or -N(R N3 )-; Each B' is independently a bond, CH2, C(O), C(S), C(NR N3 ), -C=N-, S(O), S(O)2, P(O)(OH), P(S)(OH) or P(S)(SH); and Each R N3 are independently hydrogen or C 1-6 Alkyl, or the two R in the -A'-B'-A'- group N3 Together with the atoms to which they are attached, they form a 4- to 8-membered heterocyclic group; Z 0 is a member of a second reactive pair; and R T -G 0 -OR T1 ,in G 0 -D 0 -E 0 -F 0 -,in D 0 、E 0 and F 0 independently for the bond, C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, aryl, or heteroaryl, each of which is optionally substituted with 1, 2, 3, or 4 R groups; and R T1 is hydrogen, a hydroxyl protecting group, a phosphorus coupling group or -L K -S S or -L L - an oligonucleotide, wherein L K is a supporting connecting group; S S For solid support, -OR SS or -N(R SS )2 or hydrogen, wherein each R SS are independently hydrogen or C 1-6 alkyl; and L L is an oligonucleotide linking group; and each Φ is a compound of formula (XII), in: Y is O, N(H), S or CH2; R 1 is hydrogen or C 1-6 Alkyl groups (e.g., methyl); R Y for in m is 0, 1, 2, 3 or 4; and Each R 2 R is independently R, or two R on adjacent carbon atoms 2 The group, together with the atoms to which it is bonded, forms a fused 4- to 8-membered ring, which is optionally substituted with 1, 2, 3, or 4 groups independently selected from the group consisting of R and a nitrogen protecting group; and R L -N(R 3 )(R 4 )、-O(R 5 )、-S(R 5 ) or -R 5 ,in R 3 and R 4 Is any of the following: (i)R 3 is hydrogen or C 1-6 Alkyl, and R 4 R 5 ;or (ii)R 3 and R 4 Together with the nitrogen atom to which it is attached, it forms the 5 substituted 4- to 8-membered monocyclic heterocyclic group; and R 5 -L-*, where L is -L 1 -[GL 2 ] q -GL 3 -*, * is the bond to ZZ; and q is 0 or an integer selected from 1 to 25; (e.g., 1 to 20 or 1 to 15); L 1 is a bond or -BA-; Each L 2 independently -ABA-; L 3 is a bond or -ABA-; Each G is independently -DEF-, wherein D, E and F are independently a bond, C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, aryl, or heteroaryl, each of which is optionally substituted with 1, 2, 3, or 4 R groups; Each A is independently a bond, -O-, -S- or -N(R N )-; Each B is independently a bond, CH2, C(O), C(S), C(NR N ), S(O), S(O)2, P(O)(OH), P(S)(OH) or P(S)(SH); Each R N are independently hydrogen or C 1-6 Alkyl, or the two R in the -ABA- group N Together with the atoms to which it is attached, it forms a 4- to 8-membered heterocyclic group.

46. ​​The compound of claim 45 having a structure selected from the group consisting of:

47. The compound of claim 45 or 46, wherein R Y for 48. A compound as claimed in claim 45 or 46, wherein R Y for wherein p is 0, 1, 2, 3 or 4; and each R 21 are independently selected from the group consisting of R and a nitrogen protecting group.

49. A compound as claimed in claim 45 or 46, wherein R Y for where R P A nitrogen protecting group.

50. The compound of claim 45 or 46, wherein R Y for where R P A nitrogen protecting group.

51. A compound as described in any one of claims 45 to 50, wherein R T -G 0 -OR T1 , where G 0 -D 0 -E 0 -F 0 -,in D 0 and F 0 is independently a bond or C optionally substituted with 1, 2, 3 or 4 R groups 1-10 alkyl; and E 0 C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, aryl, or heteroaryl, each of which is optionally substituted with 1, 2, 3, or 4 R groups.

52. The compound of claim 51, wherein G 0 is a 3- to 10-membered heterocyclic group optionally substituted with 1 or 2 R groups.

53. The compound of claim 51, wherein G 0 for Where * indicates the bond to T, and a broken bond indicates the bond to OR T1 The bond, and R is -C 1-6 Alkyl-OR a OR a , where R a are independently hydrogen, C 1-6 Alkyl or hydroxy protecting groups.

54. The compound of claim 51, wherein G 0 for Where * indicates the bond to T, and a broken bond indicates the bond to OR T1 The bond, and R is -C 1-6 Alkyl-OR a OR a , where R a are independently hydrogen, C 1-6 Alkyl or hydroxy protecting groups.

55. The compound of claim 51, wherein G 0 for Where * indicates the bond to T, and a broken bond indicates the bond to OR T1 The bond, and R is -C 1-6 Alkyl-OR a OR a , where R a are independently hydrogen, C 1-6 Alkyl or hydroxy protecting groups.

56. The compound of any one of claims 45 to 55, wherein T is: T is a bond or **-L 6 -G 1 -[L 5 -G 1 ] q1 -L 4 -, where ** is connected to Z 0 or R T Keys; q1 is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; Each L 4 , L 5 and L 6 independently a bond or -A 1 -B 1 -A 1 -; Each G 1 Independently -D 1 -E 1 -F 1 -, where D 1 、E 1 and F 1 independently for the bond, C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, aryl, or heteroaryl, each of which is optionally substituted with 1, 2, or 3 R groups; Each A 1 are independently a bond, -O-, -S- or -N(R N1 )-; Each B 1 are independently a bond, C(O), C(S), C(NR N1 ), S(O), S(O)2, P(O)(OH), P(S)(OH) or P(S)(SH); Each R N1 are independently hydrogen or C 1-6 Alkyl, or -A 1 -B 1 -A 1 - Two R's in the group N1 Together with the atoms to which it is attached, it forms a 4- to 8-membered heterocyclic group.

57. A compound as described in any one of claims 45 to 55, wherein T is **-L 6 -G 1 -L 5 -G 1 -L 4 -, where ** is connected to Z 0 or R T Keys; L 4 and L 6 Independently -A 1 -B 1 -or-B 1 -A 1 -; Each L 5 For key or -A 1 -B 1 -A 1 -(e.g. key, -B 1 -A 1 -or-A 1 -B 1 -; or key); Each G 1 Independently C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C 3-10 Cycloalkyl, 3- to 10-membered heterocyclic group, aryl or heteroaryl (e.g., C 1-10 Alkyl or C 2-10 alkenyl); Each A 1 are independently a bond, -O-, -S- or -N(R N1 )-, where R N1 are independently hydrogen or C 1-6 alkyl; and Each B 1 is independently a bond, C(O), C(S), S(O)2, P(O)(OH) or P(S)(OH).

58. A compound as described in any one of claims 45 to 55, wherein T is **-L 6 -G 1 -L 5 -G 1 -L 4 -, where ** is connected to Z 0 or R T Keys; L 4 and L 6 Independently -B 1 -A 1 -or-A 1 -B 1 -; Each L 5 For key or -A 1 -B 1 -A 1 -(e.g. key, -B 1 -A 1 -or-A 1 -B 1 -; or key); Each G 1 Independently C 1-10 Alkyl or C 2-10 alkenyl; Each A 1 are independently a bond, -O-, -S- or -N(R N1 )-, where R N1 are independently hydrogen or C 1-6 alkyl; and Each B 1 is independently a bond, C(O), C(S), S(O)2, P(O)(OH) or P(S)(OH).

59. A compound as described in any one of claims 45 to 55, wherein T is **-C(O)-C 2-20 Alkyl-C(O)N(H)-, where ** is attached to Z 0 or R T key.

60. The compound of any one of claims 45 to 55, wherein T is **-C(O)-[CH2CH2-O] q5 -G 5 -L 4 -, where ** is connected to Z 0 or R T Keys; q5 is an integer selected from 1 to 20; L 4 -A 1 -B 1 -A 1 -,in Each A 1 are independently a bond, -O-, -S- or -N(R N1 )-, where R N1 is hydrogen or C 1-6 alkyl; Each B 1 is independently a bond, C(O), C(S), S(O)2, P(O)(OH) or P(S)(OH); G 5 C 1-10 alkyl.

61. A compound as described in any one of claims 45 to 60, wherein Δ is #-[G 2 -L 7 ] q2 -* or #-G 3 -([L 7 -G 4 ] q3 -*) y , in # is the key connected to T; y is 1, 2, 3, 4, or 5; q2 is 1, 2, 3, 4, 5, 6, 7, or 8; q3 is 0, 1, 2, 3, 4, 5, 6, 7, or 8; Each G 2 , G 3 and G 4 Independently -D 2 -E 2 -F 2 -,in D 2 、E 2 and F 2 independently for the bond, C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, aryl, or heteroaryl, each of which is optionally substituted with 1, 2, 3, 4, or 5 R B group substitution, and Among them, each G 2 and G 4 optionally containing at least one bond to ZZ (e.g., one bond to ZZ); or G 3 is N and y is 2; Each L 7 Independently -A 2 -B 2 -A 2 -;in Each A 2 are independently a bond, -O-, -S- or -N(R N2 )-; Each B 2 are independently a bond, C(O), C(S), C(NR N2 ), S(O), S(O)2, P(O)(OH), P(S)(OH) or P(S)(SH); and Each R N2 are independently hydrogen, C 1-6 Alkyl, a bond to ZZ, or -A 2 -B 2 -A 2 - Two R's in the group N2 Together with the atoms to which they are attached, they form a 4- to 8-membered heterocyclic group; and Each R B are independently halogen, cyano, azido, nitro, -N(R 10 )2、-O(R 10 )、-S(R 10 )、-C(O)OR 10 、-C(O)R 10 、-C(O)N(R 10 )2、-C(NR 10 )OR 10 、-C(NR 10 )R 10 、-C(NR 10 )N(R 10 )2、-C(S)OR 10 、-C(S)R 10 、-C(S)N(R 10 )2、-S(O)2R 10 、-S(O)2OR 10 、-S(O)2N(R 10 )2、-N(R 10 )C(O)OR 10 、-N(R 10 )C(O)R 10 、-N(R 10 )C(O)N(R 10 )2、-N(R 10 )S(O)2R 10 、-N(R 10 )S(O)2OR 10 、-N(R 10 )S(O)2N(R 10 )2、-OC(O)OR 10 、-OC(O)R 10 、-OC(O)N(R 10 )2、-OS(O)2R 10 、-OS(O)2OR 10 、-OS(O)2N(R 10 )2 or -SC(O)R 10 , where each R 10 are independently hydrogen, C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C 3-10 Cycloalkyl, 3- to 10-membered heterocyclyl, aryl or heteroaryl, with the proviso that Δ contains x bonds to ZZ.

62. The compound of claim 61, wherein Δ is #-[G 2 -L 7 ] q2 -*,in # is the key connected to T; q2 is 1, 2, 3, 4, 5, 6, 7, or 8; q3 is 0, 1, 2, 3, 4, 5, 6, 7, or 8; Each G 2 , G 3 and G 4 Independently -D 2 -E 2 -F 2 -,in D 2 、E 2 and F 2 independently for the bond, C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, aryl, or heteroaryl, each of which is optionally substituted with 1, 2, 3, 4, or 5 R B group substitution, and Among them, each G 2 optionally containing at least one bond to ZZ; Each L 7 Independently -A 2 -B 2 -A 2 -; Each A 2 are independently a bond, -O-, -S- or -N(R N2 )-; Each B 2 are independently a bond, C(O), C(S), C(NR N2 ), S(O), S(O)2, P(O)(OH), P(S)(OH) or P(S)(SH); Each R N2 are independently hydrogen, C 1-6 Alkyl, a bond to ZZ, or -A 2 -B 2 -A 2 - Two R's in the group N2 Together with the atoms to which they are attached, they form a 4- to 8-membered heterocyclic group; Each R B are independently halogen, cyano, azido, nitro, -N(R 10 )2、-O(R 10 )、-S(R 10 )、-C(O)OR 10 、-C(O)R 10 、-C(O)N(R 10 )2、-C(NR 10 )OR 10 、-C(NR 10 )R 10 、-C(NR 10 )N(R 10 )2、-C(S)OR 10 、-C(S)R 10 、-C(S)N(R 10 )2、-S(O)2R 10 、-S(O)2OR 10 、-S(O)2N(R 10 )2、-N(R 10 )C(O)OR 10 、-N(R 10 )C(O)R 10 、-N(R 10 )C(O)N(R 10 )2、-N(R 10 )S(O)2R 10 、-N(R 10 )S(O)2OR 10 、-N(R 10 )S(O)2N(R 10 )2、-OC(O)OR 10 、-OC(O)R 10 、-OC(O)N(R 10 )2、-OS(O)2R 10 、-OS(O)2OR 10 、-OS(O)2N(R 10 )2 or -SC(O)R 10 ,in Each R 10 are independently hydrogen, C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclic group, aryl or heteroaryl.

63. The compound of claim 62, wherein Δ is selected from the group consisting of: Wherein each * is a bond connected to ZZ; # is a bond connected to T, Each G 2 Independently C 1-10 Alkyl, C 2-10 Alkenyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, aryl, or heteroaryl, each of which is optionally substituted with 1, 2, 3, 4, or 5 R B Group substituted; and each L 7 Independently -A 2 -B 2 -A 2 -, where each A 2 are independently a bond, -O-, -S- or -N(R N2 )-; each B 2 is independently a bond, C(O), S(O)2, P(O)(OH) or P(S)(OH).

64. The compound of claim 63, wherein each G 2 Independently C 1-10 Alkyl, each of which is optionally substituted with 1 or 2 R B Group substitution.

65. The compound of claim 64, wherein each G 2 Independently C 1-10 alkyl.

66. A compound as described in any one of claims 62 to 65, wherein each L 7 Independently -A 2 -B 2 -A 2 -, where each A 2 are independently a bond, -O-, -S- or -N(R N2 )-;and each B 2 is independently a bond, C(O) or S(O)2, with the proviso that at least one A 2 Not a key.

67. A compound as described in any one of claims 62 to 65, wherein each L 7 Independently -A 2 -B 2 -or-B 2 -A 2 -, where each A 2 are independently -O-, -S- or -N(R N2 )-;and each B 2 is independently a bond, C(O) or S(O)2.

68. The compound of any one of claims 62 to 67, wherein Δ is selected from the group consisting of: wherein # is a bond to T and each * is a bond to a ZZ group; and each G 2 Independently C 1-10 alkyl.

69. The compound of any one of claims 62 to 67, wherein Δ is selected from the group consisting of: wherein # is a bond to T and each * is a bond to the ZZ group.

70. The compound of claim 62, wherein Δ is -#-G 3 -([L 7 -G 4 ] q3 -*) y , where # is the bond to T and * is the bond to the ZZ group.

71. The compound of claim 70, wherein Δ is selected from the group consisting of: where # is a bond to T, each * is a bond to a ZZ group, and Each L 7 selected from the group consisting of: -O-, -S-, -N(H)-, -C(O)O-, -OC(O)-, -C(O)N(H)-, -OC(O)O-, -N(H)C(O)O-, -OC(O)N(H)-, -OP(O)(OH)O-, or -OP(S)(OH)O-; And each G 4 Independently -D 2 -E 2 -F 2 -,in Each D 2 and F 2 independently a bond or C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, aryl, or heteroaryl, each of which is optionally substituted with 1, 2, 3, 4, or 5 R B group substitution, and Each E2 is independently a bond, C 1-10 Alkyl, C 2-10 Alkenyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, aryl, or heteroaryl, each of which is optionally substituted with 1, 2, 3, 4, or 5 R B Group substitution, the limiting condition is E 2 Not a key and D 2 and F 2 Each is a key.

72. The compound of claim 71, wherein Each L 7 Selected from the group consisting of: -O-, -S-, -N(H)-, -N(H)C(O)-, -C(O)N(H)-, -OP(O)(OH)O-, and -OP(S)(OH)O-; And each G 4 Independently -D 2 -E 2 -F 2 -,in Each D 2 and F 2 independently a bond or C 1-10 alkyl; Each E 2 Independently C 1-10 Alkyl, C 2-10 Alkenyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, aryl, or heteroaryl, each of which is optionally substituted with 1, 2, 3, 4, or 5 R B Group substitution.

73. The compound of claim 71, wherein Each L 7 Selected from the group consisting of: -O-, -S-, -N(H)-, -N(H)C(O)-, -C(O)N(H)-, -OP(O)(OH)O-, and -OP(S)(OH)O-; And each G 4 Independently C 1-10 Alkyl, C 2-10 Alkenyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, aryl, or heteroaryl, each of which is optionally substituted with 1, 2, 3, 4, or 5 R B replace.

74. The compound of claim 71, wherein Each L 7 Selected from the group consisting of: -O-, -S-, -N(H)-, -N(H)C(O)-, -C(O)N(H)-, -OP(O)(OH)O-, and -OP(S)(OH)O-; And each G 4 are independently optionally 1 or 25 R B C 1-10 alkyl.

75. The compound of claim 71, wherein Each L 7 Selected from the group consisting of: -O-, -S-, -N(H)-, -N(H)C(O)-, C(O)N(H)-, -OP(O)(OH)O-, and -OP(S)(OH)O-; And each G 4 Independently C 1-10 alkyl.

76. The compound of claim 71, wherein Δ is selected from the group consisting of: wherein # is a bond to T and each * is a bond to the ZZ group.

77. A compound as described in any one of claims 45 to 76, wherein ZZ comprises a group selected from the group consisting of:

78. The compound of any one of claims 45 to 76, wherein ZZ is -A'-B'-A'-, wherein Each A' is independently a bond, -O-, -S- or -N(R N3 )-, where R N3 are independently hydrogen or C 1-6 Alkyl, and Each B' is independently CH2, C(O), C(S), S(O)2, P(O)(OH) or P(S)(OH).

79. The compound of any one of claims 45 to 76, wherein ZZ is -A'-B'- or -B'-A'-, wherein Each A' is independently -O- or -N(R N3 )-, where R N3 are independently hydrogen or C 1-6 alkyl, each B′ is independently CH 2 , C(O), S(O) 2 , P(O)(OH) or P(S)(OH); and Each R N3 are independently hydrogen or C 1-6 alkyl.

80. The compound of any one of claims 45 to 76, wherein ZZ is -CH2O-, -OCH2-, -SS-, -C=N-, -C=NO-, -C=NN(R N3 )-, -N=C-, -ON=C-, -N(R N3 )-N=C-、-C(O)N(R N3 )-、-N(R N3 )C(O)-, -C(O)O-, -OC(O)-, -OC(O)N(R N3 )-、-N(R N3 )C(O)O-、-N(R N3 )C(O)N(R N3 )-、-S(O)2N(R N3 )-、-N(R N3 )S(O)2-, -OP(O)(OH)O-, -OP(S)(OH)O-, -OP(O)(OH)-, -OP(S)(OH)-, -P(O)(OH)O- or -P(S)(OH)O-, where R N3 are independently hydrogen or C 1-6 alkyl.

81. A compound as described in any one of claims 45 to 76, wherein ZZ is -C(O)N(R N3 )-or-N(R N3 )C(O)-, where R N3 are independently hydrogen or C 1-6 alkyl.

82. A compound as described in any one of claims 45 to 81, wherein formula (XII) is selected from the group consisting of: Where p is 0, 1, 2 or 3; Each R 21 are independently selected from the group consisting of R and a nitrogen protecting group, and R P A nitrogen protecting group.

83. A compound as described in any one of claims 45 to 82, wherein L is -L 1 -[GL 2 ] q -GL 3 -*, where q is 0, 1, 2, 3, 4, or 5.

84. A compound as described in any one of claims 45 to 82, wherein L is -L 1 -G-*, where * is the bond to ZZ; G is C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C 3-10 cycloalkyl, 3- to 10-membered heterocyclyl, aryl, or heteroaryl, each of which is optionally substituted with 1, 2, 3, or 4 R groups; L 1 is -BA-, where A is a bond, -O-, -S- or -N(R N )-, where each R N are independently hydrogen or C 1-6 alkyl; and B is a bond, C(O), C(S), S(O)2, P(O)(OH) or P(S)(OH).

85. A compound as described in any one of claims 45 to 82, wherein L is Where * is a bond to ZZ; k is an integer from 1 to 10; L 1 for bonds, C(O), C(S), C(NR N ), S(O)2, P(O)(OH) or P(S)(OH); and R N is hydrogen or C 1-6 alkyl.

86. A compound as described in any one of claims 45 to 82, wherein L is a group selected from: (a) Wherein * is a bond to ZZ; t is an integer from 0 to 10; (b) L is wherein * is a bond to ZZ, t is an integer from 0 to 10; a is an integer from 1 to 3; and s and s′ are each independently an integer from 1 to 24; (c) wherein * is a bond to ZZ; a is 1, 2 or 3; and each of s, s′ and s″ is independently an integer from 1 to 24; (d) wherein * is a bond to ZZ; and s, s′, and s″ are independently integers from 1 to 24; (e) wherein * is a bond to ZZ, and each of s, s′, and s″ is independently an integer from 1 to 24; and (f) wherein * is a bond to ZZ; s and k are independently integers from 1 to 20; and w is an integer from 1 to 10.

87. A compound as described in any one of claims 45 to 82, wherein L is wherein * is a bond to ZZ, and w is an integer from 1 to 20.

88. A compound as described in any one of claims 45 to 82, wherein L is wherein * is a bond to ZZ; and k is an integer from 1 to 10.

89. A compound as described in any one of claims 45 to 88, wherein R T1 -L L - oligonucleotide, wherein L L It is a divalent linker that is attached to the 3' end of an oligonucleotide, the 5' end of an oligonucleotide, an internal 2' or 3' position on an internal nucleotide, or an internucleotide linkage.

90. The compound of claim 89, wherein L L Attached to the oxygen atom on the nucleoside and is P(O)(OH)-, -P(S)(OH)-, or -P(S)(SH).

91. The compound of claim 90, wherein L L It is P(O)(OH)-.

92. The compound of claim 90, wherein L L It is P(S)(OH)-.

93. A compound as described in any one of claims 1 to 25, wherein RT is R T1 , where R T1 -L L - oligonucleotide, wherein L L The oxygen atom attached to the nucleoside of the oligonucleotide, L L is a bond, and the nucleoside has formula (XV-f), wherein B is an optionally modified nucleobase and * represents a bond to Δ.

94. The compound of claim 93, wherein -T- is -C 4-10 Alkyl-*, where * is a bond to Δ.

95. as claimed in claim 93 compound, wherein nucleoside has the formula selected from the group consisting of: in B is an optionally modified nucleobase; each n is independently 0 or an integer selected from 1 to 10; and Each m is independently an integer selected from 1 to 20.

96. A compound as described in any one of claims 45 to 88, wherein R T1 -L L -oligonucleotide and binds to the 5' end of the oligonucleotide.

97. The compound of claim 96 having the formula: wherein Y′ is O or S.

98. A compound as described in any one of claims 45 to 88, wherein R T1 -L L -oligonucleotide and binds to the 3' end of the oligonucleotide.

99. The compound of claim 98 having the formula: wherein Y′ is O or S.

100. The compound of claim 99, having the formula: Wherein Y′ is O or S; each ZZ is N(H)C(O) or C(O)N(H); Each Φ is wherein m is an integer selected from 1 to 10; L 4 and L 6 Independently -B 1 -A 1 -or-A 1 -B 1 -; Each L 5 For key or -A 1 -B 1 -A 1 -(e.g. key, -B 1 -A 1 -or-A 1 -B 1 -; or key); Each G 1 Independently C 1-10 Alkyl or C 2-10 alkenyl; Each A 1 are independently a bond, -O-, -S- or -N(R N1 )-, where R N1 are independently hydrogen or C 1-6 alkyl; and Each B 1 is independently a bond, C(O), C(S), S(O)2, P(O)(OH) or P(S)(OH).

101. The compound of claim 99 having the formula: Wherein Y′ is O or S; each ZZ is N(H)C(O) or C(O)N(H); Each Φ is wherein m is an integer selected from 1 to 10; L 5 For key or -A 1 -B 1 -A 1 -(e.g. key, -B 1 -A 1 -or-A 1 -B 1 -; or key); Each G 1 Independently C 1-10 alkyl; Each A 1 are independently a bond, -O-, -S- or -N(R N1 )-, where R N1 are independently hydrogen or C 1-6 alkyl; and Each B 1 is independently a bond, C(O), C(S), S(O)2, P(O)(OH) or P(S)(OH).

102. The compound of claim 101, wherein each Φ is wherein m is an integer selected from 1 to 10.

103. The compound of claim 101, wherein each Φ is wherein m is an integer selected from 1 to 10.

104. The compound of claim 101, wherein each Φ is wherein m is an integer selected from 1 to 10.

105. The compound of claim 101, wherein each Φ is wherein m is an integer selected from 1 to 10.

106. The compound of claim 101, wherein each Φ is wherein m is an integer selected from 1 to 10.

107. A compound as described in any one of claims 99 to 106, wherein R P is hydrogen, and R 1 For hydrogen.

108. The compound of claim 107, wherein Y' is O.

109. The compound of claim 107, wherein Y' is S.

110. A dsRNA medicament comprising the oligonucleotide of any one of claims 1 to 109.

111. The dsRNA agent of claim 110, wherein the αvβ6 integrin targeting ligand is bound to the sense strand.

112. The dsRNA agent of claim 111, wherein the αvβ6 integrin targeting ligand is bound to the 3' end of the sense strand.

113. The dsRNA agent of claim 111, wherein the αvβ6 integrin targeting ligand is bound to the 5' end of the sense strand.

114. The dsRNA agent of claim 111, wherein the αvβ6 integrin targeting ligand is bound to both the 5' end and the 3' end of the sense strand.

115. The dsRNA agent of claim 111, wherein the αvβ6 integrin targeting ligand binds to any internal position of the sense strand.

116. The dsRNA agent of claim 110, wherein the αvβ6 integrin targeting ligand is bound to the antisense strand.

117. The dsRNA agent of claim 116, wherein the αvβ6 integrin targeting ligand is bound to the 3' end of the antisense strand.

118. The dsRNA agent of claim 116, wherein the αvβ6 integrin targeting ligand binds to an internal position of the antisense strand.

119. The dsRNA agent of any one of claims 109 to 118, wherein the target gene is selected from the group consisting of: adrenergic receptor beta 1 (ADRB1); calcium voltage-gated channel subunit alpha 1C (CACNA1C); calcium voltage-gated channel subunit alpha 1G (CACNA1G) (T-type calcium c channel); angiotensin II receptor type 1 (AGTR1); sodium voltage-gated channel alpha subunit 2 (SCN2A); hyperpolarization-activated cyclic nucleotide-gated potassium channel 1 (HCN1); hyperpolarization-activated cyclic nucleotide-gated potassium channel 4 (HCN4); hyperpolarization-activated cyclic nucleotide-gated potassium channel 3 (HCN3); potassium voltage-gated channel subfamily A member 5 (KCNA5); potassium inward rectifier channel subfamily J member 3 (KCNJ3); potassium inward rectifier channel subfamily J member 4 (KCNJ4); phospholamban (PLN); calcium / calmodulin-dependent protein kinase II delta ( 1 (CAMK2D); phosphodiesterase 1 (PDE1); myostatin (MSTN); cholinergic receptor nicotinic alpha 1 subunit (CHRNA1); cholinergic receptor nicotinic beta 1 subunit (CHRNB1); cholinergic receptor nicotinic delta subunit (CHRND); cholinergic receptor nicotinic epsilon subunit (CHRNE); cholinergic receptor nicotinic gamma subunit (CHRNG); collagen type XIII alpha 1 chain (COL13A1); docking protein 7 (DOK7); LDL receptor-related protein 4 (LRP4); muscle-associated receptor tyrosine kinase (MUSK); synaptic receptor-associated protein (RAPSN); sodium voltage-gated channel alpha subunit 4 (SCN4A); double homeobox 4 (DUX4); dystrophic myotonic protein kinase (DMPK); glycogen synthase 1 (GYS1); survival motor neuron 1 (SMN1); and alpha-glucosidase (GAA).

120. A cell comprising the dsRNA agent of any one of claims 109 to 118.

121. A pharmaceutical composition for inhibiting the expression of a target gene, comprising the dsRNA agent according to any one of claims 109 to 118.

122. A method for inhibiting the expression of a target gene in a skeletal muscle cell and / or a cardiomyocyte, comprising contacting the cell with a dsRNA agent according to any one of claims 109 to 118, thereby inhibiting the expression of the target gene in the skeletal muscle cell and / or the cardiomyocyte.

123. The method of claim 122, wherein the cell is within an individual.

124. The method of claim 123, wherein the individual is a human.

125. A method of treating a subject suffering from a skeletal muscle disorder and / or a cardiac muscle disorder, comprising administering to the subject a therapeutically effective amount of a dsRNA agent according to any one of claims 109 to 118, thereby treating the subject.

126. The method of claim 125, wherein the skeletal muscle disorder and / or cardiac muscle disorder is selected from the group consisting of myostatin-associated muscle hypertrophy, congenital myasthenic syndrome, facioscapulohumeral muscular dystrophy (FSHD), spinal muscular atrophy (SMA), myotonic dystrophy type 1 (DM1), Pompe disease, PLN cardiomyopathy, spasticity, obstructive hypertrophic cardiomyopathy (HOCM); familial hypertrophic cardiomyopathy (FHC); heart failure with preserved ejection fraction (HFPEF); atrial fibrillation (AFIB); ventricular fibrillation (VFIB); angina; myocardial infarction (MI); heart failure or heart failure with reduced ejection fraction (HFREF); supraventricular tachycardia (SVT); hypertrophic cardiomyopathy (HCM), dilated cardiomyopathy (DCM), arrhythmias, and congestive heart failure (CHF).

127. A method of inhibiting expression of a target gene in a lung cell, comprising contacting the cell with a dsRNA agent according to any one of claims 109 to 118, thereby inhibiting expression of the target gene in the lung cell.

128. The method of claim 127, wherein the cell is within a subject.

129. The method of claim 128, wherein the individual is a human.

130. A method of treating a subject having a pulmonary disorder, comprising administering to the subject a therapeutically effective amount of a dsRNA agent according to any one of claims 109 to 118, thereby treating the subject.

131. The method of claim 130, wherein the pulmonary disorder is selected from the group consisting of idiopathic pulmonary fibrosis, asthma, asthma and chronic sinusitis, nasal polyps and chronic sinusitis.

132. The method of any one of claims 122 to 131, wherein the dsRNA agent is administered subcutaneously to the individual.

133. The method of any one of claims 122 to 126, wherein the dsRNA agent is administered intramuscularly to the individual.

134. The method of any one of claims 122 to 131, wherein the dsRNA agent is administered intravenously to the subject.

135. The method of any one of claims 126 to 131, wherein the dsRNA agent is administered to the subject via inhalation.

136. An RNA-induced silencing complex (RISC) comprising the antisense strand of any of the dsRNA agents of claims 109 to 118.

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