Compounds and methods for modulation of SMN2

Abstract

To provide compounds, compositions and methods for modulating splicing of SMN2.SOLUTION: In certain embodiments, the present disclosure provides oligomeric compounds comprising or consisting of modified oligonucleotides having one or more 2'-O-(N-alkyl acetamide) or 2'-O-(N-methyl acetamide) modified sugar moieties and complementary to SMN pre-mRNA. Also provided are uses of the compounds and compositions in the manufacture of a medicament for treatment of spinal muscular atrophy.SELECTED DRAWING: None

Description

[Technical field]

[0001] Sequence Listing This application is filed with an electronic sequence listing, which is submitted as a file entitled BIOL0304WOSEQ_ST25.txt, created on July 17, 2017, and having a size of 88 Kb. The information in this electronic sequence listing is incorporated herein by reference in its entirety.

[0002] Provided herein are methods, compounds and compositions for the modulation of SMN2. [Background technology]

[0003] Proximal spinal muscular atrophy (SMA) is an inherited neurodegenerative disease characterized by the loss of motor neurons in the spinal cord. SMA is an early-onset autosomal recessive disorder and is currently the leading cause of death in infants and young children. The severity of SMA varies from patient to patient and is classified into three types. Type I SMA is the most severe form, occurring at birth or within 6 months, and is usually fatal within 2 years. Children with Type I SMA are unable to sit or walk. Type II SMA is an intermediate form, in which patients can sit but cannot stand or walk. Patients with Type III SMA, the chronic form of the disease, usually develop SMA after 18 months of age (Lefebvre et al., 2003). et al., Hum. Mol. Genet., 1998, 7, 1531-1536).

[0004] The molecular basis of SMA results from the deletion of both copies of the survival motor neuron gene 1 (SMN1). SMN1, also known as SMN Telomeric, is part of a multiprotein complex thought to be involved in snRNP biogenesis and recycling. A nearly identical gene, SMN2, also known as SMN Centromeric, resides in a duplicated region on chromosome 5q13 and modulates disease severity. Normal SMN1 gene expression results in expression of only the survival motor neuron (SMN) protein. Although SMN1 and SMN2 may code for the same protein, SMN2 contains a translationally silent mutation at position +6 of exon 7, resulting in poor inclusion of exon 7 in the SMN2 transcript. Thus, the predominant form of SMN2 is a truncated form lacking exon 7, which is unstable and inactive (Cartegni and Krainer, Nat. Genet., 2002, 30, 377-384). Expression of the SMN2 gene results in approximately 10-20% SMN protein and 80-90% unstable / non-functional SMN delta 7 protein. The SMN protein has a well-established role in spliceosome assembly and may also mediate mRNA transport in neuronal axons and nerve terminals.

[0005] Antisense technology is an effective means for regulating the expression of one or more specific gene products, including alternative splicing products, and is uniquely useful in many therapeutic, diagnostic, and research applications. The principle behind antisense technology is that antisense compounds hybridize to target nucleic acids and regulate gene expression activities such as transcription, splicing, or translation by one of many antisense mechanisms. The sequence specificity of antisense compounds makes them attractive as tools for target validation and gene functioning, and as therapeutic agents for selectively regulating the expression of genes involved in disease. Summary of the Invention

[0006] The present disclosure provides compounds, methods and compositions for the modulation of SMN2 pre-mRNA. The present disclosure also provides compounds, methods and compositions useful, for example, for treating, preventing or ameliorating one or more symptoms of spinal muscular atrophy.

[0007] In certain embodiments, the disclosure provides oligomeric compounds comprising or consisting of modified oligonucleotides having one or more 2'-O-(N-alkylacetamido) or 2'-O-(N-methylacetamido) modified sugar moieties and complementary to SMN pre-mRNA. Modified oligonucleotides having one or more 2'-O-(N-alkylacetamido) or 2'-O-(N-methylacetamido) modified sugar moieties have improved cellular uptake and / or pharmacological activity in muscle tissue. Since SMN2 is expressed in muscle tissue, modified oligonucleotides having one or more 2'-O-(N-alkylacetamido) modifications will have improved activity in muscle tissue.

[0008] Provided herein are compounds and methods useful for modulating SMN2 pre-mRNA processing. The compounds include modified oligonucleotides comprising 2'-O-(N-alkylacetamido) or 2'-O-(N-methylacetamido) modified sugar moieties. In certain embodiments, the modified oligonucleotides comprise 2'-O-(N-methylacetamido) or 2'-O-(N-methylacetamido) modified sugar moieties. In certain embodiments, the compounds of the invention modulate pre-mRNA splicing. Modified oligonucleotides having one or more 2'-O-(N-alkylacetamido) or 2'-O-(N-methylacetamido) modified sugar moieties also have enhanced pharmacological activity for modulating SMN2 pre-mRNA. For example, in certain embodiments, modified oligonucleotides bearing one or more 2'-O-(N-alkylacetamido) or 2'-O-(N-methylacetamido) modified sugar moieties targeted to the SMN2 pre-mRNA exhibit a greater increase in inclusion of exon 7 than modified oligonucleotides lacking one or more 2'-O-(N-alkylacetamido) or 2'-O-(N-methylacetamido) modified sugar moieties.

[0009] Further provided herein are methods for enhancing cellular uptake, enhancing pharmacological activity, and modulating tissue distribution of modified oligonucleotides and oligomeric compounds that comprise or consist of conjugate groups that contain 2'-O-(N-alkylacetamido) or 2'-O-(N-methylacetamido) modified sugar moieties. Also provided are compounds comprising modified oligonucleotides that contain 2'-O-(N-alkylacetamido) or 2'-O-(N-methylacetamido) modified sugar moieties for use in therapy. Also provided are compounds for preparing medicaments for modulating the processing of selected precursor transcripts in cells or tissues.

[0010] The present disclosure provides the following non-limiting numbered embodiments.

[0011] Embodiment 1: An oligomeric compound comprising a modified oligonucleotide consisting of 14 to 25 linked nucleosides, wherein the modified oligonucleotide is complementary to SMN2 pre-mRNA, and at least one nucleoside of the modified oligonucleotide has the structure of Formula I: [ka] In the formula, Bx is a nucleobase; For each nucleoside of formula I, R 1 is independently selected from methyl, ethyl, propyl, and isopropyl.

[0012] Embodiment 2: The oligomeric compound of embodiment 1, wherein Bx is selected from adenine, guanine, cytosine, thymine, uracil and 5-methylcytosine.

[0013] Embodiment 3: R 1 The oligomeric compound of embodiment 1 or 2, wherein is methyl.

[0014] Embodiment 4: The oligomeric compound of any of embodiments 1-3, wherein each of at least two nucleosides of the modified oligonucleotide has a structure independently selected from Formula I.

[0015] Embodiment 5: The oligomeric compound of any of embodiments 1-3, wherein each of the seven nucleosides of the modified oligonucleotide has a structure independently selected from Formula I.

[0016] Embodiment 6: The oligomeric compound of any of embodiments 1-3, wherein each of the eight nucleosides of the modified oligonucleotide has a structure independently selected from Formula I.

[0017] Embodiment 7: The oligomeric compound of any of embodiments 1-3, wherein each of the nine nucleosides of the modified oligonucleotide has a structure independently selected from Formula I.

[0018] Embodiment 8: The oligomeric compound of any of embodiments 1-3, wherein each of the ten nucleosides of the modified oligonucleotide has a structure independently selected from Formula I.

[0019] Embodiment 9: The oligomeric compound of any of embodiments 1-3, wherein each of the eleven nucleosides of the modified oligonucleotide has a structure independently selected from Formula I.

[0020] Embodiment 10: The oligomeric compound of any of embodiments 1-3, wherein each of the 12 nucleosides of the modified oligonucleotide has a structure independently selected from Formula I.

[0021] Embodiment 11: The oligomeric compound of any of embodiments 1-3, wherein each of the 13 nucleosides of the modified oligonucleotide has a structure independently selected from Formula I.

[0022] Embodiment 12: The oligomeric compound of any of embodiments 1-3, wherein each of the 14 nucleosides of the modified oligonucleotide has a structure independently selected from Formula I.

[0023] Embodiment 13: The oligomeric compound of any of embodiments 1-3, wherein each of the 15 nucleosides of the modified oligonucleotide has a structure independently selected from Formula I.

[0024] Embodiment 14: The oligomeric compound of any of embodiments 1-3, wherein each of the 16 nucleosides of the modified oligonucleotide has a structure independently selected from Formula I.

[0025] Embodiment 15: The oligomeric compound of any of embodiments 1-3, wherein each of the 17 nucleosides of the modified oligonucleotide has a structure independently selected from Formula I.

[0026] Embodiment 16: The oligomeric compound of any of embodiments 1-3, wherein each of the 18 nucleosides of the modified oligonucleotide has a structure independently selected from Formula I.

[0027] Embodiment 17: The oligomeric compound of any of embodiments 1-3, wherein each of the 19 nucleosides of the modified oligonucleotide has a structure independently selected from Formula I.

[0028] Embodiment 18: The oligomeric compound of any of embodiments 1-3, wherein each of the 20 nucleosides of the modified oligonucleotide has a structure independently selected from Formula I.

[0029] Embodiment 19: R of at least one nucleoside having the structure of Formula I 1 The oligomeric compound according to any one of embodiments 1-18, wherein is methyl.

[0030] Embodiment 20: R 1 is the same for all of the nucleosides having the structure of Formula I.

[0031] Embodiment 21: An oligomeric compound comprising a modified oligonucleotide consisting of 14 to 25 linked nucleosides, wherein the modified oligonucleotide is complementary to SMN2 pre-mRNA, and at least one nucleoside of the modified oligonucleotide comprises a 2'-O-(N-alkylacetamido) modified sugar moiety.

[0032] Embodiment 22: The oligomeric compound of embodiment 21, wherein the sugar moiety of each nucleoside comprising a 2'-O-(N-alkylacetamide) modified sugar moiety is selected from 2'-O-(N-methylacetamide) and 2'-O-(N-ethylacetamide).

[0033] Embodiment 23: The oligomeric compound of embodiment 21 or 22, wherein each of the seven nucleosides of said modified oligonucleotide comprises an independently selected 2'-O-(N-alkylacetamido) modified sugar moiety.

[0034] Embodiment 24: The oligomeric compound of embodiment 21 or 22, wherein each of the eight nucleosides of said modified oligonucleotide comprises an independently selected 2'-O-(N-alkylacetamido) modified sugar moiety.

[0035] Embodiment 25: The oligomeric compound of embodiment 21 or 22, wherein each of the nine nucleosides of said modified oligonucleotide comprises an independently selected 2'-O-(N-alkylacetamido) modified sugar moiety.

[0036] Embodiment 26: The oligomeric compound of embodiment 21 or 22, wherein each of the ten nucleosides of said modified oligonucleotide comprises an independently selected 2'-O-(N-alkylacetamido) modified sugar moiety.

[0037] Embodiment 27: The oligomeric compound of embodiment 21 or 22, wherein each of the eleven nucleosides of said modified oligonucleotide comprises an independently selected 2'-O-(N-alkylacetamido) modified sugar moiety.

[0038] Embodiment 28: The oligomeric compound of embodiment 21 or 22, wherein each of the 12 nucleosides of said modified oligonucleotide comprises an independently selected 2'-O-(N-alkylacetamido) modified sugar moiety.

[0039] Embodiment 29: The oligomeric compound of embodiment 21 or 22, wherein each of the 13 nucleosides of said modified oligonucleotide comprises an independently selected 2'-O-(N-alkylacetamido) modified sugar moiety.

[0040] Embodiment 30: The oligomeric compound of embodiment 21 or 22, wherein each of the 14 nucleosides of said modified oligonucleotide comprises an independently selected 2'-O-(N-alkylacetamido) modified sugar moiety.

[0041] Embodiment 31: The oligomeric compound of embodiment 21 or 22, wherein each of the 15 nucleosides of said modified oligonucleotide comprises an independently selected 2'-O-(N-alkylacetamido) modified sugar moiety.

[0042] Embodiment 32: The oligomeric compound of embodiment 21 or 22, wherein each of the 16 nucleosides of said modified oligonucleotide comprises an independently selected 2'-O-(N-alkylacetamido) modified sugar moiety.

[0043] Embodiment 33: The oligomeric compound of embodiment 21 or 22, wherein each of the 17 nucleosides of said modified oligonucleotide comprises an independently selected 2'-O-(N-alkylacetamido) modified sugar moiety.

[0044] Embodiment 34: The oligomeric compound of embodiment 21 or 22, wherein each of the 18 nucleosides of said modified oligonucleotide comprises an independently selected 2'-O-(N-alkylacetamido) modified sugar moiety.

[0045] Embodiment 35: The oligomeric compound of embodiment 21 or 22, wherein each of the 19 nucleosides of said modified oligonucleotide comprises an independently selected 2'-O-(N-alkylacetamido) modified sugar moiety.

[0046] Embodiment 36: The oligomeric compound of embodiment 21 or 22, wherein each of the 20 nucleosides of said modified oligonucleotide comprises an independently selected 2'-O-(N-alkylacetamido) modified sugar moiety.

[0047] Embodiment 37: Nucleosides containing 2'-O-(N-alkylacetamido) modified sugar moieties 37. The oligomeric compound according to any of embodiments 21 to 36, wherein at least one sugar moiety of the oligosaccharide is a 2'-O-(N-methylacetamide) modified sugar moiety.

[0048] Embodiment 38: The oligomeric compound of any of embodiments 21-37, wherein each N-alkyl group of said 2'-O-(N-alkylacetamido) modified sugar moiety is the same N-alkyl group.

[0049] Embodiment 39: The oligomeric compound according to any of embodiments 21 to 38, wherein each of said 2'-O-(N-alkylacetamido) modified sugar moieties is a 2'-O-(N-methylacetamido) sugar moiety.

[0050] Embodiment 40: The oligomeric compound according to any of embodiments 21 to 39, wherein each sugar moiety of each nucleoside of said modified oligonucleotide is a 2'-O-(N-methylacetamido) modified sugar moiety.

[0051] Embodiment 41: The oligomeric compound according to any of embodiments 1 to 40, wherein said modified oligonucleotide consists of 16 to 23 linked nucleosides.

[0052] Embodiment 42: The oligomeric compound according to any of embodiments 1 to 40, wherein said modified oligonucleotide consists of 18 to 20 linked nucleosides.

[0053] Embodiment 43: The oligomeric compound according to any of embodiments 1 to 41, wherein said modified oligonucleotide consists of 16 nucleosides.

[0054] Embodiment 44: The oligomeric compound according to any of embodiments 1 to 41, wherein said modified oligonucleotide consists of 17 nucleosides.

[0055] Embodiment 45: The oligomeric compound according to any of embodiments 1 to 41, wherein said modified oligonucleotide consists of 18 nucleosides.

[0056] Embodiment 46: The oligomeric compound according to any of embodiments 1 to 41, wherein said modified oligonucleotide consists of 19 nucleosides.

[0057] Embodiment 47: The oligomeric compound according to any of embodiments 1 to 41, wherein said modified oligonucleotide consists of 20 nucleosides.

[0058] Embodiment 48: The oligomeric compound according to any of embodiments 1 to 47, wherein said modified oligonucleotide comprises at least one modified internucleoside linkage.

[0059] Embodiment 49: The oligomeric compound according to any of embodiments 1 to 48, wherein the modified oligonucleotide comprises at least one phosphorothioate internucleoside linkage.

[0060] Embodiment 50: The oligomeric compound according to embodiment 49, wherein each internucleoside linkage of said modified oligonucleotide is selected from a phosphorothioate internucleoside linkage and a phosphate internucleoside linkage.

[0061] Embodiment 51: The oligomeric compound of embodiment 50, wherein said phosphate internucleoside linkage is a phosphodiester internucleoside linkage.

[0062] Embodiment 52: Each internucleoside linkage of the modified oligonucleotide is a phosphorothioate. The oligomeric compound according to any one of embodiments 1 to 50, wherein the linkage is a mononucleoside linkage.

[0063] Embodiment 53: The oligomeric compound according to any of embodiments 1 to 52, wherein said modified oligonucleotide comprises at least one modified nucleobase.

[0064] Embodiment 54: The oligomeric compound according to any of embodiments 1 to 53, wherein said modified oligonucleotide comprises at least one 5-methylcytosine.

[0065] Embodiment 55: The oligomeric compound according to any of embodiments 1 to 54, wherein each nucleobase of said modified oligonucleotide is selected from thymine, 5-methylcytosine, cytosine, adenine, uracil and guanine.

[0066] Embodiment 56: The oligomeric compound according to any of embodiments 1 to 55, wherein each cytosine in said modified oligonucleotide is a 5-methylcytosine.

[0067] Embodiment 57: The oligomeric compound according to any of embodiments 1 to 56, wherein each nucleobase of said modified oligonucleotide is selected from thymine, 5-methylcytosine, adenine and guanine.

[0068] Embodiment 58: The oligomeric compound of any of embodiments 1 to 57, wherein the modified oligonucleotide is at least 70% complementary to the SMN2 pre-mRNA.

[0069] Embodiment 59: The oligomeric compound of any of embodiments 1 to 57, wherein the modified oligonucleotide is at least 75% complementary to the SMN2 pre-mRNA.

[0070] Embodiment 60: The oligomeric compound according to any of embodiments 1 to 57, wherein the modified oligonucleotide is at least 80% complementary to the SMN2 pre-mRNA.

[0071] Embodiment 61: The oligomeric compound of any of embodiments 1 to 57, wherein the modified oligonucleotide is at least 85% complementary to the SMN2 pre-mRNA.

[0072] Embodiment 62: The oligomeric compound of any of embodiments 1 to 57, wherein the modified oligonucleotide is at least 90% complementary to the SMN2 pre-mRNA.

[0073] Embodiment 63: The oligomeric compound of any of embodiments 1 to 57, wherein the modified oligonucleotide is at least 95% complementary to the SMN2 pre-mRNA.

[0074] Embodiment 64: The oligomeric compound of any of embodiments 1 to 57, wherein the modified oligonucleotide is at least 100% complementary to the SMN2 pre-mRNA.

[0075] Embodiment 65: The oligomeric compound according to any of embodiments 1 to 64, wherein the modified oligonucleotide is complementary to intron 7 of SMN2 pre-mRNA.

[0076] Embodiment 66: The oligomeric compound according to any of embodiments 1 to 64, wherein the modified oligonucleotide is complementary to ISS-N1 of SMN2 pre-mRNA.

[0077] Embodiment 67: The oligomeric compound according to any of embodiments 1 to 64, wherein the sequence of the nucleobases of the modified oligonucleotide comprises a sequence selected from SEQ ID NO: 1, 2 or 3.

[0078] Embodiment 68: The oligomeric compound according to any of embodiments 1 to 64, wherein the sequence of the nucleobases of the modified oligonucleotide comprises a sequence selected from any of SEQ ID NOs: 12 to 239.

[0079] Embodiment 69: The oligomeric compound according to any of embodiments 1 to 64, wherein the sequence of the nucleobases of the modified oligonucleotide consists of a sequence selected from any of SEQ ID NOs: 1, 2, 3 or 12 to 239.

[0080] Embodiment 70: The oligomeric compound according to any of embodiments 1 to 64, wherein the modified oligonucleotide is complementary to exon 7 of SMN2 pre-mRNA.

[0081] Embodiment 71: The oligomeric compound of any of embodiments 1 to 64, wherein the modified oligonucleotide is complementary to intron 6 of SMN2 pre-mRNA.

[0082] Embodiment 72: The oligomeric compound according to any of embodiments 1 to 64, wherein the modified oligonucleotide is complementary to an exonic splicing enhancer in exon 7 of SMN2 pre-mRNA.

[0083] Embodiment 73: The oligomeric compound according to any of embodiments 1 to 64, wherein the modified oligonucleotide is complementary to an aberrant exonic splicing silencer in exon 7 of SMN2 pre-mRNA.

[0084] Embodiment 74: The oligomeric compound of any of embodiments 1 to 73, wherein the oligomeric compound comprises a conjugate group.

[0085] Embodiment 75: The oligomeric compound according to embodiment 74, wherein the conjugate group comprises a lipid or a lipophilic group.

[0086] Embodiment 76: The lipid or lipophilic group is cholesterol, C 10 ~C 26 Saturated fatty acids, C 10 ~C 26 unsaturated fatty acids, C 10 ~C 26 The oligomeric compound according to embodiment 75, which is selected from alkyl, triglyceride, tocopherol or cholic acid.

[0087] Embodiment 77: The oligomeric compound according to embodiment 76, wherein said lipid or lipophilic group is a saturated or unsaturated hydrocarbon chain.

[0088] Embodiment 78: The lipid or lipophilic group is C 16 78. The oligomeric compound according to any one of embodiments 75 to 77, which is a lipid.

[0089] Embodiment 79: The lipid or lipophilic group is C 18 78. The oligomeric compound according to any one of embodiments 75 to 77, which is a lipid.

[0090] Embodiment 80: The lipid or lipophilic group is C 16 78. The oligomeric compound according to any one of embodiments 75 to 77, wherein said alkyl is aryl.

[0091] Embodiment 81: The lipid or lipophilic group is C 18 78. The oligomeric compound according to any one of embodiments 75 to 77, wherein said alkyl is aryl.

[0092] Embodiment 82: The oligomeric compound according to embodiment 76, wherein said lipid or lipophilic group is cholesterol.

[0093] Embodiment 83: The oligomeric compound according to embodiment 76, wherein said lipid or lipophilic group is tocopherol.

[0094] Embodiment 84: The lipid or lipophilic group is a saturated C 16 77. The oligomeric compound of embodiment 76, wherein

[0095] Embodiment 85: The oligomeric compound according to any of embodiments 74 to 84, wherein the conjugate group is attached to the modified oligonucleotide at the 5' end of the modified oligonucleotide.

[0096] Embodiment 86: The oligomeric compound according to any of embodiments 74 to 85, wherein the conjugate group is attached to the modified oligonucleotide at the 3' end of the modified oligonucleotide.

[0097] Embodiment 87: The oligomeric compound according to any of embodiments 74 to 86, wherein the conjugate group comprises a cleavable linker.

[0098] Embodiment 88: The oligomeric compound of embodiment 87, wherein said cleavable linker comprises one or more linker nucleosides.

[0099] Embodiment 89: An oligomeric compound according to any of embodiments 1 to 73, consisting of said modified oligonucleotide.

[0100] Embodiment 90: An oligomeric compound according to any of embodiments 74 to 88, consisting of said modified oligonucleotide and said conjugate group.

[0101] Embodiment 91: The oligomeric compound of any of embodiments 1 to 90, wherein said oligomeric compound modulates splicing of SMN2 pre-mRNA.

[0102] Embodiment 92: The oligomeric compound of any of embodiments 1 to 91, wherein said oligomeric compound is single-stranded.

[0103] Embodiment 93: The oligomeric compound of any of embodiments 1 to 91, wherein said oligomeric compound is paired with a complementary oligomeric compound to form a double-stranded compound.

[0104] Embodiment 94: The oligomeric compound of embodiment 93, wherein said complementary oligomeric compound comprises a conjugate group.

[0105] Embodiment 95: A pharmaceutical composition comprising an oligomeric compound according to any of embodiments 1 to 94 and at least one pharma- ceutically acceptable carrier or diluent.

[0106] Embodiment 96: A method of modulating splicing of SMN2 pre-mRNA comprising contacting a cell, tissue or organ with an oligomeric compound or composition according to any of embodiments 1 to 95.

[0107] Embodiment 97: A method for promoting inclusion of exon 7 of SMN2 mRNA in a cell, tissue or organ, comprising contacting said cell, tissue or organ with an oligomeric compound described in any of embodiments 1 to 91 or a composition described in embodiment 95.

[0108] Embodiment 98: The method of embodiment 96 or 97, wherein the cell, tissue or organ is in an animal.

[0109] Embodiment 99: The method of embodiment 96 or 97, wherein the cell, tissue or organ is in a human.

[0110] Embodiment 100: A method of treating spinal muscular atrophy in a patient, comprising administering to a patient in need thereof an oligomeric compound according to any of embodiments 1 to 94 or a composition according to embodiment 95.

[0111] Embodiment 101: A method of treating spinal muscular atrophy type I in a patient, comprising administering to a patient in need thereof an oligomeric compound according to any of embodiments 1 to 94 or a composition according to embodiment 95.

[0112] Embodiment 102: A method of treating spinal muscular atrophy type II in a patient, comprising administering to a patient in need thereof an oligomeric compound according to any of embodiments 1 to 94 or a composition according to embodiment 95.

[0113] Embodiment 103: A method of treating spinal muscular atrophy type III in a patient, comprising administering to a patient in need thereof an oligomeric compound according to any of embodiments 1 to 94 or a composition according to embodiment 95.

[0114] Embodiment 104: A method of treating spinal muscular atrophy type IV in a patient, comprising administering to a patient in need thereof an oligomeric compound according to any of embodiments 1 to 94 or a composition according to embodiment 95.

[0115] Embodiment 105: The method of any of embodiments 100-104, wherein the oligomeric compound is administered to the central nervous system.

[0116] Embodiment 106: The method of embodiment 105, wherein the oligomeric compound is administered intrathecally.

[0117] Embodiment 107: The method of any of embodiments 100-104, wherein the oligomeric compound is administered systemically.

[0118] Embodiment 108: The method of embodiment 107, wherein the oligomeric compound is administered subcutaneously.

[0119] Embodiment 109: The method of embodiment 107, wherein the oligomeric compound is administered intramuscularly.

[0120] Embodiment 110: The method of any of embodiments 100-109, wherein the oligomeric compound is administered to the central nervous system and systemically.

[0121] Embodiment 111: The method of any of embodiments 100-110, wherein the oligomeric compound is co-administered with a second compound.

[0122] Embodiment 112: The method of embodiment 111, wherein the second compound is selected from a small molecule and an oligomeric compound.

[0123] Embodiment 113: The method of embodiment 112, wherein the second compound is an oligomeric compound according to any one of embodiments 1 to 94.

[0124] Embodiment 114: The method of any of embodiments 111-113, wherein the second compound is administered to the CSF.

[0125] Embodiment 115: The method of embodiment 114, wherein said second compound is administered intrathecally.

[0126] Embodiment 116: The method of any of embodiments 111 to 115, wherein the second compound is a second oligomeric compound comprising a second modified oligonucleotide having at least one modified nucleoside comprising a modified sugar moiety.

[0127] Embodiment 117: The method of embodiment 116, wherein the modified sugar moiety of the modified nucleoside of the second modified oligonucleotide of said second oligomeric compound is selected from 2'-OMe and 2'-MOE.

[0128] Embodiment 118: The method of embodiment 116 or 117, wherein each nucleoside of said second modified oligonucleotide comprises a modified sugar moiety.

[0129] Embodiment 119: The method of embodiment 118, wherein the modified sugar moiety of said second oligonucleotide is a 2'-MOE sugar moiety.

[0130] Embodiment 120: The method of embodiment 111 or 112, wherein said second compound is an oligomeric compound comprising a morpholino oligonucleotide.

[0131] Embodiment 121: The method of any of embodiments 111 to 120, wherein said second oligomeric compound has the nucleobase sequence of SEQ ID NO:1.

[0132] Embodiment 122: The method of embodiment 121, wherein said second oligomeric compound has the nucleobase sequence and chemical modification motif of Isis396443.

[0133] Embodiment 123: The method of any of embodiments 111 to 122, wherein said second oligomeric compound consists of a modified oligonucleotide.

[0134] Embodiment 124: The method of any of embodiments 111-123, wherein said second compound is administered prior to administration of the oligomeric compound of any of embodiments 1-94 or the composition of embodiment 95.

[0135] Embodiment 125: The method of any of embodiments 111-123, wherein said second compound is administered following administration of the oligomeric compound of any of embodiments 1-94 or the composition of embodiment 95.

[0136] Embodiment 126: The method of any of embodiments 111-123, wherein the second compound is administered less frequently than the administration of the oligomeric compound of any of embodiments 1-94 or the composition of embodiment 95.

[0137] Embodiment 127: An oligomeric compound according to any of embodiments 1 to 94 or a composition according to embodiment 95 for use in therapy.

[0138] Embodiment 128: Use of an oligomeric compound according to any of embodiments 1 to 94 or a composition according to embodiment 95 for the preparation of a medicament for the treatment of spinal muscular atrophy type I.

[0139] Embodiment 129: Use of an oligomeric compound according to any of embodiments 1 to 94 or a composition according to embodiment 95 for the preparation of a medicament for the treatment of spinal muscular atrophy type II.

[0140] Embodiment 130: Use of an oligomeric compound according to any of embodiments 1 to 94 or a composition according to embodiment 95 for the preparation of a medicament for the treatment of spinal muscular atrophy type III.

[0141] Embodiment 131: Use of an oligomeric compound according to any of embodiments 1 to 94 or a composition according to embodiment 95 for the preparation of a medicament for the treatment of spinal muscular atrophy type IV.

[0142] Embodiment 132: Use of an oligomeric compound according to any of embodiments 1 to 95 for the preparation of a medicament for the treatment of spinal muscular atrophy.

[0143] Embodiment 133: A method comprising: (i) systemically administering a compound comprising an oligomeric compound as described in any one of embodiments 1-73; and (ii) administering to the CNS a compound comprising an oligomeric compound as described in any one of embodiments 1-73.

[0144] Embodiment 134: A method comprising: (i) systemically administering a compound comprising an oligomeric compound or composition described in any of embodiments 1-73; and (ii) administering to the CNS a compound comprising a modified oligonucleotide having at least one modified nucleoside comprising a modified sugar moiety selected from 2'-OMe and 2'-MOE.

[0145] Embodiment 135: A method comprising (i) systemically administering a compound comprising an oligomeric compound according to any of embodiments 74-94, and (ii) administering to the CNS a compound comprising an oligomeric compound according to any of embodiments 1-73. A method comprising (i) systemically administering a compound comprising an oligomeric compound according to any of embodiments 74-94, and (ii) administering to the CNS a compound comprising a modified oligonucleotide having at least one modified nucleoside comprising a modified sugar moiety selected from 2'-OMe and 2'-MOE.

[0146] Embodiment 136: A method comprising: (i) systemically administering a modified oligonucleotide having a conjugate with at least one modified nucleoside that comprises a modified sugar moiety selected from 2'-OMe and 2'-MOE; and (ii) administering to the CNS a compound comprising an oligomeric compound described in any of embodiments 1-73.

[0147] Embodiment 137: A method comprising: (i) systemically administering a modified oligonucleotide having at least one modified nucleoside comprising a modified sugar moiety selected from 2'-OMe and 2'-MOE and a conjugate group; and (ii) administering to the CNS a compound comprising a modified oligonucleotide having at least one modified nucleoside comprising a modified sugar moiety selected from 2'-OMe and 2'-MOE.

[0148] Embodiment 138: The method according to embodiment 137, wherein the conjugate group comprises a lipid or lipophilic group selected from cholesterol, C10-C26 saturated fatty acid, C10-C26 unsaturated fatty acid, C10-C26 alkyl, triglyceride, tocopherol or cholic acid. method.

[0149] Embodiment 139: The compound or method of any of embodiments 1 to 138, wherein the SMN2 pre-mRNA is a nucleobase sequence selected from any of SEQ ID NOs: 10, 11, or 240. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0150] It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the claimed embodiments. As used herein, the use of the singular includes the plural unless specifically stated otherwise. As used herein, the use of "or" means "and / or" unless specifically stated otherwise. Furthermore, the use of the term "including" as well as other forms such as "includes" and "included" are not limiting.

[0151] The section headings used herein are for organizational purposes only and are not intended to limit the subject matter described. All documents or portions of documents cited in this application, including, but not limited to, patents, patent applications, articles, books, papers, and GenBank and NCBI reference sequence records, are expressly incorporated herein by reference in their entirety for the portions of the documents discussed herein.

[0152] Unless otherwise indicated, the following terms have the following meanings.

[0153] As used herein, "SMN2 pre-mRNA" refers to an RNA sequence including all exons, introns and untranslated regions transcribed from DNA encoding human SMN2. In certain embodiments, the DNA encoding SMN2 includes the human SMN2 genomic sequence truncated from nucleotides 19939708-19967777 set forth in GENBANK Accession No. NT_006713.14, which is incorporated herein by reference as SEQ ID NO:240. In certain embodiments, the SMN2 pre-mRNA includes SEQ ID NO:10. Nucleotides 1-60 of SEQ ID NO:10 represent a portion of intron 6, nucleotides 61-114 of SEQ ID NO:10 represent exon 7, and nucleotides 115-174 of SEQ ID NO:10 represent a portion of intron 7.

[0154] As used herein, "ISS-N1" refers to an intronic splice silencing domain in intron 7. In certain embodiments, ISS-N1 comprises the nucleobase sequence of SEQ ID NO:11.

[0155] As used herein, "2'-deoxyribonucleoside" refers to a nucleoside containing a 2'-H(H) furanosyl sugar moiety found in naturally occurring deoxyribonucleic acid (DNA). In certain embodiments, 2'-deoxyribonucleosides may contain modified nucleobases or may contain RNA nucleobases (uracil).

[0156] As used herein, "2'-substituted nucleoside" or "2-modified nucleoside" refers to a nucleoside that includes a 2'-substituted or 2'-modified sugar moiety. As used herein, "2'-substituted" or "2-modified" with respect to a sugar moiety means a sugar moiety that includes at least one 2'-substituent group other than H or OH.

[0157] As used herein, "antisense activity" refers to any detectable and / or measurable change that results from hybridization of an antisense compound to its target nucleic acid. In certain embodiments, antisense activity refers to a change in the amount or expression of a target nucleic acid or a protein encoded by the target nucleic acid that is greater than or equal to the amount or expression of the target nucleic acid in the absence of the antisense compound. The target nucleic acid or protein level is decreased as compared to the target nucleic acid or protein level in the presence of the antibody.

[0158] As used herein, "antisense compound" means a compound comprising an antisense oligonucleotide and, optionally, one or more additional features, such as a conjugate group or a terminal group.

[0159] As used herein, "antisense oligonucleotide" means an oligonucleotide having a nucleobase sequence that is at least partially complementary to a target nucleic acid.

[0160] As used herein, "ameliorating" in reference to treatment means that at least one symptom is improved compared to the same symptom in the absence of treatment. In certain embodiments, amelioration is reducing the severity or frequency of a symptom, or delaying the onset of a symptom, or slowing the progression of the severity or frequency of a symptom.

[0161] As used herein, "bicyclic nucleoside" or "BNA" refers to a nucleoside that includes a bicyclic sugar moiety. As used herein, "bicyclic sugar" or "bicyclic sugar moiety" refers to a modified sugar moiety that includes two rings, with the second ring formed by a bridge connecting two of the atoms of the first ring to form a bicyclic structure. In certain embodiments, the first ring of the bicyclic sugar moiety is a furanosyl moiety. In certain such embodiments, the furanosyl moiety is a ribosyl moiety. In certain embodiments, the bicyclic sugar moiety does not include a furanosyl moiety.

[0162] As used herein, "branching group" refers to a grouping of atoms having at least three positions capable of forming covalent bonds with at least three groups. In certain embodiments, the branching group provides multiple reactive sites for connecting tethered ligands to the oligonucleotide via conjugate linkers and / or cleavable moieties.

[0163] As used herein, a "cell targeting moiety" means a conjugate group or a portion of a conjugate group that improves uptake into a particular cell type and / or distribution to a particular tissue compared to an oligomeric compound that does not have a cell targeting moiety.

[0164] As used herein, "cleavable moiety" means a bond or group that is cleaved under physiological conditions, eg, inside a cell, animal or human.

[0165] As used herein, "complementary" with respect to an oligonucleotide means that at least 70% of the nucleobases of the oligonucleotide or one or more regions thereof and the nucleobases of another nucleic acid or one or more regions thereof can hydrogen bond with each other when the nucleobase sequence of the oligonucleotide and the other nucleic acid are aligned in reverse orientation. Complementary nucleobases refer to nucleobases that can form hydrogen bonds with each other. Complementary nucleobase pairs include adenine (A) and thymine (T), adenine (A) and uracil (U), cytosine (C) and guanine (G), 5-methylcytosine (C) and cytosine (D) and cytosine (E). m Examples of complementary bases include amino acids 1 (a, b, c) and guanine (G). Complementary oligonucleotides and / or nucleic acids need not have nucleobase complementarity at every nucleoside. Rather, some mismatches are permitted. As used herein, "fully complementary" or "100% complementary" with respect to an oligonucleotide means that the oligonucleotide is complementary to another oligonucleotide or nucleic acid at every nucleoside of the oligonucleotide.

[0166] As used herein, "conjugate group" refers to a group of atoms that is directly or indirectly attached to an oligonucleotide. A conjugate group includes a conjugate moiety and a conjugate linker that attaches the conjugate moiety to the oligonucleotide. Includes.

[0167] As used herein, "conjugate linker" means a group of atoms that includes at least one bond that connects a conjugate moiety to an oligonucleotide.

[0168] As used herein, "conjugate moiety" means a group of atoms that is attached to an oligonucleotide via a conjugate linker.

[0169] As used herein, "contiguous" in the context of an oligonucleotide refers to nucleosides, nucleobases, sugar moieties, or internucleoside linkages that are immediately adjacent to one another. For example, "contiguous nucleobases" refers to nucleobases that are immediately adjacent to one another in a sequence.

[0170] As used herein, a "double-stranded antisense compound" refers to an antisense compound that contains two oligomeric compounds that are complementary to each other and form a duplex, wherein one of the two oligomeric compounds contains an antisense oligonucleotide.

[0171] As used herein, "fully modified" in reference to modified oligonucleotide refers to modified oligonucleotide in which each sugar moiety is modified. "Uniformly modified" in reference to modified oligonucleotide refers to fully modified oligonucleotide in which each sugar moiety is identical. For example, the nucleosides of a uniformly modified oligonucleotide each have a 2'-MOE modification, but may have different nucleobase modifications and may have different internucleoside linkages.

[0172] As used herein, a "gapmer" means a modified oligonucleotide that includes an inner region having a plurality of nucleosides that include unmodified sugar moieties, disposed between outer regions having one or more nucleosides that include modified sugar moieties, each of which nucleosides in the outer regions that are adjacent to an inner region include a modified sugar moiety. The inner region may be referred to as the "gap" and the outer regions may be referred to as "wings."

[0173] As used herein, "hybridization" refers to the pairing or annealing of complementary oligonucleotides and / or nucleic acids. Although not limited to a particular mechanism, the most common hybridization mechanism involves hydrogen bonding between complementary nucleobases, which can be Watson-Crick, Hoogsteen, or reversed Hoogsteen hydrogen bonding.

[0174] As used herein, "inhibiting expression or activity" refers to a reduction or blocking of expression or activity compared to the expression of the activity in an untreated or control sample, and does not necessarily indicate a complete elimination of expression or activity.

[0175] As used herein, the term "internucleoside linkage" refers to a group or bond that forms a covalent bond between adjacent nucleosides in an oligonucleotide. As used herein, "modified internucleoside linkage" refers to any internucleoside linkage other than a naturally occurring phosphate internucleoside linkage. Non-phosphate linkages are referred to herein as modified internucleoside linkages. "Phosphorothioate linkage" refers to a modified phosphate linkage in which one of the non-bridging oxygen atoms of the phosphate linkage is replaced with a sulfur atom. A phosphorothioate internucleoside linkage is a modified internucleoside linkage. Modified internucleoside linkages include linkages that contain abasic nucleosides. As used herein, "abasic nucleoside" refers to a sugar moiety in an oligonucleotide or oligomeric compound that is not directly connected to a nucleobase. In certain embodiments, an abasic nucleoside is one or more nucleoside moieties in an oligonucleotide. is flanked by two nucleosides.

[0176] As used herein, "Isis396443" refers to an oligonucleotide having the following structure: Tes m Ces Aes m Ces Tes Tes Tes m Ces Aes Tes Aes Aes Tes Ges m Ces Tes Ges Ge In the array " m "C" indicates 5-methylcytosine, "e" indicates 2'-MOE modification, "C" indicates cytidine, "T" indicates thymidine, "A" indicates adenosine, "G" indicates guanosine, and "s" indicates a phosphorothioate bond. Isis396443 is also known in the art as nusinersen and Ionis-SMNRx.

[0177] As used herein, "linker nucleoside" refers to a nucleoside that directly or indirectly connects an oligonucleotide to a conjugate moiety. The linker nucleoside is located within the conjugate linker of an oligomeric compound. The linker nucleoside is not considered to be part of the oligonucleotide moiety of an oligomeric compound, even if it is contiguous with the oligonucleotide.

[0178] As used herein, "non-bicyclic modified sugar" or "non-bicyclic modified sugar moiety" means a modified sugar moiety that contains a modification such as a substituent that does not form a bridge between two atoms of the sugar to form a second ring.

[0179] As used herein, "linked nucleosides" are nucleosides that are joined in a contiguous sequence (ie, there are no additional nucleosides between the linked nucleosides).

[0180] As used herein, "mismatch" or "non-complementary" means that the nucleobases of a first oligonucleotide are not complementary to the corresponding nucleobases of a second oligonucleotide or a target nucleic acid when the first oligomeric compound and second oligomeric compound are aligned.

[0181] As used herein, "MOE" means methoxyethyl. "2'-MOE" means an -OCH group at the 2' position of the furanosyl ring. 2 CH 2 OCH 3 means a group.

[0182] As used herein, "motif" means a pattern of unmodified and / or modified sugar moieties, nucleobases and / or internucleoside linkages in an oligonucleotide.

[0183] As used herein, "naturally occurring" means found in nature.

[0184] As used herein, "nucleobase" refers to a naturally occurring nucleobase or a modified nucleobase. As used herein, "naturally occurring nucleobase" refers to adenine (A), thymine (T), cytosine (C), uracil (U) and guanine (G). As used herein, a modified nucleobase is an atomic group that can pair with at least one naturally occurring nucleobase. A universal base is a nucleobase that can pair with any one of the five unmodified nucleobases. As used herein, "nucleobase sequence" refers to the order of consecutive nucleobases in a nucleic acid or oligonucleotide, regardless of any sugar or internucleoside linkage modifications.

[0185] As used herein, "nucleoside" refers to a compound that includes a nucleobase and a sugar moiety. The nucleobase and sugar moieties can each independently be unmodified or modified. As used herein, "modified nucleoside" means a nucleoside that includes a modified nucleobase and / or a modified sugar moiety.

[0186] As used herein, "2'-O-(N-alkylacetamido)" refers to an -O-CH 2 means a -C(O)-NH-alkyl group.

[0187] As used herein, "2'-O-(N-methylacetamide)" or "2'-NMA" refers to an -O-CH 2 -C(O)-NH-CH 3 means a group.

[0188] As used herein, "oligomeric compound" means a compound consisting of an oligonucleotide and, optionally, one or more additional features, such as a conjugate group or a terminal group.

[0189] As used herein, "oligonucleotide" refers to a chain of linked nucleosides connected via internucleoside bonds, where each nucleoside and internucleoside bond may be modified or unmodified. Unless otherwise specified, an oligonucleotide consists of 8 to 50 linked nucleosides. As used herein, "modified oligonucleotide" refers to an oligonucleotide in which at least one nucleoside or internucleoside bond is modified. As used herein, "unmodified oligonucleotide" refers to an oligonucleotide that does not contain any nucleoside or internucleoside modifications.

[0190] As used herein, "a pharma- ceutically acceptable carrier or diluent" refers to any substance suitable for use in administration to an animal. Such a particular carrier allows for the formulation of pharmaceutical compositions for oral ingestion by a subject, such as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions, and lozenges. In a particular embodiment, the pharma- ceutically acceptable carrier or diluent is sterile water, sterile saline, or a sterile buffer solution.

[0191] As used herein, "pharmaceutically acceptable salts" refers to physiologically and pharma- ceutically acceptable salts of compounds, such as oligomeric compounds, i.e., salts that retain the desired biological activity of the parent compound and do not impart undesired toxicological effects.

[0192] As used herein, "pharmaceutical composition" refers to a mixture of substances suitable for administration to a subject. For example, a pharmaceutical composition may include an antisense compound and a sterile aqueous solution. In certain embodiments, the pharmaceutical composition exhibits activity in a free uptake assay in a particular cell line.

[0193] As used herein, "phosphorus moiety" refers to a group of atoms that includes a phosphorus atom. In certain embodiments, the phosphorus moiety includes a monophosphate, diphosphate, or triphosphate, or a mono-, di-, or triphosphorothioate.

[0194] As used herein, "phosphodiester internucleoside linkage" means a phosphate group covalently linking two adjacent nucleosides of a modified oligonucleotide.

[0195] As used herein, "pre-transcript" refers to a coding or non-coding RNA that is processed to form the processed or mature form of a transcript. Pre-transcripts include, but are not limited to, pre-mRNA, long non-coding RNA, pri-miRNA, and intronic RNA.

[0196] As used herein, "processing" with respect to a precursor transcript means that the precursor transcript is converted to form a corresponding processed transcript. Processing of a precursor transcript includes, but is not limited to, a nuclease cleavage event at the processing site of the precursor transcript.

[0197] As used herein, "prodrug" refers to a form of a therapeutic agent that is converted into a different form within the body or cells. Typically, the conversion of a prodrug within the body is facilitated by the action of enzymes (e.g., endogenous or viral enzymes) or chemicals present in cells or tissues and / or by physiological conditions.

[0198] As used herein, "RNAi compounds" refers to antisense compounds that act, at least in part, through RISC or Ago2 to regulate target nucleic acids and / or proteins encoded by target nucleic acids. RNAi compounds include, but are not limited to, double-stranded siRNAs, single-stranded RNAs (ssRNAs), and microRNAs (including microRNA mimics). In certain embodiments, RNAi compounds regulate the amount, activity, and / or splicing of target nucleic acids. The term RNAi compounds does not include antisense oligonucleotides that act through RNase H.

[0199] As used herein, the term "single-stranded" in relation to antisense compounds refers to a compound that is composed of one oligomeric compound, and does not pair with a second oligomeric compound to form a double strand.The term "self-complementary" in relation to oligonucleotides refers to an oligonucleotide that at least partially hybridizes with itself.A compound that is composed of one oligomeric compound, and the oligonucleotide of the oligomeric compound is self-complementary, is a single-stranded compound.A single-stranded antisense compound or oligomeric compound can be bound to a complementary oligomeric compound to form a double strand.

[0200] As used herein, "splicing" refers to the process by which a pre-mRNA is processed to form the corresponding mRNA, including, but not limited to, the removal of introns from the pre-mRNA and the joining of exons.

[0201] As used herein, "sugar moiety" refers to an unmodified sugar moiety or a modified sugar moiety. As used herein, "unmodified sugar moiety" refers to a 2'-OH(H) furanosyl moiety as found in RNA (an "unmodified RNA sugar moiety") or a 2'-H(H) moiety as found in DNA (an "unmodified DNA sugar moiety"). An unmodified sugar moiety has one hydrogen each at the 1', 3', and 4' positions, one oxygen at the 3' position, and two hydrogens at the 5' position. As used herein, "modified sugar moiety" or "modified sugar" refers to a modified furanosyl sugar moiety or sugar surrogate. As used herein, modified furanosyl sugar moiety refers to a furanosyl sugar that includes a non-hydrogen substituent in place of at least one hydrogen of the unmodified sugar moiety. In certain embodiments, the modified furanosyl sugar moiety is a 2'-substituted sugar moiety. Such modified furanosyl sugar moieties include bicyclic and non-bicyclic sugars. As used herein, "sugar surrogate" refers to a modified sugar moiety, other than a furanosyl moiety, that can attach a nucleobase to another group, such as an internucleoside linkage, a conjugate group, or a terminal group of an oligonucleotide. Modified nucleosides that include sugar surrogates can be incorporated at one or more positions within an oligonucleotide, and such oligonucleotides can hybridize to complementary oligomeric compounds or nucleic acids.

[0202] As used herein, "target precursor transcript" refers to a precursor transcript to which an oligonucleotide is designed to hybridize. In certain embodiments, the target precursor transcript is a target pre-mRNA. As used herein, "target processed transcript" refers to an RNA resulting from processing of a corresponding target precursor transcript. In certain embodiments, the target processed transcript is a target mRNA. As used herein, "target pre-mRNA" refers to a pre-mRNA to which an oligonucleotide is designed to hybridize. As used herein, "target mRNA" refers to an mRNA resulting from splicing of a corresponding target pre-mRNA.

[0203] As used herein, "terminal group" means a chemical group or group of atoms covalently attached to the end of an oligonucleotide.

[0204] Spinal muscular atrophy SMA is a genetic disease characterized by degeneration of motor neurons in the spinal cord. SMA results from homozygous deletion of both functional copies of the SMN1 gene. However, the SMN2 gene encodes the same protein as SMN1, and may thus overcome the genetic defect in SMA patients. SMN2 contains a translationally silent mutation (C→T) at position +6 of exon 7, which results in poor inclusion of exon 7 in SMN2 transcripts. Thus, the predominant form of SMN2 lacks exon 7 and is unstable and inactive. Therefore, therapeutic compounds that can modulate SMN2 splicing to increase the proportion of SMN2 transcripts that contain exon 7 would be useful in treating SMA. In certain embodiments, modified oligonucleotides with one or more 2'-O-(N-alkylacetamido) modified sugar moieties have improved pharmacological activity in modulating SMN2 pre-mRNA. This includes increasing the proportion of SMN2 transcripts that contain exon 7. In certain embodiments, modified oligonucleotides bearing one or more 2'-O-(N-methylacetamido) modified sugar moieties have improved pharmacological activity in modulating SMN2 pre-mRNA, including increasing the proportion of SMN2 transcripts that contain exon 7.

[0205] Although SMA is typically characterized by degeneration of motor neurons in the spinal cord, SMN2 is ubiquitously expressed in cells throughout the body, including muscle cells. In certain embodiments, improving SMN activity in muscle cells provides a therapeutic benefit. In certain embodiments, modified oligonucleotides having one or more 2'-O-(N-alkylacetamido) modified sugar moieties have improved cellular distribution to tissues throughout the body, including muscle tissue. In certain embodiments, modified oligonucleotides having one or more 2'-O-(N-methylacetamido) modified sugar moieties have improved cellular distribution to tissues throughout the body, including muscle tissue.

[0206] In certain embodiments, modified oligonucleotides having one or more 2'-O-(N-alkylacetamido) modified sugar moieties include conjugates that may further enhance cellular distribution and / or pharmacological activity. In certain embodiments, the conjugate enhances distribution to muscle tissue. In certain embodiments, the conjugate is a lipid.

[0207] In certain embodiments, modified oligonucleotides having one or more 2'-O-(N-methylacetamido) modified sugar moieties include conjugates that may further enhance cellular distribution and / or pharmacological activity. In certain embodiments, the conjugate enhances distribution to muscle tissue. In certain embodiments, the conjugate is a lipid.

[0208] Specific nucleobase sequences targeted to SMN2 pre-mRNA are exemplified in the non-limiting table below. Each nucleobase sequence in the table below contains six or more 2'-O-(N- Any of the nucleobase sequences in the table below can be modified with six or more 2'-O-(N-methylacetamido) modified sugar moieties.

[0209] Any of the nucleobase sequences in the table below can be modified with six or more 2'-O-(N-alkylacetamido) modified sugar moieties and can include a conjugate moiety. Any of the nucleobase sequences in the table below can be modified with six or more 2'-O-(N-methylacetamido) modified sugar moieties and can include a conjugate moiety. [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5] [Table 1-6] [Table 1-7] [Table 2]

[0210] In the above table, the subscript "x" represents an internucleoside linkage selected from a phosphorothioate internucleoside linkage or a phosphate internucleoside linkage, and "n" represents a 2'-O-(N-methylacetamido) modified nucleoside. The superscript "m" before the C represents 5-methylcysteine.

[0211] In certain embodiments, the present disclosure provides the nucleic acid sequences of SEQ ID NOs: 1, 2, 3, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82 , 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 8, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 2 In one embodiment, modified oligonucleotides are provided that comprise a sequence selected from any of the following: 10, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264 or 265.

[0212] In certain embodiments, the present disclosure provides the nucleic acid sequences of SEQ ID NOs: 1, 2, 3, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 11 9, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 15 0, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 1 81, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 2 In one embodiment, modified oligonucleotides are provided that consist of a sequence selected from any of the following: 12, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264 or 265.

[0213] In certain embodiments, the present disclosure provides the nucleic acid sequences of SEQ ID NOs: 1, 2, 3, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 109, 109, 109, 102, 104, 105, 106, 107, 108, 109, 109, 110, 111, 112, 113, 114, 9, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 2 13, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 311, 312, 313, 314, 315, 3 7, 258, 259, 260, 261, 262, 263, 264 or 265, wherein at least one, at least two, at least three, at least four, at least five or at least six nucleosides of the modified oligonucleotide comprise a 2'-O-(N-methylacetamido) modified sugar moiety.

[0214] In certain embodiments, the present disclosure provides the nucleic acid sequences of SEQ ID NOs: 1, 2, 3, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 1 5, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181 , 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212 , 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264 or 265, whereinAt least one, at least two, at least three, at least four, at least five, or at least six nucleosides of the modified oligonucleotide contain a 2'-O-(N-methylacetamido) modified sugar moiety.

[0215] Specific combinations In certain embodiments, the first agent comprising a compound described herein is administered in combination with one or more second In certain embodiments, the second agent is co-administered with the first agent. In certain embodiments, such a second agent is designed to treat the same disease, disorder or condition as the first agent described herein. In certain embodiments, the second agent is co-administered with the first agent to provide a combined effect. In certain embodiments, the second agent is co-administered with the first agent to provide a synergistic effect. In certain embodiments, the co-administration of the first agent and the second agent allows for the use of lower doses than would be required to achieve a therapeutic or prophylactic effect when these agents are administered as independent therapies.

[0216] In certain embodiments, the present disclosure provides for administration of a first antisense compound into the CSF in combination with systemic delivery of a second antisense compound. Systemic administration and CSF administration can be performed simultaneously, separately, or sequentially. In certain embodiments, the subject receives a first administration of the antisense compound into the CSF followed by a second systemic administration of the antisense compound. In certain embodiments, the antisense compound administered into the CSF is Isis396443. In certain embodiments, the antisense compound administered into the CSF is a modified oligonucleotide having one or more 2'-O-(N-alkylacetamido) modified sugar moieties. In certain embodiments, the antisense compound administered into the CSF is an unconjugated modified oligonucleotide having one or more 2'-O-(N-alkylacetamido) modified sugar moieties.

[0217] In certain embodiments, systemically administered antisense compounds are modified oligonucleotides having one or more 2'-O-(N-alkylacetamido) modified sugar moieties. In certain embodiments, systemically administered antisense compounds are modified oligonucleotides having one or more 2'-O-(N-methylacetamido) modified sugar moieties. In certain embodiments, systemically administered antisense compounds are modified oligonucleotides having one or more 2'-O-(N-alkylacetamido) modified sugar moieties and conjugate groups. In certain embodiments, systemically administered antisense compounds are modified oligonucleotides having one or more 2'-O-(N-methylacetamido) modified sugar moieties and conjugate groups.

[0218] The administration can be in any order and at different frequencies. For example, a subject can receive systemic administration of a modified oligonucleotide having one or more 2'-O-(N-alkylacetamido) or 2'-O-(N-methylacetamido) modified sugar moieties, followed by administration of an antisense compound (e.g., ISIS 396443) into the CSF. Alternatively, a subject can receive systemic administration of a modified oligonucleotide having one or more 2'-O-(N-alkylacetamido) or 2'-O-(N-methylacetamido) modified sugar moieties, followed by administration of an antisense compound (e.g., ISIS 396443) into the CSF. Alternatively, a subject can receive CSF administration of an antisense compound (e.g., ISIS 396443) and systemic administration of a modified oligonucleotide having one or more 2'-O-(N-alkylacetamido) or 2'-O-(N-methylacetamido) modified sugar moieties simultaneously. Administration into the CSF and systemic administration can be performed at different times and at different frequencies. For example, a subject can receive systemic administration of modified oligonucleotides having one or more 2'-O-(N-alkylacetamido) or 2'-O-(N-methylacetamido) modified sugar moieties on a weekly or monthly basis, and CSF administration of an antisense compound (e.g., ISIS 396443) into the CSF once every four months or once every six months. In certain embodiments in which the CSF administration of an antisense compound (e.g., ISIS 396443) and systemic administration of modified oligonucleotides having one or more 2'-O-(N-alkylacetamido) or 2'-O-(N-methylacetamido) modified sugar moieties are administered separately, sequentially, or sequentially, the administrations can be separated by various time periods, such as daily, weekly, or monthly.

[0219] I. Specific Oligonucleotides In certain embodiments, the present invention provides oligonucleotides consisting of linked nucleosides. The oligonucleotides may be unmodified oligonucleotides (unmodified RNA or DNA) or modified oligonucleotides. Modified oligonucleotides contain at least one modification relative to unmodified RNA or DNA (i.e., at least one modified nucleoside (containing a modified sugar moiety and / or modified nucleobase) and / or at least one modified internucleoside linkage).

[0220] A. Certain Modified Nucleosides A modified nucleoside comprises a modified sugar moiety or a modified nucleobase or both a modified sugar moiety and a modified nucleobase.

[0221] 1. Specific sugar moieties In certain embodiments, the modified sugar moiety is a non-bicyclic modified sugar moiety. In certain embodiments, the modified sugar moiety is a bicyclic or tricyclic sugar moiety. In certain embodiments, the modified sugar moiety is a sugar surrogate. Such sugar surrogates can contain one or more substitutions that correspond to the substitutions of other types of modified sugar moieties.

[0222] In certain embodiments, the modified sugar moiety is a non-bicyclic modified sugar moiety, including a furanosyl ring having one or more acyclic substituents, including but not limited to, substituents at the 2', 4', and / or 5' positions. In certain embodiments, one or more of the acyclic substituents of the non-bicyclic modified sugar moiety is branched. Examples of suitable 2'-substituents for non-bicyclic modified sugar moieties include, but are not limited to, 2'-O-(N-alkylacetamido), e.g., 2'-O-(N-methylacetamido). See, e.g., US 6,147,200 and Prakash et al., Org. Lett., 5,403-6 (2003). A "2'-O-(N-methylacetamido)" or "2'-NMA" modified nucleoside is shown below. [ka]

[0223] In certain embodiments, the 2'-substituent is 2'-F, 2'-OCH 3 ("OMe" or "O-methyl"), 2'-O(CH 2 ) 2 OCH 3 ("MOE"), halo, aryl, amino, azido, SH, CN, OCN, CF 3 , OCF 3 , O.C. 1 ~C 10 Alkoxy, OC 1 ~C 10 Substituted alkoxy, OC 1 ~C 10 Alkyl, OC 1 ~C 10 Substituted alkyl, S-alkyl, N(R m )-Alkyl, O-Alkenyl, S-Alkenyl, N(R m )-alkenyl, O-alkynyl, S-alkynyl, N(R m )-alkynyl, O-alkylenyl-O-alkyl, alkynyl, alkaryl, aralkyl, O-alkaryl, O-aralkyl, O(CH 2 ) 2 SCH 3 , O(CH 2 ) 2 ON(R m )(R n ) or OCH 2 C(=O)-N(R m )(R n )(In the formula, each R m and R n are independently H, an amino protecting group, or a substituted or unsubstituted C 1 ~C 10 alkyl) and the 2'-substituents described in Cook et al., US 6,531,584; Cook et al., US 5,859,221; and Cook et al., US 6,005,087. Particular embodiments of these 2'-substituents are selected from hydroxyl, , amino, alkoxy, carboxy, benzyl, phenyl, nitro(NO 2), thiol, thioalkoxy, thioalkyl, halogen, alkyl, aryl, alkenyl, and alkynyl. Examples of suitable 4'-substituents for non-bicyclic modified sugar moieties include, but are not limited to, alkoxy (e.g., methoxy), alkyl, and those described in Manoharan et al., WO 2015 / 106128. Examples of suitable 5'-substituents for non-bicyclic modified sugar moieties include, but are not limited to, 5'-methyl (R or S), 5'-vinyl, and 5'-methoxy. In certain embodiments, non-bicyclic modified sugars comprise two or more non-bridging sugar substituents, such as 2'-F-5'-methyl sugar moieties and modified sugar moieties and modified nucleosides described in Migawa et al., WO 2008 / 101157 and Rajeev et al., US 2013 / 0203836).

[0224] In certain embodiments, the 2'-substituted nucleoside or 2'-non-bicyclic modified nucleoside is selected from the group consisting of F, NH 2 , N 3 , OCF 3 , O.C.H. 3 , O(CH 2 ) 3 NH 2 , C.H. 2 CH=CH 2 , O.C.H. 2 CH=CH 2 , O.C.H. 2 CH 2 OCH 3 , O(CH 2 ) 2 SCH 3 , O(CH 2 ) 2 ON(R m )(R n ), O(CH 2 ) 2 O(CH 2 ) 2 N(CH 3 ) 2 and N-substituted acetamides (OCH 2 C(=O)-N(R m )(R n ))(In the formula, each R m and Rn are independently H, an amino protecting group, or a substituted or unsubstituted C 1 ~C 10 In certain embodiments, each R m and R n are independently H or C 1 ~C 3 In certain embodiments, each R m and R n is independently H or methyl.

[0225] In certain embodiments, the 2'-substituted nucleoside or 2'-non-bicyclic modified nucleoside is F, OCF 3 , O.C.H. 3 , O.C.H. 2 CH 2 OCH 3 , O(CH 2 ) 2 SCH 3 , O(CH 2 ) 2 ON(CH 3 ) 2 , O(CH 2 ) 2 O(CH 2 ) 2 N(CH 3 ) 2 and OCH 2 C(=O)-N(H)CH 3 The sugar moiety comprises a non-bridging 2'-substituent selected from:

[0226] In certain embodiments, the 2'-substituted nucleoside or 2'-non-bicyclic modified nucleoside is F, OCH 3 , O.C.H. 2 CH 2 OCH 3 and OCH 2 C(=O)-N(H)CH 3 The sugar moiety comprises a non-bridging 2'-substituent selected from:

[0227] Nucleosides that include modified sugar moieties, such as non-bicyclic modified sugar moieties, can be referred to by the position(s) of the substitution(s) on the sugar moiety of the nucleoside. For example, nucleosides that include a 2'-substituted or 2-modified sugar moiety are referred to as 2'-substituted nucleosides or 2-modified nucleosides.

[0228] Certain modified sugar moieties include bridged sugar substituents that form a second ring to provide a bicyclic sugar moiety. In certain such embodiments, the bicyclic sugar moiety includes a bridge between the 4'-furanose ring atom and the 2'-furanose ring atom. Examples of such 4'-2' bridged sugar substituents include 4'-CH 2 -2',4'-(CH 2 ) 2 -2',4'-(CH 2 ) 3 -2',4'-CH 2 -O-2' ("LNA"), 4'-CH 2 -S-2',4'-(CH 2 ) 2 -O-2' ("ENA"), 4'-CH(CH 3 )-O-2' (also called "constrained ethyl" or "cEt" when in the S configuration), 4'-CH 2 -O-CH 2 -2',4'-CH 2 -N(R)-2',4'-CH(CH 2 OCH 3 )-O-2' ("constrained MOE" or "cMOE") and its analogs (see, e.g., Seth et al., US 7,399,845; Bhat et al., US 7,569,686; Swayze et al., US 7,741,457; and Swayze et al., US 8,022,193), 4'-C(CH 3 )(CH 3 )-O-2' and its analogs (see, e.g., Seth et al., US8,278,283), 4'-CH 2 -N(OC H 3 )-2' and its analogs (see, e.g., Prakash et al., US 8,278,425), 4'-CH 2-ON(CH 3 )-2' (see, e.g., Allerson et al., US 7,696,345 and Allerson et al., US 8,124,745), 4'-CH 2 -C(H)(CH 3 )-2' (see, for example, Zhou, et al., J. Org. Chem., 2009, 74, 118-134), 4'-CH 2 -C(=CH 2 )-2' and its analogs (see, e.g., Seth et al. al., US 8,278,426), 4'-C(R a R b )-N(R)-O-2',4'-C(R a R b )-ON(R)-2',4'-CH 2 -ON(R)-2' and 4'-CH 2 -N(R)-O-2' (in the formula, each R, R a and R b are independently H, a protecting group, or C 1 ~C 12 alkyl) (see, for example, Imanishi et al., US Pat. No. 7,427,672).

[0229] In certain embodiments, such a 4'-2' bridge is independently -[C(R a )(R b )] n -,-[C(R a )(R b )] n -O-, C(R a )=C(R b )-, C(R a )=N-, C(=NR a )-, -C(=O)-, -C(=S)-, -O-, -Si(R a ) 2 -, -S(=O) x - and N(R a )-, During the ceremony, x is 0, 1 or 2; n is 1, 2, 3 or 4; Each R a and R b are independently H, a protecting group, a hydroxyl, C 1 ~C 12 Alkyl, substituted C 1 ~C 12 Alkyl, C 2 ~C 12 Alkenyl, substituted C 2 ~C 12 Alkenyl, C 2 ~C 12 Alkynyl, Substituted C 2 ~C 12 Alkynyl, C 5 ~C 20 Aryl, Substituted C 5 ~C 20 Aryl, heterocyclic group, substituted heterocyclic group, heteroaryl, substituted heteroaryl, C 5 ~C 7 Alicyclic group, substituted C 5 ~C 7 Alicyclic groups, halogens, OJ 1 , N.J. 1 J 2 , S.J. 1 , N 3 , COOJ 1 , acyl (C(=O)-H), substituted acyl, CN, sulfonyl (S(=O) 2 -J 1 ), or sulfoxyl (S(=O)-J 1 ) and each J 1 and J 2 are independently H, C 1 ~C 12 Alkyl, substituted C 1 ~C 12 Alkyl, C 2 ~C 12 Alkenyl, substituted C 2 ~C 12 Alkenyl, C 2 ~C 12 Alkynyl, Substituted C 2 ~C 12 Alkynyl, C 5 ~C 20 Aryl, Substituted C 5 ~C 20Aryl, acyl (C(=O)-H), substituted acyl, heterocyclic group, substituted heterocyclic group, C 1 ~C 12 Aminoalkyl, substituted C 1 ~C 12 It is an aminoalkyl or a protecting group.

[0230] Further bicyclic sugar moieties are known in the art, see, for example, Freier et al., Nucleic Acids Research, 1997, 25(22), 4429-4443; Albaek et al., J. Org. Chem., 2006, 71, 7731-7740; Singh et al., Chem. Commun., 1998, 4, 455-456; Koshkin et al., Tetrahedron, 1998, 54, 3607-3630; Kumar et al., Bioorg. Med. Chem. Lett., 1998, 8, 2219-2222; Singh et al., J. Org. Chem., 1998, 63, 10035-10039; Srivastava et al. al.,J.Am.Chem.Soc.,20017,129,8362-8379;Wengel et a.,US7,053,207;Imanishi et al.,US6,268,490;Imanishi et al. al.US6,770,748;Imanishi et al.,USRE44,779;Wengel et al.,US6,794,499;Wengel et al.,US6,670,461;Wengel et al.,US7,034,133;Wengel et al.,US8,080,644;Wengel et al. al.,US8,034,909;Wengel et al.,US8,153,365;Wengel et al.,US7,572,5 82; and Ramasamy et al.,US6,525,191;;Torsten et al.,WO2004 / 106356;Wengel et al.,WO1999 / 014226;Seth et al.,WO2007 / 134181;Seth et al.,US7,547,684;Seth et al. al.,US7,666,854;Seth et al.,US8,088,746;Seth et al.,US7,750,131;Seth et al.,US8,030,467;Seth et al.,US8,268,980;Seth et al.,US8,546,556;Seth et al. al.,US8,530,640;Migawa et al.,US9,012,421;Seth et al. al., US8,501,805; and U.S. Patent Application Publications Allerson et al., US2008 / 0039618 and Migawa et al., US2015 / 0191727.

[0231] In certain embodiments, bicyclic sugar moieties and nucleosides incorporating such bicyclic sugar moieties are further defined by their isomeric configuration, for example, LNA nucleosides (as described herein) can be in the α-L or β-D configuration. [ka] α-L-Methyleneoxy(4'-CH 2 -O-2') or α-L-LNA bicyclic nucleosides have been incorporated into antisense oligonucleotides, which have shown antisense activity (Frieden et al., Nucleic Acids Research, 2003, 21, 6365-6372). References herein to bicyclic nucleosides include both isomeric configurations. When the positions of specific bicyclic nucleosides (e.g., LNA or cEt) are specified in the embodiments exemplified herein, they are in the β-D configuration unless otherwise stated.

[0232] In certain embodiments, the modified sugar moiety comprises one or more non-bridging sugar substituents and one or more bridging sugar substituents (eg, 5'-substituted and 4'-2' bridged sugars).

[0233] In certain embodiments, the modified sugar moiety is a sugar surrogate. In certain such embodiments, the oxygen atom of the sugar moiety is replaced with, for example, a sulfur atom, a carbon atom, or a nitrogen atom. In certain such embodiments, such modified sugar moieties also include bridging and / or non-bridging substituents as described herein. For example, certain sugar surrogates include a 4'-sulfur atom and substitutions at the 2' position (see, e.g., Bhat et al., US 7,875,733 and Bhat et al., US 7,939,677) and / or substitutions at the 5' position.

[0234] In certain embodiments, the sugar surrogate comprises a ring with more than five atoms. For example, in certain embodiments, the sugar surrogate comprises a six-membered tetrahydropyran ("THP"). Such tetrahydropyrans may be further modified or substituted. Nucleosides containing such modified tetrahydropyrans include hexitol nucleic acid ("HNA"), anitol nucleic acid ("ANA"), mannitol nucleic acid ("MNA") (see, e.g., Leumann, CJ. Bioorg. & Med. Chem. 2002, 10, 841-854), fluoroHNA: [ka] ("F-HNA", see, e.g., Swayze et al., US8,088,904; Swayze et al., US8,440,803; Swayze et al., US8,796,437; and Swayze et al., US9,005,906; F-HNA can also be referred to as F-THP or 3'-fluorotetrahydropyran) and nucleosides, including further modified THP compounds having the formula: [ka] wherein, independently, for each of the modified THP nucleosides: Bx is a nucleobase moiety, T 3 and T 4 are each independently an internucleoside linking group that connects the modified THP nucleoside to the remainder of the oligonucleotide; or 3 and T 4 is an internucleoside linking group that connects the modified THP nucleoside to the remainder of the oligonucleotide, and T 3 and T 4 the other is H, a hydroxyl protecting group, an attached conjugate group or a 5'- or 3'-terminal group; q 1 , q 2 , q 3 , q 4 , q 5 , q 6 and q 7 are independently H, C 1 ~C 6 Alkyl, substituted C 1 ~C 6 Alkyl, C 2 ~C 6 Alkenyl, substituted C 2 ~C 6 Alkenyl, C 2 ~C 6 Alkynyl or substituted C 2 ~C 6 is alkynyl, R 1 and R 2 each independently represents hydrogen, halogen, substituted or unsubstituted alkoxy, NJ 1 J 2 , S.J. 1 , N 3 , OC(=X)J 1 ,OC(=X)NJ 1 J 2 , N.J. 3 C(=X)NJ 1 J 2 and CN, where X is O, S or NJ 1 and each J 1 , J 2 and J 3are independently H or C 1 ~C 6 Non-limiting examples include those selected from the group consisting of aryl, aryl, and alkyl.

[0235] In certain embodiments, wherein q 1 , q 2 , q 3 , q 4 , q 5 , q 6 and q 7 and H. In certain embodiments, q 1 , q 2 , q 3 , q 4 , q 5 , q 6 and q 7 At least one of q is other than H. In certain embodiments, 1 , q 2 , q 3 , q 4 , q 5 , q 6 and q 7 At least one of R 1 and R 2 and R is F. In certain embodiments, modified THP nucleosides are provided where R 1 is F and R 2 is H, and in certain embodiments, R 1 is methoxy and R 2 is H, and in certain embodiments, R 1 is methoxyethoxy, and R 2 is H.

[0236] In certain embodiments, the sugar surrogate has six or more atoms and includes a ring having two or more heteroatoms. For example, nucleosides containing morpholino sugar moieties and their use in oligonucleotides have been reported (e.g., Braasch et al., Biochemistry, 2002, 41, 4503-4510 and Summerton et al., Biochemistry, 2002, 41, 4503-4510). (See, e.g., US 5,698,685; Summerton et al., US 5,166,315; Summerton et al., US 5,185,444; and Summerton et al., US 5,034,506.) As used herein, the term "morpholino" refers to a sugar surrogate having the following structure: [ka]

[0237] In certain embodiments, morpholinos may be modified, for example, by adding or changing various substituents to the morpholino structures described above. Such sugar surrogates are also referred to herein as "modified morpholinos."

[0238] In certain embodiments, the sugar surrogate comprises an acyclic moiety. Examples of nucleosides and oligonucleotides that comprise such acyclic sugar surrogates include, but are not limited to, peptide nucleic acids ("PNAs"), acyclic butyl nucleic acids (see, e.g., Kumar et al., Org. Biomol. Chem., 2013, 11, 5853-5865), and the nucleosides and oligonucleotides described in Manoharan et al., WO2011 / 133876.

[0239] Many other bicyclic and tricyclic sugar and sugar surrogate ring systems that can be used in modified nucleosides are known in the art.

[0240] 2. Certain modified nucleobases In certain embodiments, modified oligonucleotides comprise one or more nucleosides comprising an unmodified nucleobase.In certain embodiments, modified oligonucleotides comprise one or more nucleosides comprising a modified nucleobase.

[0241] In certain embodiments, the modified nucleobase is selected from 5-substituted pyrimidines, 6-azapyrimidines, alkyl- or alkynyl-substituted pyrimidines, alkyl-substituted purines, and N-2, N-6, and O-6 substituted purines. In certain embodiments, the modified nucleobase is selected from 2-aminopropyladenine, 5-hydroxymethylcytosine, xanthine, hypoxanthine, 2-aminoadenine, 6-N-methylguanine, 6-N-methyladenine, 2-propyladenine, 2-thiouracil, 2-thiothymine, and 2-thiocytosine, 5-propynyl (-C≡C-CH 3 ) uracil, 5-propynylcytosine, 6-azouracil, 6-azocytosine, 6-azothymine, 5-ribosyluracil (pseudouracil), 4-thiouracil, 8-halo, 8-amino, 8-thiol, 8-thioalkyl, 8-hydroxyl, 8-aza and other 8-substituted purines, 5-halo, especially 5-bromo, 5-trifluoromethyl, 5-halouracil and 5-halocytosine, 7-methylguanine, 7-methyladenine, 2-F-a The bases are selected from 1,3-diazaphenoxazin-2-one, 1,3-diazaphenothiazin-2-one, and 9-(2-aminoethoxy)-1,3-diazaphenoxazin-2-one. Tricyclic pyrimidines such as noxazin-2-one (G-clamp) are included. Modified nucleobases also include those in which the purine or pyrimidine base is replaced with other heterocycles, such as 7-deaza-adenine, 7-deazaguanosine, 2-aminopyridine, and 2-pyridone. Additional nucleobases include those disclosed in Merigan et al., US 3,687,808; The Concise Encyclopedia Of Polymer Science And Engineering, Kroschwitz, JI, Ed., John Wiley & Sons, 1990, 858-859; Englisch et al., Angewandte Chemie, International Edition, 1991, 30, 613; Sanghvi, YS, Chapter 15, Antisense Research and Applications, Crooke, ST and Lebleu, B., Eds., CRC Press, 1993, 273-288; and Chapters 6 and 15, Antisense Drug Technology, Crooke ST, Ed., CRC Press, 2008, 163-166. and 442-443.

[0242] Publications that teach the preparation of some of the above and other modified nucleobases include, but are not limited to, Manoharan et al., US2003 / 0158403; Manoharan et al., US2003 / 0175906; Dinh et al., US4,845,205; Spielvogel et al., US5,130,302; Rogers et al., US5,134,066; Bischofberger et al., US5,175,273; Urdea et al., US5,367,066; Benner et al., US5,432,272; Matteucci et al., US5,434,257; Gmeiner et al., US5,457,187; Cook et al., US5,459,255; Froehler et al., US5,459,255; al.,US5,484,908;Matteucci et al.,US5,502,177;Hawkins et al.,US5,525,711;Haralambidis et al.,US5,552,540;Cook et al.,US5,587,469;Froehler et al.,US5,594,121;Switzer et al. al.,US5,596,091;Cook et al.,US5,614,617;Froehler et al.,US5,645,985;Cook et al.,US5,681,941;Cook et al.,US5,811,534;Cook et al.,US5,750,692;Cook et al.,US5,948,903;Cook et al. al.,US5,587,470;Cook et al.,US5,457,191;Matteucci et al.,US5,763,588;Froehler et al.,US5,830,653;Cook et al.,US5,808,027;Cook et al.,6,166,199;and Matteucci et al. al., US6,005,096 included.

[0243] B. Certain Modified Internucleoside Linkages In certain embodiments, the nucleosides of the modified oligonucleotides can be linked together using any internucleoside linkage. Two main types of internucleoside linkage groups are defined by the presence or absence of a phosphorus atom. Representative phosphorus-containing internucleoside linkages include, but are not limited to, phosphodiester linkages ("P=O") (also referred to as unmodified or naturally occurring linkages), phosphotriesters, methylphosphonates, phosphoramidates, and phosphate containing phosphorothioates ("P=S") and phosphorodithioates ("HS-P=S"). Representative non-phosphorus-containing internucleoside linkages include methylenemethylimino (-CH 2 -N(CH 3 )-O-CH 2 -), thiodiester, thio Onocarbamate (-OC(=O)(NH)-S-); Siloxane (-O-SiH 2 -O-); and N,N'-dimethylhydrazine (-CH 2 -N(CH 3 )-N(CH 3 )-). Modified internucleoside linkages can be used to alter, typically increase, the nuclease resistance of oligonucleotides compared to naturally occurring phosphate linkages. In certain embodiments, internucleoside linkages having chiral atoms can be prepared as racemic mixtures or as separate enantiomers. Representative chiral internucleoside linkages include, but are not limited to, alkylphosphonates and phosphorothioates. Methods for preparing phosphorus-containing and non-phosphorus-containing internucleoside linkages are well known to those skilled in the art.

[0244] Neutral internucleoside linkages include, but are not limited to, phosphotriester, methylphosphonate, MMI (3'-CH 2 -N(CH 3 )-O-5'), Amide-3 (3'-CH 2 -C(=O)-N(H)-5'), amide-4 (3'-CH2 -N(H)-C(=O)-5'), formacetal (3'-O-CH 2 -O-5'), methoxypropyl and thioform acetals (3'-S-CH 2 -O-5'). Additional neutral internucleoside linkages include nonionic linkages including siloxanes (dialkylsiloxanes), carboxylates, carboxamides, sulfides, sulfonates, and amides (see, e.g., Carbohydrate Modifications in Antisense Research; YS Sanghvi and PD Cook, Eds., ACS Symposium Series 580; Chapters 3 and 4, 40-65). Additional neutral internucleoside linkages include N, O, S, and CH 2 Non-ionic bonds containing mixed moieties are included.

[0245] C. Specific motifs In certain embodiments, modified oligonucleotides include one or more modified nucleosides that include modified sugars. In certain embodiments, modified oligonucleotides include one or more modified nucleosides that include modified nucleobases. In certain embodiments, modified oligonucleotides include one or more modified internucleoside linkages. In such embodiments, the modified, unmodified and modified different sugar moieties, nucleobases and / or internucleoside linkages of modified oligonucleotides define a pattern or motif. In certain embodiments, the sugar moieties, nucleobases and internucleoside linkage patterns are each independent of each other. Thus, modified oligonucleotides can be described by their sugar motif, nucleobase motif and / or internucleoside linkage motif (as used herein, nucleobase motif describes the modification to the nucleobase, regardless of the sequence of the nucleobase).

[0246] 1. Specific glycomotifs In certain embodiments, an oligonucleotide comprises one or more modified sugar and / or unmodified sugar moieties arranged in a predetermined pattern or in a predetermined sugar motif along the oligonucleotide or region thereof, which in certain instances includes, but is not limited to, any of the sugar modifications discussed herein.

[0247] In certain embodiments, a modified oligonucleotide comprises or consists of a region having a gapmer motif that includes two outer regions, or "wings," and a central or interior region, or "gap." The three regions of the gapmer motif (the 5'-wing, the gap, and the 3'-wing) form a contiguous nucleoside sequence in which at least some of the sugar moieties of each nucleoside in the wing are different from at least some of the sugar moieties of the nucleosides in the gap. Specifically, at least the sugar moieties of the nucleosides in each wing closest to the gap (the 3'-most nucleoside of the 5'-wing and the 5'-most nucleoside of the 3'-wing) are modified sugar moieties. The 5'-wing sugar motif is different from the sugar moieties of the gap nucleosides that are adjacent, i.e., unmodified sugar moieties that define the boundary between the wing and the gap (i.e., the wing / gap boundary). In certain embodiments, the sugar moieties within the gap are the same as each other. In certain embodiments, the gap contains one or more nucleosides having a sugar moiety that is different from the sugar moieties of one or more other nucleosides in the gap. In certain embodiments, the sugar motifs of the two wings are the same as each other (symmetric gapmer). In certain embodiments, the sugar motif of the 5'-wing is different from the sugar motif of the 3'-wing (asymmetric gapmer).

[0248] In certain embodiments, the gapmer wing comprises 1-5 nucleosides. In certain embodiments, the gapmer wing comprises 2-5 nucleosides. In certain embodiments, the gapmer wing comprises 3-5 nucleosides. In certain embodiments, the nucleosides of the gapmer are all modified nucleosides.

[0249] In certain embodiments, the gapmer gap comprises 7-12 nucleosides. In certain embodiments, the gapmer gap comprises 7-10 nucleosides. In certain embodiments, the gapmer gap comprises 8-10 nucleosides. In certain embodiments, the gapmer gap comprises 10 nucleosides. In certain embodiments, each nucleoside of the gapmer gap is an unmodified 2'-deoxynucleoside.

[0250] In certain embodiments, the gapmer is a deoxy gapmer. In such embodiments, the nucleosides on the gap side of each wing / gap boundary are unmodified 2'-deoxynucleosides and the nucleosides on the wing side of each wing / gap boundary are modified nucleosides. In certain such embodiments, each nucleoside of the gap is an unmodified 2'-deoxynucleoside. In certain such embodiments, each nucleoside of each wing is a modified nucleoside.

[0251] In certain embodiments, a modified oligonucleotide comprises or consists of a region having a fully modified sugar motif. In such embodiments, each nucleoside of the fully modified region of the modified oligonucleotide comprises a modified sugar moiety. In certain such embodiments, each nucleoside throughout the modified oligonucleotide comprises a modified sugar moiety. In certain embodiments, a modified oligonucleotide comprises or consists of a region having a fully modified sugar motif, where each nucleoside within the fully modified region comprises the same modified sugar moiety, referred to herein as a uniformly modified sugar motif. In certain embodiments, a fully modified oligonucleotide is a uniformly modified oligonucleotide. In certain embodiments, each nucleoside of a uniformly modified oligonucleotide comprises the same 2'-modification. In certain embodiments, each nucleoside of a uniformly modified oligonucleotide comprises a 2'-O-(N-alkylacetamido) group. In certain embodiments, each nucleoside of a uniformly modified oligonucleotide comprises a 2'-O-(N-methylacetamido) group.

[0252] 2. Specific nucleobase motifs In certain embodiments, an oligonucleotide comprises modified and / or unmodified nucleobases arranged in a predetermined pattern or motif along the oligonucleotide or region thereof. In certain embodiments, each nucleobase is modified. In certain embodiments, none of the nucleobases are modified. In certain embodiments, each purine or each pyrimidine is modified. In certain embodiments, each adenine is modified. In certain embodiments, each guanine is modified. In certain embodiments, each nucleobase ... In certain embodiments, each thymine is modified. In certain embodiments, each uracil is modified. In certain embodiments, each cytosine is modified. In certain embodiments, some or all of the cytosine nucleobases in the modified oligonucleotide are 5-methylcytosine.

[0253] In certain embodiments, the modified oligonucleotide comprises a block of modified nucleobases. In certain such embodiments, the block is at the 3' end of the oligonucleotide. In certain embodiments, the block is within 3 nucleosides of the 3' end of the oligonucleotide. In certain embodiments, the block is at the 5' end of the oligonucleotide. In certain embodiments, the block is within 3 nucleosides of the 5' end of the oligonucleotide.

[0254] In certain embodiments, the oligonucleotide having a gapmer motif comprises a nucleoside that comprises a modified nucleobase. In certain such embodiments, one nucleoside that comprises a modified nucleobase is in the central gap of the oligonucleotide having a gapmer motif. In certain such embodiments, the sugar moiety of the nucleoside is a 2'-deoxyribosyl moiety. In certain embodiments, the modified nucleobase is selected from 2-thiopyrimidine and 5-propynepyrimidine.

[0255] 3. Specific internucleoside linkage motifs In certain embodiments, the oligonucleotide comprises modified and / or unmodified internucleoside linkages arranged in a predetermined pattern or motif along the oligonucleotide or region thereof. In certain embodiments, essentially each internucleoside linkage group is a phosphate internucleoside linkage (P=O). In certain embodiments, each internucleoside linkage group of the modified oligonucleotide is a phosphorothioate (P=S). In certain embodiments, each internucleoside linkage group of the modified oligonucleotide is independently selected from phosphorothioate and phosphate internucleoside linkage. In certain embodiments, the sugar motif of the modified oligonucleotide is a gapmer, and all of the internucleoside linkages in the gap are modified. In certain such embodiments, some or all of the internucleoside linkages in the wings are unmodified phosphate linkages. In certain embodiments, the terminal internucleoside linkage is modified.

[0256] D. A specific length In certain embodiments, oligonucleotides (including modified oligonucleotides) can have any of a variety of length ranges. In certain embodiments, oligonucleotides consist of X to Y linked nucleosides, where X represents the minimum number of nucleosides in the range and Y represents the maximum number of nucleosides in the range. In certain such embodiments, X and Y are each independently selected from 8, 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, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, and 50, with X≦Y. For example, in certain embodiments, the oligonucleotides may be 12-13, 12-14, 12-15, 12-16, 12-17, 12-18, 12-19, 12-20, 12-21, 12-22, 12-23, 12-24, 12-25, 12-26, 12-27, 12-28, 12-29, 12-30, 13-14, 13-15, 13-16, 13- 17 pieces, 13~18 pieces, 13~19 pieces, 13~20 pieces, 13~21 pieces, 13~22 pieces, 13~23 pieces, 13~24 pieces, 13~25 pieces, 13~26 pieces, 13~27 pieces, 13~28 pieces, 13~29 pieces, 1 3~30 pieces, 14~15 pieces, 14~16 pieces, 14~17 pieces, 14~18 pieces, 14~19 pieces, 14~20 pieces, 14~21 pieces, 14~22 pieces, 14~23 pieces, 14~24 pieces, 14~25 pieces, 14~26 pieces , 14~27 pieces, 14~28 pieces, 14~29 pieces, 14~30 pieces, 15~16 pieces, 15~17 pieces, 15~18 pieces, 15~19 pieces, 15~20 pieces, 15~21 pieces, 15~22 pieces, 15~23 pieces, 15~24 pieces, 15~25 pieces, 15~26 pieces, 15~27 pieces, 15~28 pieces, 15~29 pieces, 15~30 pieces, 16~17 pieces, 16~18 pieces, 16~19 pieces, 16~20 pieces, 16~21 pieces, 16~22 pieces, 16~23 pieces, 16~24 pieces, 16~25 pieces, 16~26 pieces, 16~27 pieces, 16~28 pieces, 16~2 9 pieces, 16~30 pieces, 17~18 pieces, 17~19 pieces, 17~20 pieces, 17~21 pieces, 17~22 pieces, 17~23 pieces, 17~24 pieces, 17~25 pieces, 17~26 pieces, 17~27 pieces, 17~28 pieces, 17~29 pieces, 17~30 pieces, 18~19 pieces, 18~ 20 pieces, 18~21 pieces, 18~22 pieces, 18~23 pieces, 18~24 pieces, 18~25 pieces, 18~26 pieces, 18~27 pieces, 18~28 pieces, 18~29 pieces, 18~30 pieces, 19~20 pieces, 19~21 pieces, 19~22 pieces, 19~23 pieces, 19~24 pieces, 19 ~25 pieces, 19~26 pieces, 19~29 pieces, 19~28 pieces, 19~29 pieces, 19~30 pieces, 20~21 pieces, 20~22 pieces, 20~23 pieces, 20~24 pieces, 20~25 pieces, 20~26 pieces, 20~27 pieces, 20~28 pieces, 20~29 pieces, 20~30 pieces, 2 1~22 pieces, 21~23 pieces, 21~24 pieces, 21~25 pieces, 21~26 pieces, 21~27 pieces, 21~28 pieces, 21~29 pieces, 21~30 pieces, 22~23 pieces, 22~24 pieces, 22~25 pieces, 22~26 pieces, 22~27 pieces, 22~28 pieces, 22~29 pieces, 22-30, 23-24, 23-25, 23-26, 23-27, 23-28, 23-29, 23-30, 24-25, 24-26, 24-27, 24-28, 24-29, 24-30, 25-26, 25-27, 25-28, 25-29, 25-30, 26-27, 26-28, 26-29, 26-30, 27-28, 27-29, 27-30, 28-29, 28-30, or 29-30 linked nucleosides.

[0257] E. Certain Modified Oligonucleotides In certain embodiments, the above-mentioned modifications (sugar, nucleobase, internucleoside linkage) are incorporated into modified oligonucleotides. In certain embodiments, modified oligonucleotides are characterized by their modification motif and total length. In certain embodiments, these parameters are each independent of each other. Thus, unless otherwise specified, each internucleoside linkage of an oligonucleotide having a gapmer sugar motif may be modified or unmodified, and may or may not follow the gapmer modification pattern of sugar modification. For example, the internucleoside linkages in the wing regions of a sugar gapmer may be the same or different from each other, and may be the same or different from the internucleoside linkages in the gap region of the sugar motif. Similarly, such sugar gapmer oligonucleotides may contain one or more modified nucleobases, regardless of the gapmer pattern of sugar modification. Furthermore, in certain cases, oligonucleotides are described by an overall length or range and the length or length range of two or more regions (e.g., regions of nucleosides with specified sugar modifications), and in such cases, it may be possible to select numbers in each range that result in oligonucleotides having total lengths outside the specified ranges. In such cases, both elements must be met. For example, in certain embodiments, a modified oligonucleotide is comprised of 15-20 linked nucleosides and has a sugar motif comprised of three regions, A, B, and C, where region A is comprised of 2-6 linked nucleosides having a specified sugar motif, region B is comprised of 6-10 linked nucleosides having a specified sugar motif, and region C is comprised of 2-6 linked nucleosides having a specified sugar motif. Such embodiments do not include modified oligonucleotides where A and C are each comprised of 6 linked nucleosides and B is comprised of 10 linked nucleosides (even though each number of nucleosides may be allowed within the requirements for A, B, and C). This is because the total length of the oligonucleotide is 22, which exceeds the upper limit of the total length of modified oligonucleotide (20). If the description of the oligonucleotide is not provided herein with respect to one or more parameters, such parameters are not limited. Thus, without further description, a modified oligonucleotide described as having only a gapmer sugar motif can have any length, internucleoside linkage motif, and nucleobase motif. Unless otherwise specified, all modifications are independent of the nucleobase sequence.

[0258] F. Nucleotide sequence In certain embodiments, an oligonucleotide (unmodified or modified oligonucleotide) is further described by its nucleobase sequence. In certain embodiments, an oligonucleotide has a nucleobase sequence that is complementary to a specified reference nucleic acid, such as a second oligonucleotide or a target transcript precursor. In such certain embodiments, a region of the oligonucleotide has a nucleobase sequence that is complementary to a specified reference nucleic acid, such as a second oligonucleotide or a target transcript precursor. In certain embodiments, a region or the entire length of the oligonucleotide has a nucleobase sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% or 100% complementary to a nucleic acid, such as a second oligonucleotide or a target transcript precursor.

[0259] II. Certain Oligomeric Compounds In certain embodiments, the present invention provides oligomeric compounds consisting of an oligonucleotide (modified or unmodified) and, optionally, one or more conjugate groups and / or terminal groups. A conjugate group consists of one or more conjugate moieties and a conjugate linker that connects the conjugate moieties to the oligonucleotide. A conjugate group can be attached to one or both ends of an oligonucleotide and / or can be attached to any internal position. In certain embodiments, a conjugate group is attached to the 2' position of a nucleoside of a modified oligonucleotide. In certain embodiments, a conjugate group attached to one or both ends of an oligonucleotide is a terminal group. In certain such embodiments, a conjugate group or terminal group is attached to the 3' and / or 5' end of an oligonucleotide. In certain such embodiments, a conjugate group (or terminal group) is attached to the 3' end of an oligonucleotide. In certain embodiments, a conjugate group is attached near the 3' end of an oligonucleotide. In certain embodiments, a conjugate group (or terminal group) is attached to the 5' end of an oligonucleotide. In certain embodiments, the conjugate group is attached near the 5' end of the oligonucleotide.

[0260] Examples of terminal groups include, but are not limited to, a conjugate group, a capping group, a phosphate moiety, a protecting group, an abasic nucleoside, a modified or unmodified nucleoside, and two or more nucleosides that are independently modified or unmodified.

[0261] A. Specific Conjugation Groups In certain embodiments, the oligonucleotide is covalently linked to one or more conjugate groups. In certain embodiments, the conjugate group modifies one or more properties of the oligonucleotide to which it is attached, including but not limited to pharmacodynamics, pharmacokinetics, stability, binding, absorption, tissue distribution, cellular distribution, cellular uptake, charge and clearance. In certain embodiments, the conjugate group confers new properties to the oligonucleotide to which it is attached, for example, a fluorophore or reporter group that allows detection of the oligonucleotide. Certain conjugate groups and conjugate moieties have been previously described, such as cholesterol moieties (Letsinger et al., Proc. Natl. Acad. Sci. USA, 1989, 86, 6553-6556), cholic acid (Manoharan et al., Bioorg. Med. Chem. Lett., 1994, 4, 1053-1060), thioethers, such as hexyl-S-tritylthiol (Manoharan et al., Ann. NYAca d. Sci., 1992, 660, 306-309; Manoharan et al., Bioorg. Med. Chem. Lett., 1993, 3, 2765-2770), thiocholesterol (Oberhauser et al., Nucl. Acids Res., 1992, 20, 533-538), aliphatic chains such as dodecanediol or undecyl residues (Saison-Behmoaras et al., EMBO J., 1991, 10, 1111-1118; Kabanov et al., FEBS Lett., 1990, 259, 327-330; Svinarchuk et al., Biochimie, 1993, 75, 49-54), phospholipids such as hexadecyl-rac-glycerol or triethyl-ammonium. 1,2-di-O-hexadecyl-rac-glycero-3-H-phosphonate (Manoharan et al., Tetrahedron Lett., 1995, 36, 3651-3654; Shea et al., Nucl. Acids Res., 1990, 18, 3777-3783), polyamine or polyethylene glycol chains (Manoharan et al., Nucleosides & Nucleotides, 1995, 14, 969-973) or adamantane acetic acid, palmityl moieties (Mishra et al., Biochim. Biophys. Acta, 1995, 1264, 229-237), octadecylamine or hexylamino-carbonyl-oxycholesterol moieties (Crooke et al., al., J. Pharmacol. Exp. Ther., 1996, 277, 923-937), tocopherol groups (Nishina et al., Molecular Therapy Nucleic Acids, 2015, 4, e220; and Nishina et al., Molecular Therapy, 2008, 16, 734-740) or GalNAc clusters (e.g., WO2014 / 179620).

[0262] In certain embodiments, the conjugate group may be selected from any of C22 alkyl, C20 alkyl, C16 alkyl, C10 alkyl, C21 alkyl, C19 alkyl, C18 alkyl, C15 alkyl, C14 alkyl, C13 alkyl, C12 alkyl, C11 alkyl, C9 alkyl, C8 alkyl, C7 alkyl, C6 alkyl, C5 alkyl, C22 alkenyl, C20 alkenyl, C16 alkenyl, C10 alkenyl, C21 alkenyl, C19 alkenyl, C18 alkenyl, C15 alkenyl, C14 alkenyl, C13 alkenyl, C12 alkenyl, C11 alkenyl, C9 alkenyl, C8 alkenyl, C7 alkenyl, C6 alkenyl, or C5 alkenyl.

[0263] In certain embodiments, the conjugate group may be selected from any of C22 alkyl, C20 alkyl, C16 alkyl, C10 alkyl, C21 alkyl, C19 alkyl, C18 alkyl, C15 alkyl, C14 alkyl, C13 alkyl, C12 alkyl, C11 alkyl, C9 alkyl, C8 alkyl, C7 alkyl, C6 alkyl, and C5 alkyl, wherein the alkyl chain has one or more unsaturated bonds.

[0264] 1. Conjugate part Conjugate moieties include, but are not limited to, intercalators, reporter molecules, polyamines, polyamides, peptides, carbohydrates (e.g., GalNAc), vitamin moieties, polyethylene glycols, thioethers, polyethers, cholesterol, thiocholesterol, cholic acid moieties, folic acid, lipids, lipophilic groups, phospholipids, biotin, phenazine, phenanthridine, anthraquinone, adamantane, acridine, fluoresceins, rhodamines, coumarins, fluorophores, and dyes.

[0265] In certain embodiments, the conjugate moiety is an active drug substance, such as aspirin, warfarin, phenylbutazone, ibuprofen, suprofen, fenbufen, ketoprofen, (S)-(+)-pranoprofen, carprofen, dansylsal These include: cosine, 2,3,5-triiodobenzoic acid, fingolimod, flufenamic acid, folinic acid, benzothiadiazid, chlorothiazide, diazepines, indomethicine, barbiturates, cephalosporins, sulfa drugs, antidiabetics, antibacterial substances or antibiotics.

[0266] 2. Conjugate Linker The conjugate moiety is attached to the oligonucleotide via a conjugate linker. In certain oligomeric compounds, the conjugate linker is a single chemical bond (i.e., the conjugate moiety is directly attached to the oligonucleotide via a single bond). In certain oligomeric compounds, the conjugate moiety is attached to the oligonucleotide via a more complex conjugate linker, which comprises one or more conjugate linker moieties, which are subunits that form the conjugate linker. In certain embodiments, the conjugate linker comprises a chain structure, such as a hydrocarbyl chain, or an oligomer of repeating units, such as ethylene glycol, nucleoside or amino acid units.

[0267] In certain embodiments, the conjugate linker comprises one or more groups selected from alkyl, amino, oxo, amide, disulfide, polyethylene glycol, ether, thioether, and hydroxylamino. In certain such embodiments, the conjugate linker comprises a group selected from alkyl, amino, oxo, amide, and ether groups. In certain embodiments, the conjugate linker comprises a group selected from alkyl and amide groups. In certain embodiments, the conjugate linker comprises a group selected from alkyl and ether groups. In certain embodiments, the conjugate linker comprises at least one phosphorus moiety. In certain embodiments, the conjugate linker comprises at least one phosphate group. In certain embodiments, the conjugate linker comprises at least one neutral conjugate group.

[0268] In certain embodiments, the conjugate linker, including the conjugate linkers described above, is a bifunctional linking moiety, known in the art to be useful for attaching a conjugate group to a parent compound, such as an oligonucleotide provided herein. In general, the bifunctional linking moiety comprises at least two functional groups. One of the functional groups is selected to bind to a specific site on the parent compound, and the other is selected to bind to a conjugate group. Examples of functional groups used in the bifunctional linking moiety include, but are not limited to, electrophiles that react with nucleophilic groups and nucleophiles that react with electrophilic groups. In certain embodiments, the bifunctional linking moiety comprises one or more groups selected from amino, hydroxyl, carboxylic acid, thiol, alkyl, alkenyl, and alkynyl.

[0269] Examples of conjugate linkers include, but are not limited to, pyrrolidine, 8-amino-3,6-dioxaoctanoic acid (ADO), 4-(N-maleimidomethyl)cyclohexane-1-carboxylate succinimidyl (SMCC), and 6-aminohexanoic acid (AHEX or AHA). Other conjugate linkers include, but are not limited to, substituted or unsubstituted C 1 ~C 10 Alkyl, substituted or unsubstituted C 2 ~C 10 Alkenyl, or substituted or unsubstituted C 2 ~C 10 Alkynyl is included, where a non-limiting list of preferred substituents includes hydroxyl, amino, alkoxy, carboxy, benzyl, phenyl, nitro, thiol, thioalkoxy, halogen, alkyl, aryl, alkenyl and alkynyl.

[0270] In certain embodiments, the conjugate linker comprises 1 to 10 linker nucleosides. In certain embodiments, the linker nucleosides are modified nucleosides. In certain embodiments, the linker nucleoside comprises a modified sugar moiety. In certain embodiments, the linker nucleoside is unmodified. In certain embodiments, the linker nucleoside comprises an optionally protected heterocyclic base selected from a purine, a substituted purine, a pyrimidine, or a substituted pyrimidine. In certain embodiments, the cleavable moiety is a nucleoside selected from uracil, thymine, cytosine, 4-N-benzoylcytosine, 5-methylcytosine, 4-N-benzoyl-5-methyl-cytosine, adenine, 6-N-benzoyladenine, guanine, and 2-N-isobutyrylguanine. It is usually desirable for the linker nucleoside to be cleaved from the oligomeric compound after reaching the target tissue. Thus, the linker nucleosides are typically linked to each other and to the remainder of the oligomeric compound via a cleavable bond. In certain embodiments, such a cleavable bond is a phosphodiester bond.

[0271] Herein, linker nucleosides are not considered part of the oligonucleotide. Thus, in embodiments where an oligomeric compound comprises an oligonucleotide consisting of a specified number or range of linked nucleosides and / or a specified percentage of complementarity to a reference nucleic acid, and the oligomeric compound comprises a conjugate group comprising a conjugate linker that comprises linker nucleosides, these linker nucleosides are not counted in the length of the oligonucleotide and are not used in determining the percentage of complementarity of the oligonucleotide to the reference nucleic acid. For example, an oligomeric compound may comprise (1) a modified oligonucleotide consisting of 8-30 nucleosides and (2) a conjugate group comprising 1-10 linker nucleosides contiguous to the nucleosides of the modified oligonucleotide. The total number of contiguous linked nucleosides in such an oligomeric compound is greater than 30. Alternatively, an oligomeric compound may comprise an oligonucleotide consisting of 8-30 nucleosides and does not comprise a conjugate group. The total number of consecutive linked nucleosides in such oligomeric compounds is 30 or less. Unless otherwise indicated, the conjugate linker comprises 10 or less linker nucleosides. In certain embodiments, the conjugate linker comprises 5 or less linker nucleosides. In certain embodiments, the conjugate linker comprises 3 or less linker nucleosides. In certain embodiments, the conjugate linker comprises 2 or less linker nucleosides. In certain embodiments, the conjugate linker comprises 1 or less linker nucleoside.

[0272] In certain embodiments, it is desirable for the conjugate group to be cleaved from the oligonucleotide. For example, in certain cases, oligomeric compounds that contain certain conjugate moieties are well taken up by certain cell types, but it is desirable for the conjugate group to be cleaved to release the unconjugated oligonucleotide or parent oligonucleotide when the oligomeric compound is taken up. Thus, certain conjugate linkers may contain one or more cleavable moieties. In certain embodiments, the cleavable moiety is a cleavable bond. In certain embodiments, the cleavable moiety is an atomic group that includes at least one cleavable bond. In certain embodiments, the cleavable moiety includes an atomic group that has one, two, three, four or more cleavable bonds. In certain embodiments, the cleavable moiety is selectively cleaved inside a cell or an intracellular compartment such as a lysosome. In certain embodiments, the cleavable moiety is selectively degraded by endogenous enzymes such as nucleases.

[0273] In certain embodiments, the cleavable bond is selected from an amide, an ester, an ether, one or both esters of a phosphodiester, a phosphate ester, a carbamate, or a disulfide. In certain embodiments, the cleavable bond is one or both esters of a phosphodiester. In certain embodiments, the cleavable moiety comprises a phosphate or a phosphodiester. In certain embodiments, the cleavable moiety is an oligonucleotide. The phosphate bond between the leutide and the conjugated moiety or group.

[0274] In certain embodiments, the cleavable moiety comprises or consists of one or more linker nucleosides. In certain such embodiments, one or more linker nucleosides are linked to each other and / or to the remainder of the oligomeric compound via a cleavable bond. In certain such embodiments, such cleavable bond is an unmodified phosphodiester bond. In certain such embodiments, the cleavable moiety is a 2'-deoxynucleoside that is linked to either the 3'- or 5'-terminal nucleoside of the oligonucleotide by a phosphate internucleoside bond and covalently linked to the remaining conjugate linker or conjugate moiety by a phosphate or phosphorothioate bond. In certain such embodiments, the cleavable moiety is 2'-deoxyadenosine.

[0275] 3. Specific Cell-Targeting Conjugate Moieties In certain embodiments, the conjugate group comprises a cell targeting conjugate moiety. In certain embodiments, the conjugate group has the general formula: [ka] In the formula, n is 1 to about 3; when n is 1, m is 0; when n is 2 or more, m is 1, j is 1 or 0, and k is 1 or 0.

[0276] In certain embodiments, n is 1, j is 1, and k is 0. In certain embodiments, n is 1, j is 0, and k is 1. In certain embodiments, n is 1, j is 1, and k is 1. In certain embodiments, n is 1, j is 1, and k is 1. In certain embodiments, n is 2, j is 1, and k is 0. In certain embodiments, n is 2, j is 0, and k is 1. In certain embodiments, n is 2, j is 1, and k is 1. In certain embodiments, n is 3, j is 1, and k is 0. In certain embodiments, n is 3, j is 0, and k is 1. In certain embodiments, n is 3, j is 1, and k is 1.

[0277] In certain embodiments, the conjugate group comprises a cell targeting moiety having at least one tethered ligand. In certain embodiments, the cell targeting moiety comprises two tethered ligands covalently attached to the branching group. In certain embodiments, the cell targeting moiety comprises three tethered ligands covalently attached to the branching group.

[0278] In certain embodiments, the conjugate group comprises a cell targeting moiety having the formula: [ka]

[0279] In certain embodiments, the conjugate group comprises a cell targeting moiety having the formula: [ka] wherein n is an integer selected from 1, 2, 3, 4, 5, 6, or 7. In certain embodiments, n is 1. In certain embodiments, n is 2. In certain embodiments, n is 3. In certain embodiments, n is 4. In certain embodiments, n is 5.

[0280] In certain embodiments, the conjugate group comprises a cell targeting moiety having the formula: [ka]

[0281] In certain embodiments, the conjugate group comprises a cell targeting moiety having the formula: [ka]

[0282] In certain embodiments, the conjugate group comprises a cell targeting moiety having the formula: [ka]

[0283] In certain embodiments, the conjugate group comprises a cell targeting moiety having the formula: [ka]

[0284] In certain embodiments, the conjugate group comprises a cell targeting moiety having the formula: [ka]

[0285] In certain embodiments, the oligomeric compounds comprise a conjugate group referred to herein as "LICA-1." LICA-1 has the formula: [ka]

[0286] In certain embodiments, the oligomeric compound comprising LICA-1 has the formula: [ka] In the formula, oligo is an oligonucleotide.

[0287] Representative U.S. patents, U.S. patent application publications, international patent application publications and other publications that teach the preparation of some of the oligomeric compounds containing the conjugate groups, conjugate groups, tethers, conjugate linkers, branching groups, ligands, cleavable moieties and other modifications described above include, but are not limited to, U.S. Pat. No. 5,994,517, U.S. Pat. No. 6,300,319, U.S. Pat. No. 6,660,720, U.S. Pat. No. 6,906,182, U.S. Pat. No. 7,262,177, U.S. Pat. No. 7,491,805, U.S. Pat. No. 8,106,022, U.S. Pat. No. 7,723,509, U.S. Pat. No. 5,113,621, and U.S. Pat. No. 5,113,621. al.,J.Med.Chem.1995,38,1846-1852,Lee et al.,Bioorganic&Medicinal Chemistry 2011,19,2494-2500,Rensen et al.,J.Biol.Chem.2001,276,37577-37584,Rensen et al. al., J. Med. Chem. 2004, 47, 5798-5808, Sliedregt et al., J. Med. Chem. 1999, 42, 609-618 and Valentijn et al., Tetrahedron, 1997, 53, 759-770.

[0288] In certain embodiments, the oligomeric compounds comprise a modified oligonucleotide comprising a fully modified sugar motif and a conjugate group comprising at least one, two or three GalNAc ligands. In certain embodiments, the antisense compounds and oligomeric compounds are prepared by the methods described in the following references: Lee, Carbohydr Res, 1978, 67, 509-514; Connolly et al., J Biol Chem, 1982, 257, 939-945; Pavia et al., Int J Pep Protein Res, 1983, 22, 539-548; Lee et al., Biochem, 1984, 23, 4255-4261; Lee et al., Glycoconjugate J, 1987, 4, 317-328; Toyokuni et al., Te trahedron Lett,1990,31,2673-2676;Biessen et al.,J Med Chem,1995,38,1538-1546;Valentijn et al.,Tetrahedron,1997,53,759-770;Kim et al.,Tetrahedron Lett,1997,38,3487-3490;Lee et al.,Bioconju Chem,1997,8,762-765;Kato et al.,Glycobiol,2001,11,821-829;Rensen et al.,J Biol Chem,2001,276,37577-37584;Lee et al.,Methods Enzymol-362,2033 al.,Glycoconj J,2004,21,227-241;Lee et al.,Bioorg Med Chem Lett,2006,16(19),5132-5135;Maierhofer et al.,Bioorg Med Chem,2007,Med.7661-7676; Chem,2008,16,5216-5231;Lee et al.,Bioorg Med Chem,2011,19,2494-2500;Kornilova et al.,Analyt Biochem,2012,425,43-46;Pujol et al.,EdAngew Chem Engl,2012,51,7445-7448;Biessen et al.,J Med Chem,1995,38,1846-1852;Sliedregt et al.,J Med Chem,1999,42,609-618;Rensen et al.,7804;J Med,409-618; al.,Arterioscler Thromb Vasc Biol,2006,26,169-175;van Rossenberg et al.,Gene Ther,2004,11,457-464;Sato et al.,J Am Chem Soc,2004,126,14013-14022; Chem,2012,77,7564-7571;Biessen et al.,FASEB J,2000,14,1784-1792;Rajur et al.,Bioconjug Chem,1997,8,935-940;Duff et al.,Methods Enzymol,2000,313,297-321;Maier et al.,Bioconjug Chem,2003,14,18-29;Jayaprakash et al. al.,Org Lett,2010,12,5410-5413;Manoharan,Antisense Nucleic Acid Drug Dev,2002,12,103-128;Merwin et al.,Bioconjug Chem,1994,5,612-620;Tomiya et al.,Bioorg Med Chem,2013,21,5275-5281;International Applications WO1998 / 013381;WO2011 / 038356;WO1997 / 046098;WO2008 / 098788;WO2004 / 101619;WO2012 / 037254;WO2011 / 120053;WO2011 / 100131;WO2011 / 163121;WO2012 / 1 77947;WO2013 / 033230;WO2013 / 075035;WO2012 / 083185;WO2012 / 083046;WO2009 / 082607;WO200 9 / 134487;WO2010 / 144740;WO2010 / 148013;WO1997 / 020563;WO2010 / 088537;WO2002 / 043771;WO 2010 / 129709;WO2012 / 068187;WO2009 / 126933;WO2004 / 024757;WO2010 / 054406;WO2012 / 089352;WO2012 / 089602;WO2013 / 166121;WO2013 / 165816;U.S. Patent Nos. 4,751,219; 8,552,163; 6,908,9 No. 03; No. 7,262,177; No. 5,994,517; No. 6,300,319; No. 8,106,022; No. 7,491,805; No. 7,491,805 ; Same No. 7,582,744; Same No. 8,137,695; Same No. 6,383,812; Same No. 6,525,031; Same No. 6,660,720; Same No. 7,723,509; Same No. No. 8,541,548; No. 8,344,125; No. 8,313,772; No. 8,349,308; No. 8,450,467; No. 8,501,930; No. 8,158 , No. 7,262,177; No. 6,906,182; No. 6,620,916; No. 8,435,491; No. 8,404,862; No. 7,851,615; Published U.S. Patent Applications US2011 / 0097264;US2011 / 0097265;US2013 / 0004427;US2005 / 0164235;US2006 / 0148740;US2008 / 0281044;US2010 / 0240730;US2003 / 0119724;US2006 / 0183886;US2008 / 0206869;US201 1 / 0269814;US2009 / 0286973;US2011 / 0207799;US2012 / 0136042;US2012 / 0165393;US2008 / 0281041 ;US2009 / 0203135;US2012 / 0035115;US2012 / 0095075;US2012 / 0101148;US2012 / 0128760;US2012 / 0 157509; US2012 / 0230938; US2013 / 0109817; US2013 / 0121954; US2013 / 0178512; US2013 / 0236968; US2011 / 0123520; US2003 / 0077829; US2008 / 0108801; and US2009 / 0203132.

[0289] In certain embodiments, the compounds of the invention are single-stranded. In certain embodiments, the oligomeric compounds are paired with a second oligonucleotide or oligomeric compound to form a duplex structure that is double-stranded.

[0290] III. Specific Antisense Compounds In certain embodiments, the present invention provides antisense compounds that comprise or consist of oligomeric compounds that comprise an antisense oligonucleotide having a nucleobase sequence that is complementary to the nucleobase sequence of a target nucleic acid. In certain embodiments, the antisense compounds are single-stranded. Such single-stranded antisense compounds typically comprise or consist of an oligomeric compound that comprises or consists of a modified oligonucleotide and optionally a conjugate group. In certain embodiments, the antisense compounds are double-stranded. Such double-stranded antisense compounds comprise a first oligomeric compound that has a region that is complementary to the target nucleic acid and a second oligomeric compound that has a region that is complementary to the first oligomeric compound. The first oligomeric compound of such double-stranded antisense compounds typically comprises or consists of a modified oligonucleotide and optionally a conjugate group. The oligonucleotide of the second oligomeric compound of such double-stranded antisense compounds may be modified or unmodified. One or both of the oligomeric compounds of the double-stranded antisense compounds may comprise a conjugate group. Oligomeric compounds of double-stranded antisense compounds may contain non-complementary overhanging nucleosides.

[0291] In certain embodiments, the oligomeric compound of antisense compound can hybridize with target nucleic acid, resulting in at least one antisense activity.In certain embodiments, antisense compound selectively acts on one or more target nucleic acid.Such selective antisense compound comprises the nucleobase sequence that hybridizes with one or more target nucleic acid, resulting in one or more desired antisense activity, and does not hybridize with one or more non-target nucleic acid or does not hybridize with one or more non-target nucleic acid in such a way that it produces highly undesirable antisense activity.

[0292] In certain embodiments, hybridization of an antisense compound to a target nucleic acid alters processing, e.g., splicing, of the target precursor transcript. In certain embodiments, hybridization of an antisense compound to a target precursor transcript results in inhibition of a binding interaction between the target nucleic acid and a protein or other nucleic acid. In certain such embodiments, hybridization of an antisense compound to the target precursor transcript alters translation of the target nucleic acid.

[0293] Antisense activity can be observed directly or indirectly. In certain embodiments, observing or detecting antisense activity includes observing or detecting a change in the amount of a target nucleic acid or a protein encoded by the target nucleic acid, a change in the ratio of splice variants of a nucleic acid or a protein, and / or a change in a phenotype in a cell or animal.

[0294] IV. Specific Target Nucleic Acids In certain embodiments, the antisense compound comprises or consists of an oligonucleotide comprising a region complementary to a target nucleic acid. In certain embodiments, the target nucleic acid is DNA encoding SMN2, including the human SMN2 genomic sequence truncated from nucleotides 19939708 to 19967777 set forth in GENBANK Accession No. NT_006713.14, which is incorporated herein by reference as SEQ ID NO:240. In certain embodiments, the target nucleic acid comprises SEQ ID NO:10. Nucleotides 1-60 of SEQ ID NO:10 represent a portion of intron 6 of SMN2 pre-mRNA, nucleotides 61-114 of SEQ ID NO:10 represent exon 7 of SMN2 pre-mRNA, and nucleotides 115-174 of SEQ ID NO:10 represent a portion of intron 7 of SMN2 pre-mRNA. In certain embodiments, the target nucleic acid comprises ISS-N1 comprising the nucleobase sequence of SEQ ID NO:11.

[0295] A. Complementarity / Mismatch to Target Nucleic Acid In certain embodiments, the antisense and / or oligomeric compounds comprise antisense oligonucleotides that are complementary to a target nucleic acid over the entire length of the oligonucleotide. In certain embodiments, such oligonucleotides are 99% complementary to the target nucleic acid. In certain embodiments, such oligonucleotides are 95% complementary to the target nucleic acid. In certain embodiments, such oligonucleotides are 90% complementary to the target nucleic acid. In certain embodiments, such oligonucleotides are 85% complementary to the target nucleic acid. In certain embodiments, such oligonucleotides are 80% complementary to the target nucleic acid. In certain embodiments, the antisense oligonucleotides are at least 80% complementary to the target nucleic acid over the entire length of the oligonucleotide and comprise a region that is 100% or fully complementary to the target nucleic acid. In certain such embodiments, the region of complete complementarity is 6-20 nucleobases in length. In certain such embodiments, the region of complete complementarity is 10-18 nucleobases in length. In certain such embodiments, the region of complete complementarity is 18-20 nucleobases in length.

[0296] In certain embodiments, the oligomeric compounds and / or antisense compounds contain one or more mismatched nucleobases with respect to the target nucleic acid. In certain such embodiments, the antisense activity with respect to the target is reduced by such mismatches, while the activity with respect to the non-target is greatly reduced. Thus, in certain such embodiments, the selectivity of the antisense compounds is improved. In certain such embodiments, the mismatches are specifically located within the oligonucleotides having a gapmer motif. In certain such embodiments, the mismatches are at 1, 2, 3, 4, 5, 6, 7 or 8 positions from the 5' end of the gap region. In certain such embodiments, the mismatches are at 9, 8, 7, 6, 5, 4, 3, 2 or 1 positions from the 3' end of the gap region. In certain such embodiments, the mismatches are at 1, 2, 3 or 4 positions from the 5' end of the wing region. In certain such embodiments, the mismatches are at 4, 3, 2 or 1 positions from the 3' end of the wing region.

[0297] B. Modulation of Processing of Specific Target Nucleic Acids In certain embodiments, the oligomeric compound comprises or consists of a modified oligonucleotide complementary to a target precursor transcript. In certain such embodiments, the target precursor transcript is a target pre-mRNA. In certain such embodiments, contacting a cell with a compound complementary to the target precursor transcript modulates processing of the target precursor transcript. In certain such embodiments, the resulting target processed transcript has a different nucleobase sequence than the target processed transcript produced in the absence of the compound. In certain such embodiments, the target precursor transcript is a target pre-mRNA and contacting a cell with a compound complementary to the target pre-mRNA modulates splicing of the target pre-mRNA. In certain such embodiments, the resulting target mRNA has a different nucleobase sequence than the target mRNA produced in the absence of the compound. In certain such embodiments, an exon is excluded from the target mRNA. In certain such embodiments, an exon is included in the target mRNA. In certain embodiments, exon exclusion or inclusion results in the induction or inhibition of nonsense-mediated decay of the target mRNA, the removal or addition of a premature stop codon from the target mRNA, and / or an alteration of the reading frame of the target mRNA.

[0298] C. Specific Diseases and Conditions Associated with Specific Target Nucleic Acids In certain embodiments, the target precursor transcript is associated with a disease or pathology. In certain such embodiments, oligomeric compounds comprising or consisting of modified oligonucleotides complementary to the target precursor transcript are used to treat the disease or pathology. In certain such embodiments, the compound modulates the processing of the target precursor transcript to provide a beneficial target processed transcript. In certain such embodiments, the disease or pathology is associated with aberrant processing of the precursor transcript. In certain such embodiments, the disease or pathology is associated with aberrant splicing of pre-mRNA.

[0299] V. Certain Pharmaceutical Compositions In certain embodiments, the present invention provides a pharmaceutical composition comprising one or more antisense compounds or salts thereof. In certain such embodiments, the pharmaceutical composition comprises a suitable pharma- ceutically acceptable diluent or carrier. In certain embodiments, the pharmaceutical composition comprises a sterile saline solution and one or more antisense compounds. In certain embodiments, such a pharmaceutical composition comprises a sterile saline solution and one or more antisense compounds. In certain embodiments, the sterile saline solution is pharmaceutical grade saline. In certain embodiments, the pharmaceutical composition comprises one or more antisense compounds and sterile water. In certain embodiments, the pharmaceutical composition comprises an antisense compound and sterile water. In certain embodiments, the sterile water is pharmaceutical grade water. In certain embodiments, the pharmaceutical composition comprises one or more antisense compounds and phosphate buffered saline (PBS). In certain embodiments, the pharmaceutical composition comprises one or more antisense compounds and sterile PBS. In certain embodiments, the sterile PBS is pharmaceutical grade PBS.

[0300] In certain embodiments, pharmaceutical compositions comprise one or more antisense compounds and one or more excipients, in certain such embodiments, the excipients are selected from water, saline, alcohol, polyethylene glycol, gelatin, lactose, amylase, magnesium stearate, talc, silicic acid, viscous paraffin, hydroxymethylcellulose, and polyvinylpyrrolidone.

[0301] In certain embodiments, the antisense compounds may be mixed with pharma- ceutically acceptable active and / or inactive substances for the preparation of pharmaceutical compositions or formulations. The composition and method for preparation will depend on a number of criteria, including but not limited to the route of administration, the extent of the disease, or the dosage to be administered.

[0302] In certain embodiments, the pharmaceutical composition comprising the oligomeric compound and / or the antisense compound includes any pharmaceutically acceptable salt of the antisense compound, an ester of the antisense compound, or a salt of the ester. In certain embodiments, the pharmaceutical composition comprising the antisense compound and / or the oligomeric compound, including one or more antisense oligonucleotides, can produce (directly or indirectly) a biologically active metabolite or residue thereof when administered to an animal, including a human. Thus, for example, the present disclosure also relates to pharmaceutically acceptable salts of the antisense compound, prodrugs, pharmaceutically acceptable salts of the prodrugs, and other bioequivalents. Suitable pharmaceutically acceptable salts include, but are not limited to, sodium and potassium salts. In certain embodiments, the prodrugs include one or more conjugate groups attached to the oligonucleotide, which are cleaved by endogenous nucleases in the body.

[0303] In nucleic acid therapy, lipid moieties are used in various ways. In certain such methods, nucleic acids, such as antisense compounds, are incorporated into preformed liposomes or lipoplexes consisting of a mixture of cationic lipids and neutral lipids. In certain methods, DNA complexes containing monocationic or polycationic lipids are formed without the presence of neutral lipids. In certain embodiments, lipid moieties are selected to increase the distribution of drugs to specific cells or tissues. In certain embodiments, lipid moieties are selected to increase the distribution of drugs to adipose tissue. In certain embodiments, lipid moieties are selected to increase the distribution of drugs to muscle tissue.

[0304] In certain embodiments, the pharmaceutical composition comprises a delivery system. Examples of delivery systems include, but are not limited to, liposomes and emulsions. Certain delivery systems are useful for preparing certain pharmaceutical compositions, such as pharmaceutical compositions that include hydrophobic compounds. In certain embodiments, certain organic solvents, such as dimethylsulfoxide, are used.

[0305] In certain embodiments, the pharmaceutical composition comprises one or more tissue-specific delivery molecules designed to deliver one or more agents of the invention to a particular tissue or cell type. For example, in certain embodiments, the pharmaceutical composition comprises a liposome coated with a tissue-specific antibody.

[0306] In certain embodiments, the pharmaceutical composition includes a co-solvent system. Some such co-solvent systems include, for example, benzyl alcohol, a non-polar surfactant, a water-miscible organic polymer, and an aqueous phase. In certain embodiments, such a co-solvent system is used for hydrophobic compounds. A non-limiting example of such a co-solvent system is the VPD co-solvent system, which is an absolute ethanol solution containing 3% w / v benzyl alcohol, 8% w / v of the non-polar surfactant Polysorbate 80™, and 65% w / v of polyethylene glycol 300. The ratios of such a co-solvent system can be varied widely without significantly altering its solubility and toxicity characteristics. Furthermore, the nature of the components of the co-solvent may be changed, for example, other surfactants can be used in place of Polysorbate 80™, the fraction size of the polyethylene glycol can be changed, the polyethylene glycol can be replaced by other biocompatible polymers, such as polyvinylpyrrolidone, and other sugars or polysaccharides can be used in place of dextrose.

[0307] In certain embodiments, the pharmaceutical composition is prepared for oral administration. In certain embodiments, the pharmaceutical composition is prepared for buccal administration. In certain embodiments, the pharmaceutical composition is prepared for administration by injection (e.g., intravenous, subcutaneous, intramuscular, etc.). In certain such embodiments, the pharmaceutical composition comprises a carrier and is an aqueous solution, such as water, or a Hanks' solution. The pharmaceutical compositions are formulated in physiologically compatible buffers such as sodium chloride solution, Ringer's solution or physiological saline buffer. In certain embodiments, other ingredients are included (e.g., ingredients that aid in solubility or act as preservatives). In certain embodiments, injectable suspensions are prepared using appropriate liquid carriers, suspending agents, and the like. Certain injectable pharmaceutical compositions are provided in unit dosage form, e.g., in ampoules or in multi-dose containers. Certain injectable pharmaceutical compositions are suspensions, solutions or emulsions in oily or aqueous vehicles and may contain formulating agents such as suspending agents, stabilizing agents and / or dispersing agents. Certain solvents suitable for use in injectable pharmaceutical compositions include, but are not limited to, lipophilic solvents and fatty oils such as sesame oil, synthetic fatty acid esters such as ethyl oleate or triglycerides, and liposomes. Injectable aqueous suspensions may be included.

[0308] Non-Limiting Disclosure and Incorporation by Reference All documents or portions of documents cited in this application, including but not limited to patents, patent applications, articles, books, papers, and GenBank and NCBI reference sequence records, are expressly incorporated herein by reference in their entirety. Claims in international application [Item 1] An oligomeric compound comprising a modified oligonucleotide consisting of 14 to 25 linked nucleosides, wherein the modified oligonucleotide is complementary to SMN2 pre-mRNA, and at least one nucleoside of the modified oligonucleotide is represented by the formula II. The structure is [ka] For each nucleoside of formula II: Bx is an independently selected nucleobase; R1 and R2 are each independently selected from hydrogen and methyl; Alternatively, R1 is hydrogen and R2 is independently selected from ethyl, propyl, or isopropyl. [Item 2] The oligomeric compound according to claim 1, comprising at least one nucleoside of formula II, in which one of R1 and R2 is not hydrogen. [Item 3] The oligomeric compound according to claim 1 or 2, wherein for at least one nucleoside having the structure of formula II, R1 is hydrogen and R2 is selected from methyl, ethyl, propyl or isopropyl. [Item 4] The oligomeric compound according to any one of claims 1 to 3, comprising at least one nucleoside of formula II, in which R1 is hydrogen and R2 is selected from methyl or ethyl. [Item 5] The oligomeric compound according to any one of items 1 to 4, comprising at least one nucleoside of formula II, in which at least one of R1 and R2 is methyl. [Item 6] One of R1 and R2 is hydrogen, and the other of R1 and R2 is methyl. 6. The oligomeric compound according to claim 1, comprising at least one nucleoside. [Item 7] The oligomeric compound according to any one of claims 1 to 6, wherein the selection of R1 is different for at least two of the nucleosides having the structure of formula II. [Item 8] The oligomeric compound according to any one of claims 1 to 7, wherein the selection of R2 is different for at least two of the nucleosides having the structure of formula II. [Item 9] The oligomeric compound according to any one of claims 1 to 6, wherein the selection of R1 is the same for each of the nucleosides having the structure of formula II, and the selection of R2 is the same for each of the nucleosides having the structure of formula II. [Item 10] The oligomeric compound according to any one of items 1 to 9, wherein Bx is selected from adenine, guanine, cytosine, thymine, uracil, and 5-methylcytosine. [Item 11] The oligomeric compound according to claim 1 or 2, wherein R1 is methyl. [Item 12] The oligomeric compound according to any one of claims 1 to 11, wherein each of at least two nucleosides of the modified oligonucleotide has a structure independently selected from Formula II. [Item 13] The oligomeric compound according to any one of claims 1 to 11, wherein each of the seven nucleosides of the modified oligonucleotide has a structure independently selected from Formula II. [Item 14] The oligomeric compound according to any one of claims 1 to 11, wherein each of the eight nucleosides of the modified oligonucleotide has a structure independently selected from Formula II. [Item 15] The oligomeric compound according to any one of claims 1 to 11, wherein each of the nine nucleosides of the modified oligonucleotide has a structure independently selected from Formula II. [Item 16] The oligomeric compound according to any one of claims 1 to 11, wherein each of the ten nucleosides of the modified oligonucleotide has a structure independently selected from Formula II. [Item 17] The oligomeric compound according to any one of claims 1 to 11, wherein each of the eleven nucleosides of the modified oligonucleotide has a structure independently selected from Formula II. [Item 18] The oligomeric compound according to any one of claims 1 to 11, wherein each of the 12 nucleosides of the modified oligonucleotide has a structure independently selected from Formula II. [Item 19] The oligomeric compound according to any one of claims 1 to 11, wherein each of the 13 nucleosides of the modified oligonucleotide has a structure independently selected from Formula II. [Item 20] The oligomeric compound according to any one of claims 1 to 11, wherein each of the 14 nucleosides of the modified oligonucleotide has a structure independently selected from Formula II. [Item 21] The oligomeric compound according to any one of claims 1 to 11, wherein each of the 15 nucleosides of the modified oligonucleotide has a structure independently selected from Formula II. [Item 22] The oligomeric compound according to any one of claims 1 to 11, wherein each of the 16 nucleosides of the modified oligonucleotide has a structure independently selected from Formula II. [Item 23] The oligomeric compound according to any one of claims 1 to 11, wherein each of the 17 nucleosides of the modified oligonucleotide has a structure independently selected from Formula II. [Item 24] The oligomer according to any one of claims 1 to 11, wherein each of the 18 nucleosides of the modified oligonucleotide has a structure independently selected from Formula II. compound. [Item 25] The oligomeric compound according to any one of claims 1 to 11, wherein each of the 19 nucleosides of the modified oligonucleotide has a structure independently selected from Formula II. [Item 26] The oligomeric compound according to any one of claims 1 to 11, wherein each of the 20 nucleosides of the modified oligonucleotide has a structure independently selected from Formula II. [Item 27] ​​The oligomeric compound according to any one of claims 1 to 26, wherein R1 of at least one nucleoside having a structure of formula II is methyl. [Item 28] The oligomeric compound according to any one of items 1 to 27, wherein R1 is the same for all of the nucleosides having the structure of formula II. [Item 29] An oligomeric compound comprising a modified oligonucleotide consisting of 14 to 25 linked nucleosides, wherein the modified oligonucleotide is complementary to SMN2 pre-mRNA, and at least one nucleoside of the modified oligonucleotide comprises a 2'-O-(N-alkylacetamido) modified sugar moiety. [Item 30] The oligomeric compound of claim 29, wherein the sugar moiety of each nucleoside containing a 2'-O-(N-alkylacetamide) modified sugar moiety is selected from 2'-O-(N-methylacetamide) and 2'-O-(N-ethylacetamide). [Item 31] The oligomeric compound of claim 29 or 30, wherein each of the seven nucleosides of the modified oligonucleotide contains an independently selected 2'-O-(N-alkylacetamido) modified sugar moiety. [Item 32] The oligomeric compound of claim 29 or 30, wherein each of the eight nucleosides of the modified oligonucleotide contains an independently selected 2'-O-(N-alkylacetamide) modified sugar moiety. [Item 33] The oligomeric compound of claim 29 or 30, wherein each of the nine nucleosides of the modified oligonucleotide contains an independently selected 2'-O-(N-alkylacetamide) modified sugar moiety. [Item 34] The oligomeric compound of claim 29 or 30, wherein each of the 10 nucleosides of the modified oligonucleotide contains an independently selected 2'-O-(N-alkylacetamido) modified sugar moiety. [Item 35] The oligomeric compound of claim 29 or 30, wherein each of the 11 nucleosides of the modified oligonucleotide contains an independently selected 2'-O-(N-alkylacetamide) modified sugar moiety. [Item 36] The oligomeric compound of claim 29 or 30, wherein each of the 12 nucleosides of the modified oligonucleotide contains an independently selected 2'-O-(N-alkylacetamide) modified sugar moiety. [Item 37] The oligomeric compound of claim 29 or 30, wherein each of the 13 nucleosides of the modified oligonucleotide contains an independently selected 2'-O-(N-alkylacetamide) modified sugar moiety. [Item 38] The oligomeric compound of claim 29 or 30, wherein each of the 14 nucleosides of the modified oligonucleotide contains an independently selected 2'-O-(N-alkylacetamide) modified sugar moiety. [Item 39] The oligomeric compound of claim 29 or 30, wherein each of the 15 nucleosides of the modified oligonucleotide contains an independently selected 2'-O-(N-alkylacetamide) modified sugar moiety. [Item 40] The oligomeric compound of claim 29 or 30, wherein each of the 16 nucleosides of the modified oligonucleotide contains an independently selected 2'-O-(N-alkylacetamide) modified sugar moiety. [Item 41] The method of claim 29, wherein each of the 17 nucleosides of the modified oligonucleotide comprises an independently selected 2'-O-(N-alkylacetamido) modified sugar moiety. 31. The oligomeric compound according to claim 30. [Item 42] The oligomeric compound of claim 29 or 30, wherein each of the 18 nucleosides of the modified oligonucleotide contains an independently selected 2'-O-(N-alkylacetamide) modified sugar moiety. [Item 43] The oligomeric compound of claim 29 or 30, wherein each of the 19 nucleosides of the modified oligonucleotide contains an independently selected 2'-O-(N-alkylacetamide) modified sugar moiety. [Item 44] The oligomeric compound of claim 29 or 30, wherein each of the 20 nucleosides of the modified oligonucleotide contains an independently selected 2'-O-(N-alkylacetamide) modified sugar moiety. [Item 45] The oligomeric compound according to any one of claims 29 to 44, wherein at least one of the sugar moieties of the nucleosides containing a 2'-O-(N-alkylacetamide) modified sugar moiety is a 2'-O-(N-methylacetamide) modified sugar moiety. [Item 46] The oligomeric compound according to any one of claims 29 to 45, wherein each N-alkyl group of the 2'-O-(N-alkylacetamide) modified sugar moiety is the same N-alkyl group. [Item 47] The oligomeric compound according to any one of claims 29 to 46, wherein each of the 2'-O-(N-alkylacetamide) modified sugar moieties is a 2'-O-(N-methylacetamide) sugar moiety. [Item 48] The oligomeric compound according to any one of claims 29 to 47, wherein each sugar moiety of each nucleoside of the modified oligonucleotide is a 2'-O-(N-methylacetamide) modified sugar moiety. [Item 49] The oligomeric compound according to any one of items 29 to 47, wherein at least one sugar moiety is not a 2'-O-(N-alkylacetamide) modified sugar moiety. [Item 50] The oligomeric compound according to any one of items 29 to 47, wherein at least one sugar moiety is not a 2'-O-(N-methylacetamide) modified sugar moiety. [Item 51] The oligomeric compound according to any one of claims 29 to 47, wherein at least one sugar moiety is a modified sugar moiety selected from 2'-MOE nucleosides, 2'-OMe nucleosides, 2'-F nucleosides, (4'-CH2-O-2') bicyclic nucleosides, (4'-(CH2)2-O-2') bicyclic nucleosides, (4'-C(CH3)HO-2') bicyclic nucleosides; and morpholino. [Item 52] The oligomeric compound according to any one of items 29 to 51, wherein at least one sugar moiety is an unmodified sugar moiety. [Item 53] The oligomeric compound according to any one of claims 1 to 52, wherein the modified oligonucleotide consists of 16 to 23 linked nucleosides. [Item 54] The oligomeric compound according to any one of claims 1 to 52, wherein the modified oligonucleotide consists of 18 to 20 linked nucleosides. [Item 55] The oligomeric compound according to any one of claims 1 to 53, wherein the modified oligonucleotide consists of 16 nucleosides. [Item 56] The oligomeric compound according to any one of claims 1 to 53, wherein the modified oligonucleotide consists of 17 nucleosides. [Item 57] The oligomeric compound according to any one of claims 1 to 53, wherein the modified oligonucleotide consists of 18 nucleosides. [Item 58] The oligomeric compound according to any one of claims 1 to 53, wherein the modified oligonucleotide consists of 19 nucleosides. [Item 59] The oligomeric compound according to any one of claims 1 to 53, wherein the modified oligonucleotide consists of 20 nucleosides. [Item 60] The oligomeric compound according to any one of claims 1 to 59, wherein the modified oligonucleotide comprises at least one modified internucleoside bond. [Item 61] The modified oligonucleotide comprises at least one phosphorothioate nucleotide. 61. The oligomeric compound according to any one of claims 1 to 60, comprising an interosidic bond. [Item 62] The oligomeric compound of claim 61, wherein each internucleoside linkage of the modified oligonucleotide is selected from phosphorothioate internucleoside linkages and phosphate internucleoside linkages. Item 63. The oligomeric compound of claim 62, wherein the phosphate internucleoside linkage is a phosphodiester internucleoside linkage. [Item 64] The oligomeric compound of claim 62, wherein the modified oligonucleotide has five phosphodiester internucleoside linkages. [Item 65] The oligomeric compound of claim 62, wherein the modified oligonucleotide has six phosphodiester internucleoside linkages. [Item 66] The oligomeric compound of claim 62, wherein the modified oligonucleotide has at least six phosphodiester internucleoside linkages. [Item 67] The oligomeric compound according to any one of items 1 to 66, wherein each internucleoside bond of the modified oligonucleotide is a phosphorothioate internucleoside bond. [Item 68] The oligomeric compound according to any one of claims 1 to 67, wherein the modified oligonucleotide comprises at least one modified nucleic acid base. [Item 69] The oligomeric compound according to any one of claims 1 to 68, wherein the modified oligonucleotide contains at least one 5-methylcytosine. [Item 70] The oligomeric compound according to any one of claims 1 to 69, wherein each nucleic acid base of the modified oligonucleotide is selected from thymine, 5-methylcytosine, cytosine, adenine, uracil and guanine. [Item 71] The oligomeric compound according to any one of items 1 to 70, wherein each cytosine in the modified oligonucleotide is a 5-methylcytosine. [Item 72] The oligomeric compound according to any one of claims 1 to 65, wherein each nucleobase of the modified oligonucleotide is selected from thymine, 5-methylcytosine, adenine and guanine. [Item 73] The oligomeric compound of any one of claims 1 to 72, wherein the modified oligonucleotide is at least 70% complementary to SMN2 pre-mRNA. [Item 74] The oligomeric compound of any one of claims 1 to 72, wherein the modified oligonucleotide is at least 75% complementary to SMN2 pre-mRNA. [Item 75] The oligomeric compound of any one of claims 1 to 72, wherein the modified oligonucleotide is at least 80% complementary to SMN2 pre-mRNA. [Item 76] The oligomeric compound of any one of claims 1 to 72, wherein the modified oligonucleotide is at least 85% complementary to SMN2 pre-mRNA. [Item 77] The oligomeric compound of any one of claims 1 to 72, wherein the modified oligonucleotide is at least 90% complementary to SMN2 pre-mRNA. [Item 78] The oligomeric compound of any one of claims 1 to 72, wherein the modified oligonucleotide is at least 95% complementary to SMN2 pre-mRNA. [Item 79] The oligomeric compound of any one of claims 1 to 72, wherein the modified oligonucleotide is at least 100% complementary to SMN2 pre-mRNA. [Item 80] The oligomeric compound of any one of claims 1 to 72, wherein the modified oligonucleotide is complementary to intron 7 of the SMN2 pre-mRNA. [Item 81] The oligomeric compound of any one of claims 1 to 80, wherein the modified oligonucleotide is complementary to ISS-N1 of the SMN2 pre-mRNA. [Item 82] The oligomeric compound according to any one of claims 1 to 79, wherein the nucleic acid base sequence of the modified oligonucleotide comprises a sequence selected from SEQ ID NO: 1, 2 or 3. [Item 83] The oligomeric compound according to any one of claims 1 to 79, wherein the nucleic acid base sequence of the modified oligonucleotide comprises a sequence selected from any one of SEQ ID NOs: 12 to 239 or 241 to 265. [Item 84] The oligomeric compound according to any one of claims 1 to 79, wherein the nucleic acid base sequence of the modified oligonucleotide comprises a sequence selected from any one of SEQ ID NOs: 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264 or 265. [Item 85] The oligomeric compound according to any one of claims 1 to 79, wherein the nucleic acid base sequence of the modified oligonucleotide consists of a sequence selected from any one of SEQ ID NOs: 1, 2, 3, or 12 to 239, or 241 to 265. [Item 86] The nucleic acid base sequence of the modified oligonucleotide is selected from any one of SEQ ID NOs: 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, or 265. The oligomeric compound according to any one of claims 1 to 79. [Item 87] The oligomeric compound of any one of claims 1 to 79, wherein the modified oligonucleotide is complementary to exon 7 of the SMN2 pre-mRNA. [Item 88] The oligomeric compound of any one of claims 1 to 79, wherein the modified oligonucleotide is complementary to intron 6 of the SMN2 pre-mRNA. [Item 89] The oligomeric compound of any one of claims 1 to 79, wherein the modified oligonucleotide is complementary to an exonic splicing enhancer in exon 7 of the SMN2 pre-mRNA. [Item 90] The oligomeric compound of any one of claims 1 to 79, wherein the modified oligonucleotide is complementary to an aberrant exonic splicing silencer in exon 7 of the SMN2 pre-mRNA. [Item 91] The oligomeric compound according to any one of claims 1 to 90, wherein the oligomeric compound comprises a conjugate group. [Item 92] The oligomeric compound of claim 91, wherein the conjugate group comprises a lipid or a lipophilic group. [Item 93] The oligomeric compound according to claim 92, wherein the lipid or lipophilic group is selected from cholesterol, C10 to C26 saturated fatty acid, C10 to C26 unsaturated fatty acid, C10 to C26 alkyl, triglyceride, tocopherol or cholic acid. [Item 94] The oligomeric compound of claim 93, wherein the lipid or lipophilic group is a saturated or unsaturated hydrocarbon chain. [Item 95] The oligomeric compound according to any one of claims 92 to 94, wherein the lipid or lipophilic group is a C16 lipid. [Item 96] The oligomeric compound according to any one of claims 92 to 94, wherein the lipid or lipophilic group is a C18 lipid. [Item 97] The oligomeric compound according to any one of claims 92 to 94, wherein the lipid or lipophilic group is a C16 alkyl. [Item 98] The oligomeric compound according to any one of claims 92 to 94, wherein the lipid or lipophilic group is a C18 alkyl. [Item 99] The oligomeric compound of claim 93, wherein the lipid or lipophilic group is cholesterol. [Item 100] The oligomeric compound of claim 93, wherein the lipid or lipophilic group is tocopherol. [Item 101] The oligomeric compound of claim 93, wherein the lipid or lipophilic group is saturated C16. [Item 102] The oligomeric compound according to any one of items 91 to 101, wherein the conjugate group is attached to the modified oligonucleotide at the 5' end of the modified oligonucleotide. [Item 103] The conjugate group is a 3'-terminus of the modified oligonucleotide. 102. The oligomeric compound of any one of claims 91 to 101, which is linked to a modified oligonucleotide. [Item 104] The oligomeric compound of any one of claims 91 to 103, wherein the conjugate group comprises a cleavable linker. [Item 105] The oligomeric compound described in claim 105, wherein the cleavable linker comprises one or more linker nucleosides. [Item 106] The oligomeric compound according to any one of items 1 to 105, which consists of a modified oligonucleotide. [Item 107] The oligomeric compound according to any one of items 91 to 105, comprising the modified oligonucleotide and the conjugate group. [Item 108] The oligomeric compound of any one of claims 1 to 108, wherein the oligomeric compound regulates splicing of SMN2 pre-mRNA. [Item 109] The oligomeric compound according to any one of items 1 to 108, wherein the oligomeric compound is single-stranded. [Item 110] The oligomeric compound of any one of items 1 to 108, wherein the oligomeric compound is paired with a complementary oligomeric compound to form a double-stranded compound. [Item 111] The oligomeric compound of claim 110, wherein the complementary oligomeric compound comprises a conjugate group. [Item 112] A pharmaceutical composition comprising the oligomeric compound according to any one of claims 1 to 111 and at least one pharma- ceutically acceptable carrier or diluent. [Item 113] A method for modulating splicing of SMN2 pre-mRNA, comprising contacting a cell, tissue or organ with the oligomeric compound or composition according to any one of claims 1 to 112. [Item 114] A method for promoting inclusion of exon 7 of SMN2 mRNA in a cell, tissue or organ, the method comprising contacting the cell, tissue or organ with an oligomeric compound described in any one of claims 1 to 111 or a composition described in claim 112. [Item 115] The method of claim 113 or 114, wherein the cell, tissue or organ is within an animal. [Item 116] The method of claim 113 or 114, wherein the cell, tissue or organ is in a human. [Item 117] A method comprising administering to a subject the oligomeric compound described in any one of claims 1 to 111 or the composition described in claim 112. [Item 118] The method of claim 117, wherein the subject has SMA. [Item 119] The method of claim 117, wherein the subject has type I SMA. [Item 120] The method of claim 117, wherein the subject has SMA type II. [Item 121] The method of claim 117, wherein the subject has SMA type III. [Item 122] The method of claim 117, wherein the subject has type IV SMA. [Item 123] A method for treating spinal muscular atrophy in a patient, comprising administering to a patient in need thereof an oligomeric compound according to any one of claims 1 to 111 or a composition according to claim 112. [Item 124] A method for treating spinal muscular atrophy type I in a patient, comprising administering to a patient in need thereof an oligomeric compound according to any one of claims 1 to 111 or a composition according to claim 112. [Item 125] A method for treating spinal muscular atrophy type II in a patient, comprising administering to a patient in need thereof an oligomeric compound according to any one of claims 1 to 111 or a composition according to claim 112. [Item 126] A method for treating spinal muscular atrophy type III in a patient, comprising administering to a patient in need thereof an oligomeric compound according to any one of claims 1 to 111 or a composition according to claim 112. [Item 127] A method for treating spinal muscular atrophy type IV in a patient, comprising administering to a patient in need thereof an oligomeric compound according to any one of claims 1 to 111 or a composition according to claim 112. [Item 128] The method of any one of claims 122 to 127, wherein the oligomeric compound is administered to the central nervous system. [Item 129] The method of claim 128, wherein the oligomeric compound is administered intrathecally. [Item 130] The method of any one of claims 123 to 127, wherein the oligomeric compound is administered systemically. [Item 131] The method of claim 130, wherein the oligomeric compound is administered subcutaneously. [Item 132] The method of claim 130, wherein the oligomeric compound is administered intramuscularly. [Item 133] The method of any one of claims 122 to 132, wherein the oligomeric compound is administered to the central nervous system and systemically. [Item 134] The method of any one of claims 122 to 132, wherein the oligomeric compound is co-administered with a second compound. [Item 135] The method of claim 134, wherein the second compound is selected from a small molecule and an oligomeric compound. [Item 136] The method according to item 135, wherein the second compound is an oligomeric compound according to any one of items 1 to 111. [Item 137] The method of any one of claims 134 to 136, wherein the second compound is administered to the CSF. [Item 138] The method of claim 137, wherein the second compound is administered intrathecally. [Item 139] The method of any one of claims 134 to 138, wherein the second compound is a second oligomeric compound comprising a second modified oligonucleotide having at least one modified nucleoside comprising a modified sugar moiety. [Item 140] The method of claim 139, wherein the modified sugar moiety of the modified nucleoside of the second modified oligonucleotide of the second oligomeric compound is selected from 2'-OMe and 2'-MOE. [Item 141] The method of claim 139 or 140, wherein each nucleoside of the second modified oligonucleotide comprises a modified sugar moiety. [Item 142] The method of claim 141, wherein the modified sugar moiety of the second oligonucleotide is a 2'-MOE sugar moiety. 140. The method of claim 139, wherein the second compound is an oligomeric compound comprising a morpholino oligonucleotide. [Item 144] The method of any one of items 134 to 143, wherein the second oligomeric compound has the nucleic acid base sequence of SEQ ID NO:1. [Item 145] The method of claim 144, wherein the second oligomeric compound has a nucleic acid base sequence and a chemical modification motif of Isis396443. [Item 146] The method of any one of claims 134 to 145, wherein the second oligomeric compound consists of the modified oligonucleotide. [Item 147] The method of any one of items 134 to 146, wherein the second compound is administered prior to administration of the oligomeric compound of any one of items 1 to 111 or the composition of item 112. [Item 148] The method of any one of items 134 to 146, wherein the second compound is administered after administration of the oligomeric compound of any one of items 1 to 111 or the composition of item 112. [Item 149] The method of any one of items 134 to 146, wherein the second compound is administered less frequently than the administration of the oligomeric compound of any one of items 1 to 111 or the composition of item 112. [Item 150] An oligomeric compound according to any one of items 1 to 111 or a composition according to item 112 for use in a method of treatment. [Item 151] Use of an oligomeric compound according to any one of claims 1 to 111 or a composition according to claim 112 for the preparation of a medicament for treating spinal muscular atrophy type I. [Item 152] Use of the oligomeric compound according to any one of claims 1 to 111 or the composition according to claim 112 for the preparation of a medicament for treating spinal muscular atrophy type II. [Item 153] Use of the oligomeric compound according to any one of claims 1 to 111 or the composition according to claim 112 for the preparation of a medicament for treating spinal muscular atrophy type III. [Item 154] Use of the oligomeric compound according to any one of claims 1 to 111 or the composition according to claim 112 for the preparation of a medicament for treating spinal muscular atrophy type IV. [Item 155] Use of the oligomeric compound according to any one of claims 1 to 112 for the preparation of a medicament for treating spinal muscular atrophy. [Item 156] (i) systemically administering a compound comprising an oligomeric compound according to any one of claims 1 to 112; and (ii) administering a compound comprising an oligomeric compound according to any one of claims 1 to 112 to the CNS. A method comprising the steps of: [Item 157] A method comprising: (i) systemically administering a compound comprising an oligomeric compound or composition according to any one of claims 1 to 112; and (ii) administering to the CNS a compound comprising a modified oligonucleotide having at least one modified nucleoside comprising a modified sugar moiety selected from 2'-OMe and 2'-MOE. [Item 158] (i) systemically administering a compound comprising an oligomeric compound according to any one of claims 1 to 112; and (ii) administering a compound comprising an oligomeric compound according to any one of claims 1 to 112 to the CNS. A method comprising the steps of: [Item 159] A method comprising: (i) systemically administering a compound comprising an oligomeric compound according to any one of claims 85 to 106; and (ii) administering to the CNS a compound comprising a modified oligonucleotide having at least one modified nucleoside comprising a modified sugar moiety selected from 2'-OMe and 2'-MOE. [Item 160] A method comprising: (i) systemically administering a modified oligonucleotide having at least one modified nucleoside conjugate comprising a modified sugar moiety selected from 2'-OMe and 2'-MOE; and (ii) administering to the CNS a compound comprising an oligomeric compound described in any one of claims 1 to 112. [Item 161] A method comprising: (i) systemically administering a modified oligonucleotide having at least one modified nucleoside comprising a modified sugar moiety selected from 2'-OMe and 2'-MOE and a conjugate group; and (ii) administering to the CNS a compound comprising a modified oligonucleotide having at least one modified nucleoside comprising a modified sugar moiety selected from 2'-OMe and 2'-MOE. [Item 162] The method of claim 160, wherein the conjugate group comprises a lipid or lipophilic group selected from cholesterol, C10 to C26 saturated fatty acid, C10 to C26 unsaturated fatty acid, C10 to C26 alkyl, triglyceride, tocopherol or cholic acid. [Item 163] The compound or method according to any one of claims 1 to 162, wherein the SMN2 pre-mRNA is a nucleic acid base sequence selected from SEQ ID NO: 10, 11, or 240.

[0309] While certain compounds, compositions and methods described herein have been specifically described in accordance with certain embodiments, the following examples are provided only to illustrate, but not to limit, the compounds described herein.

[0310] Although the sequence listing accompanying this application identifies each sequence as either "RNA" or "DNA," as appropriate, in practice these sequences may be modified with any combination of chemical modifications. Those of skill in the art will readily recognize that the designations "RNA" or "DNA," as the case may be, are arbitrary when describing modified oligonucleotides. For example, oligonucleotides containing nucleosides containing 2'-OH sugar moieties and thymine bases may be modified with any combination of chemical modifications. can be described as DNA with modified sugars (2'-OH instead of one 2'-H in DNA) or RNA with modified bases (thymine (methylated uracil) instead of uracil in RNA). Thus, the nucleic acid sequences described herein, including but not limited to those in the sequence listing, are intended to encompass nucleic acids containing any combination of natural or modified RNA and / or DNA, such as, but not limited to, nucleic acids with modified nucleobases. By way of further example and without limitation, an oligomeric compound having the nucleobase sequence "ATCGATCG" encompasses any oligomeric compound having that nucleobase sequence, whether modified or unmodified, and includes, but is not limited to, compounds that contain RNA bases, such as those having the sequence "AUCGAUCG" and those having some DNA bases and some RNA bases, such as "AUCGATCG", as well as those having the sequence "ATCGATCG". m CGAUCG" (in the sequence, m C denotes a cytosine base containing a methyl group at the 5-position).

[0311] Certain compounds described herein (e.g., modified oligonucleotides) have one or more asymmetric centers, resulting in enantiomers, diastereomers, and other stereoisomeric structures. These may be defined in terms of absolute stereochemistry, such as (R) or (S), α or β (such as in the case of sugar anomers), or (D) or (L) (such as in the case of amino acids). The compounds provided herein include all possible isomers, including their racemic and optically pure forms, unless otherwise specified. Similarly, all cis and trans isomers and tautomeric forms are included unless otherwise specified. The oligomeric compounds described herein include racemic mixtures as well as chirally pure or enriched mixtures. For example, oligomeric compounds having a plurality of phosphorothioate internucleoside linkages include compounds in which the chirality of the phosphorothioate internucleoside linkages is controlled or the chirality of the phosphorothioate internucleoside linkages is random.

[0312] Unless otherwise specified, all compounds, including oligomeric compounds, described herein include pharma- ceutically acceptable salts thereof.

[0313] The compounds described herein include variations in which one or more atoms are replaced with non-radioactive or radioactive isotopes of the specified elements. For example, compounds described herein that contain hydrogen atoms include 1 Isotopic substitutions encompassed by the compounds herein include all possible deuterium substitutions for each H hydrogen atom. 1 Instead of H 2 H or 3 H, 12 Instead of C 13 C or 14 C. 14 Instead of N 15 N, 16 Instead of O 17 O or 18 O. 32 Instead of S 33 S, 34 S, 35 S or 36 In certain embodiments, non-radioactive isotope substitution can impart new properties to the oligomeric compound that are beneficial for use as a therapeutic or research tool. In certain embodiments, radioactive isotope substitution can make the compound suitable for research or diagnostic purposes, such as imaging. EXAMPLES

[0314] Non-Limiting Disclosure and Incorporation by Reference While certain compounds, compositions and methods described herein have been specifically described according to certain embodiments, the following examples are provided solely to illustrate, but not to limit, the compounds described herein. Each of the references cited in this application is incorporated herein by reference in its entirety.

[0315] Example 1: In vitro effects of modified oligonucleotides targeting SMN2 The effect of modified oligonucleotides containing the 2'-MOE or 2'-NMA modifications shown in the table below on the splicing of exon 7 in SMN2 was investigated in vitro. Tested with.

[0316] Spinal muscular atrophy (SMA) patient fibroblast cell line (GM03813: Cornell Institute) was plated at a density of 25,000 cells / well and transfected with modified oligonucleotides at the concentrations listed in the table below using electroporation at 120V. After a treatment period of approximately 24 hours, cells were washed with DPBS buffer and lysed. RNA was extracted using Qiagen RNeasy purification method and mRNA levels were measured by qRT-PCR. Levels of SMN2 with exon 7 were measured using primer / probe set hSMN2vd#4_LTS00216_MGB, levels of SMN2 without exon 7 were measured using hSMN2va#4_LTS00215_MGB, and total levels of SMN2 were measured using HTS4210. The amounts of SMN2 with exon 7 and SMN2 without exon 7 were normalized to total SMN2. The results are shown in the table below as levels of exon 7-containing (+exon 7) SMN2 relative to total SMN2 and levels of exon 7-free (-exon 7) SMN2 relative to total SMN2. As shown in the table below, treatment with modified oligonucleotides containing 2'-NMA modifications exhibited greater exon 7 inclusion (reduced exon 7 exclusion) in SMA patient fibroblasts compared to modified oligonucleotides containing 2'-MOE modifications. [Table 3]

[0317] In the above table, subscripts: "s" represents a phosphorothioate internucleoside linkage, "e" represents a 2'-MOE modified nucleoside, and "n" represents a 2'-O-(N-methylacetamide) modified nucleoside. Superscript: "m" before C represents 5-methylcytosine. [Table 4]

[0318] Example 2: Effects of modified oligonucleotides targeting SMN2 in transgenic mice Taiwanese strain SMA type III human transgenic mice (Jackson Laboratory, Bar Harbor, Maine) lack mouse SMN and are homozygous for human SMN2. These mice are described in Hsieh-Li et al., Nature Genet. 24, 66-70 (2000). Each mouse received a single intracerebroventricular (ICV) bolus of saline (PBS) or compound 396443 or compound 443305 (see Example 1) on day 1. Each treatment group consisted of 3-4 mice. Mice were sacrificed on day 7 after 7 days. Total RNA from spinal cord and brain was extracted and analyzed by RT-qPCR as described in Example 1. The ratios of exon 7-containing SMN2 to total SMN2 and exon 7-free SMN2 to total SMN2 were set to 1.0 for the PBS-treated control group. Normalized results for all treatment groups are shown in the table below. As shown in the table below, modified oligonucleotides containing 2'-NMA modifications exhibited more exon 7 inclusion and less exon 7 exclusion in vivo than modified oligonucleotides containing 2'-MOE modifications. [Table 5]

[0319] Example 3: Effects of modified oligonucleotides targeting SMN2 following systemic administration in transgenic mice Taiwanese type III human transgenic mice were administered intraperitoneal (IP) injections of saline (PBS), compound #396443, or compound #443305 (see Example 1) once every 48 hours for a total of four injections. Each treatment group consisted of 3-4 mice. Mice were sacrificed 72 hours after the last dose. Various tissues, including liver, diaphragm, quadriceps, and heart, were harvested and total RNA was isolated. Exon 7-containing and -free SMN2 as well as total SMN2 levels were measured by RT-qPCR as described in Examples 1 and 2, except that the primer / probe sets for this experiment were those described in Tiziano, et al., Eur J Humn Genet, 2010. The results are shown in the table below. These results indicate that systemic administration of modified oligonucleotides containing 2'-NMA modifications resulted in more exon 7 inclusion and less exon 7 exclusion than modified oligonucleotides containing 2'-MOE modifications. [Table 6] [Table 7]

[0320] Example 4: Effect of modified oligonucleotides targeting SMN2 in transgenic mice Taiwanese type III human transgenic mice were administered an ICV bolus of saline (PBS) or the modified oligonucleotides listed in the table below. Each treatment group consisted of 3-4 mice. Mice were sacrificed 2 weeks after administration. The brain and spinal cord of each mouse were harvested and total RNA was isolated from each tissue. Exon 7-containing and -free SMN2 as well as total SMN2 levels were measured by RT-qPCR as described in Examples 1 and 2. The results are shown in the table below. These results indicate that modified oligonucleotides containing 2'-NMA modifications resulted in more exon 7 inclusion and less exon 7 exclusion than modified oligonucleotides containing 2'-MOE modifications. [Table 8]

[0321] In the above table, subscripts: "s" represents a phosphorothioate internucleoside linkage, "e" represents a 2'-MOE modified nucleoside, and "n" represents a 2'-O-(N-methylacetamide) modified nucleoside. Superscript: "m" before C represents 5-methylcytosine. [Table 9]

[0322] Example 5: Effects of modified oligonucleotides targeting SMN2 following systemic administration in transgenic mice Taiwanese type III human transgenic mice were administered subcutaneous injections of either saline (PBS) or modified oligonucleotides listed in Example 4, once every 48-72 hours, for a total of 10-150 mg / kg / week for 3 weeks. Each treatment group consisted of 4 mice. Mice were sacrificed 72 hours after the last dose. Various tissues were harvested and total RNA was isolated from each tissue. Exon 7-containing and -free SMN2 as well as total SMN2 levels were measured by RT-qPCR as described in Examples 1 and 2. The results are shown in the table below. These results indicate that systemic administration of modified oligonucleotides containing 2'-NMA modifications resulted in more exon 7 inclusion and less exon 7 exclusion than modified oligonucleotides containing 2'-MOE modifications. [Table 10] [Table 11] "nd" indicates no data, ED 50 was not calculated.

[0323] Example 6: Effects of compounds containing conjugate groups and modified oligonucleotides targeting SMN2 following systemic administration in transgenic mice Taiwanese type III human transgenic mice were treated for 3 weeks by subcutaneous administration of 10-300 mg / kg / week of modified oligonucleotides listed in the table below or saline (PBS) alone and sacrificed 48-72 hours after the last administration. There were 3-4 mice in each group. Total RNA from various tissues was extracted and analyzed by RT-qPCR as described in Examples 1 and 2. The results shown in the table below indicate that oligomeric compounds containing C16 conjugates and 2'-NMA modifications exhibited more exon 7 inclusion and less exon 7 exclusion than other tested compounds. [Table 12] In the above table, subscripts: "s" represents a phosphorothioate internucleoside linkage, "o" represents a phosphate internucleoside linkage, "d" represents a 2'-deoxynucleoside, "e" represents a 2'-MOE modified nucleoside, and "n" represents a 2'-O-(N-methylacetamide) modified nucleoside. Superscript: "m" before C represents 5-methylcysteine. The structure of C16-HA is: [ka] [Table 13]

[0324] Example 7: In vivo dose-response effects of oligomeric compounds containing lipophilic conjugate groups The oligomeric compounds listed in the table below are complementary to both human and mouse MALAT-1 transcripts. Their effects on MALAT-1 expression were tested in vivo. Male diet-induced obese (DIO) mice were each given weekly intravenous injections of the oligomeric compounds listed in the table below or saline vehicle alone via the tail vein for two weeks. Each treatment group consisted of three or four mice. Animals were sacrificed three days after the last injection. MALAT-1 RNA expression in the heart analyzed by RT-qPCR using RiboGreen (Thermo Fisher Scientific, Carlsbad, CA) and normalized to total RNA is shown below. The average results for each group are shown as a percentage of MALAT-1 RNA levels normalized to the average results of vehicle-treated animals. The data below show that oligomeric compounds containing lipophilic conjugate groups are more potent in the heart compared to the parent compound without the lipophilic conjugate group. [Table 14] The subscript "k" represents a cEt modified bicyclic sugar moiety. See table above for additional subscripts and superscripts. The structure of "C16-HA-" is shown in Example 2. The structure of "Ole-HA-" is: [ka]

[0325] Example 8: Effects of oligomeric compounds containing lipophilic conjugate groups after different routes of administration in vivo The effect of Isis #556089 and #812134 (see Example 9) on MALAT-1 expression was tested in vivo. Male wild-type C57bl / 6 mice were each administered either intravenous (IV) or subcutaneous (SC) injections of Isis #556089, Isis #812134, or saline vehicle alone via the tail vein. Each treatment group consisted of 4 mice. Animals were sacrificed 3 days after injection. MALAT-1 RNA expression from the heart analyzed by RT-qPCR using RiboGreen (Thermo Fisher Scientific, Carlsbad, CA) and normalized to total RNA is shown below. The average results for each group are shown as a percentage of MALAT-1 RNA levels normalized to the average results of vehicle-treated animals. The data below show that oligomeric compounds containing lipophilic conjugate groups are more potent in the heart compared to the parent compound without lipophilic conjugate groups. [Table 15]

[0326] Example 9: Effects of oligomeric compounds containing lipophilic conjugate groups after different routes of administration in vivo The compounds listed in the table below are complementary to CD36. They were tested in vivo. Female wild-type C57bl / 6 mice were each given weekly intravenous or intraperitoneal injections of either compound or saline vehicle alone for three weeks. Each treatment group consisted of four mice. Animals were sacrificed three days after the last injection. CD36 mRNA expression from heart and quadriceps analyzed by RT-qPCR using RiboGreen (Thermo Fisher Scientific, Carlsbad, CA) and normalized to total RNA is shown below. The average results for each group are shown as a percentage of CD36 RNA levels normalized to the average results of vehicle-treated animals. The data below show that in both heart and quadriceps, oligomeric compounds containing lipophilic conjugate groups are more potent than the parent compound without the lipophilic conjugate group. [Table 16] See table above for legend.

[0327] Example 10: In vivo effects of oligomeric compounds containing lipophilic conjugate groups The oligomeric compounds listed in the table below are complementary to both human and mouse myotonic dystrophy protein kinase (DMPK) transcripts. Their effects on DMPK expression were tested in vivo. Wild-type Balb / c mice were each administered an intravenous injection of either the oligomeric compounds at the doses listed in the table below or saline vehicle alone. Each animal received a total of four doses, once a week for three and a half weeks. Each treatment group consisted of three or four mice. Animals were sacrificed two days after the last dose. DMPK mRNA expression from quadriceps muscle analyzed by RT-qPCR using RiboGreen (Thermo Fisher Scientific, Carlsbad, CA) and normalized to total RNA is shown below. The average results for each group are presented as a percentage of DMPK RNA levels normalized to the average results of vehicle-treated animals. Entries marked "nd" mean no data. The data below show that in quadriceps muscle, oligomeric compounds containing a lipophilic conjugate group are more potent than the parent compound without the lipophilic conjugate group. [Table 17] See table above for legend. The structures of "Chol-TEG-" and "Toco-TEG-" are shown in Examples 1 and 2, respectively. "HA-Chol" is the 2'-modification shown below. [ka] "HA-C10" and "HA-C16" are 2'-modifications shown below. [ka] In the formula, the subscript n is 1 in "HA-C10" and 7 in "HA-C16."

[0328] Example 11: In vivo effects of oligomeric compounds The oligomeric compounds listed in the table below are complementary to both human and mouse MALAT-1 transcripts. Their effects on MALAT-1 expression were tested in vivo. Male wild-type C57bl / 6 mice were each administered a subcutaneous injection of the oligomeric compounds at the doses listed in the table below or saline vehicle alone on days 0, 4, and 10 of the treatment period. Each treatment group consisted of 3 mice. Animals were sacrificed 4 days after the last injection. MALAT-1 RNA expression from the heart analyzed by RT-qPCR using RiboGreen (Thermo Fisher Scientific, Carlsbad, Calif.) and normalized to total RNA is shown below. The average results for each group are shown as a percentage of MALAT-1 RNA levels normalized to the average results of vehicle-treated animals. The data below show that in the heart, compounds containing a lipophilic conjugate group are more potent compared to the parent compound without a lipophilic conjugate group. [Table 18] See table above for legend. The structure of "C16-HA-" is shown in Example 2. The structures of “C16-2x-C6-” and “C16-2x-C3-” are as follows. [ka] In the formula, m=2 for “C16-2x-C6-”, and m=1 for “C16-2x-C3-”. The structure of “C16-C6-” is as follows. [ka] The structure of “C16-C3-Ab-” is as follows: [ka]

Claims

1. An oligomeric compound comprising a modified oligonucleotide consisting of 14 to 25 linked nucleosides, said modified oligonucleotide being complementary to SMN2 pre-mRNA, and at least one nucleoside of said modified oligonucleotide having the structure of formula II: 【Chemistry 1】 For each nucleoside of formula II: Bx is an independently selected nucleobase; R １ and R ２ are each independently selected from hydrogen and methyl; Or, R １ is hydrogen, R ２ is independently selected from ethyl, propyl or isopropyl.

2. R １ and R ２ 2. The oligomeric compound of claim 1, comprising at least one nucleoside of formula II, wherein one of said nucleosides is not hydrogen.

3. For at least one nucleoside having the structure of Formula II, R １ is hydrogen, R ２ 3. The oligomeric compound of claim 1 or 2, wherein is selected from methyl, ethyl, propyl or isopropyl.

4. R １ is hydrogen, R ２ 4. The oligomeric compound of any one of claims 1 to 3, comprising at least one nucleoside of Formula II, wherein is selected from methyl or ethyl.

5. R １ and R ２ 5. The oligomeric compound of claim 1, comprising at least one nucleoside of formula II, at least one of which is methyl.

6. R １ and R ２ is hydrogen, and R １ and R ２ The oligomeric compound according to any one of claims 1 to 5, comprising at least one nucleoside of formula II, wherein the other of said nucleosides is methyl.

7. R １ The oligomeric compound of any one of claims 1 to 6, wherein the selection of is different for at least two of said nucleosides having the structure of Formula II.

8. R ２ The oligomeric compound of any one of claims 1 to 7, wherein the selection of is different for at least two of said nucleosides having the structure of Formula II.

9. R １ is the same for each of said nucleosides having the structure of Formula II; ２ The oligomeric compound of any one of claims 1 to 6, wherein the selection of is the same for each of said nucleosides having the structure of Formula II.

10. The oligomeric compound according to any one of claims 1 to 9, wherein Bx is selected from adenine, guanine, cytosine, thymine, uracil and 5-methylcytosine.

11. R １ The oligomeric compound of claim 1 or 2, wherein is methyl.

12. The oligomeric compound of any one of claims 1 to 11, wherein each of at least two nucleosides of the modified oligonucleotide has a structure independently selected from Formula II.

13. The oligomeric compound of any one of claims 1 to 11, wherein each of the seven nucleosides of the modified oligonucleotide has a structure independently selected from Formula II.

14. The oligomeric compound of any one of claims 1 to 11, wherein each of the eight nucleosides of the modified oligonucleotide has a structure independently selected from Formula II.

15. The oligomeric compound of any one of claims 1 to 11, wherein each of the nine nucleosides of the modified oligonucleotide has a structure independently selected from Formula II.

16. The oligomeric compound of any one of claims 1 to 11, wherein each of the ten nucleosides of the modified oligonucleotide has a structure independently selected from Formula II.

17. The oligomeric compound of any one of claims 1 to 11, wherein each of the eleven nucleosides of the modified oligonucleotide has a structure independently selected from Formula II.

18. The oligomeric compound of any one of claims 1 to 11, wherein each of the 12 nucleosides of the modified oligonucleotide has a structure independently selected from Formula II.

19. The oligomeric compound of any one of claims 1 to 11, wherein each of the 13 nucleosides of the modified oligonucleotide has a structure independently selected from Formula II.

20. The oligomeric compound of any one of claims 1 to 11, wherein each of the 14 nucleosides of the modified oligonucleotide has a structure independently selected from Formula II.

21. The oligomeric compound of any one of claims 1 to 11, wherein each of the 15 nucleosides of the modified oligonucleotide has a structure independently selected from Formula II.

22. The oligomeric compound of any one of claims 1 to 11, wherein each of the 16 nucleosides of the modified oligonucleotide has a structure independently selected from Formula II.

23. The oligomeric compound of any one of claims 1 to 11, wherein each of the 17 nucleosides of the modified oligonucleotide has a structure independently selected from Formula II.

24. The oligomeric compound of any one of claims 1 to 11, wherein each of the 18 nucleosides of the modified oligonucleotide has a structure independently selected from Formula II.

25. The oligomeric compound of any one of claims 1 to 11, wherein each of the 19 nucleosides of the modified oligonucleotide has a structure independently selected from Formula II.

26. The oligomeric compound of any one of claims 1 to 11, wherein each of the 20 nucleosides of the modified oligonucleotide has a structure independently selected from Formula II.

27. R of at least one nucleoside having the structure of Formula II １ is methyl.

27. The oligomeric compound according to any one of claims 1 to 26.

28. R １ 28. The oligomeric compound of any one of claims 1 to 27, wherein is the same for all of said nucleosides having the structure of Formula II.

29. 1. An oligomeric compound comprising a modified oligonucleotide consisting of 14 to 25 linked nucleosides, said modified oligonucleotide being complementary to an SMN2 pre-mRNA, and at least one nucleoside of said modified oligonucleotide comprising a 2'-O-(N-alkylacetamido) modified sugar moiety.

30. 30. The oligomeric compound of claim 29, wherein the sugar moiety of each nucleoside comprising a 2'-O-(N-alkylacetamide) modified sugar moiety is selected from 2'-O-(N-methylacetamide) and 2'-O-(N-ethylacetamide).

31. 31. The oligomeric compound of claim 29 or 30, wherein each of the seven nucleosides of said modified oligonucleotide comprises an independently selected 2'-O-(N-alkylacetamido) modified sugar moiety.

32. 31. The oligomeric compound of claim 29 or 30, wherein each of the eight nucleosides of said modified oligonucleotide comprises an independently selected 2'-O-(N-alkylacetamido) modified sugar moiety.

33. 31. The oligomeric compound of claim 29 or 30, wherein each of the nine nucleosides of said modified oligonucleotide comprises an independently selected 2'-O-(N-alkylacetamido) modified sugar moiety.

34. 31. The oligomeric compound of claim 29 or 30, wherein each of the ten nucleosides of said modified oligonucleotide comprises an independently selected 2'-O-(N-alkylacetamido) modified sugar moiety.

35. 31. The oligomeric compound of claim 29 or 30, wherein each of the eleven nucleosides of said modified oligonucleotide comprises an independently selected 2'-O-(N-alkylacetamido) modified sugar moiety.

36. 31. The oligomeric compound of claim 29 or 30, wherein each of the 12 nucleosides of said modified oligonucleotide comprises an independently selected 2'-O-(N-alkylacetamido) modified sugar moiety.

37. 31. The oligomeric compound of claim 29 or 30, wherein each of the 13 nucleosides of said modified oligonucleotide comprises an independently selected 2'-O-(N-alkylacetamido) modified sugar moiety.

38. 31. The oligomeric compound of claim 29 or 30, wherein each of the 14 nucleosides of said modified oligonucleotide comprises an independently selected 2'-O-(N-alkylacetamido) modified sugar moiety.

39. 31. The oligomeric compound of claim 29 or 30, wherein each of the 15 nucleosides of said modified oligonucleotide comprises an independently selected 2'-O-(N-alkylacetamido) modified sugar moiety.

40. 31. The oligomeric compound of claim 29 or 30, wherein each of the 16 nucleosides of said modified oligonucleotide comprises an independently selected 2'-O-(N-alkylacetamido) modified sugar moiety.

41. 31. The oligomeric compound of claim 29 or 30, wherein each of the 17 nucleosides of said modified oligonucleotide comprises an independently selected 2'-O-(N-alkylacetamido) modified sugar moiety.

42. 31. The oligomeric compound of claim 29 or 30, wherein each of the 18 nucleosides of said modified oligonucleotide comprises an independently selected 2'-O-(N-alkylacetamido) modified sugar moiety.

43. 31. The oligomeric compound of claim 29 or 30, wherein each of the 19 nucleosides of said modified oligonucleotide comprises an independently selected 2'-O-(N-alkylacetamido) modified sugar moiety.

44. 31. The oligomeric compound of claim 29 or 30, wherein each of the 20 nucleosides of said modified oligonucleotide comprises an independently selected 2'-O-(N-alkylacetamido) modified sugar moiety.

45. 45. The oligomeric compound of any one of claims 29 to 44, wherein the sugar moiety of at least one of said nucleosides comprising a 2'-O-(N-alkylacetamido) modified sugar moiety is a 2'-O-(N-methylacetamido) modified sugar moiety.

46. 46. ​​The oligomeric compound of any one of claims 29-45, wherein each N-alkyl group of said 2'-O-(N-alkylacetamido) modified sugar moiety is the same N-alkyl group.

47. 47. The oligomeric compound of any one of claims 29 to 46, wherein each of said 2'-O-(N-alkylacetamido) modified sugar moieties is a 2'-O-(N-methylacetamido) sugar moiety.

48. The oligomeric compound of any one of claims 29 to 47, wherein each sugar moiety of each nucleoside of said modified oligonucleotide is a 2'-O-(N-methylacetamido) modified sugar moiety.

49. 48. The oligomeric compound of any one of claims 29 to 47, wherein at least one sugar moiety is not a 2'-O-(N-alkylacetamido) modified sugar moiety.

50. 48. The oligomeric compound of any one of claims 29 to 47, wherein at least one sugar moiety is not a 2'-O-(N-methylacetamido) modified sugar moiety.

51. At least one sugar moiety is a 2'-MOE nucleoside, a 2'-OMe nucleoside, a 2'-F nucleoside, (4'-CH ２ -O-2') bicyclic nucleosides, (4'-(CH ２ ) ２ -O-2') bicyclic nucleosides, (4'-C(CH ３ 48. The oligomeric compound of any one of claims 29-47, wherein the modified sugar moiety is selected from: H-O-2') a bicyclic nucleoside; and morpholino.

52. 52. The oligomeric compound according to any one of claims 29 to 51, wherein at least one sugar moiety is an unmodified sugar moiety.

53. The oligomeric compound according to any one of claims 1 to 52, wherein said modified oligonucleotide consists of 16 to 23 linked nucleosides.

54. The oligomeric compound according to any one of claims 1 to 52, wherein said modified oligonucleotide consists of 18 to 20 linked nucleosides.

55. The oligomeric compound according to any one of claims 1 to 53, wherein said modified oligonucleotide consists of 16 nucleosides.

56. The oligomeric compound according to any one of claims 1 to 53, wherein said modified oligonucleotide consists of 17 nucleosides.

57. The oligomeric compound according to any one of claims 1 to 53, wherein said modified oligonucleotide consists of 18 nucleosides.

58. The oligomeric compound according to any one of claims 1 to 53, wherein said modified oligonucleotide consists of 19 nucleosides.

59. The oligomeric compound according to any one of claims 1 to 53, wherein said modified oligonucleotide consists of 20 nucleosides.

60. 60. The oligomeric compound according to any one of claims 1 to 59, wherein said modified oligonucleotide comprises at least one modified internucleoside linkage.

61. The oligomeric compound according to any one of claims 1 to 60, wherein said modified oligonucleotide comprises at least one phosphorothioate internucleoside linkage.

62. 62. The oligomeric compound of claim 61, wherein each internucleoside linkage of said modified oligonucleotide is selected from a phosphorothioate internucleoside linkage and a phosphate internucleoside linkage.

63. 63. The oligomeric compound of claim 62, wherein said phosphate internucleoside linkage is a phosphodiester internucleoside linkage.

64. 63. The oligomeric compound of claim 62, wherein the modified oligonucleotide has five phosphodiester internucleoside linkages.

65. 63. The oligomeric compound of claim 62, wherein the modified oligonucleotide has six phosphodiester internucleoside linkages.

66. 63. The oligomeric compound of claim 62, wherein the modified oligonucleotide has at least six phosphodiester internucleoside linkages.

67. 67. The oligomeric compound according to any one of claims 1 to 66, wherein each internucleoside linkage of said modified oligonucleotide is a phosphorothioate internucleoside linkage.

68. 68. The oligomeric compound according to any one of claims 1 to 67, wherein said modified oligonucleotide comprises at least one modified nucleobase.

69. The oligomeric compound according to any one of claims 1 to 68, wherein said modified oligonucleotide comprises at least one 5-methylcytosine.

70. 70. The oligomeric compound of any one of claims 1 to 69, wherein each nucleobase of said modified oligonucleotide is selected from thymine, 5-methylcytosine, cytosine, adenine, uracil and guanine.

71. The oligomeric compound according to any one of claims 1 to 70, wherein each cytosine in said modified oligonucleotide is a 5-methylcytosine.

72. 66. The oligomeric compound according to any one of claims 1 to 65, wherein each nucleobase of said modified oligonucleotide is selected from thymine, 5-methylcytosine, adenine and guanine.

73. The oligomeric compound of any one of claims 1 to 72, wherein said modified oligonucleotide is at least 70% complementary to the SMN2 pre-mRNA.

74. The oligomeric compound of any one of claims 1 to 72, wherein said modified oligonucleotide is at least 75% complementary to SMN2 pre-mRNA.

75. The oligomeric compound of any one of claims 1 to 72, wherein said modified oligonucleotide is at least 80% complementary to the SMN2 pre-mRNA.

76. The oligomeric compound of any one of claims 1 to 72, wherein said modified oligonucleotide is at least 85% complementary to SMN2 pre-mRNA.

77. The oligomeric compound of any one of claims 1 to 72, wherein said modified oligonucleotide is at least 90% complementary to SMN2 pre-mRNA.

78. The oligomeric compound of any one of claims 1 to 72, wherein said modified oligonucleotide is at least 95% complementary to SMN2 pre-mRNA.

79. The oligomeric compound of any one of claims 1 to 72, wherein said modified oligonucleotide is at least 100% complementary to the SMN2 pre-mRNA.

80. The oligomeric compound of any one of claims 1 to 72, wherein said modified oligonucleotide is complementary to intron 7 of said SMN2 pre-mRNA.

81. The oligomeric compound of any one of claims 1 to 80, wherein said modified oligonucleotide is complementary to ISS-N1 of said SMN2 pre-mRNA.

82. 80. The oligomeric compound of any one of claims 1 to 79, wherein the sequence of the nucleobases of the modified oligonucleotide comprises a sequence selected from SEQ ID NO: 1, 2 or 3.

83. 80. The oligomeric compound of any one of claims 1 to 79, wherein the sequence of the nucleobases of the modified oligonucleotide comprises a sequence selected from any of SEQ ID NOs: 12-239 or 241-265.

84. The nucleic acid base sequences of the modified oligonucleotides are selected from SEQ ID NOs: 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 80. The oligomeric compound of any one of claims 1 to 79, comprising a sequence selected from any of: 3, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264 or 265.

85. 80. The oligomeric compound of any one of claims 1 to 79, wherein the sequence of said nucleobases of said modified oligonucleotide consists of a sequence selected from any of SEQ ID NOs: 1, 2, 3 or 12 to 239 or 241 to 265.

86. 80. The oligomeric compound of any one of claims 1 to 79, wherein the sequence of said nucleobases of said modified oligonucleotide consists of a sequence selected from any of SEQ ID NOs: 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264 or 265.

87. 80. The oligomeric compound of any one of claims 1 to 79, wherein said modified oligonucleotide is complementary to exon 7 of said SMN2 pre-mRNA.

88. 80. The oligomeric compound of any one of claims 1 to 79, wherein said modified oligonucleotide is complementary to intron 6 of said SMN2 pre-mRNA.

89. The oligomeric compound of any one of claims 1 to 79, wherein said modified oligonucleotide is complementary to an exonic splicing enhancer in exon 7 of the SMN2 pre-mRNA.

90. 80. The oligomeric compound of any one of claims 1 to 79, wherein said modified oligonucleotide is complementary to an aberrant exonic splicing silencer in exon 7 of the SMN2 pre-mRNA.

91. 91. The oligomeric compound of any one of claims 1 to 90, wherein the oligomeric compound comprises a conjugate group.

92. 92. The oligomeric compound of claim 91, wherein the conjugate group comprises a lipid or a lipophilic group.

93. The lipid or lipophilic group is cholesterol, C １０ ~C ２６ saturated fatty acids, C １０ ~C ２６ unsaturated fatty acids, C １０ ~C ２６ 93. The oligomeric compound of claim 92, selected from alkyl, triglyceride, tocopherol or cholic acid.

94. 94. The oligomeric compound of claim 93, wherein the lipid or lipophilic group is a saturated or unsaturated hydrocarbon chain.

95. The lipid or lipophilic group is C １６ 95. The oligomeric compound according to any one of claims 92 to 94, which is a lipid.

96. The lipid or lipophilic group is C １８ 95. The oligomeric compound according to any one of claims 92 to 94, which is a lipid.

97. The lipid or lipophilic group is C １６ The oligomeric compound of any one of claims 92 to 94, which is alkyl.

98. The lipid or lipophilic group is C １８ The oligomeric compound of any one of claims 92 to 94, which is alkyl.

99. 94. The oligomeric compound of claim 93, wherein said lipid or lipophilic group is cholesterol.

100. 94. The oligomeric compound of claim 93, wherein said lipid or lipophilic group is tocopherol.

101. The lipid or lipophilic group is a saturated C １６ 94. The oligomeric compound of claim 93, wherein

102. The oligomeric compound according to any one of claims 91 to 101, wherein the conjugate group is attached to the modified oligonucleotide at the 5' end of the modified oligonucleotide.

103. The oligomeric compound according to any one of claims 91 to 101, wherein the conjugate group is attached to the modified oligonucleotide at the 3' end of the modified oligonucleotide.

104. The oligomeric compound according to any one of claims 91 to 103, wherein the conjugate group comprises a cleavable linker.

105. 106. The oligomeric compound of claim 105, wherein the cleavable linker comprises one or more linker nucleosides.

106. The oligomeric compound according to any one of claims 1 to 105, consisting of said modified oligonucleotide.

107. 106. The oligomeric compound of any one of claims 91 to 105, consisting of said modified oligonucleotide and said conjugate group.

108. 109. The oligomeric compound of any one of claims 1 to 108, wherein said oligomeric compound modulates splicing of SMN2 pre-mRNA.

109. The oligomeric compound of any one of claims 1 to 108, wherein the oligomeric compound is single stranded.

110. 109. The oligomeric compound of any one of claims 1 to 108, wherein said oligomeric compound pairs with a complementary oligomeric compound to form a double-stranded compound.

111. The oligomeric compound of claim 110, wherein the complementary oligomeric compound comprises a conjugate group.

112. A pharmaceutical composition comprising an oligomeric compound according to any one of claims 1 to 111 and at least one pharma- ceutically acceptable carrier or diluent.

113. 113. A method of modulating splicing of SMN2 pre-mRNA comprising contacting a cell, tissue or organ with an oligomeric compound or composition according to any one of claims 1-112.

114. 113. A method of promoting inclusion of exon 7 in SMN2 mRNA in a cell, tissue or organ, said method comprising contacting said cell, tissue or organ with an oligomeric compound according to any one of claims 1 to 111 or a composition according to claim 112.

115. The method of claim 113 or 114, wherein the cell, tissue or organ is in an animal.

116. 115. The method of claim 113 or 114, wherein the cell, tissue or organ is in a human.

117. 112. A method comprising administering to a subject an oligomeric compound according to any one of claims 1 to 111 or a composition according to claim 112.

118. The method of claim 117, wherein the subject has SMA.

119. 118. The method of claim 117, wherein the subject has Type I SMA.

120. 118. The method of claim 117, wherein the subject has SMA type II.

121. 118. The method of claim 117, wherein the subject has SMA type III.

122. 118. The method of claim 117, wherein the subject has type IV SMA.

123. 112. A method of treating spinal muscular atrophy in a patient in need thereof comprising administering to a patient an oligomeric compound according to any one of claims 1 to 111 or a composition according to claim 112.

124. 114. A method of treating spinal muscular atrophy type I in a patient in need thereof comprising administering to the patient an oligomeric compound according to any one of claims 1 to 111 or a composition according to claim 112.

125. 114. A method of treating spinal muscular atrophy type II in a patient comprising administering to a patient in need thereof an oligomeric compound according to any one of claims 1 to 111 or a composition according to claim 112.

126. 114. A method of treating spinal muscular atrophy type III in a patient comprising administering to a patient in need thereof an oligomeric compound according to any one of claims 1 to 111 or a composition according to claim 112.

127. 114. A method of treating spinal muscular atrophy type IV in a patient comprising administering to a patient in need thereof an oligomeric compound according to any one of claims 1 to 111 or a composition according to claim 112.

128. The method of any one of claims 122-127, wherein the oligomeric compound is administered to the central nervous system.

129. The method of claim 128, wherein the oligomeric compound is administered intrathecally.

130. The method of any one of claims 123 to 127, wherein the oligomeric compound is administered systemically.

131. The method of claim 130, wherein the oligomeric compound is administered subcutaneously.

132. 131. The method of claim 130, wherein the oligomeric compound is administered intramuscularly.

133. 133. The method of any one of claims 122-132, wherein the oligomeric compound is administered to the central nervous system and systemically.

134. The method of any one of claims 122-132, wherein the oligomeric compound is co-administered with a second compound.

135. 135. The method of claim 134, wherein the second compound is selected from a small molecule and an oligomeric compound.

136. The method of claim 135, wherein the second compound is an oligomeric compound according to any one of claims 1 to 111.

137. 137. The method of any one of claims 134-136, wherein the second compound is administered to the CSF.

138. 138. The method of claim 137, wherein the second compound is administered intrathecally.

139. 139. The method of any one of claims 134-138, wherein the second compound is a second oligomeric compound comprising a second modified oligonucleotide having at least one modified nucleoside that comprises a modified sugar moiety.

140. 140. The method of claim 139, wherein said modified sugar moiety of said modified nucleoside of said second modified oligonucleotide of said second oligomeric compound is selected from 2'-OMe and 2'-MOE.

141. 141. The method of claim 139 or 140, wherein each nucleoside of the second modified oligonucleotide comprises a modified sugar moiety.

142. 142. The method of claim 141, wherein the modified sugar moiety of the second oligonucleotide is a 2'-MOE sugar moiety.

143. 140. The method of claim 139, wherein the second compound is an oligomeric compound comprising a morpholino oligonucleotide.

144. 144. The method of any one of claims 134-143, wherein said second oligomeric compound has the nucleobase sequence of SEQ ID NO:

1.

145. 145. The method of claim 144, wherein the second oligomeric compound has the nucleobase sequence and chemical modification motif of Isis396443.

146. 146. The method of any one of claims 134-145, wherein said second oligomeric compound consists of said modified oligonucleotide.

147. 147. The method of any one of claims 134 to 146, wherein said second compound is administered prior to administration of the oligomeric compound of any one of claims 1 to 111 or the composition of claim 112.

148. 147. The method of any one of claims 134-146, wherein said second compound is administered following administration of the oligomeric compound of any one of claims 1-111 or the composition of claim 112.

149. 147. The method of any one of claims 134-146, wherein said second compound is administered less frequently than the administration of the oligomeric compound of any one of claims 1-111 or the composition of claim 112.

150. An oligomeric compound according to any one of claims 1 to 111 or a composition according to claim 112 for use in therapy.

151. Use of an oligomeric compound according to any one of claims 1 to 111 or a composition according to claim 112 for the preparation of a medicament for the treatment of spinal muscular atrophy type I.

152. Use of an oligomeric compound according to any one of claims 1 to 111 or a composition according to claim 112 for the preparation of a medicament for the treatment of spinal muscular atrophy type II.

153. Use of an oligomeric compound according to any one of claims 1 to 111 or a composition according to claim 112 for the preparation of a medicament for the treatment of spinal muscular atrophy type III.

154. Use of an oligomeric compound according to any one of claims 1 to 111 or a composition according to claim 112 for the preparation of a medicament for the treatment of spinal muscular atrophy type IV.

155. Use of an oligomeric compound according to any one of claims 1 to 112 for the preparation of a medicament for the treatment of spinal muscular atrophy.

156. 112。 A method comprising: (i) systemically administering a compound comprising an oligomeric compound according to any one of claims 1 to 112; and (ii) administering to the CNS a compound comprising an oligomeric compound according to any one of claims 1 to 112.

157. 113. A method comprising: (i) systemically administering a compound comprising the oligomeric compound or composition of any one of claims 1-112; and (ii) administering to the CNS a compound comprising a modified oligonucleotide having at least one modified nucleoside comprising a modified sugar moiety selected from 2'-OMe and 2'-MOE.

158. 112。 A method comprising: (i) systemically administering a compound comprising an oligomeric compound according to any one of claims 1 to 112; and (ii) administering to the CNS a compound comprising an oligomeric compound according to any one of claims 1 to 112.

159. 107. A method comprising: (i) systemically administering a compound comprising the oligomeric compound of any one of claims 85-106; and (ii) administering to the CNS a compound comprising a modified oligonucleotide having at least one modified nucleoside comprising a modified sugar moiety selected from 2'-OMe and 2'-MOE.

160. 113. A method comprising: (i) systemically administering a modified oligonucleotide having at least one modified nucleoside conjugate comprising a modified sugar moiety selected from 2'-OMe and 2'-MOE; and (ii) administering to the CNS a compound comprising an oligomeric compound according to any one of claims 1-112.

161. (i) at least one containing a modified sugar moiety selected from 2'-OMe and 2'-MOE; and (ii) administering to the CNS a compound comprising a modified oligonucleotide having at least one modified nucleoside comprising a modified sugar moiety selected from 2'-OMe and 2'-MOE.

162. 161. The method of claim 160, wherein the conjugate group comprises a lipid or lipophilic group selected from cholesterol, C10-C26 saturated fatty acids, C10-C26 unsaturated fatty acids, C10-C26 alkyl, triglycerides, tocopherol, or cholic acid.

163. 163. The compound or method of any one of claims 1-162, wherein the SMN2 pre-mRNA is a nucleobase sequence selected from any of SEQ ID NOs: 10, 11 or 240.

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