Splicing converter antisense oligonucleotides with modified backbone chemistry

Abstract

Disclosed herein are oligonucleotides having one or more spacers or having no spacer. In various embodiments, the STMN2, KCNQ2, UNC13A, or SMN2 oligonucleotides with spacers reduce STMN2 transcripts with concealment exons or reduce misspliced KCNQ2, UNC13A, or SMN2 transcripts and increase full length STMN2, KCNQ2, UNC13A, SMN transcripts, thereby conferring therapeutic efficacy against neurological diseases such as amyotrophic lateral sclerosis (ALS), frontotemporal dementia (FTD), Alzheimer's disease (AD), or spinal muscular atrophy (SMA).

Description

[0001] Cross-reference to related applications

[0002] This application claims the benefit of and priority to U.S. Provisional Patent Application No. 63 / 285,933 filed on December 3, 2021, U.S. Provisional Patent Application No. 63 / 285,628 filed on December 3, 2021, U.S. Provisional Patent Application No. 63 / 285,786 filed on December 3, 2021, U.S. Provisional Patent Application No. 63 / 285,631 filed on December 3, 2021, U.S. Provisional Patent Application No. 63 / 350,206 filed on June 8, 2022, U.S. Provisional Patent Application No. 63 / 398,987 filed on August 18, 2022, and U.S. Provisional Patent Application No. 63 / 398,992 filed on August 18, 2022, the entire disclosure of each of which is hereby incorporated by reference in its entirety for all purposes. Background Art

[0003] Antisense oligonucleotide is a compound based on nucleic acid, which can be used to suppress the expression of certain genes related to disease. Although antisense oligonucleotide is usually designed to hybridize with target gene, conventional antisense oligonucleotide usually shows poor effect. Therefore, there is a need for developing modified antisense oligonucleotides showing improved performance and effect for preventing, improving and treating disease (examples thereof include neurological diseases).

[0004] Therefore, there is an urgent need to identify compounds and / or compositions that can prevent, ameliorate and treat neurological diseases such as amyotrophic lateral sclerosis (ALS), frontotemporal dementia (FTD), ALS with FTD, Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease, progressive supranuclear palsy (PSP), brain trauma, spinal cord injury, corticobasal degeneration (CBD), nerve injury (e.g., brachial plexus injury), neuropathies (e.g., chemotherapy-induced neuropathy), TDP43 proteinopathies (e.g., chronic traumatic encephalopathy, Perry syndrome, Lewy body dementia associated with Alzheimer's disease, Parkinson's disease with or without dementia and limbic-predominant age-related TDP-43 encephalopathy (LATE)), brain age-related TDP-43 with sclerosis (CARTS), facial onset sensory and motor neuron disease (FAP) and encephalopathy. neuronopathy, Guam Parkinson-dementia complex, multisystem proteinopathies, CTE, spinal muscular atrophy (SMA), and synaptic disorders such as autism. Summary of the Invention

[0005] Disclosed herein are compounds comprising a splice-switching oligonucleotide, and wherein the splice-switching oligonucleotide comprises a spacer.Additionally disclosed herein are splice-switching oligonucleotides, wherein the splice-switching oligonucleotide comprises a spacer.

[0006] Additionally disclosed herein are compounds comprising a splice-switching oligonucleotide comprising a sequence that is at least 85% complementary to an equal length portion of a sequence from a transcript whose mis-splicing results in a neurological disease, or whose splicing can be modulated to treat a neurological disease, wherein at least one (i.e., one or more) nucleoside bond of the oligonucleotide is a non-natural linkage.

[0007] Additionally disclosed herein are compounds comprising a splice-switching oligonucleotide comprising a sequence that is at least 85% complementary to an equal length portion of a sequence from a transcript regulated by TDP-43, wherein at least one (i.e., one or more) nucleoside linkage of the oligonucleotide is a non-natural linkage.

[0008] In various embodiments, the splice-switching oligonucleotide comprises a sequence that is at least 85% complementary to an equal length portion of the sequence from a transcript of any one of SEQ ID NO: 1339 or SEQ ID NO: 1341, SEQ ID NOs: 3032-3043, SEQ ID NOs: 9587-9595, or SEQ ID NOs: 9698-9707.

[0009] In various embodiments, the splice-switching oligonucleotide comprises a sequence that is at least 85% complementary to an equal length portion of any one of SEQ ID NO: 1339 or SEQ ID NO: 1341, SEQ ID NOs: 3032-3043, SEQ ID NOs: 9587-9595, or SEQ ID NOs: 9698-9707.

[0010] Additionally disclosed herein are compounds comprising a modified splice-switching oligonucleotide comprising a sequence that is at least 85% complementary to an equal length portion of a sequence from a transcript of any of STMN2, KCNQ2, UNC13A, or SMN2, wherein at least one (i.e., one or more) nucleoside linkage of the oligonucleotide is a non-natural linkage.

[0011] In various embodiments, the oligonucleotide comprises an equal length portion of SEQ ID NO: 1339 or SEQ ID NO: 1341, SEQ ID NOs: 3032-3043, SEQ ID NOs: 9587-9595, or SEQ ID NOs: 9698-9707, any of the sequences with 90% identity thereof, or a sequence that is at least 85% complementary to a 15 to 50 contiguous nucleobase portion thereof.

[0012] Additionally disclosed herein are compounds comprising a splice-switching oligonucleotide, wherein the splice-switching oligonucleotide comprises a spacer.

[0013] In various embodiments, the splice-switching oligonucleotide comprises a sequence that is at least 85% complementary to an equal length portion of a sequence from a transcript whose mis-splicing results in a neurological disease, or whose splicing can be modulated to treat a neurological disease.

[0014] In various embodiments, the splice-switching oligonucleotide comprises a sequence that is at least 85% complementary to an equal length portion of a sequence from a transcript regulated by TDP-43, wherein at least one (i.e., one or more) nucleoside linkage of the oligonucleotide is a non-natural linkage, and further wherein the oligonucleotide comprises a spacer.

[0015] In various embodiments, the oligonucleotide comprises a sequence that is at least 85% complementary to an equal length portion of a sequence from a transcript of any of STMN2, KCNQ2, UNC13A, or SMN2. In various embodiments, the modified oligonucleotide comprises a sequence that is at least 85% complementary to an equal length portion of any of SEQ ID NO: 1339, or SEQ ID NO: 1341, SEQ ID NOs: 3032-3043, SEQ ID NOs: 9587-9595, or SEQ ID NOs: 9698-9707.

[0016] Additionally disclosed herein are compounds comprising a modified oligonucleotide comprising a sequence that is at least 85% complementary to an equal length portion of a sequence from a transcript of any of STMN2, KCNQ2, UNC13A, or SMN2, wherein at least one (i.e., one or more) nucleoside linkage of the oligonucleotide is a non-natural linkage, and further wherein the oligonucleotide comprises a spacer.

[0017] In various embodiments, the modified oligonucleotide comprises an equal length portion of SEQ ID NO: 1339 or SEQ ID NO: 1341, SEQ ID NOs: 3032-3043, SEQ ID NOs: 9587-9595, or SEQ ID NOs: 9698-9707, any of the sequences with 90% identity thereof, or a sequence that is at least 85% complementary to a 15 to 50 contiguous nucleobase portion thereof.

[0018] Additionally disclosed herein are splice-switching oligonucleotides comprising a sequence that is at least 85% complementary to an equal length portion of a sequence from a transcript whose mis-splicing results in a neurological disease, or whose splicing can be modulated to treat a neurological disease, wherein at least one (i.e., one or more) nucleoside bond of the oligonucleotide is a non-natural linkage.

[0019] Additionally disclosed herein are splice-switching oligonucleotides comprising a sequence that is at least 85% complementary to an equal length portion of a sequence from a transcript regulated by TDP-43, wherein at least one (ie, one or more) nucleoside linkage of the oligonucleotide is a non-natural linkage.

[0020] In various embodiments, the oligonucleotide comprises a sequence that is at least 85% complementary to an equal length portion of a sequence from a transcript of any of STMN2, KCNQ2, UNC13A, or SMN2. In various embodiments, the modified oligonucleotide comprises a sequence that is at least 85% complementary to an equal length portion of any of SEQ ID NO: 1339, or SEQ ID NO: 1341, SEQ ID NOs: 3032-3043, SEQ ID NOs: 9587-9595, or SEQ ID NOs: 9698-9707.

[0021] Additionally disclosed herein are splice-switching oligonucleotides comprising a sequence that is at least 85% complementary to an equal length portion of a sequence from a transcript of any of STMN2, KCNQ2, UNC13A, or SMN2, wherein at least one (i.e., one or more) nucleoside linkage of the oligonucleotide is a non-natural linkage.

[0022] In various embodiments, the oligonucleotide comprises a sequence that is at least 85% complementary to an equal length portion of any one of SEQ ID NO: 1339 or SEQ ID NO: 1341, SEQ ID NOs: 3032-3043, SEQ ID NOs: 9587-9595, or SEQ ID NOs: 9698-9707.

[0023] Additionally disclosed herein are splice-switching oligonucleotides comprising a sequence that is at least 85% complementary to an equal length portion of any one of SEQ ID NO: 1339 or SEQ ID NO: 1341, SEQ ID NOs: 3032-3043, SEQ ID NOs: 9587-9595, or SEQ ID NOs: 9698-9707, wherein at least one (i.e., one or more) nucleoside linkage of the oligonucleotide is a non-natural linkage. In various embodiments, the oligonucleotide further comprises a spacer.

[0024] In various embodiments, the oligonucleotide comprises a sequence that is at least 85% complementary to an equal length portion of a sequence from a transcript whose mis-splicing results in a neurological disease, or whose splicing can be modulated to treat a neurological disease.

[0025] In various embodiments, the oligonucleotide comprises a sequence that is at least 85% complementary to an equal length portion of a sequence from a transcript regulated by TDP-43, wherein at least one (i.e., one or more) nucleoside linkage of the oligonucleotide is a non-natural linkage, and further wherein the oligonucleotide comprises a spacer.

[0026] In various embodiments, the oligonucleotide comprises a sequence that is at least 85% complementary to an equal length portion of a sequence from a transcript of any of STMN2, KCNQ2, UNC13A, or SMN2. In various embodiments, the modified oligonucleotide comprises a sequence that is at least 85% complementary to an equal length portion of any of SEQ ID NO: 1339, or SEQ ID NO: 1341, SEQ ID NOs: 3032-3043, SEQ ID NOs: 9587-9595, or SEQ ID NOs: 9698-9707.

[0027] Additionally disclosed herein are splice-switching oligonucleotides comprising a sequence that is at least 85% complementary to an equal length portion of a sequence from a transcript of any of STMN2, KCNQ2, UNC13A, or SMN2, wherein at least one (i.e., one or more) nucleoside linkages of the oligonucleotide are non-natural linkages, and further wherein the oligonucleotide comprises a spacer.

[0028] In various embodiments, the oligonucleotide comprises a sequence that is at least 85% complementary to an equal length portion of any one of SEQ ID NO: 1339 or SEQ ID NO: 1341, SEQ ID NOs: 3032-3043, SEQ ID NOs: 9587-9595, or SEQ ID NOs: 9698-9707.

[0029] Additionally disclosed herein are oligonucleotides comprising a sequence that is at least 85% complementary to an equal length portion of any one of SEQ ID NO: 1339 or SEQ ID NO: 1341, SEQ ID NOs: 3032-3043, SEQ ID NOs: 9587-9595, or SEQ ID NOs: 9698-9707, a sequence with 90% identity thereof, or a 15 to 50 contiguous nucleobase portion thereof, wherein at least one (i.e., one or more) nucleoside linkage of the oligonucleotide is a non-natural linkage, and further wherein the oligonucleotide comprises a spacer.

[0030] In various embodiments, the oligonucleotide comprises a segment having at most 11 connected nucleosides. In various embodiments, the oligonucleotide comprises a segment having at most 10, 9 or 8 connected nucleosides. In various embodiments, the oligonucleotide comprises a segment having at most 7 connected nucleosides. In various embodiments, the oligonucleotide comprises a segment having at most 6, 5, 4, 3 or 2 connected nucleosides. In various embodiments, each segment of the oligonucleotide comprises at most 7 connected nucleosides.

[0031] In various embodiments, the oligonucleotide comprises the oligonucleotide sequence of SEQ ID NO: 1-466, SEQ ID NO: 893-1338, SEQ ID NO: 1342-1366, SEQ ID NO: 1392-1664, or SEQ ID NO: 10655-10669, SEQ ID NO: 1676-1851, SEQ ID NO: 2028-2529, SEQ ID NO: 3046-3221, SEQ ID NO: 3398-3899, and SEQ ID NO: 4402-4530, SEQ ID NO: 4531-5794, SEQ ID NO: 7059-8322, SEQ ID NO: 9596-9603, SEQ ID NO: 9710-10141, SEQ ID NO: 10574-10651, SEQ ID NO: 10670-10779, SEQ ID NO: Equal length portions of any one of SEQ ID NOs: 10783-10808 and SEQ ID NOs: 10814-10820 share a sequence that is at least 85% identical.

[0032] In various embodiments, the oligonucleotide comprises the sequences of SEQ ID NOs: 1-466, SEQ ID NOs: 893-1338, SEQ ID NOs: 1342-1366, SEQ ID NOs: 1392-1664, or SEQ ID NOs: 10655-10669, SEQ ID NOs: 1676-1851, SEQ ID NOs: 2028-2529, SEQ ID NOs: 3046-3221, SEQ ID NOs: 3398-3899, and SEQ ID NOs: 4402-4530, SEQ ID NOs: 4531-5794, SEQ ID NOs: 7059-8322, SEQ ID NOs: 9596-9603, SEQ ID NOs: 9710-10141, SEQ ID NOs: 10574-10651, SEQ ID NOs: 10670-10779, SEQ ID NOs: Equal length portions of any one of SEQ ID NOs: 10783-10808 and SEQ ID NOs: 10814-10820 share a sequence that is at least 90% identical.

[0033] In various embodiments, the oligonucleotide comprises the sequences of SEQ ID NOs: 1-466, SEQ ID NOs: 893-1338, SEQ ID NOs: 1342-1366, SEQ ID NOs: 1392-1664, or SEQ ID NOs: 10655-10669, SEQ ID NOs: 1676-1851, SEQ ID NOs: 2028-2529, SEQ ID NOs: 3046-3221, SEQ ID NOs: 3398-3899, and SEQ ID NOs: 4402-4530, SEQ ID NOs: 4531-5794, SEQ ID NOs: 7059-8322, SEQ ID NOs: 9596-9603, SEQ ID NOs: 9710-10141, SEQ ID NOs: 10574-10651, SEQ ID NOs: 10670-10779, SEQ ID NOs: Equal length portions of any one of SEQ ID NOs: 10783-10808 and SEQ ID NOs: 10814-10820 share a sequence that is at least 95% identical.

[0034] In various embodiments, the oligonucleotide comprises the sequences of SEQ ID NOs: 1-466, SEQ ID NOs: 893-1338, SEQ ID NOs: 1342-1366, SEQ ID NOs: 1392-1664, or SEQ ID NOs: 10655-10669, SEQ ID NOs: 1676-1851, SEQ ID NOs: 2028-2529, SEQ ID NOs: 3046-3221, SEQ ID NOs: 3398-3899, and SEQ ID NOs: 4402-4530, SEQ ID NOs: 4531-5794, SEQ ID NOs: 7059-8322, SEQ ID NOs: 9596-9603, SEQ ID NOs: 9710-10141, SEQ ID NOs: 10574-10651, SEQ ID NOs: 10670-10779, SEQ ID NOs: Equal length portions of any one of SEQ ID NOs: 10783-10808 and SEQ ID NOs: 10814-10820 share a sequence with 100% identity.

[0035] In various embodiments, the oligonucleotide comprises a segment having up to 11 linked nucleosides or up to 7 linked nucleosides, and wherein the oligonucleotide comprises a segment corresponding to SEQ ID NO: 1-466, SEQ ID NO: 893-1338, SEQ ID NO: 1342-1366, SEQ ID NO: 1392-1664, or SEQ ID NO: 10655-10669, SEQ ID NO: 1676-1851, SEQ ID NO: 2028-2529, SEQ ID NO: 3046-3221, SEQ ID NO: 3398-3899, and SEQ ID NO: 4402-4530, SEQ ID NO: 4531-5794, SEQ ID NO: 7059-8322, SEQ ID NO: 9596-9603, SEQ ID NO: 9710-10141, SEQ ID NO: 10574-10651, SEQ ID Equal length portions of any of SEQ ID NOs: 10670-10779, SEQ ID NOs: 10783-10808, and SEQ ID NOs: 10814-10820 share sequences that are at least 85% identical. In various embodiments, the oligonucleotide comprises a fragment having up to 6, 5, 4, 3, or 2 linked nucleosides, wherein the oligonucleotide comprises a fragment having up to 6, 5, 4, 3, or 2 linked nucleosides, wherein the oligonucleotide comprises a fragment having up to 6, 5, 4, 3, or 2 linked nucleosides, wherein the oligonucleotide comprises a fragment having up to 6, 5, 4, 3, or 2 linked nucleosides, wherein the oligonucleotide comprises a fragment having up to 6, 5, 4, 3, or 2 linked nucleosides, wherein the oligonucleotide comprises a fragment having up to 6, 5, 4, 3, or 2 linked nucleosides, wherein the oligonucleotide comprises a fragment having up to 6, 5, 4, 3, or 2 linked nucleosides, wherein the oligonucleotide comprises a fragment having up to 6, 5, 4, 3, or 2 linked nucleosides, wherein the oligonucleotide comprises a fragment having up to 6, 5, 4, 3, or 2 linked nucleosides, wherein the oligonucleotide comprises a fragment having up to 6, 5, 4, 3, or 2 linked nucleosides, wherein the oligonucleotide comprises a fragment having up to 6, 5, 4, 3, or 2 linked nucleosides, wherein the oligonucleotide comprises a fragment having up to 6, 5, 4, 3, or 2 linked nucleosides, wherein the oligonucleotide comprises a fragment having up to 6, 5, 4, 3, or 2 linked nucleosides, wherein the oligonucleotide comprises a fragment having up to 6, 5, 4, 3, or 2 linked nucleosides, wherein the oligonucleotide comprises a fragment having up to 6, Equal length portions of any of SEQ ID NOs: 10670-10779, SEQ ID NOs: 10783-10808, and SEQ ID NOs: 10814-10820 share sequences that are at least 85% identical.In various embodiments, the oligonucleotide comprises a fragment having up to 6, 5, 4, 3, or 2 linked nucleosides, wherein the oligonucleotide comprises a fragment having up to 6, 5, 4, 3, or 2 linked nucleosides, wherein the oligonucleotide comprises a fragment having up to 6, 5, 4, 3, or 2 linked nucleosides, wherein the oligonucleotide comprises a fragment having up to 6, 5, 4, 3, or 2 linked nucleosides, wherein the oligonucleotide comprises a fragment having up to 6, 5, 4, 3, or 2 linked nucleosides, wherein the oligonucleotide comprises a fragment having up to 6, 5, 4, 3, or 2 linked nucleosides, wherein the oligonucleotide comprises a fragment having up to 6, 5, 4, 3, or 2 linked nucleosides, wherein the oligonucleotide comprises a fragment having up to 6, 5, 4, 3, or 2 linked nucleosides, wherein the oligonucleotide comprises a fragment having up to 6, 5, 4, 3, or 2 linked nucleosides, wherein the oligonucleotide comprises a fragment having up to 6, 5, 4, 3, or 2 linked nucleosides, wherein the oligonucleotide comprises a fragment having up to 6, 5, 4, 3, or 2 linked nucleosides, wherein the oligonucleotide comprises a fragment having up to 6, 5, 4, 3, or 2 linked nucleosides, wherein the oligonucleotide comprises a fragment having up to 6, 5, 4, 3, or 2 linked nucleosides, wherein the oligonucleotide comprises a fragment having up to 6, 5, 4, 3, or 2 linked nucleosides, wherein the oligonucleotide comprises a fragment having up to 6, Equal length portions of any of SEQ ID NOs: 10670-10779, SEQ ID NOs: 10783-10808, and SEQ ID NOs: 10814-10820 share sequences that are at least 90% identical.

[0036] In various embodiments, the oligonucleotide is at least 18, at least 19, at least 20, at least 21, at least 22, at least 23, at least 24, or at least 25 oligonucleotide units in length. In various embodiments, the oligonucleotide is at least 19 oligonucleotide units in length.

[0037] In various embodiments, the spacer is a nucleoside replacement group comprising a non-sugar substituent that is incapable of attaching to a nucleotide base. In various embodiments, the spacer is located between positions 10 and 15 of the oligonucleotide. In various embodiments, the spacer is located between positions 7 and 11 of the oligonucleotide.

[0038] In various embodiments, the oligonucleotide further comprises a second spacer, wherein the second spacer is located between positions 14 and 22 of the oligonucleotide. In various embodiments, the spacer and the second spacer are separated by at least 5 core bases, at least 6 core bases, or at least 7 core bases in the oligonucleotide. In various embodiments, the spacer is located between positions 7 and 9 of the oligonucleotide, and wherein the second spacer is located between positions 15 and 18 of the oligonucleotide. In various embodiments, the spacer is located at position 8 of the oligonucleotide, and wherein the second spacer is located at position 16 of the oligonucleotide.

[0039] In various embodiments, the oligonucleotide further comprises a third spacer, wherein the third spacer is located between positions 21 and 24 of the oligonucleotide. In various embodiments, the spacer is located between positions 2 and 5 of the oligonucleotide.

[0040] In various embodiments, the oligonucleotide further comprises a second spacer, wherein the second spacer is located between positions 8 and 12 of the oligonucleotide. In various embodiments, the oligonucleotide further comprises a third spacer, wherein the third spacer is located between positions 18 and 22 of the oligonucleotide. In various embodiments, the oligonucleotide further comprises a second spacer and a third spacer, wherein the three spacers are located at positions in the oligonucleotide such that each segment of the oligonucleotide has at most 7 linked nucleosides.

[0041] In various embodiments, at least two of the three spacers are adjacent to a guanine nucleobase. In various embodiments, each of at least two of the three spacers is immediately preceding a guanine nucleobase. In various embodiments, each of the first, second, or third spacers is a nucleoside replacement group comprising a non-sugar substituent, wherein the non-sugar substituent does not contain a ketone, aldehyde, ketal, hemiketal, acetal, hemiacetal, aminal, or hemiaminal moiety and is not capable of forming a covalent bond with a nucleotide base.

[0042] In various embodiments, each of the first, second, or third spacers is independently represented by formula (X), wherein:

[0043]

[0044] Ring A is an optionally substituted 4-8 membered monocyclic cycloalkyl or 4-8 membered monocyclic heterocyclyl, wherein the heterocyclyl contains 1 or 2 heteroatoms selected from O, S and N, with the proviso that A is not capable of forming a covalent bond with a nucleobase; and The symbol represents the point of attachment of the internucleoside linkage.

[0045] In various embodiments, each of the first, second, or third spacers is independently represented by formula (Xa), wherein:

[0046]

[0047] In various embodiments, Ring A is an optionally substituted 4-8 membered monocyclic cycloalkyl or 4-8 membered monocyclic heterocyclyl, wherein the 4-8 membered monocyclic cycloalkyl is selected from cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl; and the 4-8 membered monocyclic heterocyclyl is selected from oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, 1,4-dioxanyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl and azepanyl.

[0048] In various embodiments, Ring A is tetrahydrofuranyl.In various embodiments, Ring A is tetrahydropyranyl.

[0049] In various embodiments, each of the first, second, or third spacers is independently represented by Formula I, wherein:

[0050]

[0051] X is selected from -CH2- and -O-; and

[0052] n is 0, 1, 2 or 3.

[0053] In various embodiments, each of the first, second, or third spacers is independently represented by Formula I', wherein:

[0054]

[0055] X is selected from -CH2- and -O-; and

[0056] n is 0, 1, 2 or 3.

[0057] In various embodiments, each of the first, second, or third spacers is independently represented by Formula (Ia), wherein:

[0058] and

[0059] n is 0, 1, 2 or 3.

[0060] In various embodiments, each of the first, second, or third spacers is independently represented by Formula (Ia'), wherein:

[0061] and

[0062] n is 0, 1, 2 or 3.

[0063] In various embodiments, each of the first, second, or third spacers is independently represented by Formula II, wherein:

[0064] and

[0065] X is selected from -CH2- and -O-.

[0066] In various embodiments, each of the first, second, or third spacers is independently represented by Formula II', wherein:

[0067] and

[0068] X is selected from -CH2- and -O-.

[0069] In various embodiments, each of the first, second, or third spacers is independently represented by Formula (Iia), wherein:

[0070]

[0071] In various embodiments, each of the first, second, or third spacers is independently represented by Formula (Iia'), wherein:

[0072]

[0073] In some embodiments, the spacer is represented by formula (III), wherein:

[0074]

[0075] X is selected from -CH2- and -O-.

[0076] In some embodiments, the spacer is represented by formula (III'), wherein:

[0077]

[0078] X is selected from -CH2- and -O.

[0079] In some embodiments, the spacer is represented by formula (IIIb), wherein:

[0080]

[0081] In some embodiments, the spacer is represented by formula (Iiib'), wherein:

[0082]

[0083] In various embodiments, each of the first, second, or third spacers is independently represented by Formula III, wherein:

[0084] and

[0085] X is selected from -CH2- and -O-.

[0086] In various embodiments, each of the first, second, or third spacers is independently represented by Formula III', wherein:

[0087] and

[0088] X is selected from -CH2- and -O-.

[0089] In various embodiments, each of the first, second, or third spacers is independently represented by formula (IIIa), wherein:

[0090]

[0091] In various embodiments, each of the first, second, or third spacers is independently represented by formula (IIIa'), wherein:

[0092]

[0093] In various embodiments, the oligonucleotide comprising a spacer has a GC content of at least 10%. In various embodiments, the oligonucleotide comprising a spacer has a GC content of at least 20%. In various embodiments, the oligonucleotide comprising a spacer has a GC content of at least 25%. In various embodiments, the oligonucleotide comprising a spacer has a GC content of at least 30%. In various embodiments, the oligonucleotide comprising a spacer has a GC content of at least 40%. In various embodiments, the oligonucleotide comprising a spacer has a GC content of at least 50%. In various embodiments, the length of the oligonucleotide is between 12 and 40 oligonucleotide units.

[0094] In various embodiments, at least one (i.e., one or more) nucleoside linkages of the oligonucleotide are independently selected from the group consisting of a phosphodiester linkage, a phosphorothioate linkage, an alkylphosphate linkage, a phosphorodithioate linkage, a phosphotriester linkage, an alkylphosphonate linkage, a 3-methoxypropylphosphonate linkage, a methylphosphonate linkage, an aminoalkylphosphotriester linkage, an alkylenephosphonate linkage, a phosphinate linkage, a phosphoamidate linkage, a phosphorothioateamidate linkage, a phosphorodiamidate linkage, a phosphorodiamidate linkage, an aminoalkylphosphoamidate linkage, a phosphorothioateamidate linkage, a thioalkylphosphonate linkage, a thioalkylphosphotriester linkage, a thiophosphate linkage, a phosphoroselenoate linkage, and a boronic acid phosphate linkage.

[0095] In various embodiments, one or more nucleoside linkages of the base at position 3 or position 4 of the oligonucleotide are phosphodiester linkages. In various embodiments, only one nucleoside linkage of the base at position 3 or position 4 of the oligonucleotide is phosphodiester linkage. In various embodiments, the nucleoside linkages of the base at both position 3 and position 4 of the oligonucleotide are phosphodiester linkages. In various embodiments, the one or more bases immediately before the spacer in the oligonucleotide are connected by phosphodiester bonds. In various embodiments, only the base immediately before the spacer in the oligonucleotide is connected to the spacer by a phosphodiester bond. In various embodiments, the base immediately before the spacer in the oligonucleotide is further connected to the further preceding base by a phosphodiester bond.

[0096] In various embodiments, the oligonucleotide comprises a second spacer, wherein the base immediately before the second spacer is connected to the further preceding base by a phosphodiester bond. In various embodiments, the one or more bases immediately after the spacer in the oligonucleotide are connected by a phosphodiester bond. In various embodiments, only the base immediately after the spacer in the oligonucleotide is connected to the spacer by a phosphodiester bond. In various embodiments, the two bases immediately before the spacer in the oligonucleotide are connected by a phosphodiester bond.

[0097] In various embodiments, the one or more bases immediately before the spacer in the oligonucleotide are linked by a phosphodiester bond, and wherein the one or more bases immediately after the spacer in the oligonucleotide are linked by a phosphodiester bond. In various embodiments, a base immediately before the spacer and a base immediately after the spacer are linked by a phosphodiester bond.

[0098] In various embodiments, the oligonucleotide comprises a second spacer, and wherein the one or more bases immediately preceding the second spacer in the oligonucleotide are linked by a phosphodiester bond, and wherein the one or more bases immediately following the second spacer in the oligonucleotide are linked by a phosphodiester bond. In various embodiments, a base immediately preceding the second spacer and a base immediately following the second spacer are linked by a phosphodiester bond.

[0099] In various embodiments, the oligonucleotide comprises a series of bases connected by a phosphodiester bond, and the series of bases comprises at least two bases. In various embodiments, the oligonucleotide comprises a series of bases connected by a phosphodiester bond, and the series of bases comprises at least five bases. In various embodiments, the oligonucleotide comprises two or more spacers, and wherein the series of bases is located between at least two spacers.

[0100] In various embodiments, one or more internucleoside linkages of an oligonucleotide are modified internucleoside linkages. In various embodiments, the modified internucleoside linkages of an oligonucleotide are phosphorothioate linkages. In various embodiments, all internucleoside linkages of an oligonucleotide are phosphorothioate linkages. In various embodiments, the phosphorothioate linkages are in one of the Rp configuration or the Sp configuration.

[0101] In various embodiments, the oligonucleotide comprises at least one modified sugar moiety. In various embodiments, the modified sugar moiety is one of: a 2'-OMe modified sugar moiety, a bicyclic sugar moiety, 2'-O-(2-methoxyethyl) (2'-MOE), a 2'-deoxy-2'-fluoro nucleoside, a 2'-fluoro-β-D-arabinoside, a locked nucleic acid (LNA), a constrained ethyl 2'-4'-bridged nucleic acid (cEt), S-cEt, tcDNA, a hexitol nucleic acid (HNA), and a tricyclic analog (e.g., tcDNA).

[0102] In various embodiments, the oligonucleotides exhibit at least a 30%, 40%, 50%, 60%, 70%, 80%, or 90% increase in the full-length STMN2, KCNQ2, UNC13A, or SMN2 protein. In various embodiments, the oligonucleotides exhibit at least a 100% increase in the full-length protein. In various embodiments, the oligonucleotides exhibit at least a 200% increase in the full-length protein. In various embodiments, the oligonucleotides exhibit at least a 300% increase in the full-length protein. In various embodiments, the oligonucleotides exhibit at least a 400% increase in the full-length protein.

[0103] In various embodiments, the increase in full-length protein is measured compared to the level of reduction in full-length protein achieved using a TDP43 antisense oligonucleotide. In various embodiments, the oligonucleotide demonstrates at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% rescue of the full-length protein. In various embodiments, the oligonucleotide demonstrates at least 50%, 60%, 70%, 80%, or 90% reduction in mis-spliced ​​transcripts.

[0104] Additionally disclosed herein are methods of treating a neurological disease and / or neurological disorder in a patient in need thereof, the methods comprising administering to the patient a compound or oligonucleotide disclosed herein.

[0105] In various embodiments, the neurological disease is selected from the group consisting of amyotrophic lateral sclerosis (ALS), frontotemporal dementia (FTD), ALS with FTD, Alzheimer's disease (AD), Parkinson's disease (PD), Parkinson's disease with dementia, dementia with Lewy bodies, synucleinopathy, Huntington's disease, brachial plexus injury, peripheral nerve injury, progressive supranuclear palsy (PSP), brain trauma, spinal cord injury, tuberous sclerosis complex, Pick's disease, tauopathy, primary age-related tauopathy, Down syndrome, epilepsy / seizure disorder, depression, traumatic brain injury (TBI), chronic traumatic encephalopathy (CTE), HIV-associated neurocognitive disorder (HAND), multiple system atrophy, amnestic mild cognitive impairment, Corticobasal degeneration (CBD) and / or neuropathies such as chemotherapy-induced neuropathy, spinocerebellar ataxia (SCA), type 2 SCA, spinal muscular atrophy (SMA), Parkinson's disease, Niemann-Pick disease type C (NPC), Charcot-Marie-Tooth disease (CMT), mucopolysaccharidosis type II (MPSIIA), mucolipidosis IV, GM1 gangliosidosis, sporadic inclusion body myositis (sIBM), Kennock-Scholin purpura (HSP), limbic predominant age-related TDP-43 encephalopathy (late stage), brain age-related TDP-43 with sclerosis (CARTS), Gaucher disease and facial onset sensory and motor neuron disease, Guam Parkinson-dementia complex, multisystem proteinopathy, Perry disease and synaptic diseases such as autism. In various embodiments, the neurological disease is ALS. In various embodiments, the neurological disease is FTD. In various embodiments, the neurological disease is ALS with FTD. In various embodiments, the neurological disease is AD. In various embodiments, the neurological disease is PD. In various embodiments, the neurological disease is spinal muscular atrophy (SMA). In various embodiments, the neuropathy is chemotherapy-induced neuropathy.

[0106] Additionally disclosed herein are methods of restoring axonal growth and / or regeneration of neurons, the methods comprising exposing the neurons to a compound or oligonucleotide disclosed herein.

[0107] Additionally disclosed herein are methods for increasing, promoting, stabilizing, or maintaining the expression and / or function of any one of STMN2, KCNQ2, UNC13A, or SMN2 in neurons, the methods comprising exposing cells to a compound or oligonucleotide disclosed herein. In various embodiments, the neurons are motor neurons. In various embodiments, the neurons are spinal neurons. In various embodiments, the neurons are neurons in a patient in need of treatment for a neurological disease and / or neuropathy. In various embodiments, the neuropathy is a chemotherapy-induced neuropathy. In various embodiments, the exposure is performed in vivo or ex vivo. In various embodiments, the exposure comprises administering an oligonucleotide to a patient in need thereof.

[0108] In various embodiments, the oligonucleotide is administered topically, parenterally, intrathecally, intrathalamically, intracisternalally, orally, rectally, buccally, sublingually, vaginally, pulmonary, intratracheally, intranasally, transdermally, intraduodenally, or intracerebroventricularly. In various embodiments, the oligonucleotide is administered orally. In various embodiments, a therapeutically effective amount of the oligonucleotide is administered intrathecally, intrathalamically, intracerebroventricularly, or intracisternalally. In various embodiments, the patient is a human.

[0109] Additionally disclosed herein are pharmaceutical compositions comprising an oligonucleotide disclosed herein, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0110] In various embodiments, the pharmaceutical composition is suitable for topical, intrathecal, intrathalamic, intracisternal, intracerebroventricular, parenteral, oral, pulmonary, intratracheal, intranasal, transdermal, rectal, buccal, sublingual, vaginal, or intraduodenal administration.

[0111] Additionally disclosed herein are methods of treating a neurological disease or neuropathy in a patient in need thereof, the methods comprising administering to the patient in need thereof a therapeutically effective amount of a pharmaceutical composition disclosed herein.

[0112] In various embodiments, the neurological disease is selected from the group consisting of amyotrophic lateral sclerosis (ALS), frontotemporal dementia (FTD), ALS with FTD, Alzheimer's disease (AD), Parkinson's disease (PD), Parkinson's disease with dementia, dementia with Lewy bodies, synucleinopathy, Huntington's disease, brachial plexus injury, peripheral nerve injury, progressive supranuclear palsy (PSP), brain trauma, spinal cord injury, tuberous sclerosis complex, Pick's disease, tauopathy, primary age-related tauopathy, Down syndrome, epilepsy / seizure disorder, depression, traumatic brain injury (TBI), chronic traumatic encephalopathy (CTE), HIV-associated neurocognitive disorder (HAND), multiple system atrophy, amnestic mild cognitive impairment, Corticobasal degeneration (CBD) and / or neuropathies such as chemotherapy-induced neuropathy, spinocerebellar ataxia (SCA), SCA type 2, spinal muscular atrophy (SMA), Parkinson's disease, Niemann-Pick disease type C (NPC), Charcot-Marie-Tooth disease (CMT), mucopolysaccharidosis type II (MPSIIA), mucolipidosis IV, GM1 gangliosidosis, sporadic inclusion body myositis (sIBM), Kennock-Scholin purpura (HSP), limbic-predominant age-related TDP-43 encephalopathy (late stage), cerebral age-related TDP-43 with sclerosis (CARTS), Gaucher disease and facial-onset sensory and motor neuronopathy, Guam Parkinsonism-dementia complex, multisystem proteinopathy, Perry disease, and synaptic disorders such as autism.

[0113] In various embodiments, the neurological disease is ALS. In various embodiments, the neurological disease is FTD. In various embodiments, the neurological disease is ALS with FTD. In various embodiments, the neurological disease is AD. In various embodiments, the neurological disease is PD. In various embodiments, the neurological disease is spinal muscular atrophy (SMA). In various embodiments, the neuropathy is chemotherapy-induced neuropathy.

[0114] In various embodiments, the pharmaceutical composition is administered topically, parenterally, orally, pulmonary, rectal, buccal, sublingual, vaginally, intratracheally, intranasally, intracisternal, intrathecal, intrathalamic, transdermal, intraduodenally, or intracerebroventricularly. In various embodiments, the pharmaceutical composition is administered intrathecally, intrathalamically, or intracisternal. In various embodiments, a therapeutically effective amount of the oligonucleotide is administered intrathecally, intrathalamically, or intracisternal. In various embodiments, the patient is a human.

[0115] Additionally disclosed herein are methods for treating a neurological disease in a subject in need thereof, the method comprising administering to the subject an oligonucleotide, or a pharmaceutically acceptable salt thereof, the oligonucleotide comprising a segment of up to 7 linked nucleosides, and wherein the oligonucleotide is at least 85% complementary to an equal length portion of a sequence from a transcript whose mis-splicing results in a neurological disease, or a transcript whose splicing can be modulated to treat a neurological disease;

[0116] wherein at least one (i.e., one or more) nucleoside linkages of the oligonucleotide are independently selected from the group consisting of a phosphodiester linkage, a phosphorothioate linkage, an alkylphosphate linkage, a phosphorodithioate linkage, a phosphotriester linkage, an alkylphosphonate linkage, a 3-methoxypropylphosphonate linkage, a methylphosphonate linkage, an aminoalkylphosphotriester linkage, an alkylenephosphonate linkage, a phosphinate linkage, a phosphoamidate linkage, a phosphorothioateamidate linkage, a phosphorodiamidate linkage, a phosphorodiamidate linkage, an aminoalkylphosphoamidate linkage, a phosphorothioateamidate linkage, a thioalkylphosphonate linkage, a thioalkylphosphotriester linkage, a phosphorothioate linkage, a phosphoroselenoate linkage, and a phosphoroboroate linkage, and / or

[0117] wherein at least one (i.e., one or more) nucleosides are substituted with a member selected from the group consisting of 2'-O-(2-methoxyethyl) nucleosides, 2'-O-methyl nucleosides, 2'-deoxy-2'-fluoro nucleosides, 2'-fluoro-β-D-arabinoside, locked nucleic acids (LNA), tricyclic nucleic acids, constrained methoxyethyl (cMOE), constrained ethyl (cET), and peptide nucleic acids (PNA)

[0118] Optionally, wherein the oligonucleotide further comprises a spacer.

[0119] Additionally disclosed herein are methods for treating a neurological disease in a subject in need thereof, the method comprising administering to the subject an oligonucleotide, or a pharmaceutically acceptable salt thereof, the oligonucleotide comprising a segment of up to 7 linked nucleosides, and wherein the oligonucleotide is at least 85% complementary to a sequence from a transcript of any of STMN2, KCNQ2, UNC13A, or SMN2;

[0120] wherein at least one (i.e., one or more) nucleoside linkages of the oligonucleotide are independently selected from the group consisting of a phosphodiester linkage, a phosphorothioate linkage, an alkylphosphate linkage, a phosphorodithioate linkage, a phosphotriester linkage, an alkylphosphonate linkage, a 3-methoxypropylphosphonate linkage, a methylphosphonate linkage, an aminoalkylphosphotriester linkage, an alkylenephosphonate linkage, a phosphinate linkage, a phosphoamidate linkage, a phosphorothioateamidate linkage, a phosphorodiamidate linkage, a phosphorodiamidate linkage, an aminoalkylphosphoamidate linkage, a phosphorothioateamidate linkage, a thioalkylphosphonate linkage, a thioalkylphosphotriester linkage, a phosphorothioate linkage, a phosphoroselenoate linkage, and a phosphoroboroate linkage, and / or

[0121] wherein at least one (i.e., one or more) nucleosides are substituted with a member selected from the group consisting of 2'-O-(2-methoxyethyl) nucleosides, 2'-O-methyl nucleosides, 2'-deoxy-2'-fluoro nucleosides, 2'-fluoro-β-D-arabinoside, locked nucleic acids (LNA), tricyclic nucleic acids, constrained methoxyethyl (cMOE), constrained ethyl (cET), and peptide nucleic acids (PNA)

[0122] Optionally, wherein the oligonucleotide further comprises a spacer.

[0123] Additionally disclosed herein are methods for treating a neurological disease in a subject in need thereof, the method comprising administering to the subject an oligonucleotide or a pharmaceutically acceptable salt thereof, the oligonucleotide comprising a segment having up to 7 linked nucleosides, and wherein the oligonucleotide is conjugated to a nucleic acid sequence of SEQ ID NOs: 1-466, SEQ ID NOs: 893-1338, SEQ ID NOs: 1342-1366, SEQ ID NOs: 1392-1664, or SEQ ID NOs: 10655-10669, SEQ ID NOs: 1676-1851, SEQ ID NOs: 2028-2529, SEQ ID NOs: 3046-3221, SEQ ID NOs: 3398-3899, and SEQ ID NOs: 4402-4530, SEQ ID NOs: 4531-5794, SEQ ID NOs: 7059-8322, SEQ ID NOs: 9596-9603, SEQ ID NOs: 9710-10141, SEQ ID NOs: any of SEQ ID NOs: 10574-10651, SEQ ID NOs: 10670-10779, SEQ ID NOs: 10783-10808, and SEQ ID NOs: 10814-10820 share at least 85% identity;

[0124] wherein at least one (i.e., one or more) nucleoside linkages of the oligonucleotide are independently selected from the group consisting of a phosphodiester linkage, a phosphorothioate linkage, an alkylphosphate linkage, a phosphorodithioate linkage, a phosphotriester linkage, an alkylphosphonate linkage, a 3-methoxypropylphosphonate linkage, a methylphosphonate linkage, an aminoalkylphosphotriester linkage, an alkylenephosphonate linkage, a phosphinate linkage, a phosphoamidate linkage, a phosphorothioateamidate linkage, a phosphorodiamidate linkage, a phosphorodiamidate linkage, an aminoalkylphosphoamidate linkage, a phosphorothioateamidate linkage, a thioalkylphosphonate linkage, a thioalkylphosphotriester linkage, a phosphorothioate linkage, a phosphoroselenoate linkage, and a phosphoroboroate linkage, and / or

[0125] wherein at least one (i.e., one or more) nucleosides are substituted with a member selected from the group consisting of 2'-O-(2-methoxyethyl) nucleosides, 2'-O-methyl nucleosides, 2'-deoxy-2'-fluoro nucleosides, 2'-fluoro-β-D-arabinoside, locked nucleic acids (LNA), tricyclic nucleic acids, constrained methoxyethyl (cMOE), constrained ethyl (cET), and peptide nucleic acids (PNA)

[0126] Optionally, wherein the oligonucleotide further comprises a spacer.

[0127] Additionally disclosed herein are methods for treating amyotrophic lateral sclerosis (ALS) in a subject in need thereof, the method comprising administering to the subject an oligonucleotide or a pharmaceutically acceptable salt thereof, the oligonucleotide comprising a segment having up to 7 linked nucleosides, and wherein the oligonucleotide is conjugated to SEQ ID NO: 1-466, SEQ ID NO: 893-1338, SEQ ID NO: 1342-1366, SEQ ID NO: 1392-1664, or SEQ ID NO: 10655-10669, SEQ ID NO: 1676-1851, SEQ ID NO: 2028-2529, SEQ ID NO: 3046-3221, SEQ ID NO: 3398-3899, and SEQ ID NO: 4402-4530, or SEQ ID NO: 4531-5794, SEQ ID NO: 7059-8322, or SEQ ID NO: 1 any of NO: 9596-9603 sharing at least 85% identity;

[0128] wherein at least one (i.e., one or more) nucleoside linkages of the oligonucleotide are independently selected from the group consisting of a phosphodiester linkage, a phosphorothioate linkage, an alkylphosphate linkage, a phosphorodithioate linkage, a phosphotriester linkage, an alkylphosphonate linkage, a 3-methoxypropylphosphonate linkage, a methylphosphonate linkage, an aminoalkylphosphotriester linkage, an alkylenephosphonate linkage, a phosphinate linkage, a phosphoamidate linkage, a phosphorothioateamidate linkage, a phosphorodiamidate linkage, a phosphorodiamidate linkage, an aminoalkylphosphoamidate linkage, a phosphorothioateamidate linkage, a thioalkylphosphonate linkage, a thioalkylphosphotriester linkage, a phosphorothioate linkage, a phosphoroselenoate linkage, and a phosphoroboroate linkage, and / or

[0129] wherein at least one (i.e., one or more) nucleosides are substituted with a member selected from the group consisting of 2'-O-(2-methoxyethyl) nucleosides, 2'-O-methyl nucleosides, 2'-deoxy-2'-fluoro nucleosides, 2'-fluoro-β-D-arabinoside, locked nucleic acids (LNA), tricyclic nucleic acids, constrained methoxyethyl (cMOE), constrained ethyl (cET), and peptide nucleic acids (PNA)

[0130] Optionally, wherein the oligonucleotide further comprises a spacer.

[0131] Additionally disclosed herein are methods for treating Alzheimer's disease (AD) with frontotemporal dementia (FTD) in a subject in need thereof, the method comprising administering to the subject an oligonucleotide or a pharmaceutically acceptable salt thereof, the oligonucleotide comprising a segment having up to 7 linked nucleosides, and wherein the oligonucleotide is conjugated to a nucleic acid sequence of SEQ ID NOs: 1-466, SEQ ID NOs: 893-1338, SEQ ID NOs: 1342-1366, SEQ ID NOs: 1392-1664, or SEQ ID NOs: 10655-10669, SEQ ID NOs: 1676-1851, SEQ ID NOs: 2028-2529, SEQ ID NOs: 3046-3221, SEQ ID NOs: 3398-3899, and SEQ ID NOs: 4402-4530, or SEQ ID NOs: 4531-5794, SEQ ID NOs: 7059-8322, or SEQ ID NOs: any of NO: 9596-9603 sharing at least 85% identity;

[0132] wherein at least one (i.e., one or more) nucleoside linkages of the oligonucleotide are independently selected from the group consisting of a phosphodiester linkage, a phosphorothioate linkage, an alkylphosphate linkage, a phosphorodithioate linkage, a phosphotriester linkage, an alkylphosphonate linkage, a 3-methoxypropylphosphonate linkage, a methylphosphonate linkage, an aminoalkylphosphotriester linkage, an alkylenephosphonate linkage, a phosphinate linkage, a phosphoamidate linkage, a phosphorothioateamidate linkage, a phosphorodiamidate linkage, a phosphorodiamidate linkage, an aminoalkylphosphoamidate linkage, a phosphorothioateamidate linkage, a thioalkylphosphonate linkage, a thioalkylphosphotriester linkage, a phosphorothioate linkage, a phosphoroselenoate linkage, and a phosphoroboroate linkage, and / or

[0133] wherein at least one (i.e., one or more) nucleosides are substituted with a component selected from the group consisting of: 2'-O-(2-methoxyethyl) nucleosides, 2'-O-methyl nucleosides, 2'-deoxy-2'-fluoro nucleosides, 2'-fluoro-β-D-arabinoside, locked nucleic acids (LNA), tricyclic nucleic acids, constrained methoxyethyl (cMOE), constrained ethyl (cET) and peptide nucleic acids (PNA), optionally wherein the oligonucleotide further comprises a spacer.

[0134] Additionally disclosed herein are methods for treating frontotemporal dementia (FTD) in a subject in need thereof, the method comprising administering to the subject an oligonucleotide, or a pharmaceutically acceptable salt thereof, the oligonucleotide comprising a segment of up to 7 linked nucleosides, and wherein the oligonucleotide shares at least 85% identity with any one of SEQ ID NOs: 1-466, SEQ ID NOs: 893-1338, SEQ ID NOs: 1342-1366, SEQ ID NOs: 1392-1664, or SEQ ID NOs: 10655-10669, SEQ ID NOs: 1676-1851, SEQ ID NOs: 2028-2529, SEQ ID NOs: 3046-3221, SEQ ID NOs: 3398-3899, and SEQ ID NOs: 4402-4530, or SEQ ID NOs: 4531-5794, SEQ ID NOs: 7059-8322, or SEQ ID NOs: 9596-9603;

[0135] wherein at least one (i.e., one or more) nucleoside linkages of the oligonucleotide are independently selected from the group consisting of a phosphodiester linkage, a phosphorothioate linkage, an alkylphosphate linkage, a phosphorodithioate linkage, a phosphotriester linkage, an alkylphosphonate linkage, a 3-methoxypropylphosphonate linkage, a methylphosphonate linkage, an aminoalkylphosphotriester linkage, an alkylenephosphonate linkage, a phosphinate linkage, a phosphoamidate linkage, a phosphorothioateamidate linkage, a phosphorodiamidate linkage, a phosphorodiamidate linkage, an aminoalkylphosphoamidate linkage, a phosphorothioateamidate linkage, a thioalkylphosphonate linkage, a thioalkylphosphotriester linkage, a phosphorothioate linkage, a phosphoroselenoate linkage, and a phosphoroboroate linkage, and / or

[0136] wherein at least one (i.e., one or more) nucleosides are substituted with a member selected from the group consisting of 2'-O-(2-methoxyethyl) nucleosides, 2'-O-methyl nucleosides, 2'-deoxy-2'-fluoro nucleosides, 2'-fluoro-β-D-arabinoside, locked nucleic acids (LNA), tricyclic nucleic acids, constrained methoxyethyl (cMOE), constrained ethyl (cET), and peptide nucleic acids (PNA)

[0137] Optionally, wherein the oligonucleotide further comprises a spacer.

[0138] Additionally disclosed herein are methods for treating amyotrophic lateral sclerosis (ALS) with frontotemporal dementia (FTD) in a subject in need thereof, the method comprising administering to the subject an oligonucleotide or a pharmaceutically acceptable salt thereof, the oligonucleotide comprising a segment having up to 7 linked nucleosides, and wherein the oligonucleotide is conjugated to a nucleic acid sequence of SEQ ID NO: 1-466, SEQ ID NO: 893-1338, SEQ ID NO: 1342-1366, SEQ ID NO: 1392-1664, or SEQ ID NO: 10655-10669, SEQ ID NO: 1676-1851, SEQ ID NO: 2028-2529, SEQ ID NO: 3046-3221, SEQ ID NO: 3398-3899, and SEQ ID NO: 4402-4530, or SEQ ID NO: 4531-5794, SEQ ID NO: 7059-8322, or SEQ ID NO: 1 any of NO: 9596-9603 sharing at least 85% identity;

[0139] wherein at least one (i.e., one or more) nucleoside linkages of the oligonucleotide are independently selected from the group consisting of a phosphodiester linkage, a phosphorothioate linkage, an alkylphosphate linkage, a phosphorodithioate linkage, a phosphotriester linkage, an alkylphosphonate linkage, a 3-methoxypropylphosphonate linkage, a methylphosphonate linkage, an aminoalkylphosphotriester linkage, an alkylenephosphonate linkage, a phosphinate linkage, a phosphoamidate linkage, a phosphorothioateamidate linkage, a phosphorodiamidate linkage, a phosphorodiamidate linkage, an aminoalkylphosphoamidate linkage, a phosphorothioateamidate linkage, a thioalkylphosphonate linkage, a thioalkylphosphotriester linkage, a phosphorothioate linkage, a phosphoroselenoate linkage, and a phosphoroboroate linkage, and / or

[0140] wherein at least one (i.e., one or more) nucleosides are substituted with a member selected from the group consisting of 2'-O-(2-methoxyethyl) nucleosides, 2'-O-methyl nucleosides, 2'-deoxy-2'-fluoro nucleosides, 2'-fluoro-β-D-arabinoside, locked nucleic acids (LNA), tricyclic nucleic acids, constrained methoxyethyl (cMOE), constrained ethyl (cET), and peptide nucleic acids (PNA)

[0141] Optionally, wherein the oligonucleotide further comprises a spacer.

[0142] In various embodiments, the nucleoside linkage of the base at position 3 or position 4 of the oligonucleotide is a phosphodiester linkage. In various embodiments, only one nucleoside linkage of the base at position 3 or position 4 of the oligonucleotide is a phosphodiester linkage. In various embodiments, the nucleoside linkage of the base at both position 3 and position 4 of the oligonucleotide is a phosphodiester linkage. In various embodiments, the one or more bases immediately preceding the spacer in the oligonucleotide are connected by a phosphodiester bond. In various embodiments, only the base immediately preceding the spacer in the oligonucleotide is connected to the spacer by a phosphodiester bond. In various embodiments, the base immediately preceding the spacer in the oligonucleotide is further connected to the further preceding base by a phosphodiester bond. In various embodiments, the oligonucleotide comprises a second spacer, wherein the base immediately preceding the second spacer is connected to the further preceding base by a phosphodiester bond. In various embodiments, the one or more bases immediately following the spacer in the oligonucleotide are connected by a phosphodiester bond. In various embodiments, only the base immediately preceding the spacer in the oligonucleotide is connected to the spacer by a phosphodiester bond. In various embodiments, the two bases immediately before the spacer in the oligonucleotide are connected by a phosphodiester bond. In various embodiments, the one or more bases immediately before the spacer in the oligonucleotide are connected by a phosphodiester bond, and wherein the one or more bases immediately after the spacer are connected by a phosphodiester bond in the oligonucleotide. In various embodiments, a base immediately before the spacer and a base immediately after the spacer are connected by a phosphodiester bond. In various embodiments, the oligonucleotide comprises a second spacer, and wherein the one or more bases immediately before the second spacer in the oligonucleotide are connected by a phosphodiester bond, and wherein the one or more bases immediately after the second spacer in the oligonucleotide are connected by a phosphodiester bond. In various embodiments, a base immediately before the second spacer and a base immediately after the second spacer are connected by a phosphodiester bond.

[0143] In various embodiments, the oligonucleotide comprises a series of bases connected by a phosphodiester bond, and the series of bases comprises at least two bases. In various embodiments, the oligonucleotide comprises a series of bases connected by a phosphodiester bond, and the series of bases comprises at least five bases. In various embodiments, the oligonucleotide comprises two or more spacers, and wherein the series of bases is located between at least two spacers.

[0144] In various embodiments, the oligonucleotide is any of a 19-mer, a 21-mer, a 23-mer, or a 25-mer.

[0145] In various embodiments, at least one (ie, one or more) internucleoside linkage of an oligonucleotide is a phosphorothioate linkage. In various embodiments, all internucleoside linkages of an oligonucleotide are phosphorothioate linkages.

[0146] Additionally disclosed herein are oligonucleotides and pharmaceutically acceptable excipients, the oligonucleotide comprising an equal length portion of SEQ ID NO: 1339 or SEQ ID NO: 1341, SEQ ID NOs: 3032-3043, SEQ ID NOs: 9587-9595, or SEQ ID NOs: 9698-9707, or any of the sequences with 90% identity thereof, or a sequence that is at least 85% complementary to a 15 to 50 contiguous nucleobase portion thereof, optionally wherein the oligonucleotide comprises a spacer, and wherein the oligonucleotide is capable of increasing, restoring, or stabilizing expression of one or more of STMN2, KCNQ2, or UNC13A mRNAs capable of translating a functional protein in a cell or human patient with an immune-mediated demyelinating disease, and wherein the level of increase, restoration, or stabilization of expression and / or activity and / or function is sufficient for the oligonucleotide to be used as a drug for treating an immune-mediated demyelinating disease.

[0147] In various embodiments, the pharmaceutical composition disclosed herein or the oligonucleotide disclosed herein, wherein the oligonucleotide comprises one or more chiral centers and / or double bonds.

[0148] In various embodiments, the pharmaceutical composition disclosed herein or the oligonucleotide disclosed herein, wherein the oligonucleotide exists as a stereoisomer selected from geometric isomers, enantiomers, and diastereomers.

[0149] Additionally disclosed herein are methods of treating a neurological disease or neurologic disorder in a patient in need thereof, the methods comprising administering to the patient in need thereof a therapeutically effective amount of a pharmaceutical composition in combination with a second therapeutic agent.

[0150] In various embodiments, the second therapeutic agent is selected from riluzole (Rirudek), PrimeC, edaravone (Radicava), rivastigmine, donepezil, galantamine, selective serotonin reuptake inhibitors, antipsychotics, cholinesterase inhibitors, memantine, benzodiazepine anxiolytics, AMX0035 (ELYBRIO), ZILUCOPLAN (RA101495), pridopidine, dual AON intrathecal administration (e.g., BIIB067, BIIB078, and BIIB105), BIIB100, levodopa / carbidopa, dopaminergic agents (e.g., ropinirole, pramipexole, rotigotine), medroxyprogesterone, KCNQ2 / K CNQ3 openers (e.g., retigabine, XEN1101, or QRL-101), bioactive scaffolds, anticonvulsants and psychostimulants, therapy (e.g., selected from respiratory care, physical therapy, occupational therapy, speech therapy, nutritional support), deep brain stimulation, levodopa and carbidopa (duopa, rytary, Sinemet, inbrija), istradefylline (nourianz), safinamide (xadago), pramipexole (Mirapex), rotigotine (neupro), ropinirole (requip), amantadine (gocovri, Symmetrel, osmolex), benztropine (Cogentin), trihexyphenidyl (dapoxetine), phenoxyethanol ... Artane, selegiline (eldepryl, zelapar), rasagiline, entacapone (comtan), opicapone (ongentys), tolcapone (tasmar), apomorphine (apokyn, kynmobi), exenatide, Ganoderma lucidum, BIIB054, BIIB094, caffeine, thalizotan, embryonic dopamine cell implantation, aducanumab (Aduhlem), memantine (Namenda), donepezil (Aricept), rivastigmine (Exelon), galantamine (razadyne), Namzeric, suorexane (belsomra), lencaneluzumab (leca nemab), olanzapine (Zyprexa), quetiapine (Seroquel), SSRIs (citalopram (Cipramil), dapoxetine (Priligy), escitalopram (Cipralex), fluoxetine (Prozac or Oxactin), fluvoxamine (Faverin), paroxetine (Seroxat), sertraline (Lustral), vortioxetine (Brintellix)), divalproex sodium (Depakote), carbamazepine (Tegretol), medroxyprogesterone acetate, brivaracetam (briviact), cannabidiol (epidiolex), carbamazepine (carbatrol,Tegretol), phenbarate (xcopri), diazepam (valium), lorazepam (Ativan), clonazepam (klonopin), eslicarbazepine (aptiom), ethosuximide (zarontin), felbatol (felbatol), fenfluramine (fintepla), lacosamide (VIMPAT), lamotrigine (Lamictal), levetiracetam (Keppra), oxcarbazepine (oxtellar xr, Trileptal), perampanel (fycompa), phenobarbital, phenytoin (dilantin), pregabalin (lyrica), tiagabine (gabitril), topiramate (topamax), valproate (depakene, depakote) and / or zonisamide (zonegran), which are used to treat neurological diseases.

[0151] Additionally disclosed herein are methods of treating a neurological disease and / or a neurological disorder in a patient in need thereof, the method comprising administering to a patient in need thereof a therapeutically effective amount of a pharmaceutical composition, and wherein at least one nucleoside linkage of the oligonucleotide is a non-natural linkage, optionally wherein the oligonucleotide comprises a spacer, and wherein the oligonucleotide further comprises a targeting or conjugation moiety selected from cholesterol, lipoic acid, pantothenic acid, polyethylene glycol, and an antibody for crossing the blood-brain barrier.

[0152] In various embodiments, the spacer is a nucleoside replacement group comprising a non-sugar substituent that cannot be connected to a nucleotide base. In various embodiments, the spacer is located between positions 10 and 15 of the oligonucleotide. In various embodiments, the spacer is located between positions 7 and 11 of the oligonucleotide. In various embodiments, the oligonucleotide further comprises a second spacer, wherein the second spacer is located between positions 14 and 22 of the oligonucleotide. In various embodiments, the spacer and the second spacer are separated by at least 5 nucleobases, at least 6 nucleobases, or at least 7 nucleobases in the oligonucleotide. In various embodiments, the spacer is located between positions 7 and 9 of the oligonucleotide, and wherein the second spacer is located between positions 15 and 18 of the oligonucleotide. In various embodiments, the spacer is located at position 8 of the oligonucleotide, and wherein the second spacer is located at position 16 of the oligonucleotide. In various embodiments, the oligonucleotide further comprises a third spacer, wherein the third spacer is located between positions 21 and 24 of the oligonucleotide. In various embodiments, the spacer is located between positions 2 and 5 of the oligonucleotide. In various embodiments, the oligonucleotide further comprises a second spacer, wherein the second spacer is located between positions 8 and 12 of the oligonucleotide. In various embodiments, the oligonucleotide further comprises the 3rd spacer, and wherein the 3rd spacer is between the position 18 and 22 of the oligonucleotide. In various embodiments, the oligonucleotide also comprises the second spacer and the 3rd spacer, and wherein three spacers are positioned at each section of the oligonucleotide so that there are at most 7 positions connecting nucleosides in the oligonucleotide. In various embodiments, at least two of the three spacers are adjacent to the guanine core base. In various embodiments, at least two of the three spacers are immediately before the guanine core base. In various embodiments, each in the first, second or third spacer is a nucleoside replacement group comprising a non-sugar substituent, and wherein the non-sugar substituent does not contain ketone, aldehyde, ketal, hemiketal, acetal, hemiacetal, aminal or hemiaminal part and can not form a covalent bond with the nucleotide base.

[0153] In various embodiments, each of the first, second, or third spacers is independently represented by formula (X), wherein:

[0154]

[0155] Ring A is an optionally substituted 4-8 membered monocyclic cycloalkyl or 4-8 membered monocyclic heterocyclyl, wherein the heterocyclyl contains 1 or 2 heteroatoms selected from O, S and N, with the proviso that A is not capable of forming a covalent bond with a nucleobase; and

[0156] The symbol represents the point of attachment of the internucleoside linkage.

[0157] In various embodiments, each of the first, second, or third spacers is independently represented by formula (Xa), wherein:

[0158]

[0159] In various embodiments, Ring A is an optionally substituted 4-8 membered monocyclic cycloalkyl or 4-8 membered monocyclic heterocyclyl, wherein the 4-8 membered monocyclic cycloalkyl is selected from cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl; and the 4-8 membered monocyclic heterocyclyl is selected from oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, 1,4-dioxanyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl and azepanyl.

[0160] In various embodiments, Ring A is tetrahydrofuranyl.In various embodiments, Ring A is tetrahydropyranyl.

[0161] In various embodiments, each of the first, second, or third spacers is independently represented by formula (I), wherein:

[0162]

[0163] X is selected from -CH2- and -O-; and

[0164] n is 0, 1, 2 or 3.

[0165] In various embodiments, each of the first, second, or third spacers is independently represented by formula (I'), wherein:

[0166]

[0167] In various embodiments, each of the first, second, or third spacers is independently represented by Formula (Ia), wherein:

[0168]

[0169] In various embodiments, each of the first, second, or third spacers is independently represented by Formula (Ia'), wherein:

[0170]

[0171] In various embodiments, each of the first, second, or third spacers is independently represented by Formula II, wherein:

[0172] and

[0173] X is selected from -CH2- and -O-.

[0174] In various embodiments, each of the first, second, or third spacers is independently represented by Formula II', wherein:

[0175] and

[0176] X is selected from -CH2- and -O-.

[0177] In various embodiments, each of the first, second, or third spacers is independently represented by formula (IIa), wherein:

[0178]

[0179] In various embodiments, each of the first, second, or third spacers is independently represented by Formula (Iia'), wherein:

[0180]

[0181] In some embodiments, the spacer is represented by formula (III), wherein:

[0182]

[0183] X is selected from -CH2- and -O-.

[0184] In some embodiments, the spacer is represented by formula (III'), wherein:

[0185]

[0186] X is selected from -CH2- and -O.

[0187] In some embodiments, the spacer is represented by formula (IIIb), wherein:

[0188]

[0189] In some embodiments, the spacer is represented by formula (IIIb'), wherein:

[0190]

[0191] In various embodiments, each of the first, second, or third spacers is independently represented by Formula III, wherein:

[0192] and

[0193] X is selected from -CH2- and -O-.

[0194] In various embodiments, each of the first, second, or third spacers is independently represented by Formula III', wherein:

[0195] and

[0196] X is selected from -CH2- and -O-.

[0197] In various embodiments, each of the first, second, or third spacers is independently represented by formula (IIIa), wherein:

[0198]

[0199] In various embodiments, each of the first, second, or third spacers is independently represented by formula (IIIa'), wherein:

[0200]

[0201] In various embodiments, the oligonucleotide comprising a spacer has a GC content of at least 10%. In various embodiments, the oligonucleotide comprising a spacer has a GC content of at least 20%. In various embodiments, the oligonucleotide comprising a spacer has a GC content of at least 25%. In various embodiments, the oligonucleotide comprising a spacer has a GC content of at least 30%. In various embodiments, the oligonucleotide comprising a spacer has a GC content of at least 40%. In various embodiments, the oligonucleotide comprising a spacer has a GC content of at least 50%. BRIEF DESCRIPTION OF THE DRAWINGS

[0203] Figure 1 is a schematic depiction of portions of the STMN2 transcript and STMN2 translation oligonucleotides designed to target certain portions of the STMN2 transcript.

[0204] Figure 2 is a bar graph showing the results of RT-qPCR analysis of TDP43 and STMN2 full-length mRNA levels in the presence of TDP43 antisense, and recovery of full-length STMN2 transcripts in the presence of six different STMN2 parental oligonucleotides (SEQ ID NO: 36, SEQ ID NO: 55, SEQ ID NO: 177, SEQ ID NO: 203, SEQ ID NO: 244, and SEQ ID NO: 395).

[0205] Figure 3is a bar graph showing the results of RT-qPCR analysis of STMN2 transcript mRNA levels with cryptic exons in the presence of TDP43 antisense, and the reduction of STMN2 transcript mRNA levels with cryptic exons in the presence of 6 different STMN2 parental oligonucleotides (SEQ ID NO: 173, SEQ ID NO: 181, SEQ ID NO: 197, SEQ ID NO: 215, SEQ ID NO: 385, and SEQ ID NO: 400).

[0206] Figure 4 is a bar graph showing the results of RT-qPCR analysis of STMN2 full-length mRNA levels in the presence of TDP43 antisense, and recovery of full-length STMN2 transcripts in the presence of six different STMN2 parental oligonucleotides (SEQ ID NO: 173, SEQ ID NO: 181, SEQ ID NO: 197, SEQ ID NO: 215, SEQ ID NO: 385, and SEQ ID NO: 400).

[0207] Figure 5A is a bar graph showing the results of RT-qPCR analysis of STMN2 transcript mRNA levels with cryptic exons in the presence of TDP43 antisense, and the reduction of STMN2 transcript mRNA levels with cryptic exons in the presence of 6 different STMN2 parental oligonucleotides (SEQ ID NO: 185, SEQ ID NO: 209, SEQ ID NO: 237, SEQ ID NO: 252, SEQ ID NO: 380, and SEQ ID NO: 390).

[0208] Figure 5B is a bar graph showing the results of RT-qPCR analysis of STMN2 full-length mRNA levels in the presence of TDP43 antisense, and recovery of full-length STMN2 transcripts in the presence of six different STMN2 parental oligonucleotides (SEQ ID NO: 185, SEQ ID NO: 209, SEQ ID NO: 237, SEQ ID NO: 252, SEQ ID NO: 380, and SEQ ID NO: 390).

[0209] Figure 6Ais a bar graph showing the results of RT-qPCR analysis of STMN2 transcript mRNA levels with a cryptic exon in the presence of TDP43 antisense, and the reduction of STMN2 transcript mRNA levels with a cryptic exon in the presence of two different STMN2 parental oligonucleotides (SEQ ID NO: 144 and SEQ ID NO: 237) in two duplicate experiments.

[0210] Figure 6B is a bar graph showing the results of RT-qPCR analysis of STMN2 full-length mRNA levels in the presence of TDP43 antisense, and recovery of full-length STMN2 transcripts in the presence of two different STMN2 parental oligonucleotides (SEQ ID NO: 144 and SEQ ID NO: 237) in two duplicate experiments.

[0211] Figure 7A is a bar graph showing the results of RT-qPCR analysis of STMN2 transcript mRNA levels with cryptic exons in the presence of TDP43 antisense, and the reduction of STMN2 transcript mRNA levels with cryptic exons in the presence of five different STMN2 parental oligonucleotides (SEQ ID NO: 36, SEQ ID NO: 173, SEQ ID NO: 177, SEQ ID NO: 181, and SEQ ID NO: 185).

[0212] Figure 7B is a bar graph showing the results of RT-qPCR analysis of STMN2 full-length mRNA levels in the presence of TDP43 antisense, and the recovery of full-length STMN2 transcripts in the presence of five different STMN2 parental oligonucleotides (SEQ ID NO: 36, SEQ ID NO: 173, SEQ ID NO: 177, SEQ ID NO: 181, and SEQ ID NO: 185).

[0213] Figure 8A is a bar graph showing the results of RT-qPCR analysis of STMN2 transcript mRNA levels with cryptic exons in the presence of TDP43 antisense, and the reduction of STMN2 transcript mRNA levels with cryptic exons in the presence of five different STMN2 parental oligonucleotides (SEQ ID NO: 197, SEQ ID NO: 203, SEQ ID NO: 237, SEQ ID NO: 380, and SEQ ID NO: 395).

[0214] Figure 8Bis a bar graph showing the results of RT-qPCR analysis of STMN2 full-length mRNA levels in the presence of TDP43 antisense, and the recovery of full-length STMN2 transcripts in the presence of five different STMN2 parental oligonucleotides (SEQ ID NO: 197, SEQ ID NO: 203, SEQ ID NO: 237, SEQ ID NO: 380, and SEQ ID NO: 395).

[0215] Figure 9A is a bar graph showing the results of RT-qPCR analysis of STMN2 transcript mRNA levels with cryptic exons in the presence of TDP43 siRNA and TDP43 antisense, and the reduction of STMN2 transcript mRNA levels with cryptic exons in the presence of three different STMN2 parental oligonucleotides (SEQ ID NO: 144, SEQ ID NO: 173, and SEQ ID NO: 237).

[0216] Figure 9B is a bar graph showing the results of RT-qPCR analysis of TDP43 and STMN2 full-length mRNA levels in the presence of TDP43 siRNA and TDP43 antisense, and the recovery of full-length STMN2 transcripts in the presence of three different STMN2 parental oligonucleotides (SEQ ID NO: 144, SEQ ID NO: 173 and SEQ ID NO: 237).

[0217] Figure 10A is a bar graph showing the results of RT-qPCR analysis of STMN2 transcript mRNA levels with cryptic exons in the presence of TDP43 siRNA and TDP43 antisense, and the reduction of STMN2 transcript mRNA levels with cryptic exons between different doses of SEQ ID NO: 181 STMN2 parent oligonucleotide.

[0218] Figure 10B is a bar graph showing the results of RT-qPCR analysis of TDP43 and STMN2 full-length mRNA levels in the presence of TDP43 siRNA and TDP43 antisense, and the recovery of full-length STMN2 transcripts at different doses of SEQ ID NO: 181 STMN2 parental oligonucleotide.

[0219] Figure 11Ais a bar graph showing the results of RT-qPCR analysis of STMN2 transcript mRNA levels with cryptic exons in the presence of TDP43 siRNA and TDP43 antisense, and the reduction of STMN2 transcript mRNA levels with cryptic exons among different doses of SEQ ID NO: 185 STMN2 parental oligonucleotide.

[0220] Figure 11B is a bar graph showing the results of RT-qPCR analysis of TDP43 and STMN2 full-length mRNA levels in the presence of TDP43 siRNA and TDP43 antisense, and the recovery of full-length STMN2 transcripts at different doses of SEQ ID NO: 185 STMN2 parental oligonucleotide.

[0221] Figure 12A is a bar graph showing the results of RT-qPCR analysis of STMN2 transcript mRNA levels with cryptic exons in the presence of TDP43 siRNA and TDP43 antisense, and the reduction of STMN2 transcript mRNA levels with cryptic exons among different doses of SEQ ID NO: 197 STMN2 parental oligonucleotide.

[0222] Figure 12B is a bar graph showing the results of RT-qPCR analysis of TDP43 and STMN2 full-length mRNA levels in the presence of TDP43 siRNA and TDP43 antisense, and the recovery of full-length STMN2 transcripts at different doses of SEQ ID NO: 197 STMN2 parental oligonucleotide.

[0223] Figure 13A is a bar graph showing the results of RT-qPCR analysis of STMN2 transcript mRNA levels with cryptic exons in the presence of TDP43 siRNA and TDP43 antisense, and the reduction of STMN2 transcript mRNA levels with cryptic exons among different doses of SEQ ID NO: 144 STMN2 parental oligonucleotide.

[0224] Figure 13B is a bar graph showing the results of RT-qPCR analysis of TDP43 and STMN2 full-length mRNA levels in the presence of TDP43 siRNA and TDP43 antisense, and the recovery of full-length STMN2 transcripts at different doses of SEQ ID NO: 144 STMN2 parental oligonucleotide.

[0225] Figure 14Ais a bar graph showing the results of RT-qPCR analysis of STMN2 transcript mRNA levels with cryptic exons in the presence of TDP43 siRNA and TDP43 antisense, and the reduction of STMN2 transcript mRNA levels with cryptic exons among different doses of SEQ ID NO: 173 STMN2 parental oligonucleotide.

[0226] Figure 14B is a bar graph showing the results of RT-qPCR analysis of TDP43 and STMN2 full-length mRNA levels in the presence of TDP43 siRNA and TDP43 antisense, and the recovery of full-length STMN2 transcripts at different doses of SEQ ID NO: 173 STMN2 parental oligonucleotide.

[0227] Figure 15A is a bar graph showing the results of RT-qPCR analysis of STMN2 transcript mRNA levels with cryptic exons in the presence of TDP43 siRNA and TDP43 antisense, and the reduction of STMN2 transcript mRNA levels with cryptic exons among different doses of SEQ ID NO: 237 STMN2 parental oligonucleotide.

[0228] Figure 15B is a bar graph showing the results of RT-qPCR analysis of TDP43 and STMN2 full-length mRNA levels in the presence of TDP43 siRNA and TDP43 antisense, and the recovery of full-length STMN2 transcripts at different doses of SEQ ID NO: 237 STMN2 parental oligonucleotide.

[0229] Figure 16 is a Western blot and quantitative bar graph showing normalized quantification of STMN2 full-length mRNA levels in the presence of TDP43 siRNA and TDP43 antisense, as well as recovery of full-length STMN2 transcripts by two different STMN2 parental oligonucleotides (SEQ ID NO: 173 and SEQ ID NO: 237).

[0230] Figure 17A is a bar graph showing the results of RT-qPCR analysis of STMN2 transcript mRNA levels with cryptic exons in the presence of TDP43 siRNA and TDP43 antisense, and the reduction of STMN2 transcript mRNA levels with cryptic exons using different variants of the SEQ ID NO: 237 STMN2 parental oligonucleotide.

[0231] Figure 17Bis a bar graph showing the results of RT-qPCR analysis of TDP43 and STMN2 full-length mRNA levels in the presence of TDP43 siRNA and TDP43 antisense, and the recovery of full-length STMN2 transcripts using different variants of the SEQ ID NO: 237 STMN2 parental oligonucleotide.

[0232] Figure 18A is a bar graph showing the results of RT-qPCR analysis of STMN2 transcript mRNA levels with cryptic exons in the presence of TDP43 siRNA and TDP43 antisense, and the reduction of STMN2 transcript mRNA levels with cryptic exons using different variants of SEQ ID NO: 185 STMN2 parental oligonucleotide.

[0233] Figure 18B is a bar graph showing the results of RT-qPCR analysis of TDP43 and STMN2 full-length mRNA levels in the presence of TDP43 siRNA and TDP43 antisense, and recovery of full-length STMN2 transcripts using different variants of SEQ ID NO: 185 STMN2 parental oligonucleotide.

[0234] Figure 19A is a bar graph showing the results of RT-qPCR analysis of STMN2 transcript mRNA levels with cryptic exons in the presence of TDP43 siRNA and TDP43 antisense, and the reduction of STMN2 transcript mRNA levels with cryptic exons using different variants of SEQ ID NO: 173 STMN2 parental oligonucleotide.

[0235] Figure 19B is a bar graph showing the results of RT-qPCR analysis of TDP43 and STMN2 full-length mRNA levels in the presence of TDP43 siRNA and TDP43 antisense, and the recovery of full-length STMN2 transcripts using different variants of the SEQ ID NO: 173 STMN2 parental oligonucleotide.

[0236] Figure 20A is a bar graph showing the results of RT-qPCR analysis of STMN2 transcript mRNA levels with cryptic exons in the presence of TDP43 siRNA and TDP43 antisense, and the reduction of STMN2 transcript mRNA levels with cryptic exons using different variants of the SEQ ID NO: 237 STMN2 parental oligonucleotide.

[0237] Figure 20Bis a bar graph showing the results of RT-qPCR analysis of TDP43 and STMN2 full-length mRNA levels in the presence of TDP43 siRNA and TDP43 antisense, and the recovery of full-length STMN2 transcripts using different variants of the SEQ ID NO: 237 STMN2 parental oligonucleotide.

[0238] Figure 21A is a bar graph showing the results of RT-qPCR analysis of STMN2 transcript mRNA levels with cryptic exons in the presence of TDP43 siRNA and TDP43 antisense, and the reduction of STMN2 transcript mRNA levels with cryptic exons using different variants of SEQ ID NO: 173 STMN2 parental oligonucleotide.

[0239] Figure 21B is a bar graph showing the results of RT-qPCR analysis of TDP43 and STMN2 full-length mRNA levels in the presence of TDP43 siRNA and TDP43 antisense, and the recovery of full-length STMN2 transcripts using different variants of the SEQ ID NO: 173 STMN2 parental oligonucleotide.

[0240] Figure 22A is a bar graph showing the results of RT-qPCR analysis of STMN2 transcript mRNA levels with cryptic exons in the presence of TDP43 siRNA and TDP43 antisense, and the reduction of STMN2 transcript mRNA levels with cryptic exons using different variants of SEQ ID NO: 144 STMN2 parental oligonucleotide.

[0241] Figure 22B is a bar graph showing the results of RT-qPCR analysis of TDP43 and STMN2 full-length mRNA levels in the presence of TDP43 siRNA and TDP43 antisense, and the recovery of full-length STMN2 transcripts using different variants of the SEQ ID NO: 144 STMN2 parental oligonucleotide.

[0242] Figure 23 is a bar graph showing that use of the SEQ ID NO: 237 STMN2 parental oligonucleotide reverses cryptic exon induction, even in light of increasing proteasome inhibition.

[0243] Figure 24 Shown are dose-response curves illustrating increased recovery of full-length STMN2 transcripts with increasing concentrations of STMN2 AONs.

[0244] Figure 25AShown are Western blot assays showing a qualitative increase in full-length STMN2 protein in response to higher concentrations of STMN2 AONs.

[0245] Figure 25B Shown are the quantitative levels of full-length STMN2 protein normalized to GAPDH in response to different concentrations of STMN2 AONs.

[0246] Figure 26A is a bar graph showing the results of RT-qPCR analysis of STMN2 transcript mRNA levels with cryptic exons in the presence of TDP43 siRNA and TDP43 antisense, and the reduction in STMN2 transcript mRNA levels with cryptic exons among different doses of STMN2 AONs, including SEQ ID NO: 144 AON, SEQ ID NO: 144 AON with two spacers (SEQ ID NO: 1589), SEQ ID NO: 173 AON, SEQ ID NO: 173 with two spacers (SEQ ID NO: 1590), SEQ ID NO: 237 AON, and SEQ ID NO: 237 AON with two spacers (SEQ ID NO: 1591).

[0247] Figure 26B is a bar graph showing the results of RT-qPCR analysis of TDP43 and STMN2 full-length mRNA levels in the presence of TDP43 siRNA and TDP43 antisense, and the recovery of full-length STMN2 transcripts across different doses of STMN2 AONs, including SEQ ID NO: 144 AON, SEQ ID NO: 144 AON with two spacers (SEQ ID NO: 1589), SEQ ID NO: 173 AON, SEQ ID NO: 173 with two spacers (SEQ ID NO: 1590), SEQ ID NO: 237 AON, and SEQ ID NO: 237 AON with two spacers (SEQ ID NO: 1591).

[0248] Figure 27Ais a bar graph showing the results of RT-qPCR analysis of STMN2 transcript mRNA levels with cryptic exons in the presence of TDP43 antisense, and the reduction in STMN2 transcript mRNA levels with cryptic exons among different doses of STMN2 AONs (including SEQ ID NO: 173, SEQ ID NO: 1608, SEQ ID NO: 1609, SEQ ID NO: 1610, SEQ ID NO: 1611, SEQ ID NO: 1612, SEQ ID NO: 1613, SEQ ID NO: 1614, SEQ ID NO: 1615, SEQ ID NO: 1596, SEQ ID NO: 1597, and SEQ ID NO: 1418).

[0249] Figure 27B is a bar graph showing the results of RT-qPCR analysis of TDP43 and STMN2 full-length mRNA levels in the presence of TDP43 antisense, and the recovery of full-length STMN2 transcripts among different doses of STMN2 AONs (including SEQ ID NO: 173, SEQ ID NO: 1608, SEQ ID NO: 1609, SEQ ID NO: 1610, SEQ ID NO: 1611, SEQ ID NO: 1612, SEQ ID NO: 1613, SEQ ID NO: 1614, SEQ ID NO: 1615, SEQ ID NO: 1596, SEQ ID NO: 1597, and SEQ ID NO: 1418).

[0250] Figure 28A is a bar graph showing the results of RT-qPCR analysis of STMN2 transcript mRNA levels with cryptic exons in the presence of TDP43 antisense, and the reduction of STMN2 transcript mRNA levels with cryptic exons among different doses of STMN2 AONs (including SEQ ID NO: 173, SEQ ID NO: 1632, SEQ ID NO: 1346, SEQ ID NO: 1631, SEQ ID NO: 1353, and SEQ ID NO: 1598).

[0251] Figure 28Bis a bar graph showing the results of RT-qPCR analysis of TDP43 and STMN2 full-length mRNA levels in the presence of TDP43 antisense, and the recovery of full-length STMN2 transcripts among different doses of STMN2 AONs (including SEQ ID NO: 173, SEQ ID NO: 1632, SEQ ID NO: 1346, SEQ ID NO: 1631, SEQ ID NO: 1353, and SEQ ID NO: 1598).

[0252] Figure 29A is a bar graph showing the results of RT-qPCR analysis of STMN2 transcript mRNA levels with cryptic exons in the presence of TDP43 antisense, and the reduction of STMN2 transcript mRNA levels with cryptic exons among different doses of STMN2 AONs (including SEQ ID NO: 173 and SEQ ID NO: 1610).

[0253] Figure 29B is a bar graph showing the results of RT-qPCR analysis of TDP43 and STMN2 full-length mRNA levels in the presence of TDP43 antisense, and the recovery of full-length STMN2 transcripts at different doses of STMN2 AONs (including SEQ ID NO: 173 and SEQ ID NO: 1610).

[0254] Figure 30A is a bar graph showing the results of RT-qPCR analysis of STMN2 transcript mRNA levels with cryptic exons in the presence of TDP43 antisense, and the reduction of STMN2 transcript mRNA levels with cryptic exons among different doses of STMN2 AONs (including SEQ ID NO: 185 and SEQ ID NO: 1635).

[0255] Figure 30B is a bar graph showing the results of RT-qPCR analysis of TDP43 and STMN2 full-length mRNA levels in the presence of TDP43 antisense, and the recovery of full-length STMN2 transcripts at different doses of STMN2 AONs (including SEQ ID NO: 185 and SEQ ID NO: 1635).

[0256] Figure 31Ais a bar graph showing the results of RT-qPCR analysis of STMN2 transcript mRNA levels with cryptic exons in the presence of TDP43 antisense, and the reduction of STMN2 transcript mRNA levels with cryptic exons among different doses of STMN2 AONs (including SEQ ID NO: 1347, SEQ ID NO: 1633, and SEQ ID NO: 1634).

[0257] Figure 31B is a bar graph showing the results of RT-qPCR analysis of TDP43 and STMN2 full-length mRNA levels in the presence of TDP43 antisense, and the recovery of full-length STMN2 transcripts among different doses of STMN2 AONs (including SEQ ID NO: 1347, SEQ ID NO: 1633, and SEQ ID NO: 1634).

[0258] Figure 32A is a bar graph showing the results of RT-qPCR analysis of STMN2 transcript mRNA levels with cryptic exons in the presence of TDP43 antisense, and the reduction of STMN2 transcript mRNA levels with cryptic exons among different doses of STMN2 AONs (including SEQ ID NO: 197, SEQ ID NO: 1617, SEQ ID NO: 1618, and SEQ ID NO: 1619).

[0259] Figure 32B is a bar graph showing the results of RT-qPCR analysis of TDP43 and STMN2 full-length mRNA levels in the presence of TDP43 antisense, and the recovery of full-length STMN2 transcripts among different doses of STMN2 AONs (including SEQ ID NO: 197, SEQ ID NO: 1617, SEQ ID NO: 1618, and SEQ ID NO: 1619).

[0260] Figure 33A is a bar graph showing the results of RT-qPCR analysis of STMN2 transcript mRNA levels with cryptic exons in the presence of TDP43 antisense, and the reduction of STMN2 transcript mRNA levels with cryptic exons among different doses of STMN2 AONs (including SEQ ID NO: 252, SEQ ID NO: 1650, SEQ ID NO: 1434, SEQ ID NO: 1651, and SEQ ID NO: 1620).

[0261] Figure 33Bis a bar graph showing the results of RT-qPCR analysis of TDP43 and STMN2 full-length mRNA levels in the presence of TDP43 antisense, and the recovery of full-length STMN2 transcripts among different doses of STMN2 AONs (including SEQ ID NO: 252, SEQ ID NO: 1650, SEQ ID NO: 1434, SEQ ID NO: 1651, and SEQ ID NO: 1620).

[0262] Figure 34A is a bar graph showing the results of RT-qPCR analysis of STMN2 transcript mRNA levels with cryptic exons in the presence of TDP43 antisense, and the reduction of STMN2 transcript mRNA levels with cryptic exons among different doses of STMN2 AONs (including SEQ ID NO: 1434 and SEQ ID NO: 1620).

[0263] Figure 34B is a bar graph showing the results of RT-qPCR analysis of TDP43 and STMN2 full-length mRNA levels in the presence of TDP43 antisense, and the recovery of full-length STMN2 transcripts at different doses of STMN2 AONs (including SEQ ID NO: 1434 and SEQ ID NO: 1620).

[0264] Figure 35 is a bar graph showing normalized STMN2 protein levels after treatment with TDP43 antisense and recovery using STMN2 AONs (including SEQ ID NO: 144, SEQ ID NO: 1589, SEQ ID NO: 173, SEQ ID NO: 1616, SEQ ID NO: 237, and SEQ ID NO: 1591). Detailed Description of the Invention

[0266] The features and other details of the present disclosure will now be described in more detail. Certain terms are collected herein for use in the specification, examples, and appended claims. These definitions should be read in conjunction with the remainder of this disclosure and as understood by those skilled in the art. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art.

[0267] Disclosed herein are oligonucleotides capable of targeting regions of transcripts transcribed from genes. In various embodiments, such oligonucleotides target STMN2 transcripts. In various embodiments, such oligonucleotides target KCNQ2 transcripts. In various embodiments, such oligonucleotides target UNC13A transcripts. In various embodiments, such oligonucleotides target SMN2 transcripts. Additionally disclosed herein are oligonucleotides comprising antisense oligonucleotide sequences and methods of using such oligonucleotides for treating neurological diseases such as amyotrophic lateral sclerosis and frontotemporal dementia and / or neuropathies such as chemotherapy-induced neuropathy. In one embodiment, the oligonucleotide targets a cryptic exon sequence of the STMN2 transcript, thereby reducing the level of STMN2 transcripts having the cryptic exon sequence. In various embodiments, the oligonucleotide targets a sequence of the KCNQ2 transcript, resulting in a reduction in the level of mis-spliced ​​KCNQ2 transcripts. In various embodiments, the oligonucleotide targets a sequence of the UNC13A transcript, resulting in a reduction in the level of mis-spliced ​​KCNQ2 transcripts. In various embodiments, the oligonucleotide targets a sequence of the SMN2 transcript, resulting in a reduction in the level of mis-spliced ​​KCNQ2 transcripts. Also disclosed are pharmaceutical compositions comprising STMN2 oligonucleotides targeting a region of the STMN2 transcript containing a cryptic exon for use in treating neurological diseases and / or neurological disorders, as well as preparations for use in treating neurological diseases and / or neurological disorders containing the disclosed STMN2 oligonucleotides targeting a region of the STMN2 transcript containing a cryptic exon. Also disclosed are pharmaceutical compositions comprising KCNQ2 oligonucleotides targeting a region of the KCNQ2 transcript for use in treating neurological diseases and / or neurological disorders, as well as preparations for use in treating neurological diseases and / or neurological disorders containing the disclosed KCNQ2 oligonucleotides targeting a region of the STMN2 transcript. Also disclosed are pharmaceutical compositions comprising UNC13A oligonucleotides targeting a region of the UNC13A transcript for use in treating neurological diseases and / or neurological disorders, as well as preparations for use in treating neurological diseases and / or neurological disorders containing the disclosed UNC13A oligonucleotides targeting a region of the STMN2 transcript. Also disclosed are pharmaceutical compositions comprising SMN2 oligonucleotides targeted to a region of the SMN2 transcript for use in treating neurological diseases and / or neurological disorders, and methods for preparing medicaments containing the disclosed SMN2 oligonucleotides targeted to a region of the STMN2 transcript for use in treating neurological diseases and / or neurological disorders.

[0268] definition

[0269] The terms "treat," "treatment," "treating," and the like are generally used herein to mean obtaining a desired pharmacological and / or physiological effect. The effect can be therapeutic in the sense of partially or completely curing a disease and / or attributing adverse effects to the disease. As used herein, the term "treat" encompasses any treatment of a disease in a mammal, particularly a human, and includes: (a) inhibiting the disease, i.e., preventing the disease from increasing in severity or extent; (b) relieving the disease, i.e., causing partial or complete improvement in the disease; or (c) preventing recurrence of the disease, i.e., preventing the disease from returning to an active state after a previous successful treatment of the symptoms of the disease or treatment of the disease.

[0270] "Prevention" includes delaying the onset of clinical symptoms, complications, or biochemical indicators of a developing state, disorder, disease, or condition in a subject who may be suffering from or susceptible to a state, disorder, disease, or condition but who does not yet experience or exhibit clinical or subclinical symptoms of the state, disorder, disease, or condition. "Prevention" includes prophylactic treatment of a state, disorder, disease, or condition in or developing in a subject, including prophylactic treatment of clinical symptoms, complications, or biochemical indicators of a state, disorder, disease, or condition in or developing in a subject.

[0271] As used herein, the term "pharmaceutically acceptable carrier" or "pharmaceutically acceptable excipient" refers interchangeably to any and all solvents, dispersion media, coatings, isotonicity agents, absorption delaying agents, and the like that are compatible with pharmaceutical administration. The use of such media and agents for pharmaceutically active substances is well known in the art. The composition may also contain other active compounds that provide supplemental, additional, or enhanced therapeutic function.

[0272] As used herein, the term "pharmaceutical composition" refers to a composition comprising at least one biologically active compound disclosed herein, eg, STMN2 antisense oligonucleotide (AON), formulated together with one or more pharmaceutically acceptable excipients.

[0273] "Individual," "patient," or "subject" are used interchangeably and include any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cattle, sheep, horses, or non-human primates, and most preferably humans. The compounds of the present invention can be administered to mammals, such as humans, but can also be administered to other mammals, such as animals requiring veterinary treatment, for example, domestic animals (e.g., dogs, cats, etc.), farm animals (e.g., cattle, sheep, pigs, horses, etc.), and laboratory animals (e.g., rats, mice, guinea pigs, non-human primates, etc.). In some embodiments, the mammals treated in the methods of the present invention are ideally mammals in which it is desired to modulate STMN2 expression and / or activity. In some embodiments, the mammals treated in the methods of the present invention are ideally mammals in which it is desired to modulate KCNQ2 expression and / or activity. In some embodiments, the mammals treated in the methods of the present invention are ideally mammals in which it is desired to modulate UNC13A expression and / or activity. In some embodiments, the mammals treated in the methods of the present invention are ideally mammals in which it is desired to modulate SMN2 expression and / or activity.

[0274] As used herein, the phrase "regulated by TDP43" encompasses targets that are directly and indirectly regulated by TDP43. Examples of targets that are directly regulated by TDP43 include STMN2 and UNC13A. Examples of targets that are indirectly regulated by TDP43 include KCNQ2.

[0275] As used herein, the term "antisense oligonucleotide" or "AON" encompasses antisense oligonucleotides that target any one of the genes or gene products of STMN2, KCNQ2, UNC13, and SMN2. "Antisense oligonucleotide" or "AON" encompasses any one of a parent oligonucleotide, an oligonucleotide variant, an oligonucleotide with one or more spacers, or an oligonucleotide variant with one or more spacers. Examples of antisense oligonucleotides include oligonucleotides comprising the sequence of any of the following antisense oligonucleotides: SEQ ID NO: 1-466, SEQ ID NO: 893-1338, SEQ ID NO: 1342-1366, SEQ ID NO: 1392-1664, SEQ ID NO: 10655-10669, SEQ ID NO: 1676-1851, SEQ ID NO: 2028-2529, SEQ ID NO: 3046-3221, SEQ ID NO: 3398-3899, SEQ ID NO: 4402-4530, SEQ ID NO: 4531-5794, SEQ ID NO: 7059-8322, SEQ ID NO: 9596-9603, SEQ ID NO: 9710-10141, SEQ ID NO: 10574-10651, SEQ ID NO: Any one of SEQ ID NOs: 10670-10779, SEQ ID NOs: 10783-10808 and SEQ ID NOs: 10814-10820.

[0276] The term "parent oligonucleotide" refers to an oligonucleotide that targets a transcript (e.g., STMN2, KCNQ2, UNC13A, or SMN2 transcript) and is capable of increasing, restoring, or stabilizing full-length activity (e.g., full-length expression, e.g., full-length mRNA and / or full-length protein expression). The parent oligonucleotide does not comprise a spacer. Examples of parent oligonucleotides include oligonucleotides comprising the sequences of any one of SEQ ID NOs: 1-446 and SEQ ID NOs: 893-1338, SEQ ID NOs: 1676-1851, SEQ ID NOs: 2028-2529, and SEQ ID NOs: 4531-5794. As described below, oligonucleotides and oligonucleotide variants having a spacer are described in relation to the corresponding parent oligonucleotide.

[0277] The term "oligonucleotide variant" refers to an oligonucleotide that represents a modified version of the corresponding parent oligonucleotide. For example, an oligonucleotide variant represents a shortened version of a parent oligonucleotide. In various embodiments, the oligonucleotide variant is any one of 15-mer, 16-mer, 17-mer, 18-mer, 19-mer, 20-mer, 21-mer, 22-mer, 23-mer, 24-mer, 25-mer, 26-mer, or 27-mer. Examples of oligonucleotide variants include oligonucleotides comprising the sequence of any one of SEQ ID NOs: 1342-1366, SEQ ID NOs: 1392-1521, SEQ ID NOs: 3046-3221, SEQ ID NOs: 3398-3899, SEQ ID NOs: 7059-8322, and SEQ ID NOs: 9710-10141.

[0278] The term "oligonucleotide with one or more spacers" or "oligonucleotide comprising a spacer" refers to an oligonucleotide having at least one spacer. In various embodiments, the oligonucleotide with one or more spacers can comprise one spacer, two spacers, three spacers, four spacers, five spacers, six spacers, seven spacers, eight spacers, nine spacers, or ten spacers. In various embodiments, the oligonucleotide comprising one or more spacers comprises at least one segment having up to 7 connected nucleosides. For example, as described in the 5' to 3' direction, the oligonucleotide comprising a spacer can comprise a segment having 7 connected nucleosides, followed by a spacer, a second segment having 9 connected nucleosides, followed by a second spacer, and a third segment having 7 connected nucleosides. Herein, the first segment of 7 connected nucleosides and the third segment of 7 connected nucleosides each represent a segment having up to 7 connected nucleosides. As another example, the oligonucleotide comprising a spacer can comprise a section with 10 connection nucleosides, followed by a spacer, a second section with 10 connection nucleosides, followed by a second spacer, and a third section with 3 connection nucleosides. Here, the third section of 3 connection nucleosides represents a section with at most 7 connection nucleosides. In various embodiments, the oligonucleotide with one or more spacers comprises a plurality of sections with at most 7 connection nucleosides. In various embodiments, each section of the oligonucleotide with one or more spacers has at most 7 connection nucleosides. For example, the oligonucleotide can be a 23 aggressiveness and comprises two spacers, and these two spacers are divided into three separated sections of 7 connection nucleosides separately. Therefore, each section of the oligonucleotide has at most 7 connection nucleosides.

[0279] In general, oligonucleotides comprising one or more spacers are described with reference to the corresponding parent oligonucleotide or a corresponding oligonucleotide variant. Example oligonucleotides comprising one or more spacers include any of SEQ ID NOs: 1417-1420, SEQ ID NOs: 1451-1664, SEQ ID NOs: 4402-4468, SEQ ID NOs: 4469-4476, SEQ ID NOs: 4477-4530, SEQ ID NOs: 9596-9696, SEQ ID NOs: 10574-10640, and SEQ ID NOs: 10644-10651.

[0280] STMN2

[0281] As used herein, "STMN2" (also known as superior cervical ganglion-10 protein, Stathmin-like 2, SCGN10, SCG10, neuron growth-associated protein, neuron-specific growth-associated protein, or protein SCG10 (superior cervical ganglion NEAR neurospecific 10)) refers to the gene or gene product (e.g., a protein or mRNA transcript (including pre-mRNA) encoded by the gene) identified by Entrez Gene ID No. 11075 and its allelic variants, as well as orthologs found in non-human species (e.g., non-human primates or mice).

[0282] The term "STMN2 transcript" refers to an STMN2 transcript comprising a cryptic exon. Such an STMN2 transcript comprising a cryptic exon can be an STMN2 pre-mRNA sequence or an STMN2 mature RNA sequence. The term "STMN2 transcript comprising a cryptic exon" refers to an STMN2 transcript comprising one or more cryptic exon sequences. The STMN2 transcript sequence is shown to contain thymine (T), but those skilled in the art will understand that thymine (T) can generally be replaced with uracil (U) in the RNA sequence.

[0283] The terms "STMN2 oligonucleotide," "STMN2 antisense oligonucleotide," or "STMN2 AON" refer to an oligonucleotide that is capable of increasing, restoring, or stabilizing full-length STMN2 activity (e.g., full-length STMN2 expression, e.g., full-length STMN2 mRNA and / or full-length STMN2 protein expression). Generally, STMN2 oligonucleotides reduce the level of mature STMN2 transcripts having cryptic exons by targeting STMN2 transcripts that contain cryptic exons. For example, STMN2 oligonucleotides reduce the level of mature STMN2 transcripts having cryptic exons by inhibiting premature polyadenylation of STMN2 pre-mRNA and / or increasing, restoring, or stabilizing the activity or function of STMN2. In various embodiments, the STMN2 oligonucleotide comprises a sequence that is at least 85% complementary to a portion of a transcript comprising a sequence at least 90% identical to SEQ ID NO: 1339 or SEQ ID NO: 1341, or to a contiguous 15 to 50 nucleobase portion of SEQ ID NO: 1339 or SEQ ID NO: 1341. In various embodiments, the STMN2 oligonucleotide comprises a sequence that is 85% to 98% complementary to an equal length portion of a transcript comprising a sequence at least 90% identical to SEQ ID NO: 1339 or SEQ ID NO: 1341, or to a contiguous 15 to 50 nucleobase portion of SEQ ID NO: 1339 or SEQ ID NO: 1341. The STMN2 target sequence is shown to contain thymine (T), but one skilled in the art will understand that thymine (T) can typically be replaced with uracil (U) in the RNA sequence.

[0284] In various embodiments, the STMN2 oligonucleotide is characterized by having one or more spacers, wherein each spacer divides the STMN2 oligonucleotide into segments of connected nucleosides. In various embodiments, the STMN2 oligonucleotide has two spacers. In one embodiment, the STMN2 oligonucleotide has two segments of connected nucleosides separated by a spacer. In one embodiment, the STMN2 oligonucleotide has three segments of connected nucleosides separated by two spacers. In such embodiments, the STMN2 oligonucleotide has one segment with at most 7 connected nucleosides. For example, the STMN2 oligonucleotide may have 5 connected nucleosides from the 5' to the 3' end, followed by a spacer, 10 connected nucleosides, followed by a second spacer, and 8 connected nucleosides. Therefore, the first segment of 5 connected nucleosides satisfies one segment with at most 7 connected nucleosides. In various embodiments, the STMN2 oligonucleotide has three spacers that divide the STMN2 oligonucleotide into four segments. In various embodiments, each of the four segments of the STMN2 oligonucleotide has at most 7 connected nucleosides.

[0285] As used herein, the term "STMN2 oligonucleotide" encompasses "STMN2 parent oligonucleotide," "STMN2 oligonucleotide having one or more spacers" (e.g., STMN2 oligonucleotide having two spacers or STMN2 oligonucleotide having three spacers), and "STMN2 oligonucleotide variants having one or more spacers." Examples of STMN2 oligonucleotides include oligonucleotides comprising the sequence of any one of SEQ ID NOs: 1-466, 893-1338, 1342-1366, 1392-1664, and 10655-10669.

[0286] The term "STMN2 parent oligonucleotide" refers to an oligonucleotide that targets an STMN2 transcript having a cryptic exon and is capable of increasing, restoring, or stabilizing full-length STMN2 activity (e.g., full-length STMN2 expression, e.g., full-length STMN2 mRNA and / or full-length STMN2 protein expression). The STMN2 parent oligonucleotide does not contain a spacer. Examples of STMN2 parent oligonucleotides include oligonucleotides comprising the sequence of any one of SEQ ID NOs: 1-446 and SEQ ID NOs: 893-1338. As described below, STMN2 oligonucleotides and STMN2 oligonucleotide variants having a spacer are described in relation to the corresponding STMN2 parent oligonucleotide.

[0287] The term "STMN2 oligonucleotide variant" refers to a STMN2 oligonucleotide that represents a modified version of the corresponding STMN2 parent oligonucleotide. For example, a STMN2 oligonucleotide variant represents a shortened version of the STMN2 parent oligonucleotide. In various embodiments, the STMN2 oligonucleotide variant is any of a 15-mer, 16-mer, 17-mer, 18-mer, 19-mer, 20-mer, 21-mer, 22-mer, 23-mer, 24-mer, 25-mer, 26-mer, or 27-mer. Examples of STMN2 oligonucleotide variants include oligonucleotides comprising the sequence of any one of SEQ ID NOs: 1342-1366 or SEQ ID NOs: 1392-1521. In various embodiments, the STMN2 oligonucleotide variant comprises one or more spacers. Such STMN2 oligonucleotide variants comprise the sequence of any one of SEQ ID NOs: 1342-1366 and SEQ ID NOs: 1392-1416.

[0288] The terms "STMN2 oligonucleotide having one or more spacers" or "STMN2 oligonucleotide comprising a spacer" refer to an STMN2 oligonucleotide having at least one spacer. In various embodiments, an oligonucleotide having one or more spacers can comprise one spacer, two spacers, three spacers, four spacers, five spacers, six spacers, seven spacers, eight spacers, nine spacers, or ten spacers. In various embodiments, an oligonucleotide comprising one or more spacers comprises at least one segment having up to 7 linked nucleosides. For example, as described in the 5' to 3' direction, an oligonucleotide comprising a spacer can comprise a segment having 7 linked nucleosides, followed by a spacer, a second segment having 9 linked nucleosides, followed by a second spacer, and a third segment having 7 linked nucleosides. Herein, a first segment of 7 linked nucleosides and a third segment of 7 linked nucleosides each refer to a segment having up to 7 linked nucleosides. As another example, the oligonucleotide comprising a spacer can comprise a section with 10 connection nucleosides, followed by a spacer, a second section with 10 connection nucleosides, followed by a second spacer, and a third section with 3 connection nucleosides. Here, the third section of 3 connection nucleosides represents a section with at most 7 connection nucleosides. In various embodiments, the oligonucleotide with one or more spacers comprises a plurality of sections with at most 7 connection nucleosides. In various embodiments, each section of the oligonucleotide with one or more spacers has at most 7 connection nucleosides. For example, the oligonucleotide can be a 23 aggressiveness and comprises two spacers, and these two spacers are divided into three separated sections of 7 connection nucleosides separately. Therefore, each section of the oligonucleotide has at most 7 connection nucleosides.

[0289] Generally, STMN2 oligonucleotides comprising one or more spacers are described with reference to the corresponding STMN2 parent oligonucleotide or a corresponding STMN2 oligonucleotide variant.Example STMN2 oligonucleotides comprising one or more spacers include any one of SEQ ID NOs: 1417-1420 and SEQ ID NOs: 1451-1664.

[0290] The phrase "STMN2 oligonucleotide targeting an STMN2 transcript" refers to an STMN2 oligonucleotide that binds to an STMN2 transcript. Example regions of the STMN2 transcript are shown in Table 1, which describes sequences corresponding to branch point regions (e.g., branch points 1, 2, and 3), 3' splice acceptor regions, ESE binding regions, TDP43 binding sites, cryptic exons, and Poly A regions. In various embodiments, the oligonucleotide binds to a region of the STMN2 transcript having cryptic exons that is less than 75 nucleobases upstream or downstream of any of the branch points (e.g., branch points 1, 2, and 3), 3' splice acceptor regions, ESE binding regions, TDP43 binding sites, cryptic exons, and Poly A regions.

[0291] KCNQ2

[0292] As used herein, "KCNQ2" (also known as Kv7.2, KCNA11, HNSPC, ENB1, BNFC, potassium channel, voltage-gated KQT-like subfamily Q, member 2, neuroblastoma-specific potassium channel subunit ααKvLQT2, potassium voltage-gated channel subfamily KQT member 2) refers to the gene or gene product (e.g., a protein or mRNA transcript (including pre-mRNA) encoded by the gene) identified by any one of SEQ ID NOs: 3032-3043 and allelic variants thereof, as well as orthologs found in non-human species (e.g., non-human primates or mice).

[0293] The term "KCNQ2 transcript" refers to a KCNQ2 transcript. Such a KCNQ2 transcript can be a KCNQ2 pre-mRNA sequence or a KCNQ2 mature RNA sequence. The KCNQN2 transcript sequence is shown to contain thymine (T), but those skilled in the art will understand that thymine (T) can generally be replaced with uracil (U) in the RNA sequence.

[0294] The term "KCNQ2 oligonucleotide," "KCNQ2 antisense oligonucleotide," or "KCNQ2 AON" refers to an oligonucleotide that is capable of increasing, restoring, or stabilizing full-length KCNQ2 activity (e.g., full-length KCNQ2 expression, e.g., full-length KCNQ2 mRNA and / or full-length KCNQ2 protein expression). Generally, KCNQ2 oligonucleotides reduce the level of mis-spliced ​​KCNQ2 transcripts by targeting KCNQ2 transcripts (e.g., KCNQ2 pre-mRNA or mis-spliced ​​KCNQ2 with a target sequence). In various embodiments, the KCNQ2 oligonucleotide comprises a sequence that is at least 85% complementary to an equal-length portion of a transcript comprising a sequence at least 90% identical to any one of SEQ ID NOs: 3032-3045, or a contiguous 15 to 50 nucleobase portion of any one of SEQ ID NOs: 3032-3043. In various embodiments, the KCNQ2 oligonucleotide comprises a sequence that is 85% to 98% complementary to an equal length portion of a transcript comprising a sequence at least 90% identical to any one of SEQ ID NOs: 3032-3045, or a contiguous 15 to 50 nucleobase portion of any one of SEQ ID NOs: 3032-3045. The KCQN2 target sequence is shown to contain thymine (T), but one skilled in the art will understand that thymine (T) can typically be replaced with uracil (U) in the RNA sequence.

[0295] In various embodiments, the KCNQ2 oligonucleotide is characterized by having one or more spacers, wherein each spacer divides the KCNQ2 oligonucleotide into segments of linked nucleosides. In various embodiments, the KCNQ2 oligonucleotide has two spacers. In one embodiment, the KCNQ2 oligonucleotide has two segments of linked nucleosides separated by one spacer. In one embodiment, the KCNQ2 oligonucleotide has three segments of linked nucleosides separated by two spacers. In such embodiments, the KCNQ2 oligonucleotide has one segment having up to 7 linked nucleosides. For example, the KCNQ2 oligonucleotide may have 5 linked nucleosides from the 5' to the 3' end, followed by a spacer, 10 linked nucleosides, followed by a second spacer, and 8 linked nucleosides. Thus, a first segment of 5 linked nucleosides satisfies a segment having up to 7 linked nucleosides. In various embodiments, the KCNQ2 oligonucleotide has three spacers that divide the KCNQ2 oligonucleotide into four segments. In various embodiments, each of the four segments of the KCNQ2 oligonucleotide has up to 7 linked nucleosides.

[0296] As used herein, the term "KCNQ2 oligonucleotide" encompasses a "KCNQ2 parent oligonucleotide," a "KCNQ2 oligonucleotide having one or more spacers" (e.g., a KCNQ2 oligonucleotide having two spacers or a KCNQ2 oligonucleotide having three spacers), and a "KCNQ2 oligonucleotide variant having one or more spacers." Examples of KCNQ2 oligonucleotides include oligonucleotides comprising the sequence of any one of SEQ ID NOs: 1676-1851, SEQ ID NOs: 2028-2529, SEQ ID NOs: 3046-3221, SEQ ID NOs: 3398-3899, and SEQ ID NOs: 4402-4530.

[0297] The term "KCNQ2 parent oligonucleotide" refers to an oligonucleotide that targets a KCNQ2 transcript and is capable of increasing, restoring, or stabilizing full-length KCNQ2 activity (e.g., full-length KCNQ2 expression, e.g., full-length KCNQ2 mRNA and / or full-length KCNQ2 protein expression). The KCNQ2 parent oligonucleotide does not comprise a spacer. Examples of KCNQ2 parent oligonucleotides include oligonucleotides comprising the sequence of any one of SEQ ID NOs: 1676-1851 and SEQ ID NOs: 2028-2529. As described below, KCNQ2 oligonucleotides and KCNQ2 oligonucleotide variants having a spacer are described in relation to the corresponding KCNQ2 parent oligonucleotide.

[0298] The term "KCNQ2 oligonucleotide variant" refers to a KCNQ2 oligonucleotide that represents a modified version of the corresponding KCNQ2 parent oligonucleotide. For example, a KCNQ2 oligonucleotide variant represents a shortened version of a KCNQ2 parent oligonucleotide. In various embodiments, the KCNQ2 oligonucleotide variant is any of a 15mer, a 16mer, a 17mer, a 18mer, a 19mer, a 20mer, a 21mer, a 22mer, a 23mer, a 24mer, a 25mer, a 26mer, or a 27mer. Examples of KCNQ2 oligonucleotide variants include oligonucleotides comprising the sequence of any one of SEQ ID NOs: 3046-3221 and SEQ ID NOs: 3398-3899. In various embodiments, the KCNQ2 oligonucleotide variant comprises one or more spacers. Such KCNQ2 oligonucleotide variants comprise the sequence of any one of SEQ ID NOs: 4469-4476 and SEQ ID NOs: 4477-4530.

[0299] Generally, KCNQ2 oligonucleotides comprising one or more spacers are described with reference to the corresponding KC=NQ2 parent oligonucleotide or a corresponding KCNQ2 oligonucleotide variant. Example KCNQ2 oligonucleotides comprising one or more spacers include any one of SEQ ID NOs: 4402-4468, SEQ ID NOs: 4469-4476, and SEQ ID NOs: 4477-4530.

[0300] The phrase "a KCNQ2 oligonucleotide targeted to a KCNQ2 transcript" refers to a KCNQ2 oligonucleotide that binds to a KCNQ2 transcript.

[0301] UNC13A

[0302] As used herein, "UNC13A" (also known as Unc-13 homolog A, Munc13-1, KIAA1032, unc-13 homolog A (C. elegans), or protein Unc-13 homolog A) refers to the gene or gene product (e.g., protein or mRNA transcript (including pre-mRNA) encoded by the gene) identified by Entrez Gene ID No. 23025 and its allelic variants, as well as orthologs found in non-human species (e.g., non-human primates or mice). The UNC13A transcript sequence is shown to contain thymine (T), but those skilled in the art will understand that thymine (T) can generally be replaced with uracil (U) in the RNA sequence.

[0303] The term "UNC13A transcript" refers to a UNC13A transcript, which can be a UNC13A pre-mRNA sequence or a UNC13A mature RNA sequence. The terms "UNC13A oligonucleotide," "UNC13A antisense oligonucleotide," or "UNC13A AON" refer to an oligonucleotide that is capable of increasing, restoring, or stabilizing full-length UNC13A activity (e.g., full-length UNC13A expression, e.g., full-length UNC13A mRNA and / or full-length UNC13A protein expression). In general, UNC13A oligonucleotides reduce the level of mis-spliced ​​UNC13A transcripts by targeting UNC13A transcripts (e.g., UNC13A pre-mRNA or mis-spliced ​​UNC13A with a target sequence). In various embodiments, the UNC13A oligonucleotide comprises a sequence that is at least 85% complementary to an equal length portion of a transcript comprising a sequence at least 90% identical to SEQ ID NOs: 9587-9595, or a contiguous 15 to 50 nucleobase portion of SEQ ID NOs: 9587-9595. In various embodiments, the UNC13A oligonucleotide comprises a sequence that is 85% to 98% complementary to an equal length portion of a transcript comprising a sequence at least 90% identical to any of SEQ ID NOs: 9587-9595, or a contiguous 15 to 50 nucleobase portion of any of SEQ ID NOs: 9587-9595. The UNC13A target sequence is shown to contain thymine (T), but one skilled in the art will understand that thymine (T) can typically be replaced with uracil (U) in the RNA sequence.

[0304] In various embodiments, a UNC13A oligonucleotide is characterized by having one or more spacers, wherein each spacer divides the UNC13A oligonucleotide into segments of linked nucleosides. In various embodiments, a UNC13A oligonucleotide has two spacers. In one embodiment, a UNC13A oligonucleotide has two segments of linked nucleosides separated by one spacer. In one embodiment, a UNC13A oligonucleotide has three segments of linked nucleosides separated by two spacers. In such embodiments, a UNC13A oligonucleotide has one segment having at most 7 linked nucleosides. For example, a UNC13A oligonucleotide may have, from the 5' to the 3' end, 5 linked nucleosides, followed by a spacer, 10 linked nucleosides, followed by a second spacer, and 8 linked nucleosides. Thus, a first segment of 5 linked nucleosides satisfies one segment having at most 7 linked nucleosides. In various embodiments, a UNC13A oligonucleotide has three spacers that divide the UNC13A oligonucleotide into four segments. In various embodiments, each of the four segments of the UNC13A oligonucleotide has at most 7 linked nucleosides.

[0305] As used herein, the term "UNC13A oligonucleotide" encompasses a "UNC13A parent oligonucleotide," a "UNC13A oligonucleotide having one or more spacers" (e.g., a UNC13A oligonucleotide having two spacers or a UNC13A oligonucleotide having three spacers), and a "UNC13A oligonucleotide variant having one or more spacers." Examples of UNC13A oligonucleotides include oligonucleotides comprising the sequence of any one of SEQ ID NOs: 4531-5794, SEQ ID NOs: 7059-8322, SEQ ID NOs: 9596-9696, and SEQ ID NOs: 10670-10779.

[0306] The term "UNC13A parent oligonucleotide" refers to an oligonucleotide that targets a UNC13A transcript and is capable of increasing, restoring, or stabilizing full-length UNC13A activity (e.g., full-length UNC13A expression, e.g., full-length UNC13A mRNA and / or full-length UNC13A protein expression). The UNC13A parent oligonucleotide does not comprise a spacer. Examples of UNC13A parent oligonucleotides include oligonucleotides comprising the sequence of any one of SEQ ID NOs: 4531-5794. As described below, UNC13A oligonucleotides and UNC13A oligonucleotide variants having a spacer are described in relation to the corresponding UNC13A parent oligonucleotide.

[0307] The term "UNC13A oligonucleotide variant" refers to a UNC13A oligonucleotide that represents a modified version of a corresponding UNC13A parent oligonucleotide. For example, a UNC13A oligonucleotide variant represents a shortened version of a UNC13A parent oligonucleotide. In various embodiments, the UNC13A oligonucleotide variant is any of a 15-mer, 16-mer, 17-mer, 18-mer, 19-mer, 20-mer, 21-mer, 22-mer, 23-mer, or 24-mer. Examples of UNC13A oligonucleotide variants include oligonucleotides comprising the sequence of any of SEQ ID NOs: 7059-8322. In various embodiments, the UNC13A oligonucleotide variant comprises one or more spacers. Such UNC13A oligonucleotide variants comprise the sequence of any of SEQ ID NOs: 9596-9696.

[0308] Generally, UNC13A oligonucleotides comprising one or more spacers are described with reference to the corresponding UNC13A parent oligonucleotide or a corresponding UNC13A oligonucleotide variant.Example UNC13A oligonucleotides comprising one or more spacers include any one of SEQ ID NOs: 9596-9696.

[0309] The phrase "UNC13A oligonucleotide that targets a UNC13A transcript" refers to a UNC13A oligonucleotide that binds to a UNC13A transcript.

[0310] SMN2

[0311] As used herein, "SMN2" (also known as survival of motor neuron 2, SMNC, TRD16B, BCD541, GEMINI1, survival motor neuron protein, Tudor domain containing 16B, component of Gems1, Gemin-1, C-BCD541, SMNT, SMN) refers to the gene or gene product (e.g., a protein or mRNA transcript (including pre-mRNA) encoded by the gene) identified by any one of SEQ ID NOs: 9698-9709 and allelic variants thereof, as well as orthologs found in non-human species (e.g., non-human primates or mice).

[0312] The term "SMN2 transcript" refers to an SMN2 transcript. Such an SMN2 transcript can be an SMN2 pre-mRNA sequence or an SMN2 mature RNA sequence. The SMN2 transcript sequence is shown to contain thymine (T), but those skilled in the art will understand that thymine (T) can generally be replaced with uracil (U) in the RNA sequence.

[0313] The terms "SMN2 oligonucleotide," "SMN2 antisense oligonucleotide," or "SMN2 AON" refer to an oligonucleotide that is capable of increasing, restoring, or stabilizing full-length SMN2 activity (e.g., full-length SMN2 expression, e.g., full-length SMN2 mRNA and / or full-length SMN2 protein expression). Generally, SMN2 oligonucleotides reduce the level of mis-spliced ​​SMN2 transcripts by targeting SMN2 transcripts (e.g., SMN2 pre-mRNA or mis-spliced ​​SMN2 with a target sequence). In various embodiments, the SMN2 oligonucleotide comprises a sequence that is at least 85% complementary to an equal-length portion of a transcript comprising a sequence at least 90% identical to any one of SEQ ID NOs: 9698-9709, or to a contiguous 15 to 50 nucleobase portion of any one of SEQ ID NOs: 9698-9709. In various embodiments, the SMN2 oligonucleotide comprises a sequence that is 85% to 98% complementary to an equal length portion of a transcript comprising a sequence at least 90% identical to any one of SEQ ID NOs: 9698 to 9709, or a contiguous 15 to 50 nucleobase portion of any one of SEQ ID NOs: 9698 to 9709. The SMN2 target sequence is shown to contain thymine (T), but one skilled in the art will understand that thymine (T) can typically be replaced with uracil (U) in the RNA sequence.

[0314] In various embodiments, an SMN2 oligonucleotide is characterized by having one or more spacers, wherein each spacer divides the SMN2 oligonucleotide into segments of linked nucleosides. In various embodiments, the SMN2 oligonucleotide has two spacers. In one embodiment, the SMN2 oligonucleotide has two segments of linked nucleosides separated by a spacer. In one embodiment, the SMN2 oligonucleotide has three segments of linked nucleosides separated by two spacers. In such embodiments, the SMN2 oligonucleotide has one segment with at most 7 linked nucleosides. For example, an SMN2 oligonucleotide may have 5 linked nucleosides from the 5' to the 3' end, followed by a spacer, 10 linked nucleosides, followed by a second spacer, and 8 linked nucleosides. Thus, a first segment of 5 linked nucleosides satisfies one segment with at most 7 linked nucleosides. In various embodiments, the SMN2 oligonucleotide has three spacers that divide the SMN2 oligonucleotide into four segments. In various embodiments, each of the four segments of the SMN2 oligonucleotide has at most 7 linked nucleosides.

[0315] As used herein, the term "SMN2 oligonucleotide" encompasses "SMN2 parent oligonucleotide," "SMN2 oligonucleotide having one or more spacers" (e.g., an SMN2 oligonucleotide having two spacers or an SMN2 oligonucleotide having three spacers), and "SMN2 oligonucleotide variants having one or more spacers." Examples of SMN2 oligonucleotides include oligonucleotides comprising the sequence of any one of SEQ ID NOs: 9710-10141, SEQ ID NOs: 10574-10651, and SEQ ID NOs: 10783-10808.

[0316] The term "SMN2 parent oligonucleotide" refers to an oligonucleotide that targets an SMN2 transcript and is capable of increasing, restoring, or stabilizing full-length SMN2 activity (e.g., full-length SMN2 expression, e.g., full-length SMN2 mRNA and / or full-length SMN2 protein expression). The SMN2 parent oligonucleotide does not contain a spacer. As described below, SMN2 oligonucleotides and SMN2 oligonucleotide variants having a spacer are described in relation to the corresponding SMN2 parent oligonucleotide.

[0317] The term "SMN2 oligonucleotide variant" refers to an SMN2 oligonucleotide that represents a modified version of the corresponding SMN2 parent oligonucleotide. For example, an SMN2 oligonucleotide variant represents a shortened version of the SMN2 parent oligonucleotide. In various embodiments, the SMN2 oligonucleotide variant is any of a 15-mer, 16-mer, 17-mer, 18-mer, 19-mer, 20-mer, 21-mer, 22-mer, 23-mer, or 24-mer. Examples of SMN2 oligonucleotide variants include oligonucleotides comprising the sequence of any of SEQ ID NOs: 9710-10141. In various embodiments, the SMN2 oligonucleotide variant comprises one or more spacers. In various embodiments, the SMN2 oligonucleotide variant may comprise the sequence of any of SEQ ID NOs: 10574-10643 and SEQ ID NOs: 10644-10651.

[0318] Generally, SMN2 oligonucleotides comprising one or more spacers are described with reference to the corresponding SMN2 parent oligonucleotide or a corresponding SMN2 oligonucleotide variant. Example SMN2 oligonucleotides comprising one or more spacers include any one of SEQ ID NOs: 10574-10640 and SEQ ID NOs: 10644-10651.

[0319] In various embodiments, one or more spacers can be located at one or more positions of an oligonucleotide. A spacer can be located between the first position and the second position of an oligonucleotide. As used herein, the spacer located between the first position and the second position encompasses a spacer located at any position of the oligonucleotide that is located at the first position, at the second position, or at a position that is sandwiched between the first position and the second position.

[0320] As used herein, the term "therapeutically effective amount" refers to the amount of an oligonucleotide that will elicit the biological or medical response of a tissue, system, animal, or human that is being sought by a researcher, veterinarian, physician, or other clinician. In some embodiments, the oligonucleotide comprises a sequence that is at least 85% complementary to an equal length portion of a transcript comprising a sequence at least 90% identical to SEQ ID NO: 1339 or SEQ ID NO: 1341, or to a contiguous 15 to 50 nucleobase portion of SEQ ID NO: 1339 or SEQ ID NO: 1341. In some embodiments, the oligonucleotide comprises a sequence that is at least 85% complementary to an equal length portion of a transcript comprising a sequence at least 90% identical to any one of SEQ ID NOs: 3032-3045, or to a contiguous 15 to 50 nucleobase portion of any one of SEQ ID NOs: 3032-3045. In some embodiments, the oligonucleotide comprises a sequence that is at least 85% complementary to an equal length portion of a transcript comprising a sequence at least 90% identical to SEQ ID NOs: 9587-9595 or a contiguous 15 to 50 nucleobase portion of SEQ ID NOs: 9587-9595. In some embodiments, the oligonucleotide comprises a sequence that is at least 85% complementary to an equal length portion of a transcript comprising a sequence at least 90% identical to any one of SEQ ID NOs: 9698-9709 or a contiguous 15 to 50 nucleobase portion of any one of SEQ ID NOs: 9698-9709. The oligonucleotide is administered in a therapeutically effective amount to treat and / or prevent a disease, condition, disorder, or state, e.g., a neurological disease and / or a neuropathy. Alternatively, a therapeutically effective amount of an oligonucleotide is the amount required to achieve the desired therapeutic and / or prophylactic effect, such as an amount that results in the prevention or reduction of symptoms associated with a disease associated with decreased STMN2 activity in motor neurons, an amount that results in the prevention or reduction of symptoms associated with a disease associated with decreased KCNQ2 activity in motor neurons, an amount that results in the prevention or reduction of symptoms associated with a disease associated with decreased UNC13A activity in motor neurons, and / or an amount that results in the prevention or reduction of symptoms associated with a disease associated with decreased SMN2 activity in motor neurons.

[0321] As used herein, the term "pharmaceutically acceptable salt" refers to salts of acidic or basic groups that may be present in the STMN2 oligonucleotides used in the compositions of the present invention, the KCNQ2 oligonucleotides used in the compositions of the present invention, the UNC13A oligonucleotides used in the compositions of the present invention, and / or the SMN2 oligonucleotides used in the compositions of the present invention. The STMN2 oligonucleotides, KCNQ2 oligonucleotides, UNC13A oligonucleotides, and / or SMN2 oligonucleotides contained in the compositions of the present invention, which are basic in nature, are capable of forming a variety of salts with various inorganic and organic acids. Acids that can be used to prepare pharmaceutically acceptable acid addition salts of such basic compounds are those that form non-toxic acid addition salts, i.e., salts containing pharmacologically acceptable anions, including, but not limited to, malate, oxalate, chloride, bromide, iodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, isonicotinate, acetate, lactate, salicylate, citrate, tartrate, oleate, tannate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucuronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, and pamoate (i.e., 1,1'-methylene-bis-(2-hydroxy-3-naphthoate)) salts. The STMN2 oligonucleotides, KCNQ2 oligonucleotides, UNC13A oligonucleotides, and / or SMN2 oligonucleotides comprising an amino moiety contained in the compositions of the present invention can form pharmaceutically acceptable salts with various amino acids in addition to the acids mentioned above. The intrinsically acidic compounds contained in the compositions of the present invention can form base salts with various pharmaceutically acceptable cations. Examples of such salts include alkali metal salts or alkaline earth metal salts, particularly calcium, magnesium, sodium, lithium, zinc, potassium, and iron salts.The pharmaceutically acceptable salts of the present disclosure include, for example, pharmaceutically acceptable salts of STMN2 oligonucleotides comprising the sequence of any one of SEQ ID NOs: 1-466, SEQ ID NOs: 893-1338, SEQ ID NOs: 1342-1366, SEQ ID NOs: 1392-1664, and SEQ ID NOs: 10655-10669; pharmaceutically acceptable salts of KCNQ2 oligonucleotides comprising the sequence of any one of SEQ ID NOs: 1676-1851, SEQ ID NOs: 2028-2529, SEQ ID NOs: 3046-3221, SEQ ID NOs: 3398-3899, and SEQ ID NOs: 4402-4530; pharmaceutically acceptable salts of UNC13A oligonucleotides comprising the sequence of any one of SEQ ID NOs: 4531-5794, SEQ ID NOs: 5 NO: 7059-8322, SEQ ID NO: 9596-9696 and SEQ ID NO: 10670-10779; and a pharmaceutically acceptable salt of an SMN2 oligonucleotide comprising the sequence of any one of SEQ ID NO: 9710-10141 and SEQ ID NO: 10574-10651.

[0322] STMN2 oligonucleotides, KCNQ2 oligonucleotides, UNC13A oligonucleotides and / or SMN2 oligonucleotides disclosed herein may contain one or more chiral centers, groups, linkages and / or double bonds, and therefore exist as stereoisomers, such as geometric isomers, enantiomers or diastereomers. When used herein, the term "stereoisomer" is composed of all geometric isomers, enantiomers or diastereomers. These compounds can be represented by the symbol "R" or "S" (or "Rp" or "Sp"), depending on the configuration of the substituents around the stereoisomeric atoms (e.g., stereoisomeric carbon, phosphorus or sulfur atoms). In some embodiments, one or more linkages of the compound can have Rp or Sp configuration (e.g., one or more thiophosphate linkages have Rp or Sp configuration). The configuration of each thiophosphate linkage can be independent of another thiophosphate linkage (e.g., one thiophosphate linkage has Rp configuration, and a second thiophosphate linkage has Sp configuration). In various embodiments, the STMN2 oligonucleotide, KCNQ2 oligonucleotide, UNC13A oligonucleotide, and / or SMN2 oligonucleotide may have a mixed configuration of phosphorothioate linkages. For example, the STMN2 oligonucleotide, KCNQ2 oligonucleotide, UNC13A oligonucleotide, and / or SMN2 oligonucleotide may have five phosphorothioate linkages in the Rp configuration, followed by fifteen phosphorothioate linkages in the Sp configuration, followed by five phosphorothioate linkages in the Rp configuration. The present invention encompasses various stereoisomers of these compounds and mixtures thereof. Stereoisomers include enantiomers and diastereomers. A mixture of enantiomers or diastereomers may be designated as "(±)" in nomenclature, but skilled artisans will recognize that a structure may implicitly represent a chiral center.

[0323] Individual stereoisomers of the STMN2 oligonucleotides, KCNQ2 oligonucleotides, UNC13A oligonucleotides and / or SMN2 oligonucleotides of the present invention can be synthesized from commercially available starting materials containing asymmetric or stereogenic centers, or by preparing a racemic mixture followed by resolution methods well known to those skilled in the art. These resolution methods are exemplified by: (1) attaching the enantiomeric mixture to a chiral auxiliary, separating the resulting diastereomeric mixture by recrystallization or chromatography, and liberating the optically pure product from the auxiliary, (2) forming a salt using an optically active resolving agent, or (3) directly separating the optical enantiomer mixture on a chiral chromatographic column. Stereoisomeric mixtures can also be resolved into their component stereoisomers by well-known methods, such as chiral gas chromatography, chiral supercritical fluid chromatography, chiral simulated moving bed chromatography, chiral high performance liquid chromatography, crystallized compounds as chiral salt complexes, or crystallized compounds in chiral solvents. Stereoisomers can also be obtained from stereoisomerically pure intermediates, reagents, and catalysts by well-known asymmetric synthesis methods.

[0324] The STMN2 oligonucleotides, KCNQ2 oligonucleotides, UNC13A oligonucleotides and / or SMN2 oligonucleotides disclosed herein may exist in solvated as well as unsolvated forms with pharmaceutically acceptable solvents such as water, ethanol, etc., and it is intended that the invention encompass both solvated and unsolvated forms.

[0325] The present disclosure also includes fluorescently labeled compounds of the present invention. The present disclosure also includes isotope-labeled compounds of the present invention (i.e., isotope-labeled STMN2 oligonucleotides, KCNQ2 oligonucleotides, UNC13A oligonucleotides and / or SMN2 oligonucleotides), which are identical to those listed herein, except that one or more atoms are replaced by atoms having an atomic mass or mass number different from the atomic mass or mass number found in large quantities in nature. Examples of isotopes that can be incorporated into the compounds of the present invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine and chlorine, such as, respectively 2 H. 3 H. 11 C. 13 C. 14 C. 15 N. 18 O. 17 O. 31 P. 32 P. 33 P. 35 S. 18 F and 36 Cl.

[0326] Certain isotopically labeled disclosed compounds (e.g., 3 H.14 C and 35 S-labeled) can be used in compound and / or substrate tissue distribution assays. 3 H), carbon 14 (i.e. 14 C) or 35 S isotopes are particularly preferred because of their ease of preparation and detectability. 2 H) substitutions may offer certain therapeutic advantages due to greater metabolic stability (e.g., increased in vivo half-life or reduced dosage requirements) and therefore may be preferred in some circumstances.

[0327] As used herein, "2'-O-(2-methoxyethyl)" (also 2'-MOE and 2'-O(CH2)2OCH3 and MOE) refers to an O-methoxyethyl modification of the 2' position of the furanose ring. In this disclosure, 2'-O-(2-methoxyethyl) is used interchangeably as "2'-O-methoxyethyl." The sugar moiety in a nucleoside modified with 2'-MOE is a modified sugar.

[0328] As used herein, "2'-MOE nucleoside" (also 2'-O-(2-methoxyethyl) nucleoside) means a nucleoside comprising a 2'-MOE modified sugar moiety.

[0329] As used herein, "2'-substituted nucleosides" means nucleosides that include a substituent other than H or OH at the 2'-position of the furanose ring. In certain embodiments, 2'-substituted nucleosides include nucleosides with bicyclic sugar modifications.

[0330] As used herein, "5-methylcytosine" (5-MeC) means cytosine modified with a methyl group attached to the 5 position. 5-Methylcytosine (5-MeC) is a modified nucleobase.

[0331] As used herein, "bicyclic sugar" means a furanose ring modified by a bridge of two atoms. A bicyclic sugar is a modified sugar.

[0332] As used herein, "bicyclic nucleoside" (also referred to as BNA) refers to a nucleoside having a sugar moiety comprising a bridge connecting the two carbon atoms of the sugar ring, thereby forming a bicyclic ring system. In certain embodiments, the bridge connects the 4'-carbon and the 2'-carbon of the sugar ring.

[0333] As used herein, "cap structure" or "terminal cap moiety" refers to a chemical modification that has been incorporated into either terminus of an antisense compound.

[0334] As used herein, "cEt" or "constrained ethyl" refers to a bicyclic nucleoside having a sugar moiety comprising a bridge connecting the 4'-carbon and the 2'-carbon, wherein the bridge has the formula: 4'-CH(CH3)-O-2'.

[0335] As used herein, "constrained ethyl nucleoside" (also referred to as cEt nucleoside) means a nucleoside comprising a bicyclic sugar moiety comprising a 4'-CH(CH3)-O-2' bridge. In some embodiments, cEt can be modified. In some embodiments, cEt can be S-cEt (in S-constrained ethyl 2'-4'-bridged nucleic acids). In some other embodiments, cEt can be R-cEt.

[0336] As used herein, "internucleoside linkage" refers to a covalent linkage between adjacent nucleosides in an oligonucleotide. In some embodiments, as used herein, "non-natural linkage" refers to a "modified internucleoside linkage".

[0337] As used herein, "contiguous" in the context of an oligonucleotide refers to immediately adjacent nucleosides, nucleobases, sugar moieties, or internucleoside linkages. For example, "consecutive nucleobases" means immediately adjacent nucleobases in the sequence. As a comparative example, two nucleosides separated by a spacer are not contiguous.

[0338] As used herein, "locked nucleic acid" or "LNA" or "LNA nucleoside" refers to a nucleic acid monomer having a bridge (e.g., a methylene, ethylene, aminooxy, or oxyimino bridge) connecting the two carbon atoms between the 4' and 2' positions of the nucleoside sugar unit to form a bicyclic sugar. Examples of such bicyclic sugars include, but are not limited to, (A) α-L-methyleneoxy (4'-CH2-O-2') LNA, (B) β-D-methyleneoxy (4'-CH2-O-2') LNA, (C) ethoxy (4'-(CH2)2-O-2') LNA, (D) aminooxy (4'-CH2-ON(R)-2') LNA, and (E) oxyamino (4'-CH2-N(R)-O-2') LNA; wherein R is H, C1-C 12 Alkyl group or protecting group (see US Patent No. 7,427,672, issued September 23, 2008).

[0339] As used herein, LNA compounds include, but are not limited to, compounds having at least one bridge between the 4' and 2' positions of the sugar, wherein each bridge independently comprises 1 or 2 to 4 linking groups independently selected from: -[C(R1)(R2)] n -, -C(R1)=C(R2)-, -C(R1)=N-, -C(=NR1)-, -C(=O)-, -C(=S)-, -O-, -Si(R1)2-, -S(=O) x - and -N(R1)-; wherein: x is 0, 1 or 2; n is 1, 2, 3 or 4; each R1 and R2 is independently H, a protecting group, a hydroxyl group, a C1-C 12 Alkyl, substituted C1-C12 Alkyl, C2-C 12 Alkenyl, substituted C2-C 12 Alkenyl, C2-C 12 Alkynyl, substituted C2-C 12 Alkynyl, C5-C 20 Aryl, substituted C5-C 20 aromatic, heterocyclic, substituted heterocyclic, heteroaryl, substituted heteroaryl, C5-C7 alicyclic, substituted C5-C7 alicyclic, halogen, OJ1, NJ1J2, SJ1, N3, COOJ1, acyl (C(=O)-H), substituted acyl, CN, sulfonyl (S(=O)2-J1) or sulfoxide (S(=O)-J1); and each J1 and J2 are independently H, C1-C 12 Alkyl, substituted C1-C 12 Alkyl, C2-C 12 Alkenyl, substituted C2-C 12 Alkenyl, C2-C 12 Alkynyl, substituted C2-C 12 Alkynyl, C5-C 20 Aromatic, substituted C5-C 20 Aromatic group, acyl group (C (= O) -H), substituted acyl group, heterocyclic group, substituted heterocyclic group, C1-C 12 Aminoalkyl, substituted C1-C 12 aminoalkyl or a protecting group.

[0340] Examples of 4'-2' bridging groups encompassed within the definition of LNA include, but are not limited to, one of the following formulae: -[C(R1)(R2)] n -、-[C(R1)(R2)] n -O-, -C(R1R2)-N(R1)-O- or -C(R1R2)-ON(R1)-. In addition, other bridging groups encompassed by the definition of LNA are 4'-CH2-2', 4'-(CH2)2-2', 4'-(CH2)3-2', 4'-CH2-O-2', 4'-(CH2)2-O-2', 4'-CH2-ON(R1)-2' and 4'-CH2-N(R1)-O-2'-bridges, wherein each R1 and R2 are independently H, a protecting group or a C1-C 12 alkyl.

[0341] Also included in the definition of LNA according to the present invention is that wherein the 2 '-hydroxyl of ribosyl sugar ring is connected to 4 ' carbon atom of sugar ring, thus forms bridge to form the LNA of dicyclo sugar moiety.Bridge can be the methylene (-CH2-) group connecting 2 ' oxygen atom and 4 ' carbon atom, to this, use term methyleneoxy (4 '-CH2-O-2 ') LNA.In addition, in the situation that there is the dicyclo sugar moiety of ethylene bridge group in this position, use term ethyleneoxy (4 '-CH2CH2-O-2 ') LNA.

[0342] As used herein, "spacer" refers to a nucleoside replacement group (e.g., a non-nucleoside group that replaces a nucleoside present in an STMN2 parent oligonucleotide, a KCNQ2 parent oligonucleotide, a UNC13A parent oligonucleotide, and / or an SMN2 parent oligonucleotide). The spacer is characterized by lacking a nucleotide base and replacing the nucleoside sugar moiety with a non-sugar substituent. The non-sugar substituent of the spacer lacks an aldehyde, ketone, acetal, ketal, hemiacetal, or hemiketal group. Thus, the non-sugar substituent of the spacer can be connected to the 3' and 5' positions of the nucleoside adjacent to the spacer via an internucleoside linker as described herein, but cannot form a covalent bond with the nucleotide base (i.e., the nucleobase cannot be connected to another group, such as an internucleoside linkage, a conjugated group, or an end group in an oligonucleotide). Generally speaking, an STMN2 oligonucleotide with a spacer is described with respect to an STMN2 parent oligonucleotide, wherein the spacer replaces the nucleoside of the STMN2 parent oligonucleotide. Generally, a KCNQ2 oligonucleotide with a spacer is described with respect to a KCNQ2 parent oligonucleotide, wherein the spacer replaces a nucleoside of the KCNQ2 parent oligonucleotide. Generally, a UNC13A oligonucleotide with a spacer is described with respect to a UNC13A parent oligonucleotide, wherein the spacer replaces a nucleoside of the UNC13A parent oligonucleotide. Generally, an SMN2 oligonucleotide with a spacer is described with respect to an SMN2 parent oligonucleotide, wherein the spacer replaces a nucleoside of the SMN2 parent oligonucleotide. In all embodiments of the present disclosure, the spacer is not capable of hybridizing to a nucleoside comprising the nucleobase at the corresponding position of the STMN2 transcript, KCNQ2 transcript, UNC13A transcript, and / or SMN2 transcript within the numerical order of the length of the AON oligonucleotide (i.e., if the spacer is located after the nucleoside 4 of the AON (i.e., at position 5 from the 5'-end), then the spacer is not complementary to the nucleoside (A, C, G, or U) at the same corresponding position of the target STMN2 transcript, KCNQ2 transcript, UNC13A transcript, and / or SMN2 transcript).

[0343] As used herein, "mismatch" or "non-complementary groups" refers to the situation when a group (eg, nucleobase) of a first nucleic acid is unable to pair with a corresponding group (eg, nucleobase) of a second or target nucleic acid.

[0344] As used herein, "modified internucleoside linkage" refers to a substitution or any change from a naturally occurring internucleoside linkage (eg, a phosphodiester internucleoside linkage).

[0345] As used herein, "modified nucleobase" means any nucleobase other than adenine, cytosine, guanine, thymine, or uracil. Examples of modified nucleobases include 5-methylcytosine, pseudouridine, or 5-methoxyuridine. "Unmodified nucleobase" means the purine bases adenine (A) and guanine (G), and the pyrimidine bases thymine (T), cytosine (C), and uracil (U).

[0346] As used herein, "modified nucleosides" means nucleosides independently having a modified sugar moiety and / or a modified nucleobase. A universal base is a modified nucleobase that can pair with any of the five unmodified nucleobases. Modified nucleosides include abasic nucleosides that lack a nucleobase. However, modified nucleosides do not include spacers or other groups that are unable to connect to a nucleobase.

[0347] As used herein, " connection nucleosides " are nucleosides connected in a continuous sequence (that is, there is no extra nucleoside between those nucleosides connected). In various embodiments, oligonucleotide can have the different sections of the connection nucleosides connected by a spacer. Here, spacers (that is, nucleoside substitutes) are not considered to be nucleosides, and therefore, oligonucleotide is divided into two sections connecting nucleosides. Oligonucleotide can have the first section of Y connection nucleosides (for example, Y nucleosides connected in a continuous sequence), followed by a spacer, then the second section of Z connection nucleosides. Here, Y and Z connection nucleosides are described in 5' to 3' direction or 3' to 5' direction. In various embodiments, the first section is made up of 7 or less connection nucleosides (for example, Y=7 or less), and the second section includes 8 or more connection nucleosides (for example, Z=8 or more).

[0348] As used herein, "modified oligonucleotide" means an oligonucleotide comprising at least one (ie, one or more) modified internucleoside linkage, modified sugar, and / or modified nucleobase.

[0349] As used herein, "modified sugar" or "modified sugar moiety" means a modified furanosyl sugar moiety or a modified sugar moiety having a moiety other than a furanosyl moiety that can link a nucleobase to another group, such as an internucleoside linkage, a conjugate group, or a terminal group in an oligonucleotide.

[0350] As used herein, "monomer" means a single unit of an oligomer. Monomers include, but are not limited to, nucleosides and nucleotides, whether naturally occurring or modified.

[0351] As used herein, "motif" means the pattern of unmodified and modified nucleosides in an antisense compound.

[0352] As used herein, "natural sugar moiety" means a sugar moiety found in DNA (2'-H) or RNA (2'-OH).

[0353] As used herein, "naturally occurring internucleoside linkage" means a 3' to 5' phosphodiester linkage.

[0354] As used herein, "non-complementary nucleobases" refers to a pair of nucleobases that do not hydrogen bond or otherwise support hybridization with each other.

[0355] As used herein, "nucleic acid" refers to a molecule composed of monomeric nucleotides. Nucleic acids include, but are not limited to, ribonucleic acid (RNA), deoxyribonucleic acid (DNA), single-stranded nucleic acid, double-stranded nucleic acid, non-coding RNA, small interfering RNA (siRNA), short hairpin RNA (shRNA), and microRNA (miRNA).

[0356] As used herein, "nucleobase" refers to a heterocyclic moiety capable of base pairing with a base of another nucleic acid.

[0357] As used herein, "nucleobase complementarity" refers to a nucleobase that can base pair with another nucleobase. For example, in DNA, adenine (A) is complementary to thymine (T). For example, in RNA, adenine (A) is complementary to uracil (U). In certain embodiments, a complementary nucleobase refers to a nucleobase of an antisense compound that can base pair with the corresponding nucleobase of its target nucleic acid. For example, if a nucleobase at a certain position of the antisense compound can hydrogen bond with a nucleobase at a certain position of the target nucleic acid, the hydrogen bonding position between the oligonucleotide and the target nucleic acid is considered to be complementary at the nucleobase pair.

[0358] As used herein, "nucleobase sequence" means the order of nucleobases independent of any sugar, linkage and / or nucleobase modifications.

[0359] As used herein, "nucleoside" refers to a nucleobase linked to a sugar. The term "nucleoside" also includes "modified nucleosides" independently having a modified sugar moiety and / or a modified nucleobase.

[0360] As used herein, " nucleoside mimics " comprises for replacing sugar, sugar and base or base and those structures of not necessarily linkage at one or more positions of oligomeric compound, such as the nucleoside mimics with morpholino, cyclohexenyl, cyclohexyl, tetrahydropyranyl, dicyclo or tricyclic sugar mimics (for example, non-furanose unit). Nucleotide mimics comprises for replacing nucleosides at one or more positions of oligomeric compound and those structures of linkage, such as peptide nucleic acid or morpholino (morpholino connected by phosphorodiamidate or other non-phosphodiester linkages). Sugar substitute overlaps with slightly broader term " nucleoside mimics ", but is intended to represent the displacement of only sugar unit (furanose ring). Tetrahydropyranyl ring provided herein is the illustration of sugar substitute example, and wherein furanose group is replaced by tetrahydropyranyl ring system. " analogies " refers to be substituted into the group of linkage between sugar, core base and / or nucleoside. Typically, a mimetic is used to replace the sugar or sugar-internucleoside linkage combination, and the nucleobase is maintained for hybridization to the selected target.

[0361] As used herein, "nucleotide" means a nucleoside having a phosphate group covalently linked to the sugar portion of the nucleoside.

[0362] As used herein, "oligomeric compound" or "oligomer" means a polymer of linked monomeric subunits capable of hybridizing to at least a region of a nucleic acid molecule.

[0363] As used herein, "oligonucleotide" means a polymer of one or more segments of linked nucleosides, each of which independently of the others may be modified or unmodified.

[0364] As used herein, a "hotspot region" is a series of nucleobases on a target nucleic acid that are amenable to oligomeric compound-mediated regulation of target nucleic acid splicing.

[0365] As used herein, "hybridization" means 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, which can be Watson-Crick, Hoogsteen, or reversed Hoogsteen hydrogen bonding between complementary nucleobases.

[0366] As used herein, "increasing the amount of activity" refers to greater transcript expression, more accurate splicing resulting in full-length mature mRNA and / or protein expression, and / or greater activity relative to transcript expression or activity in an untreated or control sample.

[0367] antisense therapeutics

[0368] Antisense therapeutics are a class of nucleic acid-based compounds that can be used to regulate transcripts, such as mRNA. In various embodiments, antisense therapeutics comprise one or more spacers and can be used to regulate transcripts transcribed from genes, such as STMN2 pre-mRNA, KCNQ2 pre-mRNA, UNC13A pre-mRNA, and / or SMN2 pre-mRNA, which contain cryptic exons.

[0369] Antisense therapeutics can be based on single-stranded or double-stranded deoxyribonucleic acid (DNA), based on ribonucleic acid (RNA) or DNA / RNA chemical analog compounds. Generally speaking, antisense therapeutics are designed to include sequences complementary or almost complementary to mRNA or pre-mRNA sequences transcribed from a given gene to promote the binding between antisense therapeutics and pre-mRNA or mRNA. In certain embodiments, antisense therapeutics work by binding to mRNA or pre-mRNA, thereby inhibiting protein translation, changing pre-mRNA splicing into mature mRNA (e.g., by preventing suitable proteins such as splicing activating proteins from binding) and / or causing destruction of mRNA. In certain embodiments, the antisense therapeutic sequence is complementary to a portion of the sense sequence of the target gene or mRNA. In certain embodiments, the antisense therapeutics described herein are oligonucleotide-based compounds comprising oligonucleotide sequences complementary to pre-mRNA sense or its portion, and one or more spacers. In certain embodiments, the antisense therapeutics described herein can also be compounds based on nucleotide chemical analogs.

[0370] In certain embodiments, an oligonucleotide (such as disclosed herein) can be an oligonucleotide sequence of 5 to 100 oligonucleotide units in length, e.g., 10 to 60 oligonucleotide units in length, e.g., 12 to 50 oligonucleotide units in length, 14 to 40 oligonucleotide units in length, 10 to 30 oligonucleotide units in length, e.g., 14 to 30 oligonucleotide units in length, e.g., 14 to 25 or 15 to 22 oligonucleotide units in length, or 18, 19, 20, 21, 22, 23, 24, 25, 26, or 27 oligonucleotide units in length. As used herein, an "oligonucleotide unit" refers to a nucleoside (e.g., a nucleoside comprising a sugar and / or a nucleobase) or a nucleoside replacement group (e.g., a spacer) of an oligonucleotide.

[0371] In a specific embodiment, the oligonucleotide is 25 oligonucleotide units in length. In a specific embodiment, the oligonucleotide is 23 oligonucleotide units in length. In a specific embodiment, the oligonucleotide is 21 oligonucleotide units in length. In a specific embodiment, the oligonucleotide is 19 oligonucleotide units in length. In various embodiments, the oligonucleotide is at least 18, at least 19, at least 20, at least 21, at least 22, at least 23, at least 24, or at least 25 oligonucleotide units in length. In various embodiments, the oligonucleotide is at least 18 oligonucleotide units in length. In various embodiments, the oligonucleotide is at least 19 oligonucleotide units in length. In various embodiments, the oligonucleotide is at least 20 oligonucleotide units in length. In various embodiments, the oligonucleotide is at least 21 oligonucleotide units in length. In various embodiments, the oligonucleotide is at least 22 oligonucleotide units in length. In various embodiments, the oligonucleotide is at least 23 oligonucleotide units in length. In various embodiments, the oligonucleotide is at least 24 oligonucleotide units in length. In various embodiments, the oligonucleotide is at least 25 oligonucleotide units in length. In various embodiments, the oligonucleotide is at least 26 oligonucleotide units in length. In various embodiments, the oligonucleotide is at least 27 oligonucleotide units in length.

[0372] In certain embodiments, the AON may comprise chemically modified nucleosides (e.g., 2'-O-methylated nucleosides or 2'-O-(2-methoxyethyl) nucleosides) and modified internucleoside linkages (e.g., phosphorothioate linkages). In certain embodiments, the AON described herein comprises an oligonucleotide sequence complementary to an RNA sequence (e.g., STMN2 mRNA sequence). In certain embodiments, the AON described herein may comprise chemically modified nucleosides and modified internucleoside linkages (e.g., phosphorothioate linkages). In specific embodiments, the AON described herein comprises one or more spacers.

[0373] In various embodiments, oligonucleotide comprises one or more spacers. In specific embodiments, oligonucleotide comprises a spacer. In various embodiments, oligonucleotide comprises two spacers. For example, oligonucleotide comprises 23 oligonucleotide units, which have 21 core bases and two nucleoside replacement groups (for example, two spacers). Further embodiments of oligonucleotides with a spacer and oligonucleotides with two spacers are described herein. In various embodiments, oligonucleotide comprises three spacers.

[0374] In some embodiments, the antisense oligonucleotide can be, but is not limited to, an inhibitor of a gene transcript (e.g., shRNA, siRNA, PNA, LNA, 2'-O-methyl (2'OMe) antisense oligonucleotide (AON), 2'-O-(2-methoxyethyl) (MOE) AON, or morpholino oligomer (e.g., phosphorodiamidate morpholino (PMO))), or a composition comprising such a compound. In some embodiments, the oligonucleotide is an antisense oligonucleotide (AON) comprising 2'OMe (e.g., an AON comprising one or more 2'OMe-modified sugars), MOE (e.g., an AON comprising one or more MOE-modified sugars), a peptide nucleic acid (e.g., an AON comprising one or more N-(2-aminoethyl)-glycine units linked by amide bonds or carbonylmethylene bonds as repeating units instead of a sugar-phosphate backbone), a locked nucleic acid (e.g., an AON comprising one or more locked ribose sugars, and which can be a mixture of 2'-deoxynucleotides or 2'OMe nucleotides), a c-ET (e.g., an AON comprising one or more cET sugars), or a composition comprising a 2'OMe-modified sugar. ), constrained methoxyethyl (cMOE) (e.g., an AON comprising one or more cMOE sugars), a morpholino oligomer (e.g., an AON comprising a backbone including one or more PMOs), a deoxy-2'-fluoro nucleoside (e.g., an AON comprising one or more 2'-fluoro-β-D-arabinoside), a tricyclic DNA (tcDNA) (e.g., an AON comprising one or more tcDNA-modified sugars), a 2'-O,4'-C-ethylene-bridged nucleic acid (ENA) (e.g., an AON comprising one or more ENA-modified sugars), or a hexitol nucleic acid (HNA) (e.g., an AON comprising one or more HNA-modified sugars). In some embodiments, the AON comprises one or more internucleoside linkages independently selected from the group consisting of phosphorothioate linkages, phosphodiester linkages, phosphotriester linkages, methylphosphonate linkages, phosphoamide linkages, phosphorothioamide linkages, phosphorodiamidate linkages, phosphorodiamidate morpholino (PMO) (morpholino) linkages, PNA linkages, or any combination of phosphorothioate linkages, phosphodiester linkages, phosphotriester linkages, methylphosphonate linkages, phosphoamide linkages, phosphorothioamide linkages, phosphorodiamidate linkages, phosphorodiamidate morpholino (PMO) (morpholino) linkages, and PNA linkages. In some embodiments, the STMN2 AON, KCNQ2 AON, UNC13A AON, or SMN2 AON comprises one or more phosphorothioate linkages, phosphodiester linkages, or a combination of phosphorothioate and phosphodiester linkages.

[0375] Peptide nucleic acids (PNAs) are short, synthetic polymers with a structure that mimics DNA or RNA. PNAs comprise a backbone composed of repeating N-(2-aminoethyl)-glycine units linked by peptide bonds. In certain embodiments, the PNAs described herein can be used as antisense therapeutics that bind to RNA sequences with high specificity and increase, restore, and / or stabilize levels (e.g., full-length UNC13A mRNA or protein levels) and / or activity (e.g., biological activity, such as UNC13A activity).

[0376] Locked nucleic acids (LNA) are oligonucleotide sequences containing one or more modified RNA nucleotides in which the ribose moiety is modified with an additional bridge connecting the 2' oxygen and the 4' carbon. LNAs are believed to have a higher Tm than similar oligonucleotide sequences. In certain embodiments, the LNAs described herein can be used as antisense therapeutics that bind to RNA sequences with high specificity. For example, LNA can bind to STMN2 pre-RNA and inhibit premature polyadenylation of STMN2 pre-mRNA, and increase, restore and / or stabilize STMN2 levels (e.g., STMN2 mRNA or protein levels) and / or activity (e.g., biological activity, e.g., STMN2 activity); LNA can bind to KCNQ2 pre-RNA and reduce mis-splicing of KCNQ2 pre-mRNA, and increase, restore and / or stabilize KCNQ2 levels (e.g., KCNQ2 mRNA or protein levels) and / or activity (e.g., biological activity, e.g., KCNQ2 activity); LNA can bind to UNC13A pre-mRNA and prevent mis-splicing of UNC13A pre-mRNA, and increase, restore and / or stabilize UNC13A levels (e.g., UNC13A mRNA or protein levels) and / or activity (e.g., biological activity, e.g., UNC13A activity); and / or LNA can bind to SMN2 pre-RNA and reduce mis-splicing of SMN2 pre-mRNA, and increase, restore and / or stabilize SMN2 levels (e.g., SMN2 mRNA or protein levels) and / or activity (e.g., biological activity, e.g., SMN2 activity).

[0377] Morpholino oligomers are oligonucleotide compounds comprising a base attached to a backbone of methylenemorpholine rings linked through phosphorodiamidate groups. In certain embodiments, the morpholino oligomers of the invention can be designed to bind to a specific pre-mRNA sequence of interest.

[0378] STMN2 oligonucleotides complementary to STMN2 transcripts with cryptic exons

[0379] In some embodiments, the STMN2 AON comprises a sequence that is at least 85% complementary to a sequence that shares at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with a region of the STMN2 transcript comprising a cryptic exon (e.g., SEQ ID NO: 1339 or SEQ ID NO: 1341). In some embodiments, the STMN2 AON comprises a sequence that is 85% to 98% complementary to a sequence that shares at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with a region of the STMN2 transcript comprising a cryptic exon (e.g., SEQ ID NO: 1339 or SEQ ID NO: 1341). In some embodiments, the STMN2 AON comprises a sequence that is 90%-95% complementary to a sequence that shares at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with a region of the STMN2 transcript comprising a cryptic exon (e.g., SEQ ID NO: 1339 or SEQ ID NO: 1341). In specific embodiments, the STMN2 AON comprises a sequence that is at least 85% complementary to a sequence that shares at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with a region of the STMN2 transcript comprising a cryptic exon (e.g., SEQ ID NO: 1339 or SEQ ID NO: 1341). In specific embodiments, the STMN2 AON comprises a sequence that is 84% ​​to 88% complementary to a sequence that shares at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with a region of the STMN2 transcript comprising the cryptic exon (e.g., SEQ ID NO: 1339 or SEQ ID NO: 1341). In specific embodiments, the STMN2 AON comprises a sequence that is 89% to 92% complementary to a sequence that shares at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with a region of the STMN2 transcript comprising the cryptic exon (e.g., SEQ ID NO: 1339 or SEQ ID NO: 1341).In specific embodiments, the STMN2 AON comprises a sequence that is 94% to 96% complementary to a sequence that shares at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with a region of the STMN2 transcript comprising a cryptic exon (e.g., SEQ ID NO: 1339 or SEQ ID NO: 1341).

[0380] In various embodiments, the STMN2 AON comprises a sequence that shares at least 85% identity with an equal length portion of any one of SEQ ID NOs: 1-466, SEQ ID NOs: 893-1338, SEQ ID NOs: 1342-1366, SEQ ID NOs: 1392-1664, or SEQ ID NOs: 10655-10669. In various embodiments, the STMN2 AON comprises a sequence that shares at least 90% identity with an equal length portion of any one of SEQ ID NOs: 1-466, SEQ ID NOs: 893-1338, SEQ ID NOs: 1342-1366, SEQ ID NOs: 1392-1664, or SEQ ID NOs: 10655-10669.

[0381] In various embodiments, the region of the STMN2 transcript targeted by the STMN2 AON is a cryptic exon sequence. In various embodiments, the region of the STMN2 transcript targeted by the STMN2 AON is a sequence located upstream or downstream (e.g., 100 or 200 bases upstream or downstream) of the cryptic exon sequence. In some embodiments, the STMN2 AON comprises a spacer and has a segment containing up to 7 linked nucleosides. In some embodiments, the STMN2 AON comprises a spacer and has a segment containing up to 6, 5, 4, 3, or 2 linked nucleosides.

[0382] STMN2 AON binding specificity can be assessed via measuring changes in parameters such as dissociation constant, melting temperature, or other criteria such as protein or RNA expression levels, or other assays that measure STMN2 activity or expression.

[0383] In some embodiments, the STMN2 AON may comprise a non-duplex oligonucleotide. In some embodiments, the STMN2 AON may comprise a duplex of two oligonucleotides, wherein the first oligonucleotide comprises a nucleobase sequence that is completely or almost completely complementary to the STMN2 pre-mRNA sequence, and the second oligonucleotide comprises a nucleobase sequence that is complementary to the nucleobase sequence of the first oligonucleotide.

[0384] In some embodiments, the STMN2 AON can target STMN2 pre-mRNA, which contains cryptic exons produced from the STMN2 gene of one or more species. For example, the STMN2 AON can target the STMN2 pre-mRNA of a mammalian STMN2 gene, such as the STMN2 pre-mRNA of a human (i.e., Homo sapiens) STMN2 gene, which contains cryptic exons. In specific embodiments, the STMN2 AON targets the human STMN2 pre-mRNA containing cryptic exons. In some embodiments, the STMN2 AON comprises a nucleobase sequence complementary to the nucleobase sequence of the STMN2 gene or STMN2 pre-mRNA, or a portion thereof, which contains cryptic exons.

[0385] The STMN2 AONs described herein include antisense oligonucleotides comprising the oligonucleotide sequences listed in Table 1 below:

[0386] Table 1. STMN2 AON sequences, in each of which one or more spacers described in this disclosure were incorporated to generate oligonucleotides of the invention.

[0387]

[0388]

[0389]

[0390]

[0391]

[0392]

[0393]

[0394]

[0395]

[0396]

[0397]

[0398]

[0399]

[0400]

[0401]

[0402] *At least one (i.e., one or more) nucleoside linkages of the oligonucleotide sequence are independently selected from phosphorothioate linkages, alkylphosphate linkages, phosphorodithioate linkages, phosphotriester linkages, alkylphosphonate linkages, 3-methoxypropylphosphonate linkages, methylphosphonate linkages, aminoalkylphosphotriester linkages, alkylenephosphonate linkages, phosphinate linkages, phosphoamidate linkages, phosphorothioateamidate linkages, phosphorodiamidate linkages, phosphorodiamidate (e.g., comprising phosphorodiamidate morpholino (PMO), 3' aminoribose, or 5' aminoribose) linkages, aminoalkylphosphoamidate linkages, phosphorothioateamidate linkages, thioalkylphosphonate linkages, thioalkylphosphotriester linkages, phosphorothioate linkages, phosphoroselenoate linkages, and phosphoroboronate linkages.

[0403] Table 2 below identifies additional STMN2 AON sequences:

[0404] Table 2. Additional STMN2 AON sequences (corresponding to SEQ ID NOs: 1-446, but with uracil bases substituted for thymine bases)

[0405]

[0406]

[0407]

[0408]

[0409]

[0410]

[0411]

[0412]

[0413]

[0414]

[0415]

[0416]

[0417] *At least one (i.e., one or more) nucleoside linkages of the oligonucleotide sequence are independently selected from the group consisting of phosphorothioate linkages, alkylphosphate linkages, phosphorodithioate linkages, phosphotriester linkages, alkylphosphonate linkages, 3-methoxypropylphosphonate linkages, methylphosphonate linkages, aminoalkylphosphotriester linkages, alkylenephosphonate linkages, phosphinate linkages, phosphoamidate linkages, phosphorothioateamidate linkages, phosphorodiamidate linkages, phosphorodiamidate (e.g., comprising phosphorodiamidate morpholino (PMO), 3' aminoribose, or 5' aminoribose) linkages, aminoalkylphosphoamidate linkages, phosphorothioateamidate linkages, thioalkylphosphonate linkages, thioalkylphosphotriester linkages, phosphorothioate linkages, phosphoroselenoate linkages, and phosphoroboronate linkages.

[0418] Table 3 below identifies exemplary STMN2 AON sequences:

[0419] Table 3. Exemplary STMN2 AON sequences, in each of which one or more spacers described in this disclosure are incorporated to generate oligonucleotides of the invention.

[0420]

[0421]

[0422] *At least one (i.e., one or more) nucleoside linkages of the oligonucleotide sequence are independently selected from phosphorothioate linkages, alkylphosphate linkages, phosphorodithioate linkages, phosphotriester linkages, alkylphosphonate linkages, 3-methoxypropylphosphonate linkages, methylphosphonate linkages, aminoalkylphosphotriester linkages, alkylenephosphonate linkages, phosphinate linkages, phosphoamidate linkages, phosphorothioateamidate linkages, phosphorodiamidate linkages, phosphorodiamidate (e.g., comprising phosphorodiamidate morpholino (PMO), 3' aminoribose, or 5' aminoribose) linkages, aminoalkylphosphoamidate linkages, phosphorothioateamidate linkages, thioalkylphosphonate linkages, thioalkylphosphotriester linkages, phosphorothioate linkages, phosphoroselenoate linkages, and phosphoroboronate linkages.

[0423] In some embodiments, all internucleoside linkages of the STMN2 AON oligonucleotides listed in Table 3 are phosphorothioate linkages (except when a spacer is present, which linkage may or may not be a phosphorothioate linkage), and each linked nucleoside of the oligonucleotide is a 2'-O-(2-methoxyethyl) (2'-MOE) nucleoside, and each "C" is replaced with 5-MeC. In some embodiments, all internucleoside linkages of the STMN2 AON oligonucleotides listed in Table 3 are phosphorothioate linkages, and each linked nucleoside of the oligonucleotide is a 2'-O-(2-methoxyethyl) (2'-MOE) nucleoside, and not all or no "C"s are replaced with 5-MeC.

[0424] Table 4 below identifies additional exemplary STMN2 AON sequences:

[0425] Table 4. Additional exemplary STMN2 AON sequences (corresponding to the AONs shown in Table 3, but with uracil bases substituted for thymine bases)

[0426]

[0427]

[0428] STMN2 transcripts with cryptic exons

[0429] In one embodiment, the STMN2 AON targets a region of the STMN2 transcript comprising a cryptic exonic sequence, the STMN2 transcript comprising the sequence provided as SEQ ID NO:1339.

[0430]

[0431] The cryptic exon sequence within the STMN2 transcript is provided as SEQ ID NO:1340.

[0432]

[0433] (SEQ ID NO: 1340) (Source: NCBI Reference Sequence: NC_000008.11).

[0434] In various embodiments, the STMN2 transcript with the cryptic exon is 90-100% identical to SEQ ID NO: 1339. In various embodiments, the STMN2 transcript with the cryptic exon is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 1341.

[0435] In one embodiment, the STMN2 transcript having a cryptic exon may comprise a pre-mRNA STMN2 transcript. In one embodiment, the STMN2 transcript having a cryptic exon may comprise the sequence provided as SEQ ID NO: 1341.

[0436]

[0437]

[0438]

[0439]

[0440]

[0441]

[0442]

[0443]

[0444]

[0445]

[0446]

[0447]

[0448]

[0449]

[0450]

[0451]

[0452]

[0453]

[0454]

[0455]

[0456]

[0457]

[0458]

[0459]

[0460]

[0461]

[0462]

[0463]

[0464]

[0465]

[0466]

[0467] STMN2 oligonucleotides targeting a region of the STMN2 transcript

[0468] In various embodiments, the STMN2 AON disclosed herein is complementary to a specific region of a STMN2 transcript (e.g., STMN2 pre-mRNA comprising a cryptic exon) comprising a sequence that shares at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 1341. In some embodiments, the STMN2 AON comprises a sequence that is complementary to a specific region of a STMN2 transcript (e.g., STMN2 pre-mRNA comprising a cryptic exon) comprising a sequence that shares at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 1339 or SEQ ID NO: 1341. In some embodiments, the STMN2 AON comprises a sequence that is at least 85% complementary to a specific region of the STMN2 transcript. In some embodiments, the STMN2 AON comprises a sequence that is 90% to 95% complementary to a specific region of the STMN2 transcript.

[0469] In some embodiments, the STMN2 AON (e.g., STMN2 AON) has a segment containing up to 7 linked nucleosides. In some embodiments, the STMN2 AON has a segment containing up to 6, 5, 4, 3, or 2 linked nucleosides. The segment of the STMN2 AON can be separated from other segments of the STMN2 AON by a spacer. The segment of the STMN2 AON is complementary to a specific region of the STMN2 transcript (e.g., a STMN2 transcript containing a cryptic exon) that comprises a sequence that shares at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 1339 or SEQ ID NO: 1341.

[0470] In some embodiments, the STMN2 AON targets a specific portion of the STMN2 transcript comprising any one of positions 144-168, 173-197, 185-209, or 237-261 of SEQ ID NO: 1339. In some embodiments, the STMN2 AON targets a specific portion of the STMN2 transcript comprising any one of positions 121-144, 146-170, 150-170, 150-172, 150-170, 150-172, 150-174, 169-193, 170-194, 171-195, 172-196, 197-221, 249-273, 252-276, or 276-300. In some embodiments, the STMN2 AON targets a specific portion of the STMN2 transcript comprising any one of positions 144-164, 144-166, 145-167, 146-166, 146-168, 147-165, or 148-168 of SEQ ID NO: 1339. In some embodiments, the STMN2 AON targets a specific portion of the STMN2 transcript comprising any one of positions 173-191, 173-193, 173-195, 173-197, 175-195, 175-197, 177-197, or 179-197 of SEQ ID NO: 1339. In some embodiments, the STMN2 AON targets a specific portion of the STMN2 transcript comprising any one of positions 185-205, 187-209, 189-209, or 191-209 of SEQ ID NO: 1339. In some embodiments, the STMN2 AON targets a specific portion of the STMN2 transcript comprising any one of positions 237-255, 237-259, 239-259, 239-261, 241-261, or 243-261 of SEQ ID NO: 1339.

[0471] In some embodiments, the STMN2 AON targets a specific portion of the STMN2 transcript consisting of any one of positions 144-168, 173-197, 185-209, or 237-261 of SEQ ID NO: 1339. In some embodiments, the STMN2 AON targets a specific portion of the STMN2 transcript consisting of any one of positions 144-164, 144-166, 145-167, 146-166, 146-168, 147-165, or 148-168 of SEQ ID NO: 1339. In some embodiments, the STMN2 AON targets a specific portion of the STMN2 transcript consisting of any one of positions 173-191, 173-193, 173-195, 173-197, 175-195, 175-197, 177-197, or 179-197 of SEQ ID NO: 1339. In some embodiments, the STMN2 AON targets a specific portion of the STMN2 transcript consisting of any one of positions 185-205, 187-209, 189-209, or 191-209 of SEQ ID NO: 1339. In some embodiments, the STMN2 AON targets a specific portion of the STMN2 transcript consisting of any of positions 237-255, 237-259, 239-259, 239-261, 241-261, or 243-261 of SEQ ID NO: 1339.

[0472] STMN2 oligonucleotide variants

[0473] In various embodiments, the STMN2 AON includes different variants, hereinafter referred to as STMN2 AON variants. STMN2 AON variants can be oligonucleotide sequences of 5 to 100 nucleobases in length, for example, 10 to 40 nucleobases in length, for example, 14 to 40 nucleobases in length, 10 to 30 nucleobases in length, for example, 14 to 30 nucleobases in length, for example, 16 to 28 nucleobases in length, for example, 19 to 23 nucleobases in length, for example, 21 to 23 nucleobases in length, or for example, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleobases in length. STMN2 AON variants can be oligonucleotide sequences that are complementary to a portion of the STMN2 pre-mRNA sequence or the STMN2 gene sequence.

[0474] In various embodiments, the STMN2 AON variant represents a modified form of the corresponding STMN2 parent oligonucleotide comprising a nucleobase sequence selected from any one of SEQ ID NOs: 1-446 or SEQ ID NOs: 893-1338. In some embodiments, the STMN2 AON variant comprises a nucleobase sequence that represents a shortened version of the STMN2 AON nucleobase sequence selected from any one of SEQ ID NOs: 1-446 or SEQ ID NOs: 893-1338. As an example, if the STMN2 parent oligonucleotide comprises a 25-mer (e.g., 25 nucleotide bases in length), the variant (e.g., STMN2 variant) can comprise a shorter version (e.g., a 15-mer, 17-mer, 19-mer, 21-mer, or 23-mer of the 25-mer STMN2 parent oligonucleotide). In one embodiment, the nucleobase sequence of the STMN2 AON variant differs from the corresponding nucleobase sequence of the STMN2 parent oligonucleotide in that 1, 2, 3, 4, 5, or 6 nucleotide bases are removed from one or both of the 5' and 3' ends of the nucleobase sequence of the STMN2 parent oligonucleotide. In one embodiment, the corresponding STMN2 AON variant may include a 23-mer in which two nucleotide bases are removed from one of the 3' or 5' ends of the 25-mer contained in the STMN2 parent oligonucleotide. In one embodiment, the corresponding STMN2 AON variant may include a 23-mer in which one nucleotide base is removed from each of the 3' and 5' ends of the 25-mer contained in the STMN2 parent oligonucleotide. In one embodiment, the corresponding STMN2 AON variant may include a 21-mer in which two nucleotide bases are removed from each of the 3' and 5' ends of the 25-mer contained in the STMN2 parent oligonucleotide. In one embodiment, the corresponding STMN2 AON variant may include a 21-mer in which four nucleotide bases are removed from the 3' end or the 5' end of the 25-mer contained in the STMN2 parent oligonucleotide. In one embodiment, the corresponding STMN2 AON variant may include a 19-mer in which three nucleotide bases are removed from each of the 3' end and the 5' end of the 25-mer contained in the STMN2 parent oligonucleotide. In one embodiment, the corresponding STMN2 AON variant may include a 19-mer in which six nucleotide bases are removed from the 3' end or the 5' end of the 25-mer contained in the STMN2 parent oligonucleotide.

[0475] Example sequences of STMN2 AON variants are shown below in Tables 5A and 5B.

[0476] Table 5A. STMN2 oligonucleotide variant sequences

[0477]

[0478]

[0479] *The at least one nucleoside linkage of the nucleobase sequence is selected from the group consisting of phosphorothioate linkages, alkylphosphate linkages, phosphorodithioate linkages, phosphotriester linkages, alkylphosphonate linkages, 3-methoxypropylphosphonate linkages, methylphosphonate linkages, aminoalkylphosphotriester linkages, alkylenephosphonate linkages, phosphinate linkages, phosphoamidate linkages, phosphorothioateamidate linkages, phosphorodiamidate linkages, phosphorodiamidate (e.g., comprising phosphorodiamidate morpholino (PMO), 3' aminoribose, or 5' aminoribose) linkages, aminoalkylphosphoamidate linkages, phosphorothioateamidate linkages, thioalkylphosphonate linkages, thioalkylphosphotriester linkages, phosphorothioate linkages, phosphoroselenoate linkages, and phosphoroboronate linkages.

[0480] Table 5B: Additional STMN2 oligonucleotide variant sequences

[0481]

[0482]

[0483] *The at least one nucleoside linkage of the nucleobase sequence is selected from the group consisting of phosphorothioate linkages, alkylphosphate linkages, phosphorodithioate linkages, phosphotriester linkages, alkylphosphonate linkages, 3-methoxypropylphosphonate linkages, methylphosphonate linkages, aminoalkylphosphotriester linkages, alkylenephosphonate linkages, phosphinate linkages, phosphoamidate linkages, phosphorothioateamidate linkages, phosphorodiamidate linkages, phosphorodiamidate (e.g., comprising phosphorodiamidate morpholino (PMO), 3' aminoribose, or 5' aminoribose) linkages, aminoalkylphosphoamidate linkages, phosphorothioateamidate linkages, thioalkylphosphonate linkages, thioalkylphosphotriester linkages, phosphorothioate linkages, phosphoroselenoate linkages, and phosphoroboronate linkages.

[0484] Table 6 below identifies additional variants of the STMN2 AON sequence:

[0485] Table 6. Additional STMN2 oligonucleotide variant sequences

[0486]

[0487]

[0488] *The at least one nucleoside linkage of the nucleobase sequence is selected from the group consisting of phosphorothioate linkages, alkylphosphate linkages, phosphorodithioate linkages, phosphotriester linkages, alkylphosphonate linkages, 3-methoxypropylphosphonate linkages, methylphosphonate linkages, aminoalkylphosphotriester linkages, alkylenephosphonate linkages, phosphinate linkages, phosphoamidate linkages, phosphorothioateamidate linkages, phosphorodiamidate linkages, phosphorodiamidate (e.g., comprising phosphorodiamidate morpholino (PMO), 3' aminoribose, or 5' aminoribose) linkages, aminoalkylphosphoamidate linkages, phosphorothioateamidate linkages, thioalkylphosphonate linkages, thioalkylphosphotriester linkages, phosphorothioate linkages, phosphoroselenoate linkages, and phosphoroboronate linkages.

[0489] KCNQ2 oligonucleotides complementary to KCNQ2 transcripts

[0490] In some embodiments, the KCNQ2 AON comprises a sequence that is at least 85% complementary to a sequence that shares at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with a region of a KCNQ2 transcript (e.g., any one of SEQ ID NOs: 3032-3045). In some embodiments, the KCNQ2 AON comprises a sequence that is 90% to 95% complementary to a sequence that shares at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with a region of a KCNQ2 transcript (e.g., any one of SEQ ID NOs: 3032-3043). In specific embodiments, the KCNQ2 AON comprises a sequence that is 85% to 90% complementary to a sequence that shares at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with a region of a KCNQ2 transcript (e.g., any one of SEQ ID NOs: 3032-3043). In specific embodiments, the KCNQ2 AON comprises a sequence that is 84% ​​to 88% complementary to a sequence that shares at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with a region of a KCNQ2 transcript (e.g., any one of SEQ ID NOs: 3032-3043). In specific embodiments, the KCNQ2 AON comprises a sequence that is 89% to 92% complementary to a sequence that shares at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with a region of a KCNQ2 transcript (e.g., any one of SEQ ID NOs: 3032-3043). In specific embodiments, the KCNQ2 AON comprises a sequence that is 94% to 96% complementary to a sequence that shares at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with a region of a KCNQ2 transcript (e.g., SEQ ID NOs: 3032-3043).

[0491] In various embodiments, the KCNQ2 AON comprises a sequence that shares at least 85% identity with an equal length portion of any one of SEQ ID NOs: 1676-1851, SEQ ID NOs: 2028-2529, SEQ ID NOs: 3046-3221, SEQ ID NOs: 3398-3899, and SEQ ID NOs: 4402-4530. In various embodiments, the KCNQ2 AON comprises a sequence that shares at least 90% identity with an equal length portion of any one of SEQ ID NOs: 1676-1851, SEQ ID NOs: 2028-2529, SEQ ID NOs: 3046-3221, SEQ ID NOs: 3398-3899, and SEQ ID NOs: 4402-4530.

[0492] In some embodiments, the KCNQ2 AON comprises a spacer and has a segment containing up to 7 linked nucleosides. In some embodiments, the KCNQ2 AON comprises a spacer and has a segment containing up to 6, 5, 4, 3, or 2 linked nucleosides.

[0493] KCNQ2 AON binding specificity can be assessed via measuring changes in parameters such as dissociation constant, melting temperature, or other criteria such as protein or RNA expression levels, or other assays that measure KCNQ activity or expression.

[0494] In some embodiments, the KCNQ2 AON can comprise a non-duplex oligonucleotide. In some embodiments, the KCNQ2 AON can comprise a duplex of two oligonucleotides, wherein the first oligonucleotide comprises a nucleobase sequence that is completely or almost completely complementary to the KCNQ2 pre-mRNA sequence, and the second oligonucleotide comprises a nucleobase sequence that is complementary to the nucleobase sequence of the first oligonucleotide.

[0495] In some embodiments, the KCNQ2 AON can target one or more isoforms of KCNQ2 mRNA. In some embodiments, the KCNQ2 AON comprises a nucleobase sequence that is complementary to a nucleobase sequence of a KCNQ2 gene or KCNQ2 mRNA.

[0496] The KCNQ2 AONs described herein include antisense oligonucleotides comprising the oligonucleotide sequences listed in Tables 7A and 7B below:

[0497] Table 7A. Example KCNQ2 AON sequences complementary to sequences in the intron 4 region of the KCNQ2 transcript.

[0498]

[0499]

[0500]

[0501]

[0502]

[0503]

[0504]

[0505]

[0506]

[0507] *At least one (i.e., one or more) nucleoside linkages of the oligonucleotide sequence are independently selected from phosphorothioate linkages, alkylphosphate linkages, phosphorodithioate linkages, phosphotriester linkages, alkylphosphonate linkages, 3-methoxypropylphosphonate linkages, methylphosphonate linkages, aminoalkylphosphotriester linkages, alkylenephosphonate linkages, phosphinate linkages, phosphoamidate linkages, phosphorothioateamidate linkages, phosphorodiamidate linkages, phosphorodiamidate (e.g., comprising phosphorodiamidate morpholino (PMO), 3' aminoribose, or 5' aminoribose) linkages, aminoalkylphosphoamidate linkages, phosphorothioateamidate linkages, thioalkylphosphonate linkages, thioalkylphosphotriester linkages, phosphorothioate linkages, phosphoroselenoate linkages, and phosphoroboronate linkages.

[0508] Table 7B. Example KCNQ2 AON sequences complementary to the sequences of KCNQ2 transcripts.

[0509]

[0510]

[0511]

[0512]

[0513]

[0514]

[0515]

[0516]

[0517]

[0518]

[0519]

[0520]

[0521] *At least one (i.e., one or more) nucleoside linkages of the oligonucleotide sequence are independently selected from phosphorothioate linkages, alkylphosphate linkages, phosphorodithioate linkages, phosphotriester linkages, alkylphosphonate linkages, 3-methoxypropylphosphonate linkages, methylphosphonate linkages, aminoalkylphosphotriester linkages, alkylenephosphonate linkages, phosphinate linkages, phosphoamidate linkages, phosphorothioateamidate linkages, phosphorodiamidate linkages, phosphorodiamidate (e.g., comprising phosphorodiamidate morpholino (PMO), 3' aminoribose, or 5' aminoribose) linkages, aminoalkylphosphoamidate linkages, phosphorothioateamidate linkages, thioalkylphosphonate linkages, thioalkylphosphotriester linkages, phosphorothioate linkages, phosphoroselenoate linkages, and phosphoroboronate linkages.

[0522] KCNQ2 transcripts

[0523] In various embodiments, the KCNQ2 transcript comprises the sequence provided as SEQ ID NO:3032.

[0524]

[0525]

[0526]

[0527]

[0528] (SEQ ID NO: 3032 (Source NCBI Reference No.: NM_001382235.1)

[0529] In various embodiments, the KCNQ2 mRNA transcript comprises the sequence provided as SEQ ID NO:3033.

[0530]

[0531]

[0532]

[0533]

[0534] (SEQ ID NO: 3033) (Source: NCBI Reference Sequence: NM_004518.6).

[0535] In various embodiments, the KCNQ2 mRNA transcript comprises the sequence provided as SEQ ID NO:3034.

[0536]

[0537]

[0538]

[0539]

[0540] (SEQ ID NO: 3034) (Source: NCBI Reference Sequence: NM_172106.3).

[0541] In various embodiments, the KCNQ2 mRNA transcript comprises the sequence provided as SEQ ID NO:3035.

[0542]

[0543]

[0544]

[0545]

[0546] (SEQ ID NO: 3035) (Source: NCBI Reference Sequence: NM_172107.4).

[0547] In various embodiments, the KCNQ2 mRNA transcript comprises the sequence provided as SEQ ID NO:3036.

[0548]

[0549]

[0550]

[0551]

[0552] (SEQ ID NO: 3036) (Source: NCBI Reference Sequence: NM_172108.5).

[0553] In various embodiments, the KCNQ2 mRNA transcript comprises the sequence provided as SEQ ID NO:3037.

[0554]

[0555]

[0556] (SEQ ID NO: 3037) (Source: NCBI Reference Sequence: NM_172109.3).

[0557] In various embodiments, the KCNQ2 mRNA transcript comprises the sequence provided as SEQ ID NO:3038.

[0558]

[0559]

[0560]

[0561]

[0562] (SEQ ID NO: 3038) (Source: NCBI Reference Sequence: XM_011528811.2)

[0563] In various embodiments, the KCNQ2 mRNA transcript comprises the sequence provided as SEQ ID NO:3039.

[0564]

[0565]

[0566]

[0567]

[0568] (SEQ ID NO: 3039) (Source: NCBI Reference Sequence: XM_017027844.2)

[0569] In various embodiments, the KCNQ2 mRNA transcript comprises the sequence provided as SEQ ID NO:3040.

[0570]

[0571]

[0572]

[0573]

[0574] (SEQ ID NO: 3040) (Source: NCBI Reference Sequence: XM_017027841.2)

[0575] In various embodiments, the KCNQ2 mRNA transcript comprises the sequence provided as SEQ ID NO:3041.

[0576]

[0577]

[0578]

[0579]

[0580]

[0581] (SEQ ID NO: 3041) (Source: NCBI Reference Sequence: XM_017027842.2)

[0582] In various embodiments, the KCNQ2 mRNA transcript comprises the sequence provided as SEQ ID NO:3042.

[0583]

[0584]

[0585]

[0586]

[0587] (SEQ ID NO: 3042) (Source: NCBI Reference Sequence: XM_017027843.1)

[0588] In various embodiments, the KCNQ2 mRNA transcript comprises the sequence provided as SEQ ID NO:3043.

[0589]

[0590]

[0591]

[0592]

[0593] (SEQ ID NO: 3043) (Source: NCBI Reference Sequence: XM_017027845.1)

[0594] In various embodiments, the KCNQ2 transcript is a KCNQ2 pre-mRNA transcript. In various embodiments, the KCNQ2 pre-mRNA transcript comprises the sequence provided as SEQ ID NO: 3044.

[0595]

[0596]

[0597]

[0598]

[0599]

[0600]

[0601]

[0602]

[0603]

[0604]

[0605]

[0606]

[0607]

[0608]

[0609]

[0610]

[0611]

[0612]

[0613]

[0614]

[0615]

[0616]

[0617] The NCBI reference sequence NG_009004.2 is SEQ ID NO:3044.

[0618] In various embodiments, the KCNQ2 pre-mRNA transcript comprises the sequence provided as SEQ ID NO: 3045. The NCBI reference sequence NC_000020.11 references the GRCh38.pl3 primary assembly as SEQ ID NO: 3045.

[0619] In various embodiments, the KCNQ2 transcript shares between 90%-100% identity with any one of SEQ ID NOs: 3032-3043, or shares between 90%-100% identity with a KCNQ2 pre-mRNA transcript comprising SEQ ID NO: 3044 or SEQ ID NO: 3045. In various embodiments, the KCNQ2 transcript shares at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with SEQ ID NOs: 3032-3043, or shares at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with a KCNQ2 pre-mRNA transcript transcribed from SEQ ID NO: 3044 or SEQ ID NO: 3045.

[0620] In various embodiments, KCNQ2 is indirectly regulated by TDP43, and thus is encompassed by the phrase "regulated by TDP43."

[0621] KCNQ2 oligonucleotides targeting the KCNQ2 transcript region

[0622] In various embodiments, the KCNQ2 AONs disclosed herein are complementary to a specific region of a KCNQ2 transcript (e.g., a KCNQ2 transcript) comprising a sequence that shares at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with any one of SEQ ID NOs: 3032-3045. In some embodiments, the KCNQ2 AON comprises a sequence that is complementary to a specific region of a KCNQ2 transcript that comprises a sequence that shares at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with any one of SEQ ID NOs: 3032-3045. In some embodiments, the KCNQ2 AON comprises a sequence that is at least 85% complementary to a specific region of the KCNQ2 transcript. In some embodiments, the KCNQ2 AON comprises a sequence that is at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 100% complementary to a specific region of the KCNQ2 transcript. In some embodiments, the KCNQ2 AON comprises a sequence that is 90% to 99% complementary to a specific region of the KCNQ2 transcript. In some embodiments, the KCNQ2 AON comprises a sequence that is 90% to 95% complementary to a specific region of the KCNQ2 transcript. In some embodiments, the KCNQ2 AON comprises a sequence that is 95% to 99% complementary to a specific region of the KCNQ2 transcript. In various embodiments, the KCQN2 transcript is a pre-mRNA transcript and the specific region is intron 4 of the KCNQ2 pre-mRNA transcript. In various embodiments, the KCQN2 transcript is a pre-mRNA transcript and the specific region is intron 5 of the KCNQ2 pre-mRNA transcript. In various embodiments, the specific region is exon 5 of the KCNQ2 pre-mRNA transcript.

[0623] In some embodiments, the KCNQ2 AON (e.g., KCNQ2 AON) has a segment containing up to 7 linked nucleosides. In some embodiments, the KCNQ2 AON has a segment containing up to 6, 5, 4, 3, or 2 linked nucleosides. The segment of the KCNQ2 AON can be separated from other segments of the KCNQ2 AON by a spacer. The segment of the KCNQ2 AON is complementary to a specific region of a KCNQ2 transcript (e.g., a KCNQ2 transcript) that comprises a sequence that shares at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with any one of SEQ ID NOs: 3032-3045.

[0624] KCNQ2 oligonucleotide variants

[0625] In various embodiments, the KCNQ2 AON includes different variants, hereinafter referred to as KCNQ2 AON variants. The KCNQ2 AON variant can be an oligonucleotide sequence of 5 to 100 nucleobases in length, such as 10 to 40 nucleobases in length, such as 14 to 40 nucleobases in length, 10 to 30 nucleobases in length, such as 14 to 30 nucleobases in length, such as 16 to 28 nucleobases in length, such as 19 to 23 nucleobases in length, such as 21 to 23 nucleobases in length, or such as 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 nucleobases in length. The KCNQ2 AON variant can be an oligonucleotide sequence that is complementary to a portion of a KCNQ2 pre-mRNA sequence or a KCNQ2 gene sequence. Example KCNQ2 AON variants include a nucleobase sequence selected from any one of SEQ ID NOs: 4402-4468, SEQ ID NOs: 4469-4476, or SEQ ID NOs: 4477-4530.

[0626] In various embodiments, the KCNQ2 AON variant represents a modified form of the corresponding KCNQ2 parent oligonucleotide comprising a nucleobase sequence selected from any one of SEQ ID NOs: 1676-1851 and SEQ ID NOs: 2028-2529. In some embodiments, the KCNQ2 AON variant comprises a nucleobase sequence that represents a shortened version of a KCNQ2 AON nucleobase sequence selected from any one of SEQ ID NOs: 1676-1851 and SEQ ID NOs: 2028-2529. As an example, if the KCNQ2 parent oligonucleotide comprises a 25-mer (e.g., 25 nucleotide bases in length), the variant (e.g., KCNQ2 variant) can comprise a shorter version (e.g., a 15-mer, 17-mer, 19-mer, 21-mer, or 23-mer of the 25-mer KCNQ2 parent oligonucleotide). In one embodiment, the nucleobase sequence of the KCNQ2 AON variant differs from the corresponding nucleobase sequence of the KCNQ2 parent oligonucleotide in that 1, 2, 3, 4, 5, or 6 nucleotide bases are removed from one or both of the 5' and 3' ends of the nucleobase sequence of the KCNQ2 parent oligonucleotide. In one embodiment, the corresponding KCNQ2 AON variant may include a 23-mer in which two nucleotide bases are removed from one of the 3' or 5' ends of the 25-mer contained in the KCNQ2 parent oligonucleotide. In one embodiment, the corresponding KCNQ2 AON variant may include a 23-mer in which one nucleotide base is removed from each of the 3' and 5' ends of the 25-mer contained in the KCNQ2 parent oligonucleotide. In one embodiment, the corresponding KCNQ2 AON variant may include a 21-mer in which two nucleotide bases are removed from each of the 3' and 5' ends of the 25-mer contained in the KCNQ2 parent oligonucleotide. In one embodiment, the corresponding KCNQ2 AON variant may include a 21-mer in which four nucleotide bases are removed from the 3' end or the 5' end of the 25-mer contained in the KCNQ2 parent oligonucleotide. In one embodiment, the corresponding KCNQ2 AON variant may include a 19-mer in which three nucleotide bases are removed from each of the 3' end and the 5' end of the 25-mer contained in the KCNQ2 parent oligonucleotide. In one embodiment, the corresponding KCNQ2 AON variant may include a 19-mer in which six nucleotide bases are removed from the 3' end or the 5' end of the 25-mer contained in the KCNQ2 parent oligonucleotide.

[0627] Example sequences of KCNQ2 AON variants are shown below in Tables 8A and 8B.

[0628] Table 8A. KCNQ2 oligonucleotide variant sequences complementary to the sequences of KCNQ2 transcripts

[0629]

[0630]

[0631]

[0632]

[0633]

[0634] *The at least one nucleoside linkage of the nucleobase sequence is selected from the group consisting of phosphorothioate linkages, alkylphosphate linkages, phosphorodithioate linkages, phosphotriester linkages, alkylphosphonate linkages, 3-methoxypropylphosphonate linkages, methylphosphonate linkages, aminoalkylphosphotriester linkages, alkylenephosphonate linkages, phosphinate linkages, phosphoamidate linkages, phosphorothioateamidate linkages, phosphorodiamidate linkages, phosphorodiamidate (e.g., comprising phosphorodiamidate morpholino (PMO), 3' aminoribose, or 5' aminoribose) linkages, aminoalkylphosphoamidate linkages, phosphorothioateamidate linkages, thioalkylphosphonate linkages, thioalkylphosphotriester linkages, phosphorothioate linkages, phosphoroselenoate linkages, and phosphoroboronate linkages.

[0635] Table 8B: Additional KCNQ2 oligonucleotide variant sequences complementary to KCNQ2 transcripts

[0636]

[0637]

[0638]

[0639]

[0640]

[0641]

[0642]

[0643]

[0644]

[0645]

[0646]

[0647] *The at least one nucleoside linkage of the nucleobase sequence is selected from the group consisting of phosphorothioate linkages, alkylphosphate linkages, phosphorodithioate linkages, phosphotriester linkages, alkylphosphonate linkages, 3-methoxypropylphosphonate linkages, methylphosphonate linkages, aminoalkylphosphotriester linkages, alkylenephosphonate linkages, phosphinate linkages, phosphoamidate linkages, phosphorothioateamidate linkages, phosphorodiamidate linkages, phosphorodiamidate (e.g., comprising phosphorodiamidate morpholino (PMO), 3' aminoribose, or 5' aminoribose) linkages, aminoalkylphosphoamidate linkages, phosphorothioateamidate linkages, thioalkylphosphonate linkages, thioalkylphosphotriester linkages, phosphorothioate linkages, phosphoroselenoate linkages, and phosphoroboronate linkages.

[0648] UNC13A oligonucleotide complementary to UNC13A transcript

[0649] In some embodiments, the UNC13A AON comprises a sequence that is at least 85% complementary to a sequence that shares at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with a region of the UNC13A transcript (e.g., SEQ ID NOs: 9587-9595). In some embodiments, the UNC13A AON comprises a sequence that is 940% to 96% complementary to a sequence that shares at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with a region of the UNC13A transcript (e.g., SEQ ID NOs: 9587-9595). In specific embodiments, the UNC13A AON comprises a sequence that is 85% to 90% complementary to a sequence that shares at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with a region of the UNC13A transcript (e.g., SEQ ID NOs: 9587-9595). In specific embodiments, the UNC13A AON comprises a sequence that is 84% ​​to 88% complementary to a sequence that shares at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with a region of the UNC13A transcript (e.g., SEQ ID NOs: 9587-9595). In specific embodiments, the UNC13A AON comprises a sequence that is 89% to 92% complementary to a sequence that shares at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with a region of a mis-spliced ​​UNC13A transcript (e.g., SEQ ID NOs: 9587-9595). In specific embodiments, the UNC13A AON comprises a sequence that is 94% to 96% complementary to a sequence that shares at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with a region of a UNC13A transcript (e.g., SEQ ID NOs: 9587-9595).

[0650] In various embodiments, the UNC13A AON comprises a sequence that shares at least 85% identity with an equal length portion of any one of SEQ ID NOs: 4531-5794, SEQ ID NOs: 7059-8322, SEQ ID NOs: 9596-9696, and SEQ ID NOs: 10670-10779. In various embodiments, the UNC13A AON comprises a sequence that shares at least 90% identity with an equal length portion of any one of SEQ ID NOs: 4531-5794, SEQ ID NOs: 7059-8322, SEQ ID NOs: 9596-9696, and SEQ ID NOs: 10670-10779.

[0651] In some embodiments, the UNC13A AON comprises a spacer and has a segment containing up to 7 linked nucleosides. In some embodiments, the UNC13A AON comprises a spacer and has a segment containing up to 6, 5, 4, 3, or 2 linked nucleosides.

[0652] UNC13A AON binding specificity can be assessed via measuring changes in parameters such as dissociation constant, melting temperature, or other criteria such as protein or RNA expression levels, or other assays that measure UNC13A activity or expression.

[0653] In some embodiments, the UNC13A AON can comprise a non-duplex oligonucleotide. In some embodiments, the UNC13A AON can comprise a duplex of two oligonucleotides, wherein the first oligonucleotide comprises a nucleobase sequence that is completely or nearly completely complementary to the UNC13A pre-mRNA sequence, and the second oligonucleotide comprises a nucleobase sequence that is complementary to the nucleobase sequence of the first oligonucleotide.

[0654] In some embodiments, the UNC13A AON can target a UNC13A pre-mRNA produced from a UNC13A gene of one or more species. For example, the UNC13A AON can target a UNC13A pre-mRNA of a mammalian UNC13A gene, such as a human (i.e., Homo sapiens) UNC13A gene. In specific embodiments, the UNC13A AON targets a human UNC13A pre-mRNA. In some embodiments, the UNC13A AON comprises a nucleobase sequence that is complementary to a nucleobase sequence of a UNC13A gene or UNC13A pre-mRNA, or a portion thereof.

[0655] The UNC13A AONs described herein include antisense oligonucleotides comprising the oligonucleotide sequences listed in Table 9 below:

[0656] Table 9. Example UNC13A AON sequences.

[0657]

[0658]

[0659]

[0660]

[0661]

[0662]

[0663]

[0664]

[0665]

[0666]

[0667]

[0668]

[0669]

[0670]

[0671]

[0672]

[0673]

[0674]

[0675]

[0676]

[0677]

[0678]

[0679]

[0680]

[0681]

[0682]

[0683]

[0684]

[0685]

[0686]

[0687]

[0688]

[0689]

[0690]

[0691]

[0692]

[0693]

[0694]

[0695]

[0696]

[0697]

[0698]

[0699]

[0700]

[0701]

[0702]

[0703]

[0704]

[0705]

[0706]

[0707]

[0708]

[0709]

[0710]

[0711]

[0712]

[0713]

[0714]

[0715]

[0716]

[0717]

[0718]

[0719]

[0720] *At least one (i.e., one or more) nucleoside linkages of the oligonucleotide sequence are independently selected from phosphorothioate linkages, alkylphosphate linkages, phosphorodithioate linkages, phosphotriester linkages, alkylphosphonate linkages, 3-methoxypropylphosphonate linkages, methylphosphonate linkages, aminoalkylphosphotriester linkages, alkylenephosphonate linkages, phosphinate linkages, phosphoamidate linkages, phosphorothioateamidate linkages, phosphorodiamidate linkages, phosphorodiamidate (e.g., comprising phosphorodiamidate morpholino (PMO), 3' aminoribose, or 5' aminoribose) linkages, aminoalkylphosphoamidate linkages, phosphorothioateamidate linkages, thioalkylphosphonate linkages, thioalkylphosphotriester linkages, phosphorothioate linkages, phosphoroselenoate linkages, and phosphoroboronate linkages.

[0721] UNC13A transcripts

[0722] In various embodiments, the UNC13A mRNA transcript comprises the sequence provided as SEQ ID NO:9587.

[0723]

[0724]

[0725]

[0726]

[0727] (SEQ ID NO: 9587 (source NCBI reference sequence NM_001080421.3)).

[0728] In various embodiments, the UNC13A mRNA transcript comprises the sequence provided as SEQ ID NO:9588.

[0729]

[0730]

[0731]

[0732] (SEQ ID NO: 9588 (source NCBI reference sequence NM_001387021.1)).

[0733] In various embodiments, the UNC13A mRNA transcript comprises the sequence provided as SEQ ID NO:9589.

[0734]

[0735]

[0736]

[0737] (SEQ ID NO: 9589 (source NCBI reference sequence NM_001387022.1)).

[0738] In various embodiments, the UNC13A mRNA transcript comprises the sequence provided as SEQ ID NO:9590.

[0739]

[0740]

[0741]

[0742] (SEQ ID NO: 9590 (source NCBI reference sequence NM_001387023.1)).

[0743] In various embodiments, the UNC13A mRNA transcript comprises the sequence provided as SEQ ID NO:9591.

[0744]

[0745]

[0746]

[0747] (SEQ ID NO: 9591 (source NCBI reference sequence XM_011527810.2)).

[0748] In various embodiments, the UNC13A mRNA transcript comprises the sequence provided as SEQ ID NO:9592.

[0749]

[0750]

[0751]

[0752] (SEQ ID NO: 9592 (Source (NCBI Reference Sequence XM_017026502.1)).

[0753] In various embodiments, the UNC13A mRNA transcript comprises the sequence provided as SEQ ID NO:9593.

[0754]

[0755]

[0756]

[0757] (SEQ ID NO: 9593 (source NCBI reference sequence XM_011527811.2)).

[0758] In various embodiments, the UNC13A transcript is a pre-mRNA UNC13A transcript. In various embodiments, the UNC13A pre-mRNA transcript comprises the sequence provided as SEQ ID NO:9594.

[0759]

[0760]

[0761]

[0762]

[0763]

[0764]

[0765]

[0766]

[0767]

[0768]

[0769]

[0770]

[0771]

[0772]

[0773]

[0774]

[0775]

[0776]

[0777]

[0778]

[0779]

[0780]

[0781] (SEQ ID NO: 9594 (source NCBI reference sequence NG_052872.1)).

[0782] In various embodiments, the UNC13A transcript is a pre-mRNA UNC13A transcript. In various embodiments, the UNC13A pre-mRNA transcript comprises the sequence provided as SEQ ID NO: 9595. The NCBI reference sequence NC_000019.10 references the GRCh38.pl3 primary assembly is SEQ ID NO: 9595.

[0783] UNC13A oligonucleotides targeting the UNC13A transcript region

[0784] In various embodiments, the UNC13A AON disclosed herein is complementary to a specific region of a UNC13A transcript (e.g., UNC13A pre-mRNA) comprising a sequence that shares at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with any one of SEQ ID NOs: 5057-5065. In some embodiments, the UNC13A AON comprises a sequence that is complementary to a specific region of a UNC13A transcript (e.g., UNC13A pre-mRNA) comprising a sequence that shares at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with any one of SEQ ID NOs: 9587-9595. In some embodiments, the UNC13A AON comprises a sequence that is at least 85% complementary to a specific region of the UNC13A transcript. In some embodiments, the UNC13A AON comprises a sequence that is at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 100% complementary to a specific region of the UNC13A transcript. In some embodiments, the UNC13A AON comprises a sequence that is 90% to 99% complementary to a specific region of the UNC13A transcript. In some embodiments, the UNC13A AON comprises a sequence that is 90% to 95% complementary to a specific region of the UNC13A transcript. In some embodiments, the UNC13A AON comprises a sequence that is 95% to 99% complementary to a specific region of the UNC13A transcript.

[0785] In some embodiments, a UNC13A AON (e.g., a UNC13A AON) has a segment containing up to 7 linked nucleosides. In some embodiments, a UNC13A AON has a segment containing up to 6, 5, 4, 3, or 2 linked nucleosides. A segment of a UNC13A AON can be separated from other segments of a UNC13A AON by a spacer. A segment of a UNC13A AON is complementary to a specific region of a UNC13A transcript (e.g., a UNC13A transcript) comprising a sequence that shares at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with any one of SEQ ID NOs: 9587-9593 or a UNC13A pre-mRNA transcript transcribed from SEQ ID NOs: 9594 or 9595.

[0786] UNC13A oligonucleotide variants

[0787] In various embodiments, the UNC13A AON comprises different variants, hereinafter referred to as UNC13A AON variants. The UNC13A AON variant can be an oligonucleotide sequence of 5 to 100 nucleobases in length, such as 10 to 40 nucleobases in length, such as 14 to 40 nucleobases in length, 10 to 30 nucleobases in length, such as 14 to 30 nucleobases in length, such as 16 to 28 nucleobases in length, such as 19 to 23 nucleobases in length, such as 21 to 23 nucleobases in length, or such as 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleobases in length. The UNC13A AON variant can be an oligonucleotide sequence that is complementary to a portion of a UNC13A pre-mRNA sequence or a UNC13A gene sequence.

[0788] In various embodiments, the UNC13A AON variant represents a modified form of the corresponding UNC13A parent oligonucleotide comprising a nucleobase sequence selected from any one of SEQ ID NOs: 4531-5794. In some embodiments, the UNC13A AON variant comprises a nucleobase sequence that represents a shortened version of a UNC13A AON nucleobase sequence selected from any one of SEQ ID NOs: 4531-5794. As an example, if the UNC13A parent oligonucleotide comprises a 25-mer (e.g., 25 oligonucleotide units in length), the variant (e.g., UNC13A variant) can comprise a shorter version of the 25-mer UNC13A parent oligonucleotide (e.g., a 15-mer, 17-mer, 19-mer, 21-mer, or 23-mer). In one embodiment, the nucleobase sequence of a UNC13A AON variant differs from the corresponding nucleobase sequence of a UNC13A parent oligonucleotide in that one, two, three, four, five, or six oligonucleotide units are removed from one or both of the 5' and 3' ends of the nucleobase sequence of the UNC13A parent oligonucleotide. In one embodiment, the corresponding UNC13A AON variant may comprise a 23-mer in which two oligonucleotide units are removed from one of the 3' or 5' ends of a 25-mer comprised in a UNC13A parent oligonucleotide. In one embodiment, the corresponding UNC13A AON variant may comprise a 23-mer in which one nucleotide base is removed from each of the 3' and 5' ends of a 25-mer comprised in a UNC13A parent oligonucleotide. In one embodiment, the corresponding UNC13A AON variant may comprise a 21-mer in which two oligonucleotide units are removed from each of the 3' and 5' ends of a 25-mer comprised in a UNC13A parent oligonucleotide. In one embodiment, the corresponding UNC13A AON variant may comprise a 21-mer wherein four oligonucleotide units are removed from the 3' end or the 5' end of the 25-mer contained in the UNC13A parent oligonucleotide. In one embodiment, the corresponding UNC13A AON variant may comprise a 19-mer wherein three oligonucleotide units are removed from each of the 3' end and the 5' end of the 25-mer contained in the UNC13A parent oligonucleotide. In one embodiment, the corresponding UNC13A AON variant may comprise a 19-mer wherein six oligonucleotide units are removed from the 3' end or the 5' end of the 25-mer contained in the UNC13A parent oligonucleotide.

[0789] Example sequences of UNC13A AON variants are shown in Table 10 below.

[0790] Table 10. Example UNC13A oligonucleotide variant sequences

[0791]

[0792]

[0793]

[0794]

[0795]

[0796]

[0797]

[0798]

[0799]

[0800]

[0801]

[0802]

[0803]

[0804]

[0805]

[0806]

[0807]

[0808]

[0809]

[0810]

[0811]

[0812]

[0813]

[0814]

[0815]

[0816]

[0817]

[0818]

[0819]

[0820]

[0821]

[0822]

[0823]

[0824]

[0825]

[0826]

[0827]

[0828]

[0829]

[0830]

[0831]

[0832]

[0833]

[0834]

[0835]

[0836]

[0837]

[0838]

[0839]

[0840]

[0841]

[0842]

[0843]

[0844]

[0845]

[0846]

[0847]

[0848]

[0849]

[0850]

[0851]

[0852]

[0853] *The at least one nucleoside linkage of the nucleobase sequence is selected from the group consisting of phosphorothioate linkages, alkylphosphate linkages, phosphorodithioate linkages, phosphotriester linkages, alkylphosphonate linkages, 3-methoxypropylphosphonate linkages, methylphosphonate linkages, aminoalkylphosphotriester linkages, alkylenephosphonate linkages, phosphinate linkages, phosphoamidate linkages, phosphorothioateamidate linkages, phosphorodiamidate linkages, phosphorodiamidate (e.g., comprising phosphorodiamidate morpholino (PMO), 3' aminoribose, or 5' aminoribose) linkages, aminoalkylphosphoamidate linkages, phosphorothioateamidate linkages, thioalkylphosphonate linkages, thioalkylphosphotriester linkages, phosphorothioate linkages, phosphoroselenoate linkages, and phosphoroboronate linkages.

[0854] SMN2 oligonucleotide complementary to the SMN2 transcript

[0855] In some embodiments, the SMN2 AON comprises a sequence that is at least 85% complementary to a sequence that shares at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with a region of the SMN2 transcript (e.g., any one of SEQ ID NOs: 9698-9709). In some embodiments, the SMN2 AON comprises a sequence that is 90% to 95% complementary to a sequence that shares at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with a region of the SMN2 transcript (e.g., any one of SEQ ID NOs: 9698-97095). In specific embodiments, the SMN2 AON comprises a sequence that is 85% to 90% complementary to a sequence that shares at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with a region of the SMN2 transcript (e.g., any one of SEQ ID NOs: 9698-9709). In specific embodiments, the SMN2 AON comprises a sequence that is 84% ​​to 88% complementary to a sequence that shares at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with a region of the SMN2 transcript (e.g., any one of SEQ ID NOs: 9698-9709). In specific embodiments, the SMN2 AON comprises a sequence that is 89% to 92% complementary to a sequence that shares at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with a region of the SMN2 transcript (e.g., any one of SEQ ID NOs: 9698-9709). In specific embodiments, the SMN2 AON comprises a sequence that is 94% to 96% complementary to a sequence that shares at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with a region of the SMN2 transcript (e.g., SEQ ID NOs: 9698-9709).

[0856] In various embodiments, the SMN2 AON comprises a sequence that shares at least 85% identity with an equal length portion of any one of SEQ ID NOs: 9710-10141 and SEQ ID NOs: 10574-10651. In various embodiments, the SMN2 AON comprises a sequence that shares at least 90% identity with an equal length portion of any one of SEQ ID NOs: 9710-10141 and SEQ ID NOs: 10574-10651.

[0857] In some embodiments, the SMN2 AON comprises a spacer and has a segment containing up to 7 linked nucleosides. In some embodiments, the SMN2 AON comprises a spacer and has a segment containing up to 6, 5, 4, 3, or 2 linked nucleosides.

[0858] SMN2 AON binding specificity can be assessed via measuring changes in parameters such as dissociation constant, melting temperature, or other criteria such as protein or RNA expression levels, or other assays that measure SMN2 activity or expression.

[0859] In some embodiments, the SMN2 AON can include a non-duplex oligonucleotide. In some embodiments, the SMN2 AON can include a duplex of two oligonucleotides, wherein the first oligonucleotide includes a nucleobase sequence that is completely or almost completely complementary to the SMN2 pre-mRNA sequence, and the second oligonucleotide includes a nucleobase sequence that is complementary to the nucleobase sequence of the first oligonucleotide.

[0860] In some embodiments, the SMN2 AON can target one or more isoforms of SMN2 mRNA. In some embodiments, the SMN2 AON comprises a nucleobase sequence that is complementary to a nucleobase sequence of the SMN2 gene or SMN2 mRNA.

[0861] SMN2 transcript

[0862] In various embodiments, the SMN2 transcript comprises the sequence provided as SEQ ID NO:9698.

[0863]

[0864] (SEQ ID NO: 9698 (Source NCBI Reference No.: NM_017411.4)

[0865] In various embodiments, the SMN2 mRNA transcript comprises the sequence provided as SEQ ID NO:9699.

[0866]

[0867] (SEQ ID NO: 9699) (Source: NCBI Reference Sequence: NM_022875.3).

[0868] In various embodiments, the SMN2 mRNA transcript comprises the sequence provided as SEQ ID NO:9700.

[0869]

[0870]

[0871] (SEQ ID NO: 9700) (Source: NCBI Reference Sequence: NM_022876.2).

[0872] In various embodiments, the SMN2 mRNA transcript comprises the sequence provided as SEQ ID NO:9701.

[0873]

[0874]

[0875] (SEQ ID NO: 9701) (Source: NCBI Reference Sequence: NM_022877.2).

[0876] In various embodiments, the SMN2 mRNA transcript comprises the sequence provided as SEQ ID NO:9702.

[0877]

[0878] (SEQ ID NO: 9702) (Source: NCBI Reference Sequence: XM_011543602.3).

[0879] In various embodiments, the SMN2 mRNA transcript comprises the sequence provided as SEQ ID NO:9703.

[0880]

[0881]

[0882] (SEQ ID NO: 9703) (Source: NCBI Reference Sequence: XM_011543603.3).

[0883] In various embodiments, the SMN2 mRNA transcript comprises the sequence provided as SEQ ID NO:9704.

[0884]

[0885] (SEQ ID NO: 9704) (Source: NCBI Reference Sequence: XM_011543600.2

[0886] In various embodiments, the SMN2 mRNA transcript comprises the sequence provided as SEQ ID NO:9705.

[0887]

[0888]

[0889] (SEQ ID NO: 9705) (Source: NCBI Reference Sequence: XM_011543601.1)

[0890] In various embodiments, the SMN2 mRNA transcript comprises the sequence provided as SEQ ID NO:9706.

[0891]

[0892] (SEQ ID NO: 9706) (Source: NCBI Reference Sequence: XM_011543599.1)

[0893] In various embodiments, the SMN2 mRNA transcript comprises the sequence provided as SEQ ID NO:9707.

[0894]

[0895]

[0896] (SEQ ID NO: 9707) (Source: NCBI Reference Sequence: XM_017009787.1)

[0897] In various embodiments, the SMN2 transcript is an SMN2 pre-mRNA transcript. In various embodiments, the SMN2 pre-mRNA transcript comprises the sequence provided as SEQ ID NO: 9708. The NCBI Reference Sequence NG_008728.1 is SEQ ID NO: 9708.

[0898] In various embodiments, the SMN2 pre-mRNA transcript comprises the sequence provided as SEQ ID NO: 9709. NCBI Reference Sequence NC_000005.10 is SEQ ID NO:9709.

[0899] In various embodiments, the SMN2 transcript shares between 90%-100% identity with any one of SEQ ID NOs: 9698-9707, or shares between 90%-100% identity with an SMN2 pre-mRNA transcript comprising SEQ ID NO: 9708 or SEQ ID NO: 9709. In various embodiments, the SMN2 transcript shares at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with SEQ ID NOs: 9698-9707, or shares at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with an SMN2 pre-mRNA transcript transcribed from SEQ ID NO: 9708 or SEQ ID NO: 9709.

[0900] SMN2 oligonucleotides targeting the SMN2 transcript region

[0901] In various embodiments, the SMN2 AONs disclosed herein are complementary to a specific region of an SMN2 transcript (e.g., an SMN2 transcript) comprising a sequence that shares at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with any one of SEQ ID NOs: 9698-9709. In some embodiments, the SMN2 AONs comprise a sequence that is complementary to a specific region of an SMN2 transcript comprising a sequence that shares at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with any one of SEQ ID NOs: 9698-9709. In some embodiments, the SMN2 AONs comprise a sequence that is at least 85% complementary to a specific region of an SMN2 transcript. In some embodiments, the SMN2 AON comprises a sequence that is at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 100% complementary to a specific region of the SMN2 transcript. In some embodiments, the SMN2 AON comprises a sequence that is 90% to 99% complementary to a specific region of the SMN2 transcript. In some embodiments, the SMN2 AON comprises a sequence that is 90% to 95% complementary to a specific region of the SMN2 transcript. In some embodiments, the SMN2 AON comprises a sequence that is 95% to 99% complementary to a specific region of the SMN2 transcript.

[0902] In some embodiments, an SMN2 AON (e.g., an SMN2 AON) has a segment comprising up to 7 linked nucleosides. In some embodiments, an SMN2 AON has a segment comprising up to 6, 5, 4, 3, or 2 linked nucleosides. A segment of an SMN2 AON can be separated from other segments of an SMN2 AON by a spacer. A segment of an SMN2 AON is complementary to a specific region of an SMN2 transcript (e.g., an SMN2 transcript) comprising a sequence that shares at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with any one of SEQ ID NOs: 9698-9709.

[0903] SMN2 oligonucleotide variants

[0904] In various embodiments, the SMN2 AON comprises different variants, hereinafter referred to as SMN2 AON variants. The SMN2 AON variant can be an oligonucleotide sequence of 5 to 100 nucleobases in length, such as 10 to 40 nucleobases in length, such as 14 to 40 nucleobases in length, 10 to 30 nucleobases in length, such as 14 to 30 nucleobases in length, such as 16 to 28 nucleobases in length, such as 19 to 23 nucleobases in length, such as 21 to 23 nucleobases in length, or such as 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleobases in length. The SMN2 AON variant can be an oligonucleotide sequence that is complementary to a portion of the SMN2 pre-mRNA sequence or the SMN2 gene sequence. Example SMN2 AON variants include a nucleobase sequence selected from any one of SEQ ID NOs: 9710-10141.

[0905] In various embodiments, the SMN2 AON variant represents a modified version of the corresponding SMN2 parent oligonucleotide. In some embodiments, the SMN2 AON variant comprises a nucleobase sequence that represents a shortened version of the SMN2 parent AON nucleobase sequence. As an example, if the SMN2 parent oligonucleotide comprises a 25-mer (e.g., 25 nucleotide bases in length), the variant (e.g., SMN2 variant) may comprise a shorter version (e.g., a 15-mer, 17-mer, 19-mer, 21-mer, or 23-mer of the 25-mer SMN2 parent oligonucleotide). In one embodiment, the nucleobase sequence of the SMN2 AON variant differs from the corresponding nucleobase sequence of the SMN2 parent oligonucleotide in that 1, 2, 3, 4, 5, or 6 nucleotide bases are removed from one or both of the 5' and 3' ends of the nucleobase sequence of the SMN2 parent oligonucleotide. In one embodiment, the corresponding SMN2 AON variant may include a 23-mer wherein two nucleotide bases are removed from one of the 3' end or the 5' end of the 25-mer contained in the SMN2 parent oligonucleotide. In one embodiment, the corresponding SMN2 AON variant may include a 23-mer wherein one nucleotide base is removed from each of the 3' end and the 5' end of the 25-mer contained in the SMN2 parent oligonucleotide. In one embodiment, the corresponding SMN2 AON variant may include a 21-mer wherein two nucleotide bases are removed from each of the 3' end and the 5' end of the 25-mer contained in the SMN2 parent oligonucleotide. In one embodiment, the corresponding SMN2 AON variant may include a 21-mer wherein four nucleotide bases are removed from the 3' end or the 5' end of the 25-mer contained in the SMN2 parent oligonucleotide. In one embodiment, the corresponding SMN2 AON variant may include a 19-mer wherein three nucleotide bases are removed from each of the 3' end and the 5' end of the 25-mer contained in the SMN2 parent oligonucleotide. In one embodiment, the corresponding SMN2 AON variant may comprise a 19-mer in which six nucleotide bases are removed from either the 3' end or the 5' end of the 25-mer comprised in the SMN2 parent oligonucleotide.

[0906] Example sequences of SMN2 AON variants are shown in Table 11 below.

[0907] Table 11. Sequences of SMN2 oligonucleotide variants complementary to sequence SMN2 transcripts

[0908]

[0909]

[0910]

[0911]

[0912]

[0913]

[0914]

[0915]

[0916]

[0917]

[0918]

[0919]

[0920]

[0921]

[0922]

[0923]

[0924]

[0925]

[0926]

[0927]

[0928]

[0929]

[0930] *The at least one nucleoside linkage of the nucleobase sequence is selected from the group consisting of phosphorothioate linkages, alkylphosphate linkages, phosphorodithioate linkages, phosphotriester linkages, alkylphosphonate linkages, 3-methoxypropylphosphonate linkages, methylphosphonate linkages, aminoalkylphosphotriester linkages, alkylenephosphonate linkages, phosphinate linkages, phosphoamidate linkages, phosphorothioateamidate linkages, phosphorodiamidate linkages, phosphorodiamidate (e.g., comprising phosphorodiamidate morpholino (PMO), 3' aminoribose, or 5' aminoribose) linkages, aminoalkylphosphoamidate linkages, phosphorothioateamidate linkages, thioalkylphosphonate linkages, thioalkylphosphotriester linkages, phosphorothioate linkages, phosphoroselenoate linkages, and phosphoroboronate linkages.

[0931] Antisense oligonucleotides with one or more locked nucleic acids (LNA)

[0932] In various embodiments, the antisense oligonucleotides disclosed herein (e.g., AON parent oligonucleotides and / or AON oligonucleotide variants) comprise one or more locked nucleic acids (LNAs). In specific embodiments, the antisense oligonucleotide comprises one LNA. In specific embodiments, the antisense oligonucleotide comprises two LNAs. In specific embodiments, the antisense oligonucleotide comprises three LNAs. In general, LNA refers to a nucleic acid monomer having a bridge (e.g., a methylene, ethylene, aminooxy, or oxyimino bridge) connecting two carbon atoms between the 4' and 2' positions of a nucleoside sugar unit to form a bicyclic sugar. In general, LNA includes a nucleobase (e.g., one of A, C, G, or T). In various embodiments, LNAs with a cytosine nucleobase include modified cytosine nucleobases (e.g., 5-methylcytosine nucleobases).

[0933] In some embodiments, if counting from 5' to 3', the antisense oligonucleotides disclosed herein include an LNA at the 4th position of the antisense oligonucleotide. In some embodiments, if counting from 5' to 3', the antisense oligonucleotides disclosed herein include an LNA at the 7th position of the antisense oligonucleotide. In some embodiments, if counting from 5' to 3', the antisense oligonucleotides disclosed herein include an LNA at the 9th position of the antisense oligonucleotide. In some embodiments, if counting from 5' to 3', the antisense oligonucleotides disclosed herein include an LNA at the 12th position of the antisense oligonucleotide. In some embodiments, if counting from 5' to 3', the antisense oligonucleotides disclosed herein include an LNA at the 15th position of the antisense oligonucleotide. In some embodiments, if counting from 5' to 3', the antisense oligonucleotides disclosed herein include an LNA at the 17th position of the antisense oligonucleotide. In some embodiments, if counting from 5' to 3', the antisense oligonucleotides disclosed herein include an LNA at the 20th position of the antisense oligonucleotide.

[0934] In various embodiments, the antisense oligonucleotides disclosed herein (e.g., SMN2 parent oligonucleotides and / or SMN2 oligonucleotide variants) comprise two LNAs located at two different positions of the antisense oligonucleotide. In some embodiments, if counting from 5' to 3', the antisense oligonucleotides disclosed herein include a first LNA at the fourth position of the antisense oligonucleotide and a second LNA at the 20th position of the antisense oligonucleotide. In some embodiments, if counting from 5' to 3', the antisense oligonucleotides disclosed herein include a first LNA at the 7th position of the antisense oligonucleotide and a second LNA at the 15th position of the antisense oligonucleotide. In some embodiments, if counting from 5' to 3', the antisense oligonucleotides disclosed herein include a first LNA at the 7th position of the antisense oligonucleotide and a second LNA at the 17th position of the antisense oligonucleotide. In some embodiments, if counting from 5' to 3', the antisense oligonucleotides disclosed herein include a first LNA at the 9th position of the antisense oligonucleotide and a second LNA at the 17th position of the antisense oligonucleotide.

[0935] In various embodiments, the antisense oligonucleotides disclosed herein (e.g., SMN2 parent oligonucleotides and / or SMN2 oligonucleotide variants) comprise three LNAs located at three different positions of the antisense oligonucleotide. In some embodiments, if counting from 5' to 3', the antisense oligonucleotides disclosed herein include a first LNA at the 4th position of the antisense oligonucleotide, a second LNA at the 12th position of the antisense oligonucleotide, and a third LNA at the 20th position of the antisense oligonucleotide.

[0936] Antisense oligonucleotides with one or more spacers

[0937] In various embodiments, the antisense oligonucleotide comprises one or more spacers. In a specific embodiment, the antisense oligonucleotide comprises one spacer. In a specific embodiment, the antisense oligonucleotide comprises two spacers. In a specific embodiment, the antisense oligonucleotide comprises three spacers. Generally speaking, a spacer refers to a nucleoside replacement group lacking a nucleotide base, and wherein the nucleoside sugar moiety is replaced by a non-sugar substituent. The non-sugar substituent cannot be connected to the core base, but can be connected to the 3' and 5' positions of the nucleoside adjacent to the spacer through an internucleoside linking group.

[0938] In certain embodiments, an oligonucleotide having one or more spacers (such as disclosed herein) can be an oligonucleotide of 5 to 100 oligonucleotide units in length, e.g., 10 to 60 oligonucleotide units in length, e.g., 12 to 50 oligonucleotide units in length, 14 to 40 oligonucleotide units in length, 10 to 30 oligonucleotide units in length, e.g., 14 to 30 oligonucleotide units in length, e.g., 14 to 25 or 15 to 22 oligonucleotide units in length, or 18, 19, 20, 21, 22, 23, 24, or 25 oligonucleotide units in length. As used herein, an "oligonucleotide unit" refers to a nucleoside (e.g., a nucleoside comprising a sugar and / or a nucleobase) or a nucleoside replacement group (e.g., a spacer) of an oligonucleotide.

[0939] In a specific embodiment, the oligonucleotide having one or more spacers is 25 oligonucleotide units in length. In a specific embodiment, the oligonucleotide having one or more spacers is 23 oligonucleotide units in length. In a specific embodiment, the oligonucleotide having one or more spacers is 21 oligonucleotide units in length. In a specific embodiment, the oligonucleotide having one or more spacers is 19 oligonucleotide units in length. In various embodiments, the oligonucleotide having one or more spacers is at least 18, at least 19, at least 20, at least 21, at least 22, at least 23, at least 24, or at least 25 oligonucleotide units in length. In various embodiments, the oligonucleotide having one or more spacers is at least 18 oligonucleotide units in length. In various embodiments, the oligonucleotide having one or more spacers is at least 19 oligonucleotide units in length. In various embodiments, the oligonucleotide having one or more spacers is at least 20 oligonucleotide units in length. In various embodiments, the oligonucleotide having one or more spacers is at least 21 oligonucleotide units in length. In various embodiments, the oligonucleotide having one or more spacers is at least 22 oligonucleotide units in length. In various embodiments, the oligonucleotide having one or more spacers is at least 23 oligonucleotide units in length. In various embodiments, the oligonucleotide having one or more spacers is at least 24 oligonucleotide units in length. In various embodiments, the oligonucleotide having one or more spacers is at least 25 oligonucleotide units in length.

[0940] In various embodiments, the STMN2 AON comprises a sequence that shares at least 80% identity with an equal length portion of any one of SEQ ID NOs: 1451-1664. In various embodiments, the STMN2 AON comprises a sequence that shares at least 85% identity with an equal length portion of any one of SEQ ID NOs: 1451-1664. In various embodiments, the STMN2 AON comprises a sequence that shares at least 90% identity with an equal length portion of any one of SEQ ID NOs: 1451-1664. In various embodiments, the STMN2 AON comprises a sequence that shares at least 95% identity with an equal length portion of any one of SEQ ID NOs: 1451-1664. In various embodiments, the STMN2 AON comprises a sequence that shares 100% identity with an equal length portion of any one of SEQ ID NOs: 1451-1664.

[0941] In various embodiments, the KCNQ2 AON comprises a sequence that shares at least 80% identity with an equal length portion of any one of SEQ ID NOs: 4402-4468, SEQ ID NOs: 4469-4476, and SEQ ID NOs: 4477-4530. In various embodiments, the KCNQ2 AON comprises a sequence that shares at least 85% identity with an equal length portion of any one of SEQ ID NOs: 4402-4468, SEQ ID NOs: 4469-4476, and SEQ ID NOs: 4477-4530. In various embodiments, the KCNQ2 AON comprises a sequence that shares at least 90% identity with an equal length portion of any one of SEQ ID NOs: 4402-4468, SEQ ID NOs: 4469-4476, and SEQ ID NOs: 4477-4530. In various embodiments, the KCNQ2 AON comprises a sequence that shares at least 95% identity with an equal length portion of any one of SEQ ID NOs: 4402-4468, SEQ ID NOs: 4469-4476, and SEQ ID NOs: 4477-4530. In various embodiments, the KCNQ2 AON comprises a sequence that shares 100% identity with an equal length portion of any one of SEQ ID NOs: 4402-4468, SEQ ID NOs: 4469-4476, and SEQ ID NOs: 4477-4530.

[0942] In various embodiments, the UNC13A AON comprises a sequence that shares at least 80% identity with an equal length portion of any one of SEQ ID NOs: 9596-9696 or SEQ ID NOs: 10670-10779. In various embodiments, the UNC13A AON comprises a sequence that shares at least 85% identity with an equal length portion of any one of SEQ ID NOs: 9596-9696 or SEQ ID NOs: 10670-10779. In various embodiments, the UNC13A AON comprises a sequence that shares at least 90% identity with an equal length portion of any one of SEQ ID NOs: 9596-9696 or SEQ ID NOs: 10670-10779. In various embodiments, the UNC13A AON comprises a sequence that shares at least 95% identity with an equal length portion of any one of SEQ ID NOs: 9596-9696 or SEQ ID NOs: 10670-10779. In various embodiments, the UNC13A AON comprises a sequence that shares 100% identity with an equal length portion of any one of SEQ ID NOs: 9596-9696 or SEQ ID NOs: 10670-10779.

[0943] In various embodiments, the SMN2 AON comprises a sequence that shares at least 80% identity with an equal length portion of any one of SEQ ID NOs: 10574-10640 and SEQ ID NOs: 10644-10651. In various embodiments, the SMN2 AON comprises a sequence that shares at least 85% identity with an equal length portion of any one of SEQ ID NOs: 10574-10640 and SEQ ID NOs: 10644-10651. In various embodiments, the SMN2 AON comprises a sequence that shares at least 90% identity with an equal length portion of any one of SEQ ID NOs: 10574-10640 and SEQ ID NOs: 10644-10651. In various embodiments, the SMN2 AON comprises a sequence that shares at least 95% identity with an equal length portion of any one of SEQ ID NOs: 10574-10640 and SEQ ID NOs: 10644-10651. In various embodiments, the SMN2 AON comprises a sequence that shares at least 99% identity with an equal length portion of any one of SEQ ID NOs: 10574-10640 and SEQ ID NOs: 10644-10651. In various embodiments, the SMN2 AON comprises a sequence that shares 100% identity with an equal length portion of any one of SEQ ID NOs: 10574-10640 and SEQ ID NOs: 10644-10651.

[0944] In some embodiments, the spacer has formula (X):

[0945]

[0946] wherein Ring A is as defined herein.

[0947] In some embodiments, the spacer has formula (Xa):

[0948]

[0949] wherein Ring A is as defined herein and the -CH2-O- group is located on the Ring A atom adjacent to the -O- group.

[0950] As generally defined herein, Ring A of Formula (X) and (Xa) is an optionally substituted 4-8 membered monocyclic cycloalkyl (e.g., Ring A is cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl) or a 4-8 membered monocyclic heterocyclyl, wherein the heterocyclyl contains 1 or 2 heteroatoms selected from O, S, and N (e.g., Ring A is oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, 1,4-dioxanyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, azepanyl). In some embodiments, Ring A is tetrahydrofuranyl. In some embodiments, Ring A is tetrahydropyranyl. In some embodiments, Ring A is pyrrolidinyl. In some embodiments, Ring A is cyclopentyl. In some embodiments, the monocyclic cycloalkyl or monocyclic heterocyclyl is not further substituted. In some embodiments, the cycloalkyl or heterocyclyl is further substituted with 0, 1, 2, or 3 substituents selected from halogen (e.g., -F, -Cl), -OMe, -OEt-O(CH2)OMe, -O(CH2)2OMe, and CN.

[0951] In some embodiments, the tetrahydrofuranyl group is substituted with 1 or 2 substituents selected from halogen (e.g., -F, -Cl), -OMe, -OEt-O(CH2)OMe, -O(CH2)2OMe, and CN. In some embodiments, the tetrahydrofuranyl group is substituted with 2 substituents selected from halogen (e.g., -F, -Cl), -OMe, -OEt-O(CH2)OMe, -O(CH2)2OMe, and CN. In some embodiments, the tetrahydrofuranyl group is substituted with 1 substituent selected from halogen (e.g., -F, -Cl), -OMe, -OEt-O(CH2)OMe, -O(CH2)2OMe, and CN. In some embodiments, the tetrahydrofuranyl group is substituted with -O(CH2)2OMe.

[0952] In some embodiments, the spacer is represented by formula (I), wherein:

[0953]

[0954] X is selected from -CH2- and -O-; and

[0955] n is 0, 1, 2 or 3.

[0956] In some embodiments, the spacer is represented by formula (I'), wherein:

[0957]

[0958] X is selected from -CH2- and -O-; and

[0959] n is 0, 1, 2 or 3.

[0960] In some embodiments, the spacer is represented by Formula (Ia), wherein:

[0961]

[0962] And n is 0, 1, 2 or 3.

[0963] In some embodiments, the spacer is represented by formula (Ia'), wherein:

[0964]

[0965] And n is 0, 1, 2 or 3.

[0966] As generally defined herein, X is selected from -CH2- and -O-. In some embodiments, X is -CH2-. In other embodiments, X is -O-.

[0967] As generally defined herein, n is 0, 1, 2, or 3. In some embodiments, n is 0. In some embodiments, n is 1 or 2. In some embodiments, n is 1. In other embodiments, n is 2. In certain embodiments, n is 3.

[0968] In some embodiments, the spacer is represented by formula (II), wherein:

[0969]

[0970] X is selected from -CH2- and -O-.

[0971] In some embodiments, the spacer is represented by formula (II'), wherein:

[0972]

[0973] X is selected from -CH2- and -O.

[0974] In some embodiments, the spacer is represented by formula (Iia), wherein:

[0975]

[0976] In some embodiments, the spacer is represented by formula (Iia'), wherein:

[0977]

[0978] In some embodiments, the spacer is represented by formula (IIIb), wherein:

[0979]

[0980] In some embodiments, the spacer is represented by formula (IIIb'), wherein:

[0981]

[0982] In some embodiments, the spacer is represented by formula (III), wherein:

[0983]

[0984] X is selected from -CH2- and -O-.

[0985] In some embodiments, the spacer is represented by formula (III'), wherein:

[0986]

[0987] X is selected from -CH2- and -O.

[0988] In some embodiments, the spacer is represented by formula (IIIa), wherein:

[0989]

[0990] In some embodiments, the spacer is represented by formula (IIIa'), wherein:

[0991]

[0992] In some embodiments, the open positions (i.e., positions not specifically described as bearing only hydrogen atoms, -CH- groups comprising X) of Formulas (I), (I'), (Ia), (Ia'), (II), (II'), (Iia), (Iia'), (III), (III'), (IIIa), and (IIIa') are further substituted with 0-3 substituents selected from halogen (e.g., -F, -Cl), -OMe, -OEt, -O(CH)OMe, -O(CH)OMe, and CN. In some embodiments, Formulas (I), (I'), (Ia), (Ia'), (II), (II'), (Iia), (Iia'), (III), (III'), (IIIa), and (IIIa') are not further substituted.

[0993] STMN2

[0994] As further described below, STMN2 oligonucleotides having one or more spacers are described with reference to the corresponding STMN2 parent oligonucleotide. In various embodiments, the STMN2 oligonucleotide having a spacer differs from the STMN2 parent oligonucleotide in that the spacer replaces a nucleoside in the STMN2 parent oligonucleotide. As used below, "position" of an STMN2 oligonucleotide refers to a specific position counted from the 5' end of the STMN2 oligonucleotide. In various embodiments, the spacer replaces a nucleoside at any one of positions 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 of the STMN2 parent oligonucleotide. In specific embodiments, the spacer replaces a nucleoside at one of positions 7, 8, 11, 14, 16, 19, or 22 of the STMN2 parent oligonucleotide.

[0995] In various embodiments, the STMN2 oligonucleotide includes a spacer that replaces a nucleoside in the STMN2 parent oligonucleotide (e.g., a spacer replaces a nucleoside in the STMN2 parent oligonucleotide). In specific embodiments, the spacer replaces the nucleoside between positions 9 and 15 of the STMN2 parent oligonucleotide. In specific embodiments, the spacer replaces the nucleoside between positions 9 and 12 of the STMN2 parent oligonucleotide. In specific embodiments, the spacer replaces the nucleoside at position 10 of the STMN2 parent oligonucleotide. In specific embodiments, the spacer replaces the nucleoside at position 11 of the STMN2 parent oligonucleotide. In specific embodiments, the spacer replaces the nucleoside at position 12 of the STMN2 parent oligonucleotide. In specific embodiments, the spacer replaces the nucleoside between positions 12 and 16 of the STMN2 parent oligonucleotide. In specific embodiments, the spacer replaces the nucleoside at position 15 of the STMN2 parent oligonucleotide.

[0996] In various embodiments, the STMN2 oligonucleotide comprising a spacer has two segments, wherein at least one of the two segments has at most 11 connected nucleosides. For example, the length of the STMN2 oligonucleotide can be 23 oligonucleotide units, and the spacer can be located at position 12. Therefore, the STMN2 oligonucleotide has two segments separated by a spacer, wherein both of the two segments have a length of 11 nucleobases. In various embodiments, the STMN2 oligonucleotide comprising a spacer has two segments, wherein at least one of the two segments has at most 10 connected nucleosides. For example, the length of the STMN2 oligonucleotide can be 21 oligonucleotide units, and the spacer can be located at position 11. Therefore, the STMN2 oligonucleotide has two segments separated by a spacer, wherein both of the two segments have a length of 10 nucleobases. As another example, the length of the STMN2 oligonucleotide can be 25 oligonucleotide units, and the spacer can be located at position 15. Thus, the STMN2 oligonucleotide has 2 segments separated by a spacer, wherein one of the 2 segments is 14 nucleobases in length and the second of the 2 segments is 10 nucleobases in length.

[0997] In various embodiments, the STMN2 oligonucleotide comprises two spacers, each of which replaces a nucleoside in the STMN2 parent oligonucleotide (e.g., two spacers replace two separate nucleosides in the STMN2 parent oligonucleotide). In various embodiments, the first spacer and the second spacer are separated by at least 5 nucleobases, at least 6 nucleobases, at least 7 nucleobases, at least 8 nucleobases, at least 9 nucleobases, or at least 10 nucleobases in the oligonucleotide. In specific embodiments, the first spacer and the second spacer are separated by at least 5 nucleobases, at least 6 nucleobases, or at least 7 nucleobases. In specific embodiments, the first spacer and the second spacer are not adjacent to each other in the oligonucleotide.

[0998] In specific embodiments, the first spacer replaces the nucleosides between positions 7 and 11 of the STMN2 parent oligonucleotide. In various embodiments, the first spacer replaces the nucleosides between positions 8 and 11, positions 9 and 11, positions 10 and 11, positions 7 and 10, positions 7 and 9, positions 7 and 8, positions 8 and 10, positions 8 and 9, or positions 9 and 10 of the STMN2 parent oligonucleotide. In specific embodiments, the second spacer replaces the nucleosides between positions 14 and 22 of the STMN2 parent oligonucleotide. In various embodiments, the second spacer replaces the nucleosides at positions 15 and 22, 16 and 22, 17 and 22, 18 and 22, 19 and 22, 20 and 22, 21 and 22, 15 and 21, 16 and 21, 17 and 21, 18 and 21, 19 and 21, 20 and 21, 15 and 20, 16 and 20, 17 and 20, 18 and 20, 19 and 20, 15 and 19, 16 and 19, 17 and 19, 18 and 19, 15 and 18, 16 and 18, 17 and 18, 15 and 17, 16 and 17, or between positions 15 and 16 of the STMN2 parent oligonucleotide.

[0999] In a preferred embodiment, the first spacer replaces the nucleoside at position 7 of the STMN2 parent oligonucleotide and the second spacer replaces the nucleoside at position 14 of the STMN2 parent oligonucleotide. In a preferred embodiment, the first spacer replaces the nucleoside at position 8 of the STMN2 parent oligonucleotide and the second spacer replaces the nucleoside at position 16 of the STMN2 parent oligonucleotide. In a preferred embodiment, the first spacer replaces the nucleoside at position 11 of the STMN2 parent oligonucleotide and the second spacer replaces the nucleoside at position 22 of the STMN2 parent oligonucleotide. In a preferred embodiment, the first spacer replaces the nucleoside at position 9 of the STMN2 parent oligonucleotide and the second spacer replaces the nucleoside at position 19 of the STMN2 parent oligonucleotide.

[1000] In various embodiments, the STMN2 oligonucleotide comprises three spacers, each of which replaces a nucleoside in a STMN2 parent oligonucleotide (e.g., three spacers replace three separate nucleosides of a STMN2 parent oligonucleotide). In specific embodiments, a first spacer replaces a nucleoside between positions 7 and 11 of the STMN2 parent oligonucleotide. In specific embodiments, a second spacer replaces a nucleoside between positions 14 and 22 of the STMN2 parent oligonucleotide. In specific embodiments, a third spacer replaces a nucleoside between positions 21 and 24 of the STMN2 parent oligonucleotide. In some embodiments, a first spacer replaces a nucleoside between positions 2 and 5 of the STMN2 parent oligonucleotide. In specific embodiments, a second spacer replaces a nucleoside between positions 8 and 12 of the STMN2 parent oligonucleotide. In specific embodiments, a third spacer replaces a nucleoside between positions 18 and 22 of the STMN2 parent oligonucleotide.

[1001] In various embodiments, the three spacers in the STMN2 oligonucleotide are positioned so that each of the four segments of the STMN2 oligonucleotide is at most 7 linked nucleosides in length. For example, the STMN2 oligonucleotide can have a first segment having 7 linked nucleosides linked to a first spacer, followed by a second segment having 7 linked nucleosides linked at one end to the first spacer and at the other end to the second spacer, followed by a third segment having 6 linked nucleosides linked at one end to the second spacer and at the other end to the third spacer, followed by a fourth segment having 6 linked nucleosides linked to the third spacer.

[1002] In various embodiments, one or more spacers are positioned in the oligonucleotide to replace one or more adenosine or thymidine (relative to guanine or cytidine). For example, one or more spacers can replace one, two, three, four, five, six, seven, eight or nine adenosine or thymidine in the oligonucleotide. In various embodiments, one or more spacers are positioned in the oligonucleotide to replace one or more guanine or cytidine (relative to adenosine or thymidine). For example, one or more spacers can replace one, two, three, four, five, six, seven, eight or nine guanine or cytidine in the oligonucleotide. In various embodiments, spacers are positioned in the oligonucleotide to replace the same amount of adenosine / thymidine and guanine / cytidine. For example, the first spacer in the oligonucleotide can replace adenosine / thymidine, and the second spacer in the oligonucleotide can replace guanine / cytidine.

[1003] In various embodiments, one or more spacers are positioned in the oligonucleotide to control the sequence content in the oligonucleotide.For example, one or more spacers are positioned so that at least one spacer is positioned near the guanine group.In various embodiments, the oligonucleotide with spacer can include one spacer adjacent to the guanine group, two spacers adjacent to the guanine group, three spacers adjacent to the guanine group, four spacers adjacent to the guanine group or five spacers adjacent to the guanine group.In one embodiment, if counting from the 5 ' end of the oligonucleotide, the spacer is immediately before the guanine group in the sequence.Therefore, in various embodiments, the oligonucleotide with spacer can include a spacer immediately before the guanine group, two spacers immediately before the guanine group respectively, three spacers immediately before the guanine group respectively, four spacers immediately before the guanine group respectively or five spacers immediately before the guanine group respectively.In one embodiment, if counting from the 5 ' end of the oligonucleotide, the guanine group is immediately followed by a spacer. Thus, in various embodiments, an oligonucleotide with a spacer can include one spacer immediately following the guanine group, two spacers each immediately following the guanine group, three spacers each immediately following the guanine group, four spacers each immediately following the guanine group, or five spacers each immediately following the guanine group. In various embodiments, the spacers in the oligonucleotide can be positioned so as to maximize the number of spacers adjacent to the guanine group.

[1004] In various embodiments, one or more spacers are positioned in an oligonucleotide to replace one or more adenosine or thymidine nucleosides such that the one or more spacers are positioned adjacent to a guanine group. For example, two spacers can replace an adenosine or thymidine nucleoside in an oligonucleotide, with each of the two spacers being positioned adjacent to a guanine group.

[1005] In various embodiments, the STMN2 oligonucleotides with one or more spacers have a specific GC content. As used herein, GC content (or guanine-cytosine content) is the percentage of nitrogenous bases that are guanine (G) or cytosine (C) in an oligonucleotide. In various embodiments, the STMN2 oligonucleotides with one or more spacers have at least 10% GC content, at least 20% GC content, at least 25% GC content, at least 30% GC content, at least 35% GC content, at least 40% GC content, at least 45% GC content, at least 50% GC content, at least 55% GC content, at least 60% GC content, at least 65% GC content, at least 75% GC content, at least 80% GC content, at least 85% GC content, at least 90% GC content, or at least 95% GC content. In specific embodiments, the STMN2 oligonucleotides with one or more spacers have at least 30% GC content. In specific embodiments, the STMN2 oligonucleotides with one or more spacers have at least 40% GC content. In various embodiments, one or more spacers are positioned in the STMN2 oligonucleotide to maximize GC content. For example, rather than selecting guanine or cytosine for replacement by a spacer, thymine or adenine in the STMN2 oligonucleotide can be selected for replacement by a spacer.

[1006] In various embodiments, the STMN2 oligonucleotide with a spacer is designed such that 1) each segment of the STMN2 oligonucleotide has at most 7 linked nucleosides, and 2) at least two, three, or four spacers are positioned adjacent to a guanine group. In some embodiments, the STMN2 oligonucleotide with a spacer is designed such that 1) each segment of the STMN2 oligonucleotide has at most 7 linked nucleosides, and 2) each of the two spacers is positioned before a guanine group.

[1007] In various embodiments, the inclusion of one or more spacers in the STMN2 oligonucleotide does not reduce the effectiveness of the STMN2 oligonucleotide with a spacer in restoring full-length STMN2 protein or full-length STMN2 mRNA, as compared to the effectiveness of the corresponding STMN2 parent oligonucleotide. In various embodiments, the inclusion of one or more spacers in the STMN2 oligonucleotide increases the effectiveness of the STMN2 oligonucleotide with a spacer in restoring full-length STMN2 protein or full-length STMN2 mRNA, as compared to the effectiveness of the corresponding STMN2 parent oligonucleotide. In various embodiments, the inclusion of one or more spacers in the STMN2 oligonucleotide does not reduce the effectiveness of the STMN2 oligonucleotide with a spacer in reducing the number of STMN2 transcripts with cryptic exons, as compared to the effectiveness of the corresponding STMN2 parent oligonucleotide. In various embodiments, the inclusion of one or more spacers in the STMN2 oligonucleotide increases the effectiveness of the STMN2 oligonucleotide with a spacer in reducing the number of STMN2 transcripts with cryptic exons, as compared to the effectiveness of the corresponding STMN2 parent oligonucleotide.

[1008] Table 12A, 12B, 13 and 14 record the example STMN2 oligonucleotide with one or more spacers and the relationship of these one or more spacers and corresponding STMN2 parent oligonucleotide.Each STMN2 oligonucleotide distributes sequence name.As used hereinafter, the nomenclature of sequence name is expressed as " X_spA " (for STMN2 AON with one spacer), " X_spA_spB " (for STMN2 AON with two spacers) or " X_spA_spB_spC " (for STMN2 AON with three spacers).Here, " X ​​" refers to the length of STMN2 AON, " A " refers to the position of the first spacer in STMN2 AON, " B " refers to the position of the second spacer in STMN2 AON, and if present, " C " refers to the position of the third spacer in STMN AON.

[1009] In various embodiments, the STMN2 oligonucleotide comprises a spacer. In various embodiments, the STMN2 oligonucleotide is an oligonucleotide variant, such as any of a 23-mer, a 21-mer, or a 19-mer. In various embodiments, the inclusion of a spacer divides the STMN2 oligonucleotide into two separate segments, wherein at least one of the segments is at most 11 linked nucleosides in length. In various embodiments, the inclusion of a spacer divides the STMN2 oligonucleotide into two separate segments, wherein at least one of the segments is at most 10 linked nucleosides in length.

[1010] In various embodiments, the spacer is located between positions 10 and 15 of the oligonucleotide. In various embodiments, the spacer is located between positions 10 and 12 of the oligonucleotide. In specific embodiments, the spacer is located at position 10 of the oligonucleotide. In specific embodiments, the spacer is located at position 11 of the oligonucleotide. In specific embodiments, the spacer is located at position 12 of the oligonucleotide. In specific embodiments, the spacer is located at position 15 of the oligonucleotide.

[1011] Example STMN2 AONs with one spacer are recorded below in Table 12A.

[1012] Table 12A: Identification of STMN2 AONs with one spacer. Here, each STMN2 AON has 2 segments, wherein at least one of the segments has a maximum of 11 linked nucleosides.

[1013]

[1014]

[1015] *At least one nucleoside linkage of the nucleobase sequence is selected from the group consisting of phosphorothioate linkages, alkylphosphate linkages, phosphorodithioate linkages, phosphotriester linkages, alkylphosphonate linkages, 3-methoxypropylphosphonate linkages, methylphosphonate linkages, aminoalkylphosphotriester linkages, alkylenephosphonate linkages, phosphinate linkages, phosphoamidate linkages, phosphorothioateamidate linkages, phosphorodiamidate (e.g., comprising phosphorodiamidate morpholino (PMO), 3' aminoribose, or 5' aminoribose) linkages, aminoalkylphosphoamidate linkages, phosphorothioateamidate linkages, thioalkylphosphonate linkages, thioalkylphosphotriester linkages, phosphorothioate linkages, phosphoroselenoate linkages, and phosphoroboronate linkages.

[1016] In various embodiments, the STMN2 oligonucleotide comprises two spacers. In various embodiments, the inclusion of a spacer divides the STMN2 oligonucleotide into three separate segments, wherein at least one of the segments is at most 7 linked nucleosides in length. Example STMN2 AONs with two spacers are recorded in Table 12B below.

[1017] Table 12B: Identification of STMN2 AONs with two spacers. Here, each STMN2 AON has three segments, wherein at least one of the segments has up to 7 linked nucleosides.

[1018]

[1019]

[1020]

[1021]

[1022]

[1023] *At least one nucleoside linkage of the nucleobase sequence is selected from the group consisting of phosphorothioate linkages, alkylphosphate linkages, phosphorodithioate linkages, phosphotriester linkages, alkylphosphonate linkages, 3-methoxypropylphosphonate linkages, methylphosphonate linkages, aminoalkylphosphotriester linkages, alkylenephosphonate linkages, phosphinate linkages, phosphoamidate linkages, phosphorothioateamidate linkages, phosphorodiamidate (e.g., comprising phosphorodiamidate morpholino (PMO), 3' aminoribose, or 5' aminoribose) linkages, aminoalkylphosphoamidate linkages, phosphorothioateamidate linkages, thioalkylphosphonate linkages, thioalkylphosphotriester linkages, phosphorothioate linkages, phosphoroselenoate linkages, and phosphoroboronate linkages.

[1024] In various embodiments, the STMN2 oligonucleotide comprises three spacers. The inclusion of the three spacers divides the STMN2 oligonucleotide into four separate segments. In various embodiments, the three spacers are located at different positions in the STMN2 oligonucleotide such that each segment of the STMN2 oligonucleotide is at most 7 linked nucleosides in length.

[1025] In various embodiments, the length of the STMN2 AON with three spacers is 27 oligonucleotide units. For example, the STMN2 AON with three spacers can be divided into four separate segments. In various embodiments, the length of each of the four separate segments is 6 oligonucleotide units. For example, the STMN2 AON can be as follows (from the 5' end to the 3' end): a first segment of 6 connected nucleosides, a first spacer, a second segment of 6 connected nucleosides, a second spacer, a third segment of 6 connected nucleosides, a third spacer, and a fourth segment of 6 connected nucleosides.

[1026] Examples of STMN2 AONs with three spacers are recorded in Table 13 below.

[1027] Table 13: Identification of STMN2 AONs or AON variants with three spacers. Here, each STMN2 AON has four segments, wherein each segment has up to seven linked nucleosides.

[1028]

[1029]

[1030]

[1031] *At least one nucleoside linkage of the nucleobase sequence is selected from the group consisting of phosphorothioate linkages, alkylphosphate linkages, phosphorodithioate linkages, phosphotriester linkages, alkylphosphonate linkages, 3-methoxypropylphosphonate linkages, methylphosphonate linkages, aminoalkylphosphotriester linkages, alkylenephosphonate linkages, phosphinate linkages, phosphoamidate linkages, phosphorothioateamidate linkages, phosphorodiamidate (e.g., comprising phosphorodiamidate morpholino (PMO), 3' aminoribose, or 5' aminoribose) linkages, aminoalkylphosphoamidate linkages, phosphorothioateamidate linkages, thioalkylphosphonate linkages, thioalkylphosphotriester linkages, phosphorothioate linkages, phosphoroselenoate linkages, and phosphoroboronate linkages.

[1032] In various embodiments, the length of the STMN2 AON with one or more spacers is reduced compared to the STMN2 AON described in Tables 12B and 13 above. For example, such STMN2 AONs can be STMN2 oligonucleotide variants with one or more spacers. In various embodiments, the STMN2 oligonucleotide variant with one or more spacers is a 23-mer, a 21-mer, or a 19-mer. In various embodiments, the STMN2 oligonucleotide variant comprises two spacers, such that the STMN2 oligonucleotide variant comprises three segments separated by two spacers. In various embodiments, at least one of the three segments has no more than 7 linked nucleosides. In various embodiments, each of the three segments has no more than 7 linked nucleosides. Example STMN2 oligonucleotide variants with one or more spacers are shown in Table 14 below.

[1033] Table 14: STMN2 AON or AON variants with three spacers. Here, each STMN2 AON variant has three segments, wherein each segment has up to 7 linked nucleosides.

[1034]

[1035]

[1036]

[1037] *At least one nucleoside linkage of the nucleobase sequence is selected from the group consisting of phosphorothioate linkages, alkylphosphate linkages, phosphorodithioate linkages, phosphotriester linkages, alkylphosphonate linkages, 3-methoxypropylphosphonate linkages, methylphosphonate linkages, aminoalkylphosphotriester linkages, alkylenephosphonate linkages, phosphinate linkages, phosphoamidate linkages, phosphorothioateamidate linkages, phosphorodiamidate (e.g., comprising phosphorodiamidate morpholino (PMO), 3' aminoribose, or 5' aminoribose) linkages, aminoalkylphosphoamidate linkages, phosphorothioateamidate linkages, thioalkylphosphonate linkages, thioalkylphosphotriester linkages, phosphorothioate linkages, phosphoroselenoate linkages, and phosphoroboronate linkages.

[1038] In some embodiments, the antisense oligonucleotides disclosed herein (e.g., STMN2 parent oligonucleotides and / or STMN2 oligonucleotide variants) comprise one or more spacers and one or more locked nucleic acids (LNAs). In some embodiments, the antisense oligonucleotides disclosed herein (e.g., STMN2 parent oligonucleotides and / or STMN2 oligonucleotide variants) comprise two spacers and two LNAs. In some embodiments, the antisense oligonucleotides disclosed herein (e.g., STMN2 parent oligonucleotides and / or STMN2 oligonucleotide variants) comprise two spacers and three LNAs.

[1039] In various embodiments, the spacer and LNA are positioned adjacent to each other in the antisense oligonucleotide. For example, if counting from 5' to 3', the LNA can be located at the 7th position of the antisense oligonucleotide, and the spacer can be located at the 8th position of the antisense oligonucleotide. As another example, if counting from 5' to 3', the LNA can be located at the 9th position of the antisense oligonucleotide, and the spacer can be located at the 8th position of the antisense oligonucleotide. As another example, if counting from 5' to 3', the LNA can be located at the 15th position of the antisense oligonucleotide, and the spacer can be located at the 16th position of the antisense oligonucleotide. As another example, if counting from 5' to 3', the LNA can be located at the 17th position of the antisense oligonucleotide, and the spacer can be located at the 16th position of the antisense oligonucleotide.

[1040] In certain embodiments, in the antisense oligonucleotide, the first spacer is located adjacent to the first LNA, and the second spacer is located adjacent to the second LNA. For example, if counting from 5' to 3', the first LNA may be located at the 7th position of the antisense oligonucleotide, the first spacer may be located at the 8th position of the antisense oligonucleotide, the second LNA may be located at the 15th position of the antisense oligonucleotide, and the second spacer may be located at the 16th position of the antisense oligonucleotide. As another example, if counting from 5' to 3', the first LNA may be located at the 7th position of the antisense oligonucleotide, the first spacer may be located at the 8th position of the antisense oligonucleotide, the second LNA may be located at the 17th position of the antisense oligonucleotide, and the second spacer may be located at the 16th position of the antisense oligonucleotide. As another example, if counting from 5' to 3', the first LNA may be located at the 9th position of the antisense oligonucleotide, the first spacer may be located at the 8th position of the antisense oligonucleotide, the second LNA may be located at the 17th position of the antisense oligonucleotide, and the second spacer may be located at the 16th position of the antisense oligonucleotide.

[1041] In various embodiments, the one or more spacers and the one or more LNAs are not positioned adjacent to each other in the antisense oligonucleotide. For example, if counting from 5' to 3', the LNA can be located at the 4th position of the antisense oligonucleotide, and the spacer can be located at the 8th position of the antisense oligonucleotide. As an example, if counting from 5' to 3', the LNA can be located at the 20th position of the antisense oligonucleotide, and the spacer can be located at the 16th position of the antisense oligonucleotide. In a specific embodiment, if counting from 5' to 3', the first LNA can be located at the 4th position of the antisense oligonucleotide, the first spacer can be located at the 8th position of the antisense oligonucleotide, the second LNA can be located at the 20th position of the antisense oligonucleotide, and the second spacer can be located at the 16th position of the antisense oligonucleotide. In a specific embodiment, if counting from 5' to 3', the first LNA can be located at the 4th position of the antisense oligonucleotide, the first spacer can be located at the 8th position of the antisense oligonucleotide, the second LNA can be located at the 12th position of the antisense oligonucleotide, the second spacer can be located at the 16th position of the antisense oligonucleotide, and the third LNA can be located at the 20th position of the antisense oligonucleotide.

[1042] KCNQ2

[1043] As further described below, KCNQ2 oligonucleotides having one or more spacers are described with reference to the corresponding KCNQ2 parent oligonucleotide or KCNQ2 variant oligonucleotide. In various embodiments, the KCNQ2 oligonucleotide having a spacer differs from the KCNQ2 parent oligonucleotide or KCNQ2 variant oligonucleotide in that the spacer replaces a nucleoside in the KCNQ2 parent oligonucleotide or KCNQ2 variant oligonucleotide. As used below, a "position" of a KCNQ2 oligonucleotide refers to a specific position counted from the 5' end of the KCNQ2 oligonucleotide. In various embodiments, the spacer replaces a nucleoside at any of positions 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 of the KCNQ2 parent oligonucleotide or KCNQ2 variant oligonucleotide. In specific embodiments, the spacer replaces the nucleoside at one of positions 7, 8, 11, 14, 16, 19, or 22 of the KCNQ2 parent oligonucleotide or the KCNQ2 variant oligonucleotide.

[1044] In various embodiments, the KCNQ2 oligonucleotide comprises a spacer that replaces a nucleoside in the KCNQ2 oligonucleotide (e.g., a spacer replaces a nucleoside in the KCNQ2 oligonucleotide). In specific embodiments, the spacer replaces the nucleoside between positions 9 and 15 of the KCNQ2 oligonucleotide. In specific embodiments, the spacer replaces the nucleoside between positions 9 and 12 of the KCNQ2 oligonucleotide. In specific embodiments, the spacer replaces the nucleoside at position 10 of the KCNQ2 oligonucleotide. In specific embodiments, the spacer replaces the nucleoside at position 11 of the KCNQ2 oligonucleotide. In specific embodiments, the spacer replaces the nucleoside at position 12 of the KCNQ2 oligonucleotide. In specific embodiments, the spacer replaces the nucleoside between positions 12 and 16 of the KCNQ2 oligonucleotide. In specific embodiments, the spacer replaces the nucleoside at position 15 of the KCNQ2 oligonucleotide.

[1045] In various embodiments, a KCNQ2 oligonucleotide comprising a spacer has two segments, wherein at least one of the two segments has at most 11 linked nucleosides. For example, the KCNQ2 oligonucleotide can be 23 oligonucleotide units in length, and the spacer can be located at position 12. Thus, the KCNQ2 oligonucleotide has two segments separated by a spacer, wherein both of the two segments are 11 nucleobases in length. In various embodiments, a KCNQ2 oligonucleotide comprising a spacer has two segments, wherein at least one of the two segments has at most 10 linked nucleosides. For example, the KCNQ2 oligonucleotide can be 21 oligonucleotide units in length, and the spacer can be located at position 11. Thus, the KCNQ2 oligonucleotide has two segments separated by a spacer, wherein both of the two segments are 10 nucleobases in length. As another example, the KCNQ2 oligonucleotide can be 25 oligonucleotide units in length, and the spacer can be located at position 15. Thus, the KCNQ2 oligonucleotide has 2 segments separated by a spacer, wherein one of the 2 segments is 14 nucleobases in length and the second of the 2 segments is 10 nucleobases in length.

[1046] In various embodiments, the KCNQ2 oligonucleotide comprises two spacers, each of which replaces a nucleoside in the KCNQ2 oligonucleotide (e.g., two spacers replace two separate nucleosides of the KCNQ2 oligonucleotide). In various embodiments, the first spacer and the second spacer are separated by at least 5 nucleobases, at least 6 nucleobases, at least 7 nucleobases, at least 8 nucleobases, at least 9 nucleobases, or at least 10 nucleobases in the oligonucleotide. In specific embodiments, the first spacer and the second spacer are separated by at least 5 nucleobases, at least 6 nucleobases, or at least 7 nucleobases. In specific embodiments, the first spacer and the second spacer are not adjacent to each other in the oligonucleotide.

[1047] In specific embodiments, the first spacer replaces the nucleosides between positions 7 and 11 of the KCNQ2 oligonucleotide. In various embodiments, the first spacer replaces the nucleosides between positions 8 and 11, positions 9 and 11, positions 10 and 11, positions 7 and 10, positions 7 and 9, positions 7 and 8, positions 8 and 10, positions 8 and 9, or positions 9 and 10 of the KCNQ2 oligonucleotide. In specific embodiments, the second spacer replaces the nucleosides between positions 14 and 22 of the KCNQ2 oligonucleotide. In various embodiments, the second spacer replaces the nucleosides at positions 15 and 22, 16 and 22, 17 and 22, 18 and 22, 19 and 22, 20 and 22, 21 and 22, 15 and 21, 16 and 21, 17 and 21, 18 and 21, 19 and 21, 20 and 21, 15 and 20, 16 and 20, 17 and 20, 18 and 20, 19 and 20, 15 and 19, 16 and 19, 17 and 19, 18 and 19, 15 and 18, 16 and 18, 17 and 18, 15 and 17, 16 and 17, or between positions 15 and 16 of the KCNQ2 oligonucleotide.

[1048] In a preferred embodiment, the first spacer replaces the nucleoside at position 7 of the KCNQ2 oligonucleotide and the second spacer replaces the nucleoside at position 14 of the KCNQ2 oligonucleotide. In a preferred embodiment, the first spacer replaces the nucleoside at position 7 of the KCNQ2 oligonucleotide and the second spacer replaces the nucleoside at position 19 of the KCNQ2 oligonucleotide. In a preferred embodiment, the first spacer replaces the nucleoside at position 8 of the KCNQ2 oligonucleotide and the second spacer replaces the nucleoside at position 16 of the KCNQ2 oligonucleotide. In a preferred embodiment, the first spacer replaces the nucleoside at position 8 of the KCNQ2 oligonucleotide and the second spacer replaces the nucleoside at position 15 of the KCNQ2 oligonucleotide. In a preferred embodiment, the first spacer replaces the nucleoside at position 11 of the KCNQ2 oligonucleotide and the second spacer replaces the nucleoside at position 22 of the KCNQ2 oligonucleotide. In a preferred embodiment, the first spacer replaces the nucleoside at position 9 of the KCNQ2 oligonucleotide and the second spacer replaces the nucleoside at position 19 of the KCNQ2 oligonucleotide.

[1049] In various embodiments, the KCNQ2 oligonucleotide comprises three spacers, each of which replaces a nucleoside in the KCNQ2 oligonucleotide (e.g., three spacers replace three separate nucleosides of the KCNQ2 oligonucleotide). In a specific embodiment, the first spacer replaces the nucleoside between positions 7 and 11 of the KCNQ2 oligonucleotide. In a specific embodiment, the second spacer replaces the nucleoside between positions 14 and 22 of the KCNQ2 oligonucleotide. In a specific embodiment, the third spacer replaces the nucleoside between positions 21 and 24 of the KCNQ2 oligonucleotide. In some embodiments, the first spacer replaces the nucleoside between positions 2 and 5 of the KCNQ2 oligonucleotide. In a specific embodiment, the second spacer replaces the nucleoside between positions 8 and 12 of the KCNQ2 oligonucleotide. In a specific embodiment, the third spacer replaces the nucleoside between positions 18 and 22 of the KCNQ2 oligonucleotide.

[1050] In various embodiments, the three spacers in the KCNQ2 oligonucleotide are positioned so that each of the four segments of the KCNQ2 oligonucleotide is at most 7 linked nucleosides in length. For example, a KCNQ2 oligonucleotide can have a first segment having 7 linked nucleosides linked to a first spacer, followed by a second segment having 7 linked nucleosides linked at one end to the first spacer and at the other end to the second spacer, followed by a third segment having 6 linked nucleosides linked at one end to the second spacer and at the other end to the third spacer, followed by a fourth segment having 6 linked nucleosides linked to the third spacer.

[1051] In various embodiments, one or more spacers are positioned in the oligonucleotide to replace one or more adenosine or thymidine (relative to guanine or cytidine). For example, one or more spacers can replace one, two, three, four, five, six, seven, eight or nine adenosine or thymidine in the oligonucleotide. In various embodiments, one or more spacers are positioned in the oligonucleotide to replace one or more guanine or cytidine (relative to adenosine or thymidine). For example, one or more spacers can replace one, two, three, four, five, six, seven, eight or nine guanine or cytidine in the oligonucleotide. In various embodiments, spacers are positioned in the oligonucleotide to replace the same amount of adenosine / thymidine and guanine / cytidine. For example, the first spacer in the oligonucleotide can replace adenosine / thymidine, and the second spacer in the oligonucleotide can replace guanine / cytidine.

[1052] In various embodiments, one or more spacers are positioned in the oligonucleotide to control the sequence content in the oligonucleotide.For example, one or more spacers are positioned so that at least one spacer is positioned near the guanine group.In various embodiments, the oligonucleotide with spacer can include one spacer adjacent to the guanine group, two spacers adjacent to the guanine group, three spacers adjacent to the guanine group, four spacers adjacent to the guanine group or five spacers adjacent to the guanine group.In one embodiment, if counting from the 5 ' end of the oligonucleotide, the spacer is immediately before the guanine group in the sequence.Therefore, in various embodiments, the oligonucleotide with spacer can include a spacer immediately before the guanine group, two spacers immediately before the guanine group respectively, three spacers immediately before the guanine group respectively, four spacers immediately before the guanine group respectively or five spacers immediately before the guanine group respectively.In one embodiment, if counting from the 5 ' end of the oligonucleotide, the guanine group is immediately followed by a spacer. Thus, in various embodiments, an oligonucleotide with a spacer can include one spacer immediately following the guanine group, two spacers each immediately following the guanine group, three spacers each immediately following the guanine group, four spacers each immediately following the guanine group, or five spacers each immediately following the guanine group. In various embodiments, the spacers in the oligonucleotide can be positioned so as to maximize the number of spacers adjacent to the guanine group.

[1053] In various embodiments, one or more spacers are positioned in an oligonucleotide to replace one or more adenosine or thymidine nucleosides such that the one or more spacers are positioned adjacent to a guanine group. For example, two spacers can replace an adenosine or thymidine nucleoside in an oligonucleotide, with each of the two spacers being positioned adjacent to a guanine group.

[1054] In various embodiments, the KCNQ2 oligonucleotides with one or more spacers have a specific GC content. As used herein, GC content (or guanine-cytosine content) is the percentage of nitrogenous bases in an oligonucleotide that are guanine (G) or cytosine (C). In various embodiments, the KCNQ2 oligonucleotides with one or more spacers have at least 10% GC content, at least 20% GC content, at least 25% GC content, at least 30% GC content, at least 35% GC content, at least 40% GC content, at least 45% GC content, at least 50% GC content, at least 55% GC content, at least 60% GC content, at least 65% GC content, at least 75% GC content, at least 80% GC content, at least 85% GC content, at least 90% GC content, or at least 95% GC content. In specific embodiments, the KCNQ2 oligonucleotides with one or more spacers have at least 30% GC content. In specific embodiments, the KCNQ2 oligonucleotides with one or more spacers have at least 40% GC content. In various embodiments, one or more spacers are positioned in the KCNQ2 oligonucleotide to maximize GC content. For example, rather than selecting guanine or cytosine for replacement by a spacer, thymine or adenine in the KCNQ2 oligonucleotide can be selected for replacement by a spacer.

[1055] In various embodiments, the KCNQ2 oligonucleotide with a spacer is designed such that 1) each segment of the KCNQ2 oligonucleotide has at most 7 linked nucleosides, and 2) at least two, three, or four spacers are positioned adjacent to a guanine group. In some embodiments, the KCNQ2 oligonucleotide with a spacer is designed such that 1) each segment of the KCNQ2 oligonucleotide has at most 7 linked nucleosides, and 2) each of the two spacers is positioned before a guanine group.

[1056] In various embodiments, the inclusion of one or more spacers in the KCNQ2 oligonucleotide does not reduce the effectiveness of the KCNQ2 oligonucleotide with a spacer in restoring full-length KCNQ2 protein or full-length KCNQ2 mRNA, as compared to the effect of the corresponding KCNQ2 parent oligonucleotide. In various embodiments, the inclusion of one or more spacers in the KCNQ2 oligonucleotide increases the effectiveness of the KCNQ2 oligonucleotide with a spacer in restoring full-length KCNQ2 protein or full-length KCNQ2 mRNA, as compared to the effect of the corresponding KCNQ2 parent oligonucleotide. In various embodiments, the inclusion of one or more spacers in the KCNQ2 oligonucleotide does not reduce the effectiveness of the KCNQ2 oligonucleotide with a spacer in reducing the number of KCNQ2 transcripts, as compared to the effect of the corresponding KCNQ2 parent oligonucleotide. In various embodiments, the inclusion of one or more spacers in the KCNQ2 oligonucleotide increases the effectiveness of the KCNQ2 oligonucleotide with a spacer in reducing the number of KCNQ2 transcripts, as compared to the effect of the corresponding KCNQ2 parent oligonucleotide.

[1057] Tables 15A, 15B, 16, and 14 record example KCNQ2 oligonucleotides having one or more spacers and the relationship of the one or more spacers to the corresponding KCNQ2 parent oligonucleotides. Each KCNQ2 oligonucleotide is assigned a sequence name. As used below, the nomenclature of the sequence name is expressed as "X_spA" (for a KCNQ2 AON with one spacer), "X_spA_spB" (for a KCNQ2 AON with two spacers), or "X_spA_spB_spC" (for a KCNQ2 AON with three spacers). Here, "X" refers to the length of the KCNQ2 AON, "A" refers to the position of the first spacer in the KCNQ2 AON, "B" refers to the position of the second spacer in the KCNQ2 AON, and, if present, "C" refers to the position of the third spacer in the KCNQ2 AON.

[1058] In various embodiments, the KCNQ2 oligonucleotide comprises a spacer. In various embodiments, the KCNQ2 oligonucleotide is an oligonucleotide variant, such as any of a 23-mer, a 21-mer, or a 19-mer. In various embodiments, the inclusion of a spacer divides the KCNQ2 oligonucleotide into two separate segments, wherein at least one of the segments is at most 11 linked nucleosides in length. In various embodiments, the inclusion of a spacer divides the KCNQ2 oligonucleotide into two separate segments, wherein at least one of the segments is at most 10 linked nucleosides in length.

[1059] In various embodiments, the spacer is located between positions 10 and 15 of the oligonucleotide. In various embodiments, the spacer is located between positions 10 and 12 of the oligonucleotide. In specific embodiments, the spacer is located at position 10 of the oligonucleotide. In specific embodiments, the spacer is located at position 11 of the oligonucleotide. In specific embodiments, the spacer is located at position 12 of the oligonucleotide. In specific embodiments, the spacer is located at position 15 of the oligonucleotide. Example KCNQ2 AONs with one spacer are recorded in Table 15A below.

[1060] Table 15A: Identification of KCNQ2 AONs with one spacer. Here, each KCNQ2 AON has 2 segments, wherein at least one of the segments has at most 11 linked nucleosides.

[1061]

[1062]

[1063] *At least one nucleoside linkage of the nucleobase sequence is selected from the group consisting of phosphorothioate linkages, alkylphosphate linkages, phosphorodithioate linkages, phosphotriester linkages, alkylphosphonate linkages, 3-methoxypropylphosphonate linkages, methylphosphonate linkages, aminoalkylphosphotriester linkages, alkylenephosphonate linkages, phosphinate linkages, phosphoamidate linkages, phosphorothioateamidate linkages, phosphorodiamidate (e.g., comprising phosphorodiamidate morpholino (PMO), 3' aminoribose, or 5' aminoribose) linkages, aminoalkylphosphoamidate linkages, phosphorothioateamidate linkages, thioalkylphosphonate linkages, thioalkylphosphotriester linkages, phosphorothioate linkages, phosphoroselenoate linkages, and phosphoroboronate linkages.

[1064] In various embodiments, the KCNQ2 oligonucleotide comprises two spacers. In various embodiments, the inclusion of a spacer divides the KCNQ2 oligonucleotide into three separate segments, wherein at least one of the segments is at most 7 linked nucleosides in length. Example KCNQ2 AONs with two spacers are recorded in Table 15B below.

[1065] Table 15B: Identification of KCNQ2 AONs with two spacers. Here, each KCNQ2 AON has three segments, wherein at least one of the segments has up to 7 linked nucleosides.

[1066]

[1067]

[1068]

[1069]

[1070]

[1071]

[1072] *At least one nucleoside linkage of the nucleobase sequence is selected from the group consisting of phosphorothioate linkages, alkylphosphate linkages, phosphorodithioate linkages, phosphotriester linkages, alkylphosphonate linkages, 3-methoxypropylphosphonate linkages, methylphosphonate linkages, aminoalkylphosphotriester linkages, alkylenephosphonate linkages, phosphinate linkages, phosphoamidate linkages, phosphorothioateamidate linkages, phosphorodiamidate (e.g., comprising phosphorodiamidate morpholino (PMO), 3' aminoribose, or 5' aminoribose) linkages, aminoalkylphosphoamidate linkages, phosphorothioateamidate linkages, thioalkylphosphonate linkages, thioalkylphosphotriester linkages, phosphorothioate linkages, phosphoroselenoate linkages, and phosphoroboronate linkages.

[1073] In various embodiments, the KCNQ2 oligonucleotide comprises three spacers. The inclusion of three spacers divides the KCNQ2 oligonucleotide into four separate segments. In various embodiments, the three spacers are located at different positions of the KCNQ2 oligonucleotide such that each segment of the KCNQ2 oligonucleotide is at most 7 linked nucleosides in length. Examples of KCNQ2 AONs with three spacers are recorded in Table 16 below.

[1074] Table 16: Identification of KCNQ2 AONs or AON variants with three spacers. Here, each KCNQ2 AON has four segments, wherein each segment has up to seven linked nucleosides.

[1075]

[1076]

[1077]

[1078] *At least one nucleoside linkage of the nucleobase sequence is selected from the group consisting of phosphorothioate linkages, alkylphosphate linkages, phosphorodithioate linkages, phosphotriester linkages, alkylphosphonate linkages, 3-methoxypropylphosphonate linkages, methylphosphonate linkages, aminoalkylphosphotriester linkages, alkylenephosphonate linkages, phosphinate linkages, phosphoamidate linkages, phosphorothioateamidate linkages, phosphorodiamidate (e.g., comprising phosphorodiamidate morpholino (PMO), 3' aminoribose, or 5' aminoribose) linkages, aminoalkylphosphoamidate linkages, phosphorothioateamidate linkages, thioalkylphosphonate linkages, thioalkylphosphotriester linkages, phosphorothioate linkages, phosphoroselenoate linkages, and phosphoroboronate linkages.

[1079] In various embodiments, the length of the KCNQ2 AON with one or more spacers is reduced compared to the KCNQ2 AON described in Tables 15B and 16 above. For example, such KCNQ2 AON can be a KCNQ2 oligonucleotide variant with one or more spacers. In various embodiments, the KCNQ2 oligonucleotide variant with one or more spacers is a 23-mer, a 21-mer, or a 19-mer. In various embodiments, the KCNQ2 oligonucleotide variant comprises two spacers such that the KCNQ2 oligonucleotide variant comprises three segments separated by two spacers. In various embodiments, at least one of the three segments has up to 7 linked nucleosides. In various embodiments, each of the three segments has up to 7 linked nucleosides. Example KCNQ2 oligonucleotide variants with one or more spacers are shown in Table 17 below.

[1080] Table 17: KCNQ2AON variants with two spacers. Here, each KCNQ2AON variant has three segments, wherein each segment has up to 7 linked nucleosides.

[1081]

[1082]

[1083]

[1084] *At least one nucleoside linkage of the nucleobase sequence is selected from the group consisting of phosphorothioate linkages, alkylphosphate linkages, phosphorodithioate linkages, phosphotriester linkages, alkylphosphonate linkages, 3-methoxypropylphosphonate linkages, methylphosphonate linkages, aminoalkylphosphotriester linkages, alkylenephosphonate linkages, phosphinate linkages, phosphoamidate linkages, phosphorothioateamidate linkages, phosphorodiamidate (e.g., comprising phosphorodiamidate morpholino (PMO), 3' aminoribose, or 5' aminoribose) linkages, aminoalkylphosphoamidate linkages, phosphorothioateamidate linkages, thioalkylphosphonate linkages, thioalkylphosphotriester linkages, phosphorothioate linkages, phosphoroselenoate linkages, and phosphoroboronate linkages.

[1085] In some embodiments, the antisense oligonucleotides disclosed herein (e.g., KCNQ2 parent oligonucleotides and / or KCNQ2 oligonucleotide variants) comprise one or more spacers and one or more locked nucleic acids (LNAs). In some embodiments, the antisense oligonucleotides disclosed herein (e.g., KCNQ2 parent oligonucleotides and / or KCNQ2 oligonucleotide variants) comprise two spacers and two LNAs. In some embodiments, the antisense oligonucleotides disclosed herein (e.g., KCNQ2 parent oligonucleotides and / or KCNQ2 oligonucleotide variants) comprise two spacers and three LNAs.

[1086] In various embodiments, the spacer and LNA are positioned adjacent to each other in the antisense oligonucleotide. For example, if counting from 5' to 3', the LNA can be located at the 7th position of the antisense oligonucleotide, and the spacer can be located at the 8th position of the antisense oligonucleotide. As another example, if counting from 5' to 3', the LNA can be located at the 9th position of the antisense oligonucleotide, and the spacer can be located at the 8th position of the antisense oligonucleotide. As another example, if counting from 5' to 3', the LNA can be located at the 15th position of the antisense oligonucleotide, and the spacer can be located at the 16th position of the antisense oligonucleotide. As another example, if counting from 5' to 3', the LNA can be located at the 17th position of the antisense oligonucleotide, and the spacer can be located at the 16th position of the antisense oligonucleotide.

[1087] In certain embodiments, in the antisense oligonucleotide, the first spacer is located adjacent to the first LNA, and the second spacer is located adjacent to the second LNA. For example, if counting from 5' to 3', the first LNA may be located at the 7th position of the antisense oligonucleotide, the first spacer may be located at the 8th position of the antisense oligonucleotide, the second LNA may be located at the 15th position of the antisense oligonucleotide, and the second spacer may be located at the 16th position of the antisense oligonucleotide. As another example, if counting from 5' to 3', the first LNA may be located at the 7th position of the antisense oligonucleotide, the first spacer may be located at the 8th position of the antisense oligonucleotide, the second LNA may be located at the 17th position of the antisense oligonucleotide, and the second spacer may be located at the 16th position of the antisense oligonucleotide. As another example, if counting from 5' to 3', the first LNA may be located at the 9th position of the antisense oligonucleotide, the first spacer may be located at the 8th position of the antisense oligonucleotide, the second LNA may be located at the 17th position of the antisense oligonucleotide, and the second spacer may be located at the 16th position of the antisense oligonucleotide.

[1088] In various embodiments, the one or more spacers and the one or more LNAs are not positioned adjacent to each other in the antisense oligonucleotide. For example, if counting from 5' to 3', the LNA can be located at the 4th position of the antisense oligonucleotide, and the spacer can be located at the 8th position of the antisense oligonucleotide. As an example, if counting from 5' to 3', the LNA can be located at the 20th position of the antisense oligonucleotide, and the spacer can be located at the 16th position of the antisense oligonucleotide. In a specific embodiment, if counting from 5' to 3', the first LNA can be located at the 4th position of the antisense oligonucleotide, the first spacer can be located at the 8th position of the antisense oligonucleotide, the second LNA can be located at the 20th position of the antisense oligonucleotide, and the second spacer can be located at the 16th position of the antisense oligonucleotide. In a specific embodiment, if counting from 5' to 3', the first LNA can be located at the 4th position of the antisense oligonucleotide, the first spacer can be located at the 8th position of the antisense oligonucleotide, the second LNA can be located at the 12th position of the antisense oligonucleotide, the second spacer can be located at the 16th position of the antisense oligonucleotide, and the third LNA can be located at the 20th position of the antisense oligonucleotide.

[1089] UNC13A

[1090] As further described below, UNC13A oligonucleotides having one or more spacers are described with reference to the corresponding UNC13A parent oligonucleotide or the corresponding UNC13A variant oligonucleotide. In various embodiments, a UNC13A oligonucleotide having a spacer differs from a UNC13A parent oligonucleotide or a UNC13A variant oligonucleotide in that the spacer replaces a nucleoside in the UNC13A parent oligonucleotide or the UNC13A variant oligonucleotide. As used below, a "position" of a UNC13A oligonucleotide refers to a specific position counted from the 5' end of the UNC13A oligonucleotide. In various embodiments, the spacer replaces a nucleoside at any of positions 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 of a UNC13A parent oligonucleotide or a UNC13A variant oligonucleotide. In specific embodiments, the spacer replaces the nucleoside at one of positions 7, 8, 11, 14, 16, 19, or 22 of the UNC13A parent oligonucleotide or the UNC13A variant oligonucleotide.

[1091] In various embodiments, a UNC13A oligonucleotide comprises a spacer that replaces a nucleoside in a UNC13A oligonucleotide (e.g., a spacer replaces a nucleoside in a UNC13A oligonucleotide). In specific embodiments, the spacer replaces the nucleoside between positions 9 and 15 of the UNC13A oligonucleotide. In specific embodiments, the spacer replaces the nucleoside between positions 9 and 12 of the UNC13A oligonucleotide. In specific embodiments, the spacer replaces the nucleoside at position 10 of the UNC13A oligonucleotide. In specific embodiments, the spacer replaces the nucleoside at position 11 of the UNC13A oligonucleotide. In specific embodiments, the spacer replaces the nucleoside at position 12 of the UNC13A oligonucleotide. In specific embodiments, the spacer replaces the nucleoside between positions 12 and 16 of the UNC13A oligonucleotide. In specific embodiments, the spacer replaces the nucleoside at position 15 of the UNC13A oligonucleotide.

[1092] In various embodiments, a UNC13A oligonucleotide comprising a spacer has two segments, wherein at least one of the two segments has at most 11 linked nucleosides. For example, a UNC13A oligonucleotide can be 23 oligonucleotide units in length, and the spacer can be located at position 12. Thus, the UNC13A oligonucleotide has two segments separated by a spacer, wherein both of the two segments are 11 nucleobases in length. In various embodiments, a UNC13A oligonucleotide comprising a spacer has two segments, wherein at least one of the two segments has at most 10 linked nucleosides. For example, a UNC13A oligonucleotide can be 21 oligonucleotide units in length, and the spacer can be located at position 11. Thus, the UNC13A oligonucleotide has two segments separated by a spacer, wherein both of the two segments are 10 nucleobases in length. As another example, a UNC13A oligonucleotide can be 25 oligonucleotide units in length, and the spacer can be located at position 15. Thus, the UNC13A oligonucleotide has two segments separated by a spacer, wherein one of the two segments is 14 nucleobases in length and the second of the two segments is 10 nucleobases in length.

[1093] In various embodiments, the UNC13A oligonucleotide comprises two spacers, each of which replaces a nucleoside in the UNC13A oligonucleotide (e.g., the two spacers replace two separate nucleosides of the UNC13A oligonucleotide). In various embodiments, the first spacer and the second spacer are separated by at least 5 nucleobases, at least 6 nucleobases, at least 7 nucleobases, at least 8 nucleobases, at least 9 nucleobases, or at least 10 nucleobases in the oligonucleotide. In specific embodiments, the first spacer and the second spacer are separated by at least 5 nucleobases, at least 6 nucleobases, or at least 7 nucleobases. In specific embodiments, the first spacer and the second spacer are not adjacent to each other in the oligonucleotide.

[1094] In specific embodiments, the first spacer replaces the nucleosides between positions 7 and 11 of the UNC13A oligonucleotide. In various embodiments, the first spacer replaces the nucleosides between positions 8 and 11, positions 9 and 11, positions 10 and 11, positions 7 and 10, positions 7 and 9, positions 7 and 8, positions 8 and 10, positions 8 and 9, or positions 9 and 10 of the UNC13A oligonucleotide. In specific embodiments, the second spacer replaces the nucleosides between positions 14 and 22 of the UNC13A oligonucleotide. In various embodiments, the second spacer replaces the nucleosides at positions 15 and 22, positions 16 and 22, positions 17 and 22, positions 18 and 22, positions 19 and 22, positions 20 and 22, positions 21 and 22, positions 15 and 21, positions 16 and 21, positions 17 and 21, positions 18 and 21, positions 19 and 21, positions...

Claims

1. A compound comprising a splice-switching oligonucleotide, wherein the splice-switching oligonucleotide comprises a spacer.

2. The compound of claim 1, wherein the splice-switching oligonucleotide comprises a sequence that is at least 85% complementary to an equal length portion of a sequence from a transcript whose mis-splicing causes a neurological disease or whose splicing can be modulated to treat a neurological disease.

3. The compound of claim 1, wherein the splice-switching oligonucleotide comprises a sequence that is at least 85% complementary to an equal length portion of a sequence from a transcript regulated by TDP-43, wherein at least one (i.e., one or more) nucleoside linkage of the oligonucleotide is a non-natural linkage, and further wherein the oligonucleotide comprises a spacer.

4. The compound of claim 1 or 2, wherein the modified oligonucleotide comprises a sequence that is at least 85% complementary to an equal length portion of a sequence from a transcript of any of STMN2, KCNQ2, UNC13A, or SMN2.

5. The compound of claim 1-2 or 4, wherein the modified oligonucleotide comprises a sequence that is at least 85% complementary to an equal length portion of any one of SEQ ID NO: 1339 or SEQ ID NO: 1341, SEQ ID NOs: 3032-3043, SEQ ID NOs: 9587-9595, or SEQ ID NOs: 9698-9707.

6. A compound comprising a modified oligonucleotide comprising a sequence that is at least 85% complementary to an equal length portion of a sequence from a transcript of any of STMN2, KCNQ2, UNC13A or SMN2, wherein at least one (i.e., one or more) nucleoside linkages of the oligonucleotide are non-natural linkages, and further wherein the oligonucleotide comprises a spacer.

7. The compound of claim 6, wherein the modified oligonucleotide comprises a sequence that is at least 85% complementary to an equal length portion of any one of SEQ ID NO: 1339 or SEQ ID NO: 1341, SEQ ID NOs: 3032-3043, SEQ ID NOs: 9587-9595, or SEQ ID NOs: 9698-9707, or a 15 to 50 consecutive nucleobase portion thereof having 90% identity thereto.

8. A splice-switching oligonucleotide, wherein the splice-switching oligonucleotide comprises a spacer.

9. The oligonucleotide of claim 8, wherein the oligonucleotide comprises a sequence that is at least 85% complementary to an equal length portion of a sequence from a transcript whose mis-splicing results in a neurological disease or whose splicing can be modulated to treat a neurological disease.

10. The oligonucleotide of claim 8, wherein the oligonucleotide comprises an equal length portion of a sequence from a transcript regulated by TDP-43 that is at least 85% identical to the sequence of the transcript regulated by TDP-43. - The oligonucleotide may further comprise a sequence complementary to the oligonucleotide, wherein at least one (ie, one or more) nucleoside linkage of the oligonucleotide is a non-natural linkage, and further wherein the oligonucleotide comprises a spacer.

11. The oligonucleotide of claim 8 or 9, wherein the oligonucleotide comprises a sequence that is at least 85% complementary to an equal length portion of a sequence from a transcript of any of STMN2, KCNQ2, UNC13A, or SMN2.

12. The oligonucleotide of claim 8-9 or 11, wherein the modified oligonucleotide comprises a sequence that is at least 85% complementary to an equal length portion of any one of SEQ ID NO: 1339 or SEQ ID NO: 1341, SEQ ID NOs: 3032-3043, SEQ ID NOs: 9587-9595, or SEQ ID NOs: 9698-9707.

13. A splice-switching oligonucleotide comprising a sequence that is at least 85% complementary to an equal length portion of a sequence from a transcript of any of STMN2, KCNQ2, UNC13A or SMN2, wherein at least one (i.e., one or more) nucleoside linkages of the oligonucleotide are non-natural linkages, and further wherein the oligonucleotide comprises a spacer.

14. The oligonucleotide of claim 13, wherein the modified oligonucleotide comprises a sequence that is at least 85% complementary to an equal length portion of any one of SEQ ID NO: 1339 or SEQ ID NO: 1341, SEQ ID NOs: 3032-3043, SEQ ID NOs: 9587-9595, or SEQ ID NOs: 9698-9707.

15. An oligonucleotide comprising an equal length portion of SEQ ID NO: 1339 or SEQ ID NO: 1341, SEQ ID NO: 3032-3043, SEQ ID NO: 9587-9595 or SEQ ID NO: 9698-9707, any of the sequences having 90% identity thereof, or a sequence that is at least 85% complementary to a 15 to 50 consecutive nucleobase portion thereof, wherein at least one (i.e., one or more) nucleoside linkage of the oligonucleotide is a non-natural linkage, and further wherein the oligonucleotide comprises a spacer.

16. A compound comprising a splice-switching oligonucleotide comprising a sequence that is at least 85% complementary to an equal length portion of a sequence from a transcript whose mis-splicing results in a neurological disease or whose splicing can be modulated to treat a neurological disease, wherein at least one (i.e., one or more) nucleoside bond of the oligonucleotide is a non-natural linkage.

17. A compound comprising a splice-switching oligonucleotide comprising a sequence that is at least 85% complementary to an equal length portion of a sequence from a transcript regulated by TDP-43, wherein at least one (ie, one or more) nucleoside linkage of the oligonucleotide is a non-natural linkage.

18. The compound of claim 16, wherein the splice-switching oligonucleotide comprises a sequence that is at least 85% complementary to an equal length portion of a sequence from a transcript of any one of SEQ ID NO: 1339 or SEQ ID NO: 1341, SEQ ID NOs: 3032-3043, SEQ ID NOs: 9587-9595, or SEQ ID NOs: 9698-9707.

19. The compound of claim 16 or 18, wherein the modified splice-switching oligonucleotide comprises a sequence that is at least 85% complementary to an equal length portion of any one of SEQ ID NO: 1339 or SEQ ID NO: 1341, SEQ ID NOs: 3032-3043, SEQ ID NOs: 9587-9595, or SEQ ID NOs: 9698-9707.

20. A compound comprising a modified splice-switching oligonucleotide comprising a sequence that is at least 85% complementary to an equal length portion of a sequence from a transcript of any of STMN2, KCNQ2, UNC13A or SMN2, wherein at least one (i.e., one or more) nucleoside linkage of the oligonucleotide is a non-natural linkage.

21. The compound of claim 20, wherein the modified oligonucleotide comprises a sequence that is at least 85% complementary to an equal length portion of any one of SEQ ID NO: 1339 or SEQ ID NO: 1341, SEQ ID NOs: 3032-3043, SEQ ID NOs: 9587-9595, or SEQ ID NOs: 9698-9707, or a 15 to 50 contiguous nucleobase portion thereof having 90% identity thereto.

22. A splice-switching oligonucleotide comprising a sequence that is at least 85% complementary to an equal length portion of a sequence from a transcript whose mis-splicing results in a neurological disease or whose splicing can be modulated to treat a neurological disease, wherein at least one (i.e., one or more) nucleoside bond of the oligonucleotide is a non-natural linkage.

23. A splice-switching oligonucleotide comprising a sequence that is at least 85% complementary to an equal length portion of a sequence from a transcript regulated by TDP-43, wherein at least one (ie, one or more) nucleoside linkage of the oligonucleotide is a non-natural linkage.

24. The oligonucleotide of claim 23, wherein the oligonucleotide comprises a sequence that is at least 85% complementary to an equal length portion of a sequence from a transcript of any of STMN2, KCNQ2, UNC13A, or SMN2.

25. The oligonucleotide of claim 22 or 24, wherein the modified oligonucleotide comprises a sequence that is at least 85% complementary to an equal length portion of any one of SEQ ID NO: 1339 or SEQ ID NO: 1341, SEQ ID NOs: 3032-3043, SEQ ID NOs: 9587-9595, or SEQ ID NOs: 9698-9707.

26. A splice-switching oligonucleotide comprising a sequence that is at least 85% complementary to an equal length portion of a sequence from a transcript of any of STMN2, KCNQ2, UNC13A or SMN2, wherein at least one (ie, one or more) nucleoside linkage of the oligonucleotide is a non-natural linkage.

27. The oligonucleotide of claim 26, wherein the oligonucleotide comprises a sequence that is at least 85% complementary to an equal length portion of any one of SEQ ID NO: 1339 or SEQ ID NO: 1341, SEQ ID NOs: 3032-3043, SEQ ID NOs: 9587-9595, or SEQ ID NOs: 9698-9707.

28. A splice-switching oligonucleotide comprising a sequence that is at least 85% complementary to an equal length portion of any one of SEQ ID NO: 1339 or SEQ ID NO: 1341, SEQ ID NOs: 3032-3043, SEQ ID NOs: 9587-9595, or SEQ ID NOs: 9698-9707, wherein at least one (i.e., one or more) nucleoside linkage of the oligonucleotide is a non-natural linkage.

29. The oligonucleotide of claim 28, further comprising a spacer.

30. The compound or oligonucleotide of claims 1-29, wherein the oligonucleotide comprises a segment of up to 11 linked nucleosides.

31. The compound or oligonucleotide of claims 1-29, wherein the oligonucleotide comprises a segment of up to 10, 9, or 8 linked nucleosides.

32. The compound or oligonucleotide of claims 1-29, wherein the oligonucleotide comprises a segment of up to 7 linked nucleosides.

33. The compound or oligonucleotide of claims 1-29, wherein the oligonucleotide comprises a segment of at most 6, 5, 4, 3, or 2 linked nucleosides.

34. The compound or oligonucleotide of claims 1-33, wherein each segment of the oligonucleotide comprises up to 7 linked nucleosides.

35. The compound or oligonucleotide of any one of claims 30-34, wherein the oligonucleotide comprises the residues of SEQ ID NOs: 1-466, SEQ ID NOs: 893-1338, SEQ ID NOs: 1342-1366, SEQ ID NOs: 1392-1664, or SEQ ID NOs: 10655-10669, SEQ ID NOs: 1676-1851, SEQ ID NOs: 2028-2529, SEQ ID NOs: 3046-3221, SEQ ID NOs: 3398-3899, and SEQ ID NOs: 4402-4530, SEQ ID NOs: 4531-5794, SEQ ID NOs: 7059-8322, SEQ ID NOs: 9596-9603, SEQ ID NOs: 9710-10141, SEQ ID NOs: 10574-10651, SEQ ID NOs: Equal length portions of any of SEQ ID NOs: 10670-10779, SEQ ID NOs: 10783-10808, and SEQ ID NOs: 10814-10820 share a sequence that is at least 85% identical.

36. The compound or oligonucleotide of any one of claims 30-35, wherein the oligonucleotide comprises the residues of SEQ ID NOs: 1-466, SEQ ID NOs: 893-1338, SEQ ID NOs: 1342-1366, SEQ ID NOs: 1392-1664, or SEQ ID NOs: 10655-10669, SEQ ID NOs: 1676-1851, SEQ ID NOs: 2028-2529, SEQ ID NOs: 3046-3221, SEQ ID NOs: 3398-3899, and SEQ ID NOs: 4402-4530, SEQ ID NOs: 4531-5794, SEQ ID NOs: 7059-8322, SEQ ID NOs: 9596-9603, SEQ ID NOs: 9710-10141, SEQ ID NOs: 10574-10651, SEQ ID NOs: Equal length portions of any of SEQ ID NOs: 10670-10779, SEQ ID NOs: 10783-10808, and SEQ ID NOs: 10814-10820 share a sequence that is at least 90% identical.

37. The compound or oligonucleotide of any one of claims 30-36, wherein the oligonucleotide comprises the residues of SEQ ID NOs: 1-466, SEQ ID NOs: 893-1338, SEQ ID NOs: 1342-1366, SEQ ID NOs: 1392-1664, or SEQ ID NOs: 10655-10669, SEQ ID NOs: 1676-1851, SEQ ID NOs: 2028-2529, SEQ ID NOs: 3046-3221, SEQ ID NOs: 3398-3899, and SEQ ID NOs: 4402-4530, SEQ ID NOs: 4531-5794, SEQ ID NOs: 7059-8322, SEQ ID NOs: 9596-9603, SEQ ID NOs: 9710-10141, SEQ ID NOs: 10574-10651, SEQ ID NOs: Equal length portions of any of SEQ ID NOs: 10670-10779, SEQ ID NOs: 10783-10808, and SEQ ID NOs: 10814-10820 share a sequence that is at least 95% identical.

38. The compound or oligonucleotide of any one of claims 30-36, wherein the oligonucleotide comprises the residues of SEQ ID NOs: 1-466, SEQ ID NOs: 893-1338, SEQ ID NOs: 1342-1366, SEQ ID NOs: 1392-1664, or SEQ ID NOs: 10655-10669, SEQ ID NOs: 1676-1851, SEQ ID NOs: 2028-2529, SEQ ID NOs: 3046-3221, SEQ ID NOs: 3398-3899, and SEQ ID NOs: 4402-4530, SEQ ID NOs: 4531-5794, SEQ ID NOs: 7059-8322, SEQ ID NOs: 9596-9603, SEQ ID NOs: 9710-10141, SEQ ID NOs: 10574-10651, SEQ ID NOs: Equal length portions of any of SEQ ID NOs: 10670-10779, SEQ ID NOs: 10783-10808, and SEQ ID NOs: 10814-10820 share a sequence with 100% identity.

39. The compound or oligonucleotide of any one of claims 1-38, wherein the oligonucleotide comprises a segment having up to 11 linked nucleosides or up to 7 linked nucleosides, and wherein the oligonucleotide comprises a segment having up to 11 linked nucleosides or up to 7 linked nucleosides, and wherein the oligonucleotide comprises a segment having up to 11 linked nucleosides or up to 7 linked nucleosides, and wherein the oligonucleotide comprises a segment having up to 11 linked nucleosides or up to 7 linked nucleosides, and wherein the oligonucleotide comprises a segment having up to 11 linked nucleosides or up to 7 linked nucleosides, and wherein the oligonucleotide comprises a segment having up to 11 linked nucleosides or up to 7 linked nucleosides, and wherein the oligonucleotide comprises a segment having up to 11 linked nucleosides or up to 7 linked nucleosides, and wherein the oligonucleotide comprises a segment having up to 11 linked nucleosides or up to 7 linked nucleosides, and wherein the oligonucleotide comprises a segment having up to 11 linked nucleosides or up to 7 linked nucleosides, and wherein the oligonucleotide comprises a segment having up to 11 linked nucleosides or up to 7 linked nucleosides, and wherein the oligonucleotide comprises a segment having up to 11 linked nucleosides or up to 7 linked nucleosides, and wherein the oligonucleotide comprises a segment having up to 11 linked nucleosides or up to 7 linked nucleosides, Equal length portions of any of SEQ ID NOs: 10574-10651, SEQ ID NOs: 10670-10779, SEQ ID NOs: 10783-10808, and SEQ ID NOs: 10814-10820 share a sequence that is at least 85% identical.

40. The compound or oligonucleotide of claim 39, wherein the oligonucleotide comprises a fragment having up to 6, 5, 4, 3, or 2 linked nucleosides, wherein the oligonucleotide comprises a fragment having up to 6, 5, 4, 3, or 2 linked nucleosides, wherein the oligonucleotide comprises a fragment having up to 6, 5, 4, 3, or 2 linked nucleosides, wherein the oligonucleotide comprises a fragment having up to 6, 5, 4, 3, or 2 linked nucleosides, wherein the oligonucleotide comprises a fragment having up to 6, 5, 4, 3, or 2 linked nucleosides, wherein the oligonucleotide comprises a fragment having up to 6, 5, 4, 3, or 2 linked nucleosides, wherein the oligonucleotide comprises a fragment having up to 6, 5, 4, 3, or 2 linked nucleosides, wherein the oligonucleotide comprises a fragment having up to 6, 5, 4, 3, or 2 linked nucleosides, wherein the oligonucleotide comprises a fragment having up to 6, 5, 4, 3, or 2 linked nucleosides, wherein the oligonucleotide comprises a fragment having up to 6, 5, 4, 3, or 2 linked nucleosides, wherein the oligonucleotide comprises a fragment having up to 6, 5, 4, 3, or 2 linked nucleosides, wherein the oligonucleotide comprises a fragment having up to 6, 5, 4, 3, or 2 linked nucleosides, wherein the oligonucleotide comprises a fragment having up to 6, 5, 4, 3, or 2 linked nucleosides, wherein the oligonucleotide comprises a fragment having up to 6, 5, 4, Equal length portions of any of SEQ ID NOs: 10574-10651, SEQ ID NOs: 10670-10779, SEQ ID NOs: 10783-10808, and SEQ ID NOs: 10814-10820 share a sequence that is at least 85% identical.

41. The compound or oligonucleotide of claim 39 or 40, wherein the oligonucleotide comprises a fragment having up to 6, 5, 4, 3 or 2 linked nucleosides ... Equal length portions of any of SEQ ID NOs: 10574-10651, SEQ ID NOs: 10670-10779, SEQ ID NOs: 10783-10808, and SEQ ID NOs: 10814-10820 share a sequence that is at least 90% identical.

42. The compound or oligonucleotide of any one of claims 1-41, wherein the oligonucleotide is at least 18, at least 19, at least 20, at least 21, at least 22, at least 23, at least 24, or at least 25 oligonucleotide units in length.

43. The compound or oligonucleotide of claim 42, wherein the oligonucleotide is at least 19 oligonucleotide units in length.

44. The compound or oligonucleotide of any one of claims 1-43, wherein the spacer is a nucleoside replacement group comprising a non-sugar substituent that is not capable of linking to a nucleotide base.

45. The compound or oligonucleotide of claim 44, wherein the spacer is located between positions 10 and 15 of the oligonucleotide.

46. ​​The compound or oligonucleotide of claim 44, wherein the spacer is located between positions 7 and 11 of the oligonucleotide.

47. The compound or oligonucleotide of claims 44-46, wherein the oligonucleotide further comprises a second spacer, wherein the second spacer is located between positions 14 and 22 of the oligonucleotide.

48. The compound or oligonucleotide of claim 47, wherein the spacer and the second spacer are separated by at least 5 nucleobases, at least 6 nucleobases, or at least 7 nucleobases in the oligonucleotide.

49. The compound or oligonucleotide of claim 47 or 48, wherein the spacer is located between positions 7 and 9 of the oligonucleotide, and wherein the second spacer is located between positions 15 and 18 of the oligonucleotide.

50. The compound or oligonucleotide of any one of claims 47-49, wherein the spacer is located at position 8 of the oligonucleotide, and wherein the second spacer is located at position 16 of the oligonucleotide.

51. The compound or oligonucleotide of any one of claims 47-50, wherein the oligonucleotide further comprises a third spacer, wherein the third spacer is located between positions 21 and 24 of the oligonucleotide.

52. The compound or oligonucleotide of claim 44, wherein the spacer is located between positions 2 and 5 of the oligonucleotide.

53. The compound or oligonucleotide of claim 52, wherein the oligonucleotide further comprises a second spacer, wherein the second spacer is located between positions 8 and 12 of the oligonucleotide.

54. The compound or oligonucleotide of claim 53, wherein the oligonucleotide further comprises a third spacer, wherein the third spacer is located between positions 18 and 22 of the oligonucleotide.

55. The compound or oligonucleotide of claim 44, wherein the oligonucleotide further comprises a second spacer and a third spacer, wherein the three spacers are located at positions in the oligonucleotide such that each segment of the oligonucleotide has at most 7 linked nucleosides.

56. The compound or oligonucleotide of claim 55, wherein at least two of the three spacers are adjacent to a guanine nucleobase.

57. The compound or oligonucleotide of claim 56, wherein each of at least two of the three spacers is immediately preceded by a guanine nucleobase.

58. The compound or oligonucleotide of any one of claims 44-57, wherein each of the first, second or third spacers is a nucleoside replacement group comprising a non-sugar substituent, wherein the non-sugar substituent does not contain a ketone, aldehyde, ketal, hemiketal, acetal, hemiacetal, aminal or hemiaminal moiety and is not capable of forming a covalent bond with a nucleotide base.

59. The compound or oligonucleotide of any one of claims 44-57, wherein each of the first, second or third spacers is independently represented by formula (X), wherein: Ring A is an optionally substituted 4-8 membered monocyclic cycloalkyl or 4-8 membered monocyclic heterocyclyl, wherein the heterocyclyl contains 1 or 2 heteroatoms selected from O, S and N, provided that A is not capable of forming a covalent bond with a nucleobase; and The symbol represents the point of attachment of the internucleoside linkage.

60. The compound or oligonucleotide of claim 59, wherein each of the first, second or third spacers is independently represented by formula (Xa), wherein:

61. The compound or nucleotide of claim 59 or 60, wherein Ring A is an optionally substituted 4-8 membered monocyclic cycloalkyl or 4-8 membered monocyclic heterocyclyl, wherein the 4-8 membered monocyclic cycloalkyl is selected from cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl; and the 4-8 membered monocyclic heterocyclyl is selected from oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, 1,4-dioxane, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl and azepanyl.

62. The compound or nucleotide of claim 61, wherein Ring A is tetrahydrofuranyl.

63. The compound or nucleotide of claim 61, wherein Ring A is tetrahydropyranyl.

64. The compound or oligonucleotide of any one of claims 44-57, wherein each of the first, second or third spacers is independently represented by Formula I, wherein: X is selected from -CH2- and -O-; and n is 0, 1, 2 or 3.

65. The compound or oligonucleotide of any one of claims 44-57, wherein each of the first, second or third spacers is independently represented by Formula I', wherein: X is selected from -CH2- and -O-; and n is 0, 1, 2 or 3.

66. The compound or oligonucleotide of any one of claims 44-57, wherein each of the first, second or third spacers is independently represented by formula (Ia), wherein: and n is 0, 1, 2 or 3.

67. The compound or oligonucleotide of any one of claims 4457, wherein each of the first, second or third spacers is independently represented by formula (Ia'), wherein: and n is 0, 1, 2 or 3.

68. The compound or oligonucleotide of any one of claims 44-57, wherein each of the first, second or third spacers is independently represented by Formula II, wherein: and X is selected from -CH2- and -O-.

69. The compound or oligonucleotide of any one of claims 44-57, wherein each of the first, second or third spacers is independently represented by Formula II', wherein: and X is selected from -CH2- and -O-.

70. The compound or oligonucleotide of any one of claims 44-57, wherein each of the first, second or third spacers is independently represented by formula (Iia), wherein:

71. The compound or oligonucleotide of any one of claims 44-57, wherein each of the first, second or third spacers is independently represented by formula (Iia'), wherein:

72. The compound or oligonucleotide of any one of claims 44-57, wherein each of the first, second or third spacers is independently represented by formula (IIi), wherein: X is selected from -CH2- and -O-.

73. The compound or oligonucleotide of any one of claims 44-57, wherein each of the first, second or third spacers is independently represented by formula (III'), wherein: X is selected from -CH2- and -O-.

74. The compound or oligonucleotide of any one of claims 44-57, wherein each of the first, second or third spacers is independently represented by formula (IIIb), wherein:

75. The compound or oligonucleotide of any one of claims 44-57, wherein each of the first, second or third spacers is independently represented by formula (IIIb'), wherein:

76. The compound or oligonucleotide of any one of claims 44-57, wherein each of the first, second or third spacers is independently represented by Formula III, wherein: and X is selected from -CH2- and -O-.

77. The compound or oligonucleotide of any one of claims 44-57, wherein each of the first, second or third spacers is independently represented by Formula III, wherein: and X is selected from -CH2- and -O-.

78. The compound or oligonucleotide of any one of claims 44-57, wherein each of the first, second or third spacers is independently represented by formula (IIIa), wherein:

79. The compound or oligonucleotide of any one of claims 44-57, wherein each of the first, second or third spacers is independently represented by formula (IIia'), wherein:

80. The compound or oligonucleotide of any one of the preceding claims, wherein the oligonucleotide further comprises a locked nucleic acid (LNA).

81. The compound or oligonucleotide of claim 80, wherein the locked nucleic acid (LNA) is located at one of positions 4, 7, 9, 12, 15 or 20 of the oligonucleotide.

82. The compound or oligonucleotide of any of the preceding claims, wherein the oligonucleotide comprising the spacer has a GC content of at least 10%.

83. The compound or oligonucleotide of any of the preceding claims, wherein the oligonucleotide comprising the spacer has a GC content of at least 20%.

84. The compound or oligonucleotide of any of the preceding claims, wherein the oligonucleotide comprising the spacer has a GC content of at least 25%.

85. The compound or oligonucleotide of any of the preceding claims, wherein the oligonucleotide comprising the spacer has a GC content of at least 30%.

86. The compound or oligonucleotide of any of the preceding claims, wherein the oligonucleotide comprising the spacer has a GC content of at least 40%.

87. The compound or oligonucleotide of any of the preceding claims, wherein the oligonucleotide comprising the spacer has a GC content of at least 50%.

88. The compound or oligonucleotide of any of the preceding claims, wherein the oligonucleotide is between 12 and 40 oligonucleotide units in length.

89. The compound or oligonucleotide of any of the preceding claims, wherein at least one (i.e., one or more) nucleoside linkages of the oligonucleotide are independently selected from the group consisting of phosphodiester linkages, phosphorothioate linkages, alkyl phosphate linkages, phosphorodithioate linkages, phosphotriester linkages, alkylphosphonate linkages, 3-methoxypropylphosphonate linkages, methylphosphonate linkages, aminoalkylphosphotriester linkages, alkylenephosphonate linkages, phosphinate linkages, phosphoamidate linkages, phosphorothioate amide linkages, phosphorothioate diamide linkages, phosphorodiamidate linkages, aminoalkylphosphoamidate linkages, phosphorothioate amide linkages, alkylthiophosphonate linkages, alkylthiophosphotriester linkages, phosphorothioate linkages, selenophosphate linkages, and borophosphate linkages.

90. The compound or oligonucleotide of any one of claims 1-89, wherein the one or more nucleoside linkages linking the base at position 3 or position 4 of the oligonucleotide are phosphodiester linkages.

91. The compound or oligonucleotide of claim 90, wherein only one nucleoside linkage linking the base at position 3 or position 4 of the oligonucleotide is a phosphodiester linkage.

92. The compound or oligonucleotide of any one of claims 1-89, wherein the nucleoside linkage linking the bases at both position 3 and position 4 of the oligonucleotide is a phosphodiester linkage.

93. The compound or oligonucleotide of any one of claims 1-89, wherein one or more bases immediately preceding the spacer in the oligonucleotide are linked by a phosphodiester bond.

94. The compound or oligonucleotide of claim 93, wherein only the base immediately preceding the spacer in the oligonucleotide is linked to the spacer via a phosphodiester bond.

95. The compound or oligonucleotide of claim 94, wherein the base immediately preceding the spacer in the oligonucleotide is further linked to a further preceding base via a phosphodiester bond.

96. The compound or oligonucleotide of claim 94, wherein the oligonucleotide comprises a second spacer, wherein the base immediately preceding the second spacer is linked to a further preceding base via a phosphodiester bond.

97. The compound or oligonucleotide of any one of claims 1-89, wherein one or more bases immediately following the spacer in the oligonucleotide are linked by a phosphodiester bond.

98. The compound or oligonucleotide of claim 97, wherein only the base immediately following the spacer in the oligonucleotide is linked to the spacer via a phosphodiester bond.

99. The compound or oligonucleotide of claim 93, wherein the two bases immediately preceding the spacer in the oligonucleotide are linked by a phosphodiester bond.

100. The compound or oligonucleotide of any one of claims 1-89, wherein the one or more bases in the oligonucleotide immediately before the spacer are linked by a phosphodiester bond, and wherein the one or more bases in the oligonucleotide immediately after the spacer are linked by a phosphodiester bond.

101. The compound or oligonucleotide of claim 100, wherein the base immediately before the spacer and the base immediately after the spacer are linked by a phosphodiester bond.

102. The compound or oligonucleotide of claim 100 or 101, wherein the oligonucleotide comprises a second spacer, and wherein the one or more bases immediately preceding the second spacer in the oligonucleotide are linked by a phosphodiester bond, and wherein the one or more bases immediately following the second spacer in the oligonucleotide are linked by a phosphodiester bond.

103. The compound or oligonucleotide of claim 102, wherein the base immediately before the second spacer and the base immediately after the second spacer are linked by a phosphodiester bond.

104. The compound or oligonucleotide of any one of claims 189, wherein the oligonucleotide comprises a series of bases linked by phosphodiester bonds, the series of bases comprising at least two bases.

105. The compound or oligonucleotide of any one of claims 1-89, wherein the oligonucleotide comprises a series of bases linked by phosphodiester bonds, the series of bases comprising at least five bases.

106. The compound or oligonucleotide of claim 104 or 105, wherein the oligonucleotide comprises two or more spacers, and wherein the series of bases is located between the at least two spacers.

107. The compound or oligonucleotide of any of the preceding claims, wherein one or more internucleoside linkages of the oligonucleotide are modified internucleoside linkages.

108. The compound or oligonucleotide of claim 107, wherein the modified internucleoside linkage of the oligonucleotide is a phosphorothioate linkage.

109. The compound or oligonucleotide of claim 107 or 108, wherein all internucleoside linkages of the oligonucleotide are phosphorothioate linkages.

110. The compound or oligonucleotide of claim 108, wherein the phosphorothioate linkage is in one of the Rp configuration or the Sp configuration.

111. The compound or oligonucleotide of any of the preceding claims, wherein the oligonucleotide comprises at least one modified sugar moiety.

112. A compound or oligonucleotide according to claim 111, wherein the modified sugar moiety is one of the following: a 2'-OMe modified sugar moiety, a bicyclic sugar moiety, 2'-O-(2-methoxyethyl) (2'-MOE), a 2'-deoxy-2'-fluoro nucleoside, a 2'-fluoro-β-D-arabinoside, a locked nucleic acid (LNA), a constrained ethyl 2'-4'-bridged nucleic acid (cEt), S-cEt, tcDNA, a hexitol nucleic acid (HNA), and a tricyclic analog (e.g., tcDNA).

113. The compound or oligonucleotide of any of the preceding claims, wherein the oligonucleotide exhibits at least a 30%, 40%, 50%, 60%, 70%, 80% or 90% increase in full-length STMN2, KCNQ2, UNC13A or SMN2 protein.

114. The compound or oligonucleotide of any of the preceding claims, wherein the oligonucleotide exhibits at least a 100% increase in full-length protein.

115. The compound or oligonucleotide of any of the preceding claims, wherein the oligonucleotide exhibits at least a 200% increase in full-length protein.

116. The compound or oligonucleotide of any of the preceding claims, wherein the oligonucleotide exhibits at least a 300% increase in full-length protein.

117. The compound or oligonucleotide of any of the preceding claims, wherein the oligonucleotide exhibits at least a 400% increase in full-length protein.

118. The compound or oligonucleotide of any one of claims 113-117, wherein the increase in full-length protein is measured compared to the level of decrease in full-length protein achieved using a TDP43 antisense oligonucleotide.

119. The compound or oligonucleotide of any of the preceding claims, wherein the oligonucleotide exhibits at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% rescue of the full-length protein.

120. The compound or oligonucleotide of any of the preceding claims, wherein the oligonucleotide exhibits at least a 50%, 60%, 70%, 80% or 90% reduction in mis-spliced ​​transcripts.

121. A method of treating a neurological disease and / or a neurological disorder in a patient in need thereof, the method comprising administering to the patient a compound or oligonucleotide as claimed in any one of claims 1 to 120.

122. The method of claim 121, wherein the neurological disease is selected from the group consisting of amyotrophic lateral sclerosis (ALS), frontotemporal dementia (FTD), ALS with FTD, Alzheimer's disease (AD), Parkinson's disease (PD), Parkinson's disease with dementia, dementia with Lewy bodies, synucleinopathy, Huntington's disease, brachial plexus injury, peripheral nerve injury, progressive supranuclear palsy (PSP), brain trauma, spinal cord injury, tuberous sclerosis complex, Pick's disease, tauopathies, primary age-related tauopathies, Down syndrome, epilepsy / seizure disorders, depression, traumatic brain injury (TBI), chronic traumatic encephalopathy (CTE), HIV-associated neurocognitive disorder (HAND), multiple system atrophy, amnestic mild cognitive impairment Disorders, corticobasal degeneration (CBD) and / or neuropathies such as chemotherapy-induced neuropathy, spinocerebellar ataxia (SCA), SCA type 2, spinal muscular atrophy (SMA), Parkinson's disease, Niemann-Pick disease type C (NPC), Charcot-Marie-Tooth disease (CMT), mucopolysaccharidosis type II (MPSIIA), mucolipidosis IV, GM1 gangliosidosis, sporadic inclusion body myositis (sIBM), Kennock-Scholin purpura (HSP), limbic-predominant age-related TDP-43 encephalopathy (late stage), cerebral age-related TDP-43 with sclerosis (CARTS), Gaucher disease and facial-onset sensory and motor neuronopathy, Guam Parkinson-dementia complex, multisystem proteinopathy, Perry disease, and synaptic diseases such as autism.

123. The method of claim 122, wherein the neurological disease is ALS.

124. The method of claim 122, wherein the neurological disease is FTD.

125. The method of claim 122, wherein the neurological disease is ALS with FTD.

126. The method of claim 122, wherein the neurological disease is AD.

127. The method of claim 122, wherein the neurological disease is PD.

128. The method of claim 122, wherein the neurological disease is spinal muscular atrophy (SMA).

129. The method of claim 121, wherein the neuropathy is chemotherapy-induced neuropathy.

130. A method of restoring axonal growth and / or regeneration of a neuron, the method comprising exposing the neuron to a compound or oligonucleotide as described in any one of claims 1-120.

131. A method of increasing, promoting, stabilizing or maintaining the expression and / or function of any one of STMN2, KCNQ2, UNC13A or SMN2 in a neuron, the method comprising exposing the cell to a compound or oligonucleotide as described in any one of claims 1-120.

132. The method of claim 130 or 131, wherein the neuron is a motor neuron.

133. The method of claim 130 or 131, wherein the neuron is a spinal cord neuron.

134. The method of any one of claims 130-133, wherein the neuron is a neuron of a patient in need of treatment for a neurological disease and / or a neuropathy.

135. The method of claim 134, wherein the neuropathy is chemotherapy-induced neuropathy.

136. The method of any one of claims 130-135, wherein the exposing is performed in vivo or ex vivo.

137. The method of any one of claims 130-135, wherein said exposing comprises administering said oligonucleotide to a patient in need thereof.

138. The method of any one of claims 130-137, wherein the oligonucleotide is administered topically, parenterally, intrathecally, intrathalamically, intracisternal, orally, rectally, buccally, sublingually, vaginally, pulmonary, intratracheally, intranasally, transdermally, intraduodenally, or intracerebroventricularly.

139. The method of claim 138, wherein the oligonucleotide is administered orally.

140. The method of any one of claims 130-138, wherein a therapeutically effective amount of the oligonucleotide is administered intrathecally, intrathalamus, intracerebroventricularly, or intracisternal. E 141. The method of any one of E 130-140, wherein the patient is a human.

142. A pharmaceutical composition comprising the oligonucleotide of any one of claims 1-120 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

143. The pharmaceutical composition of claim 142, wherein the pharmaceutical composition is suitable for topical, intrathecal, intrathalamic, intracisternal, intraventricular, parenteral, oral, pulmonary, intratracheal, intranasal, transdermal, rectal, buccal, sublingual, vaginal, or intraduodenal administration.

144. A method of treating a neurological disease or neuropathy in a patient in need thereof, the method comprising administering to the patient in need thereof a therapeutically effective amount of the pharmaceutical composition of claim 142 or 143.

145. The method of claim 144, wherein the neurological disease is selected from the group consisting of amyotrophic lateral sclerosis (ALS), frontotemporal dementia (FTD), ALS with FTD, Alzheimer's disease (AD), Parkinson's disease (PD), Parkinson's disease with dementia, dementia with Lewy bodies, synucleinopathy, Huntington's disease, brachial plexus injury, peripheral nerve injury, progressive supranuclear palsy (PSP), brain trauma, spinal cord injury, tuberous sclerosis complex, Pick's disease, tauopathies, primary age-related tauopathies, Down syndrome, epilepsy / seizure disorders, depression, traumatic brain injury (TBI), chronic traumatic encephalopathy (CTE), HIV-associated neurocognitive disorder (HAND), multiple system atrophy, amnestic mild cognitive impairment Disorders, corticobasal degeneration (CBD) and / or neuropathies such as chemotherapy-induced neuropathy, spinocerebellar ataxia (SCA), SCA type 2, spinal muscular atrophy (SMA), Parkinson's disease, Niemann-Pick disease type C (NPC), Charcot-Marie-Tooth disease (CMT), mucopolysaccharidosis type II (MPSIIA), mucolipidosis IV, GM1 gangliosidosis, sporadic inclusion body myositis (sIBM), Kennock-Scholin purpura (HSP), limbic-predominant age-related TDP-43 encephalopathy (late stage), cerebral age-related TDP-43 with sclerosis (CARTS), Gaucher disease and facial-onset sensory and motor neuronopathy, Guam Parkinson-dementia complex, multisystem proteinopathy, Perry disease, and synaptic diseases such as autism.

146. The method of claim 145, wherein the neurological disease is ALS.

147. The method of claim 145, wherein the neurological disease is FTD.

148. The method of claim 145, wherein the neurological disease is ALS with FTD.

149. The method of claim 145, wherein the neurological disease is AD.

150. The method of claim 145, wherein the neurological disease is PD.

151. The method of claim 145, wherein the neurological disease is spinal muscular atrophy (SMA).

152. The method of claim 144, wherein the neuropathy is chemotherapy-induced neuropathy.

153. The method of any of claims 144-152, wherein the pharmaceutical composition is administered topically, parenterally, orally, pulmonary, rectal, buccally, sublingually, vaginally, intratracheally, intranasally, intracisternal, intrathecal, intrathalamically, transdermally, intraduodenally, or intracerebroventricularly.

154. The method of any one of claims 144-152, wherein the pharmaceutical composition is administered intrathecally, intrathalamus, or intracisternal.

155. The method of any one of claims 144-154, wherein a therapeutically effective amount of the oligonucleotide is administered intrathecally, intrathalamus, or intracisternal.

156. The method of any one of claims 144-155, wherein the patient is a human.

157. A method for treating a neurological disease in a subject in need thereof, the method comprising administering to the subject an oligonucleotide or a pharmaceutically acceptable salt thereof, the oligonucleotide comprising a segment having up to 7 linked nucleosides, and wherein the oligonucleotide is at least 85% complementary to an equal length portion of a sequence from a transcript whose mis-splicing results in a neurological disease or a transcript whose splicing can be modulated to treat a neurological disease; wherein at least one (i.e., one or more) nucleoside linkages of the oligonucleotide are independently selected from the group consisting of phosphodiester linkages, phosphorothioate linkages, alkyl phosphate linkages, phosphorodithioate linkages, phosphotriester linkages, alkylphosphonate linkages, 3-methoxypropylphosphonate linkages, methylphosphonate linkages, aminoalkylphosphotriester linkages, alkylenephosphonate linkages, phosphinate linkages, phosphoamidate linkages, phosphorothioate amide linkages, phosphorothioate diamide linkages, phosphorodiamidate linkages, aminoalkylphosphoamidate linkages, phosphorothioate amide linkages, alkylphosphonate linkages, alkylthioate phosphotriester linkages, phosphorothioate linkages, phosphorothioate linkages, phosphorothioate linkages, phosphorothioate linkages, phosphorothioate linkages, and phosphoroborate linkages, and / or wherein at least one (i.e., one or more) nucleosides are substituted with a component selected from the group consisting of 2'-O-(2-methoxyethyl) nucleosides, 2'-O-methyl nucleosides, 2'-O-(N-methylacetamide) nucleosides, 2'-deoxy-2'-fluoro nucleosides, 2'-fluoro-β-D-arabinoside, locked nucleic acids (LNA), tricyclic nucleic acids, constrained methoxyethyl (cMOE), constrained ethyl (cET), and peptide nucleic acids (PNA), Optionally, wherein the oligonucleotide further comprises a spacer.

158. A method for treating a neurological disease in a subject in need thereof, the method comprising administering to the subject an oligonucleotide or a pharmaceutically acceptable salt thereof, the oligonucleotide comprising a segment of up to 7 linked nucleosides, and wherein the oligonucleotide is at least 85% complementary to a sequence from a transcript of any of STMN2, KCNQ2, UNC13A or SMN2; wherein at least one (i.e., one or more) nucleoside linkages of the oligonucleotide are independently selected from the group consisting of phosphodiester linkages, phosphorothioate linkages, alkyl phosphate linkages, phosphorodithioate linkages, phosphotriester linkages, alkylphosphonate linkages, 3-methoxypropylphosphonate linkages, methylphosphonate linkages, aminoalkylphosphotriester linkages, alkylenephosphonate linkages, phosphinate linkages, phosphoamidate linkages, phosphorothioate amide linkages, phosphorothioate diamide linkages, phosphorodiamidate linkages, aminoalkylphosphoamidate linkages, phosphorothioate amide linkages, alkylphosphonate linkages, alkylthioate phosphotriester linkages, phosphorothioate linkages, phosphorothioate linkages, phosphorothioate linkages, phosphorothioate linkages, phosphorothioate linkages, and phosphoroborate linkages, and / or wherein at least one (i.e., one or more) nucleosides are substituted with a component selected from the group consisting of 2'-O-(2-methoxyethyl) nucleosides, 2'-O-methyl nucleosides, 2'-O-(N-methylacetamide) nucleosides, 2'-deoxy-2'-fluoro nucleosides, 2'-fluoro-β-D-arabinoside, locked nucleic acids (LNA), tricyclic nucleic acids, constrained methoxyethyl (cMOE), constrained ethyl (cET), and peptide nucleic acids (PNA), Optionally, wherein the oligonucleotide further comprises a spacer.

159. A method for treating a neurological disease in a subject in need thereof, the method comprising administering to the subject an oligonucleotide or a pharmaceutically acceptable salt thereof, the oligonucleotide comprising a segment having up to 7 linked nucleosides, and wherein the oligonucleotide is conjugated to SEQ ID NO: 1-466, SEQ ID NO: 893-1338, SEQ ID NO: 1342-1366, SEQ ID NO: 1392-1664, or SEQ ID NO: 10655-10669, SEQ ID NO: 1676-1851, SEQ ID NO: 2028-2529, SEQ ID NO: 3046-3221, SEQ ID NO: 3398-3899, and SEQ ID NO: 4402-4530, SEQ ID NO: 4531-5794, SEQ ID NO: 7059-8322, SEQ ID NO: 9596-9603, SEQ ID NO: 10655-10669, SEQ ID NO: 1676-1851, SEQ ID NO: 2028-2529, SEQ ID NO: 3046-3221, SEQ ID NO: 3398-3899, and SEQ ID NO: 4402-4530, SEQ ID NO: 4531-5794, SEQ ID NO: 7059-8322, SEQ ID NO: 9596-9603, SEQ ID any of SEQ ID NOs: 9710-10141, SEQ ID NOs: 10574-10651, SEQ ID NOs: 10670-10779, SEQ ID NOs: 10783-10808, and SEQ ID NOs: 10814-10820 share at least 85% identity; wherein at least one (i.e., one or more) nucleoside linkages of the oligonucleotide are independently selected from the group consisting of phosphodiester linkages, phosphorothioate linkages, alkyl phosphate linkages, phosphorodithioate linkages, phosphotriester linkages, alkylphosphonate linkages, 3-methoxypropylphosphonate linkages, methylphosphonate linkages, aminoalkylphosphotriester linkages, alkylenephosphonate linkages, phosphinate linkages, phosphoamidate linkages, phosphorothioate amide linkages, phosphorothioate diamide linkages, phosphorodiamidate linkages, aminoalkylphosphoamidate linkages, phosphorothioate amide linkages, alkylphosphonate linkages, alkylthioate phosphotriester linkages, phosphorothioate linkages, phosphorothioate linkages, phosphorothioate linkages, phosphorothioate linkages, phosphorothioate linkages, and phosphoroborate linkages, and / or wherein at least one (i.e., one or more) nucleosides are substituted with a component selected from the group consisting of 2'-O-(2-methoxyethyl) nucleosides, 2'-O-methyl nucleosides, 2'-O-(N-methylacetamide) nucleosides, 2'-deoxy-2'-fluoro nucleosides, 2'-fluoro-β-D-arabinoside, locked nucleic acids (LNA), tricyclic nucleic acids, constrained methoxyethyl (cMOE), constrained ethyl (cET), and peptide nucleic acids (PNA), Optionally, wherein the oligonucleotide further comprises a spacer.

160. A method for treating amyotrophic lateral sclerosis (ALS) in a subject in need thereof, the method comprising administering to the subject an oligonucleotide or a pharmaceutically acceptable salt thereof, the oligonucleotide comprising a segment having up to 7 linked nucleosides, and wherein the oligonucleotide is conjugated to SEQ ID NO: 1-466, SEQ ID NO: 893-1338, SEQ ID NO: 1342-1366, SEQ ID NO: 1392-1664, or SEQ ID NO: 10655-10669, SEQ ID NO: 1676-1851, SEQ ID NO: 2028-2529, SEQ ID NO: 3046-3221, SEQ ID NO: 3398-3899, and SEQ ID NO: 4402-4530, or SEQ ID NO: 4531-5794, SEQ ID NO: 7059-8322, or SEQ ID NO: Any of NO: 9596-9603 shares at least 85% identity; wherein at least one (i.e., one or more) nucleoside linkages of the oligonucleotide are independently selected from the group consisting of phosphodiester linkages, phosphorothioate linkages, alkyl phosphate linkages, phosphorodithioate linkages, phosphotriester linkages, alkylphosphonate linkages, 3-methoxypropylphosphonate linkages, methylphosphonate linkages, aminoalkylphosphotriester linkages, alkylenephosphonate linkages, phosphinate linkages, phosphoamidate linkages, phosphorothioate amide linkages, phosphorothioate diamide linkages, phosphorodiamidate linkages, aminoalkylphosphoamidate linkages, phosphorothioate amide linkages, alkylphosphonate linkages, alkylthioate phosphotriester linkages, phosphorothioate linkages, phosphorothioate linkages, phosphorothioate linkages, phosphorothioate linkages, phosphorothioate linkages, and phosphoroborate linkages, and / or wherein at least one (i.e., one or more) nucleosides are substituted with a component selected from the group consisting of 2'-O-(2-methoxyethyl) nucleosides, 2'-O-methyl nucleosides, 2'-O-(N-methylacetamide) nucleosides, 2'-deoxy-2'-fluoro nucleosides, 2'-fluoro-β-D-arabinoside, locked nucleic acids (LNA), tricyclic nucleic acids, constrained methoxyethyl (cMOE), constrained ethyl (cET), and peptide nucleic acids (PNA), Optionally, wherein the oligonucleotide further comprises a spacer.

161. A method for treating Alzheimer's disease (AD) with frontotemporal dementia (FTD) in a subject in need thereof, the method comprising administering to the subject an oligonucleotide or a pharmaceutically acceptable salt thereof, the oligonucleotide comprising a segment having up to 7 linked nucleosides, and wherein the oligonucleotide is conjugated to SEQ ID NO: 1-466, SEQ ID NO: 893-1338, SEQ ID NO: 1342-1366, SEQ ID NO: 1392-1664, or SEQ ID NO: 10655-10669, SEQ ID NO: 1676-1851, SEQ ID NO: 2028-2529, SEQ ID NO: 3046-3221, SEQ ID NO: 3398-3899, and SEQ ID NO: 4402-4530, or SEQ ID NO: 4531-5794, SEQ ID NO: 7059-8322, or SEQ ID NO: Any of NO: 9596-9603 shares at least 85% identity; wherein at least one (i.e., one or more) nucleoside linkages of the oligonucleotide are independently selected from the group consisting of phosphodiester linkages, phosphorothioate linkages, alkyl phosphate linkages, phosphorodithioate linkages, phosphotriester linkages, alkylphosphonate linkages, 3-methoxypropylphosphonate linkages, methylphosphonate linkages, aminoalkylphosphotriester linkages, alkylenephosphonate linkages, phosphinate linkages, phosphoamidate linkages, phosphorothioate amide linkages, phosphorothioate diamide linkages, phosphorodiamidate linkages, aminoalkylphosphoamidate linkages, phosphorothioate amide linkages, alkylphosphonate linkages, alkylthioate phosphotriester linkages, phosphorothioate linkages, phosphorothioate linkages, phosphorothioate linkages, phosphorothioate linkages, phosphorothioate linkages, and phosphoroborate linkages, and / or wherein at least one (i.e., one or more) nucleosides are substituted with a component selected from the group consisting of 2'-O-(2-methoxyethyl) nucleosides, 2'-O-methyl nucleosides, 2'-O-(N-methylacetamide) nucleosides, 2'-deoxy-2'-fluoro nucleosides, 2'-fluoro-β-D-arabinoside, locked nucleic acids (LNA), tricyclic nucleic acids, constrained methoxyethyl (cMOE), constrained ethyl (cET) and peptide nucleic acids (PNA), optionally wherein the oligonucleotide further comprises a spacer.

162. A method for treating frontotemporal dementia (FTD) in a subject in need thereof, the method comprising administering to the subject an oligonucleotide or a pharmaceutically acceptable salt thereof, the oligonucleotide comprising a segment having up to 7 linked nucleosides, and wherein the oligonucleotide is conjugated to SEQ ID NO: 1-466, SEQ ID NO: 893-1338, SEQ ID NO: 1342-1366, SEQ ID NO: 1392-1664, or SEQ ID NO: 10655-10669, SEQ ID NO: 1676-1851, SEQ ID NO: 2028-2529, SEQ ID NO: 3046-3221, SEQ ID NO: 3398-3899, and SEQ ID NO: 4402-4530, or SEQ ID NO: 4531-5794, SEQ ID NO: 7059-8322, or SEQ ID NO: Any of NO: 9596-9603 shares at least 85% identity; wherein at least one (i.e., one or more) nucleoside linkages of the oligonucleotide are independently selected from the group consisting of phosphodiester linkages, phosphorothioate linkages, alkyl phosphate linkages, phosphorodithioate linkages, phosphotriester linkages, alkylphosphonate linkages, 3-methoxypropylphosphonate linkages, methylphosphonate linkages, aminoalkylphosphotriester linkages, alkylenephosphonate linkages, phosphinate linkages, phosphoamidate linkages, phosphorothioate amide linkages, phosphorothioate diamide linkages, phosphorodiamidate linkages, aminoalkylphosphoamidate linkages, phosphorothioate amide linkages, alkylphosphonate linkages, alkylthioate phosphotriester linkages, phosphorothioate linkages, phosphorothioate linkages, phosphorothioate linkages, phosphorothioate linkages, phosphorothioate linkages, and phosphoroborate linkages, and / or wherein at least one (i.e., one or more) nucleosides are substituted with a component selected from the group consisting of 2'-O-(2-methoxyethyl) nucleosides, 2'-O-methyl nucleosides, 2'-O-(N-methylacetamide) nucleosides, 2'-deoxy-2'-fluoro nucleosides, 2'-fluoro-β-D-arabinoside, locked nucleic acids (LNA), tricyclic nucleic acids, constrained methoxyethyl (cMOE), constrained ethyl (cET), and peptide nucleic acids (PNA), Optionally, wherein the oligonucleotide further comprises a spacer.

163. A method for treating amyotrophic lateral sclerosis (ALS) with frontotemporal dementia (FTD) in a subject in need thereof, the method comprising administering to the subject an oligonucleotide or a pharmaceutically acceptable salt thereof, the oligonucleotide comprising a segment having up to 7 linked nucleosides, and wherein the oligonucleotide is conjugated to SEQ ID NO: 1-466, SEQ ID NO: 893-1338, SEQ ID NO: 1342-1366, SEQ ID NO: 1392-1664, or SEQ ID NO: 10655-10669, SEQ ID NO: 1676-1851, SEQ ID NO: 2028-2529, SEQ ID NO: 3046-3221, SEQ ID NO: 3398-3899, and SEQ ID NO: 4402-4530, or SEQ ID NO: 4531-5794, SEQ ID NO: 7059-8322, or SEQ ID NO: Any of NO: 9596-9603 shares at least 85% identity; wherein at least one (i.e., one or more) nucleoside linkages of the oligonucleotide are independently selected from the group consisting of phosphodiester linkages, phosphorothioate linkages, alkyl phosphate linkages, phosphorodithioate linkages, phosphotriester linkages, alkylphosphonate linkages, 3-methoxypropylphosphonate linkages, methylphosphonate linkages, aminoalkylphosphotriester linkages, alkylenephosphonate linkages, phosphinate linkages, phosphoamidate linkages, phosphorothioate amide linkages, phosphorothioate diamide linkages, phosphorodiamidate linkages, aminoalkylphosphoamidate linkages, phosphorothioate amide linkages, alkylphosphonate linkages, alkylthioate phosphotriester linkages, phosphorothioate linkages, phosphorothioate linkages, phosphorothioate linkages, phosphorothioate linkages, phosphorothioate linkages, and phosphoroborate linkages, and / or wherein at least one (i.e., one or more) nucleosides are substituted with a component selected from the group consisting of 2'-O-(2-methoxyethyl) nucleosides, 2'-O-methyl nucleosides, 2'-O-(N-methylacetamide) nucleosides, 2'-deoxy-2'-fluoro nucleosides, 2'-fluoro-β-D-arabinoside, locked nucleic acids (LNA), tricyclic nucleic acids, constrained methoxyethyl (cMOE), constrained ethyl (cET), and peptide nucleic acids (PNA), Optionally, wherein the oligonucleotide further comprises a spacer.

164. The method of any one of claims 157-163, wherein the nucleoside linkage linking the base at position 3 or position 4 of the oligonucleotide is a phosphodiester linkage.

165. The method of claim 164, wherein only one nucleoside linkage linking the base at position 3 or position 4 of the oligonucleotide is a phosphodiester linkage.

166. The method of any one of claims 157-163, wherein the nucleoside linkage linking the bases at both position 3 and position 4 of the oligonucleotide is a phosphodiester linkage.

167. The method of any one of claims 157-163, wherein one or more bases immediately preceding the spacer in the oligonucleotide are linked by a phosphodiester bond.

168. The method of claim 167, wherein only the base immediately preceding the spacer in the oligonucleotide is linked to the spacer via a phosphodiester bond.

169. The method of claim 168, wherein the base immediately preceding the spacer in the oligonucleotide is further linked to a further preceding base via a phosphodiester bond.

170. The method of claim 168, wherein the oligonucleotide comprises a second spacer, wherein the base immediately preceding the second spacer is linked to a further preceding base via a phosphodiester bond.

171. The method of any one of claims 157-163, wherein one or more bases immediately following the spacer in the oligonucleotide are linked by a phosphodiester bond.

172. The method of claim 171, wherein only the base immediately following the spacer in the oligonucleotide is linked to the spacer via a phosphodiester bond.

173. The method of any one of claims 157-163, wherein the two bases immediately preceding the spacer in the oligonucleotide are linked by a phosphodiester bond.

174. The method of any one of claims 157-163, wherein the one or more bases in the oligonucleotide immediately before the spacer are linked by a phosphodiester bond, and wherein the one or more bases in the oligonucleotide immediately after the spacer are linked by a phosphodiester bond.

175. The method of claim 174, wherein a base immediately before the spacer and a base immediately after the spacer are linked by a phosphodiester bond.

176. The method of claim 174 or 175, wherein the oligonucleotide comprises a second spacer, and wherein one or more bases in the oligonucleotide immediately preceding the second spacer are linked by a phosphodiester bond, and wherein one or more bases in the oligonucleotide immediately following the second spacer are linked by a phosphodiester bond.

177. The method of claim 176, wherein a base immediately before the second spacer and a base immediately after the second spacer are linked by a phosphodiester bond.

178. The method of any one of claims 157-163, wherein the oligonucleotide comprises a series of bases linked by phosphodiester bonds, the series of bases comprising at least two bases.

179. The method of any one of claims 157-163, wherein the oligonucleotide comprises a series of bases linked by phosphodiester bonds, the series of bases comprising at least five bases.

180. The method of claim 178 or 179, wherein the oligonucleotide comprises two or more spacers, and wherein the series of bases is located between the at least two spacers.

181. The method of any one of claims 164-180, wherein the oligonucleotide is any of a 19-mer, a 21-mer, a 23-mer, or a 25-mer.

182. The method of any one of claims 157-163, wherein at least one (ie, one or more) internucleoside linkage of the oligonucleotide is a phosphorothioate linkage.

183. The method of any one of claims 157-163, wherein all internucleoside linkages of the oligonucleotide are phosphorothioate linkages.

184. An oligonucleotide and a pharmaceutically acceptable excipient, the oligonucleotide comprising an equal length portion to SEQ ID NO: 1339 or SEQ ID NO: 1341, SEQ ID NO: 3032-3043, SEQ ID NO: 9587-9595 or SEQ ID NO: 9698-9707, any of the sequences having 90% identity thereof, or a sequence that is at least 85% complementary to a 15 to 50 consecutive nucleobase portion thereof, optionally wherein the oligonucleotide comprises a spacer, and wherein the oligonucleotide is capable of increasing, restoring or stabilizing the expression of one or more of STMN2, KCNQ2, UNC13A or SMN2 mRNAs capable of translating a functional protein in a cell or human patient with an immune-mediated demyelinating disease, and wherein the level of increase, restoration or stabilization of expression and / or activity and / or function is sufficient for the oligonucleotide to be used as a drug for treating the immune-mediated demyelinating disease.

185. The method of any one of claims 121-141 or 144-183, the pharmaceutical composition of claim 142 or 143, or the oligonucleotide of any one of claims 1-120 or 184, wherein the oligonucleotide comprises one or more chiral centers and / or double bonds.

186. The method of any one of claims 121-141, 144-183, or 185, the pharmaceutical composition of claim 142, 143, or 185, or the oligonucleotide of any one of claims 1-120 or 184-185, wherein the oligonucleotide exists as a stereoisomer selected from geometric isomers, enantiomers, and diastereomers.

187. A method of treating a neurological disease or neuropathy in a patient in need thereof, the method comprising administering to the patient in need thereof a therapeutically effective amount of the pharmaceutical composition of claim 142 or 143 in combination with a second therapeutic agent.

188. The method of claim 187, wherein the second therapeutic agent is selected from the group consisting of riluzole (Rirudek), PrimeC, edaravone (Radicava), rivastigmine, donepezil, galantamine, selective serotonin reuptake inhibitors, antipsychotics, cholinesterase inhibitors, memantine, benzodiazepine anxiolytics, AMX0035 (ELYBRIO), ZILUCOPLAN (RA101495), pridopidine, dual AON intrathecal administration (e.g., BIIB067, BIIB078, and BIIB105), BHB100, levodopa / carbidopa, dopaminergic agents (e.g., ropinirole, pramipexole, rotigotine), medroxyprogesterone, ketones, KCNQ2 / KCNQ3 openers (e.g., retigabine, XEN1101, or QRL-101), bioactive scaffolds, anticonvulsants and psychostimulants, therapy (e.g., selected from respiratory care, physical therapy, occupational therapy, speech therapy, nutritional support), deep brain stimulation, levodopa and carbidopa (duopa, rytary, Sinemet, inbrija), istradefylline (nourianz), safinamide (xadago), pramipexole (Mirapex), rotigotine (neupro), ropinirole (requip), amantadine (gocovri, Symmetrel, osmolex), benztropine (Cog entin), trihexyphenidyl (artane), selegiline (eldepryl, zelapar), rasagiline, entacapone (comtan), opicapone (ongentys), tolcapone (tasmar), apomorphine (apokyn, kynmobi), exenatide, ganoderma lucidum, BIIB054, BIIB094, caffeine, thalizotan, embryonic dopamine cell implantation, aducanumab (Aduhlem), memantine (Namenda), donepezil (Aricept), rivastigmine (Exelon), galantamine (razadyne), Namzeric, sulvorexant (belsomra) , lencanezumab, olanzapine (Zyprexa), quetiapine (Seroquel), SSRIs (citalopram (Cipramil), dapoxetine (Priligy), escitalopram (Cipralex), fluoxetine (Prozac or Oxactin), fluvoxamine (Faverin), paroxetine (Seroxat), sertraline (Lustral), vortioxetine (Brintellix)), divalproex sodium (Depakote), carbamazepine (Tegretol), medroxyprogesterone, brivaracetam (briviact), cannabidiol (epidiolex), carbamazepine (carbatrol,Tegretol), phenbarate (xcopri), diazepam (valium), lorazepam (Ativan), clonazepam (klonopin), eslicarbazepine (aptiom), ethosuximide (zarontin), felbatol (felbatol), fenfluramine (fintepla), lacosamide (VIMPAT), lamotrigine (Lamictal), levetiracetam (Keppra), oxcarbazepine (oxtellar xr, Trileptal), perampanel (fycompa), phenobarbital, phenytoin (dilantin), pregabalin (lyrica), tiagabine (gabitril), topiramate (topamax), valproate (depakene, depakote) and / or zonisamide (zonegran), which are used to treat the neurological disease. , 189. A method for treating a neurological disease and / or a neurological disorder in a patient in need thereof, the method comprising administering to a patient in need thereof a therapeutically effective amount of the pharmaceutical composition of claim 142 or 143, and wherein at least one nucleoside linkage of the oligonucleotide is a non-natural linkage, optionally wherein the oligonucleotide comprises a spacer, and wherein the oligonucleotide further comprises a targeting or conjugation moiety selected from cholesterol, lipoic acid, pantothenic acid, polyethylene glycol, and an antibody for crossing the blood-brain barrier.

190. The method of any one of claims 121-141, 144-183, or 185-189, wherein the spacer is a nucleoside replacement group comprising a non-sugar substituent that is not capable of attachment to a nucleotide base.

191. The method of claim 190, wherein the spacer is located between positions 10 and 15 of the oligonucleotide.

192. The method of claim 190, wherein the spacer is located between positions 7 and 11 of the oligonucleotide.

193. The method of claim 190 or 192, wherein the oligonucleotide further comprises a second spacer, wherein the second spacer is located between positions 14 and 22 of the oligonucleotide.

194. The method of claim 193, wherein the spacer and the second spacer are separated by at least 5 nucleobases, at least 6 nucleobases, or at least 7 nucleobases in the oligonucleotide.

195. The method of claim 193 or 194, wherein the spacer is located between positions 7 and 9 of the oligonucleotide, and wherein the second spacer is located between positions 15 and 18 of the oligonucleotide.

196. The method of any one of claims 193-195, wherein the spacer is located at position 8 of the oligonucleotide, and wherein the second spacer is located at position 16 of the oligonucleotide.

197. The method of any one of claims 193-196, wherein the oligonucleotide further comprises a third spacer, wherein the third spacer is located between positions 21 and 24 of the oligonucleotide.

198. The method of claim 190, wherein the spacer is located between positions 2 and 5 of the oligonucleotide.

199. The method of claim 198, wherein the oligonucleotide further comprises a second spacer, wherein the second spacer is located between positions 8 and 12 of the oligonucleotide.

200. The method of claim 199, wherein the oligonucleotide further comprises a third spacer, wherein the third spacer is located between positions 18 and 22 of the oligonucleotide.

201. The method of claim 190, wherein the oligonucleotide further comprises a second spacer and a third spacer, wherein the three spacers are located at positions in the oligonucleotide such that each segment of the oligonucleotide has at most 7 linked nucleosides.

202. The method of claim 201, wherein at least two of the three spacers are adjacent to a guanine nucleobase.

203. The method of claim 202, wherein each of at least two of the three spacers is immediately preceded by a guanine nucleobase.

204. The method of any one of claims 190-203, wherein each of the first, second or third spacers is a nucleoside replacement group comprising a non-sugar substituent, wherein the non-sugar substituent does not contain a ketone, aldehyde, ketal, hemiketal, acetal, hemiacetal, aminal or hemiaminal moiety and is not capable of forming a covalent bond with a nucleotide base.

205. The method of any one of claims 190-203, wherein each of the first, second, or third spacers is independently represented by formula (X), wherein: Ring A is an optionally substituted 4-8 membered monocyclic cycloalkyl or 4-8 membered monocyclic heterocyclyl, wherein the heterocyclyl contains 1 or 2 heteroatoms selected from O, S and N, provided that A is not capable of forming a covalent bond with a nucleobase; and Said The symbol represents the point of attachment of the internucleoside linkage.

206. The method of claim 205, wherein each of the first, second, or third spacers is independently represented by formula (Xa), wherein:

207. The method of claim 205 or 206, wherein Ring A is an optionally substituted 4-8 membered monocyclic cycloalkyl or 4-8 membered monocyclic heterocyclyl, wherein the 4-8 membered monocyclic cycloalkyl is selected from cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl; and the 4-8 membered monocyclic heterocyclyl is selected from oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, 1,4-dioxacyclohexyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl and azepanyl.

208. The method of claim 207, wherein Ring A is tetrahydrofuranyl.

209. The method of claim 207, wherein Ring A is tetrahydropyranyl.

210. The method of any one of claims 190-203, wherein each of the first, second, or third spacers is independently represented by formula (I), wherein: X is selected from -CH2- and -O-; and n is 0, 1, 2 or 3.

211. The method of any one of claims 190-203, wherein each of the first, second, or third spacers is independently represented by formula (I'), wherein:

212. The method of any one of claims 190-203, wherein each of the first, second, or third spacers is independently represented by formula (la), wherein:

213. The method of any one of claims 190-203, wherein each of the first, second, or third spacers is independently represented by formula (Ia'), wherein:

214. The method of any one of claims 190-203, wherein each of the first, second, or third spacers is independently represented by Formula II, wherein: and X is selected from -CH2- and -O-.

215. The method of any one of claims 190-203, wherein each of the first, second, or third spacers is independently represented by Formula II', wherein: and X is selected from -CH2- and -O-.

216. The method of any one of claims 190-203, wherein each of the first, second, or third spacers is independently represented by formula (IIa), wherein:

217. The method of any one of claims 190-203, wherein each of the first, second or third spacers is independently represented by formula (Iia'), wherein:

218. The method of any one of claims 190-203, wherein each of the first, second, or third spacers is independently represented by formula (III), wherein: X is selected from -CH2- and -O-.

219. The method of any one of claims 190-203, wherein each of the first, second, or third spacers is independently represented by formula (III'), wherein: X is selected from -CH2- and -O-.

220. The method of any one of claims 190-203, wherein each of the first, second, or third spacers is independently represented by formula (IIIb), wherein:

221. The method of any one of claims 190-203, wherein each of the first, second, or third spacers is independently represented by formula (IIIb'), wherein:

222. The method of any one of claims 190-203, wherein each of the first, second, or third spacers is independently represented by Formula III, wherein: and X is selected from -CH2- and -O-.

223. The method of any one of claims 190-203, wherein each of the first, second, or third spacers is independently represented by Formula III', wherein: and X is selected from -CH2- and -O-.

224. The method of any one of claims 190-203, wherein each of the first, second, or third spacers is independently represented by formula (IIIa), wherein:

225. The method of any one of claims 190-203, wherein each of the first, second, or third spacers is independently represented by formula (IIIa'), wherein:

226. The method of any one of claims 190-225, wherein the oligonucleotide comprising the spacer has a GC content of at least 10%.

227. The method of any one of claims 190-226, wherein the oligonucleotide comprising the spacer has a GC content of at least 20%.

228. The method of any one of claims 190-227, wherein the oligonucleotide comprising the spacer has a GC content of at least 25%.

229. The method of any one of claims 190-228, wherein the oligonucleotide comprising the spacer has a GC content of at least 30%.

230. The method of any one of claims 190-229, wherein the oligonucleotide comprising the spacer has a GC content of at least 40%.

231. The method of any one of claims 190-230, wherein the oligonucleotide comprising the spacer has a GC content of at least 50%.

232. A method for synthesizing an AON with a spacer, the method comprising: Obtained by Compound A represented by Benzoyl protection reaction of compound A is carried out to produce Compound B represented by Reductive nucleobase cleavage of compound B is performed to generate a Compound C represented by The saponification reaction of compound C is carried out to produce a compound of the formula Compound D represented by A protection reaction of compound D is carried out to generate a compound of the formula The compound E represented by and The phosphoramidite installation reaction of compound E is carried out to generate Represents compound F.

233. A composition comprising: With formula Compound F.

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