Gapmer antisense oligonucleotides with modified backbone chemistry - Patent Application 20070229933

Abstract

Disclosed herein are antisense oligonucleotides, such as gapmer antisense oligonucleotides with modified backbone chemistry (e.g., with one or more spacers).Such gapmer antisense oligonucleotides with modified backbone chemistry can be useful for treating various diseases, such as neurological diseases.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Application No. 63 / 285,705, filed December 3, 2021, U.S. Provisional Application No. 63 / 285,888, filed December 3, 2021, U.S. Provisional Application No. 63 / 285,692, filed December 3, 2021, U.S. Provisional Application No. 63 / 285,696, filed December 3, 2021, and U.S. Provisional Application No. 63 / 285,665, filed December 3, 2021, each of which is incorporated by reference herein in its entirety for all purposes.

[0002] Reference to sequence table XML This application contains a Sequence Listing that has been submitted electronically in XML format. The Sequence Listing XML is incorporated herein by reference. The XML file, created on November 17, 2022, is named QRL-011WO_SL.xml and is 418,206,230 in size. [Background technology]

[0003] Antisense oligonucleotide is a compound based on nucleic acid, which can be used to inhibit the expression of certain genes related to disease.Antisense oligonucleotide can generally be designed to hybridize with target gene, but traditional antisense oligonucleotide often shows poor efficacy.Therefore, it is necessary to develop modified antisense oligonucleotide that shows improved performance and efficacy for preventing, mitigating and treating diseases, for example, neurological diseases. Summary of the Invention

[0004] Disclosed herein are compounds comprising gapmer oligonucleotides, wherein the gapmer oligonucleotides comprise a spacer. Disclosed herein are also gapmer oligonucleotides comprising a spacer. In various embodiments, the gapmer oligonucleotides comprise a second spacer that is not adjacent to the spacer.

[0005] In various embodiments, the gapmer oligonucleotide comprises a sequence that is 85-98% complementary to an equal length portion of a transcript whose presence results in a neurological disorder. In various embodiments, the gapmer oligonucleotide comprises a sequence that is 85-98% complementary to an equal length portion of a PPM1A mRNA or pre-mRNA transcript, an ATXN2 mRNA or pre-mRNA transcript, an SOD1 mRNA or pre-mRNA transcript, or an MAPT mRNA or pre-mRNA transcript. In various embodiments, the gapmer oligonucleotide comprises SEQ ID NOs: 1909-1913, 149355-149361, 167802-167804, or 301567-301589, a sequence having 90% identity thereto, or a sequence that is 85-98% complementary to an equal length portion of a 15-50 contiguous nucleobase portion thereof.

[0006] In various embodiments, the gapmer oligonucleotide comprises a sequence that shares at least 85% identity with an equal length portion of any one of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, 167805-301566, SEQ ID NOs: 301692-301742, or the sequences in Tables 4A-4C. In various embodiments, the gapmer oligonucleotide comprises a sequence that shares at least 90% identity with an equal length portion of any one of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, 167805-301566, SEQ ID NOs: 301692-301742, or the sequences in Tables 4A-4C. In various embodiments, the gapmer oligonucleotide comprises a sequence that shares at least 95% identity with an equal length portion of any one of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, 167805-301566, SEQ ID NOs: 301692-301742, or the sequences in Tables 4A-4C. In various embodiments, the gapmer oligonucleotide comprises a sequence that shares at least 100% identity with an equal length portion of any one of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, 167805-301566, SEQ ID NOs: 301692-301742, or the sequences in Tables 4A-4C.

[0007] In various embodiments, the gapmer oligonucleotide comprises a segment having at most 11 linked nucleosides, wherein the gapmer oligonucleotide comprises a sequence that shares at least 85% identity with an equal length portion of any one of SEQ ID NOS: 1-954, 1914-149354, 149362-158581, 167805-301566, SEQ ID NOS: 301692-301742, or the sequences in Tables 4A-4C. In various embodiments, the gapmer oligonucleotide comprises a segment having at most 10, 9, 8, 7, 6, 5, 4, 3, or 2 linked nucleosides, wherein the gapmer oligonucleotide comprises a sequence that shares at least 85% identity with an equal length portion of any one of SEQ ID NOS: 301692-301742, or the sequences in Tables 4A-4C. In various embodiments, the gapmer oligonucleotide comprises a segment having at most 11 linked nucleosides, wherein the gapmer oligonucleotide comprises a sequence that shares at least 90% identity with an equal length portion of any one of SEQ ID NOs: 301692-301742 or the sequences in Tables 4A-4C. In various embodiments, the gapmer oligonucleotide comprises a segment having at most 10, 9, 8, 7, 6, 5, 4, 3, or 2 linked nucleosides, wherein the gapmer oligonucleotide comprises a sequence that shares at least 90% identity with an equal length portion of any one of SEQ ID NOs: 301692-301742 or the sequences in Tables 4A-4C.

[0008] In various embodiments, the gapmer 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, at least one (i.e., one or more) nucleoside linkage of the gapmer oligonucleotide is a non-natural linkage, and in various embodiments, the gapmer oligonucleotide is at least 19 oligonucleotide units in length.

[0009] In various embodiments, the spacer is a nucleoside substitute comprising a non-sugar surrogate that cannot be linked to a nucleotide base. In various embodiments, the spacer is positioned between positions 5 and 11 of the gapmer oligonucleotide. In various embodiments, the spacer is positioned between positions 7 and 11 of the gapmer oligonucleotide. In various embodiments, the gapmer oligonucleotide further comprises a second spacer, wherein the spacer and the second spacer are not adjacent to each other. In various embodiments, the gapmer oligonucleotide further comprises a second spacer, wherein the second spacer is positioned between positions 15 and 19 of the gapmer oligonucleotide. In various embodiments, the spacer and the second spacer are spaced apart by at least 5 nucleobases, at least 6 nucleobases, or at least 7 nucleobases in the gapmer oligonucleotide. In various embodiments, the spacer is positioned between positions 5 and 11 of the gapmer oligonucleotide, and the second spacer is positioned between positions 15 and 19 of the gapmer oligonucleotide. In various embodiments, the spacer is positioned at position 8 of the gapmer oligonucleotide and the second spacer is positioned at position 16 of the gapmer oligonucleotide. In various embodiments, the spacer is positioned at position 5 of the gapmer oligonucleotide and the second spacer is positioned at position 17 of the gapmer oligonucleotide. In various embodiments, the spacer is positioned at position 7 of the gapmer oligonucleotide and the second spacer is positioned at position 15 of the gapmer oligonucleotide. In various embodiments, the spacer is positioned at position 11 of the gapmer oligonucleotide and the second spacer is positioned at position 19 of the gapmer oligonucleotide.

[0010] In various embodiments, each of the spacer or second spacer is a nucleoside substitute that includes a non-sugar surrogate, wherein the non-sugar surrogate does not include a ketone, aldehyde, ketal, hemiketal, acetal, hemiacetal, aminal, or hemiaminal moiety and is unable to form a covalent bond with a nucleotide base. In various embodiments, each of the spacer or second spacer independently has the formula (X):

[0011] [ka] wherein: Ring A is an optionally substituted 4- to 8-membered monocyclic cycloalkyl group or a 4- to 8-membered monocyclic heterocyclyl group, the heterocyclyl group containing 1 or 2 heteroatoms selected from O, S, and N, with the proviso that A cannot form a covalent bond with a nucleobase;

[0012] [ka] The symbol represents the point of attachment to the internucleoside linkage.

[0013] In various embodiments, each of the spacer or second spacer independently has the formula (Xa):

[0014] [ka] is expressed by

[0015] In various embodiments, Ring A is an optionally substituted 4-8 membered monocyclic cycloalkyl group selected from cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl, or a 4-8 membered monocyclic heterocyclyl group selected from oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, 1,4-dioxanyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, and azepanyl. In various embodiments, Ring A is tetrahydrofuranyl. In various embodiments, Ring A is tetrahydropyranyl.

[0016] In various embodiments, each of the spacer and the second spacer independently has Formula I:

[0017] [ka] wherein: X is selected from —CH— and —O—; n is 0, 1, 2, or 3.

[0018] In various embodiments, each of the spacer and the second spacer independently has the formula I':

[0019] [ka] wherein: X is selected from —CH— and —O—; n is 0, 1, 2, or 3.

[0020] In various embodiments, each of the spacer and the second spacer independently has the formula (Ia):

[0021] [ka] wherein: n is 0, 1, 2, or 3.

[0022] In various embodiments, each of the spacer and the second spacer independently has the formula (Ia'):

[0023] [ka] wherein: n is 0, 1, 2, or 3.

[0024] In various embodiments, each of the spacer and the second spacer independently has Formula II:

[0025] [ka] wherein: X is selected from -CH2- and -O-.

[0026] In various embodiments, each of the spacer and the second spacer independently has the formula II':

[0027] [ka] wherein: X is selected from -CH2- and -O-.

[0028] In various embodiments, each of the spacer and the second spacer independently has the formula (Iia):

[0029] [ka] is expressed by

[0030] In various embodiments, each of the spacer and the second spacer independently has the formula (Iia'):

[0031] [ka] is expressed by

[0032] In various embodiments, each of the spacer and the second spacer independently has the formula III:

[0033] [ka] wherein: X is selected from -CH2- and -O-.

[0034] In various embodiments, each of the spacer and the second spacer independently has the formula III':

[0035] [ka] wherein: X is selected from -CH2- and -O-.

[0036] In various embodiments, each of the spacer and the second spacer independently has the formula (IIIa):

[0037] [ka] is expressed by

[0038] In various embodiments, each of the spacer and the second spacer independently has the formula (IIIa'):

[0039] [ka] is expressed by

[0040] In various embodiments, gapmer oligonucleotides comprising a spacer have a GC content of at least 10%. In various embodiments, gapmer oligonucleotides comprising a spacer have a GC content of at least 20%. In various embodiments, gapmer oligonucleotides comprising a spacer have a GC content of at least 25%. In various embodiments, gapmer oligonucleotides comprising a spacer have a GC content of at least 30%. In various embodiments, gapmer oligonucleotides comprising a spacer have a GC content of at least 40%. In various embodiments, gapmer oligonucleotides comprising a spacer have a GC content of at least 50%. In various embodiments, at least one (i.e., one or more) internucleoside linkages of the gapmer oligonucleotide are independently selected from the group consisting of phosphodiester linkages, phosphorothioate linkages, alkylphosphate linkages, phosphorodithioate linkages, phosphotriester linkages, alkylphosphonate linkages, 3-methoxypropylphosphonate linkages, methylphosphonate linkages, aminoalkylphosphotriester linkages, alkylenephosphonate linkages, phosphinate linkages, phosphoramidate linkages, phosphoramidothioate linkages, thiophosphorodiamidate linkages, phosphorodiamidate linkages, aminoalkylphosphoramidate linkages, thiophosphoramidate linkages, thionoalkylphosphonate linkages, thionoalkylphosphotriester linkages, thiophosphate linkages, selenophosphate linkages, and boranophosphate linkages. In various embodiments, at least one internucleoside linkage of the nucleotide sequence is a phosphorothioate linkage. In various embodiments, the phosphorothioate internucleoside linkages are in one of the Rp or Sp conformations.

[0041] In various embodiments, the gapmer oligonucleotide contains one or more chiral centers and / or double bonds. In various embodiments, the gapmer oligonucleotide exists as a stereoisomer selected from geometric isomers, enantiomers, and diastereomers. In various embodiments, all internucleoside linkages of the nucleotide sequence are phosphorothioate linkages. In various embodiments, the gapmer oligonucleotide contains at least one modified nucleobase. In various embodiments, the at least one modified nucleobase is 5-methylcytosine, pseudouridine, or 5-methoxyuridine. In various embodiments, the gapmer oligonucleotide contains at least one nucleoside having a 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), 2'-O-(N-methylacetamide), a 2'-deoxy-2'-fluoronucleoside, a 2'-fluoro-β-D-arabinonucleoside, a locked nucleic acid (LNA), a constrained ethyl 2'-4'-bridged nucleic acid (cEt), an S-cEt, a hexitol nucleic acid (HNA), and a tricyclic analog (e.g., tcDNA).

[0042] In various embodiments, the gapmer oligonucleotide comprises 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleosides having modified sugar moieties. In various embodiments, the modified sugar moieties are independently any one of 2'-OMe modified sugar moieties, bicyclic sugar moieties, 2'-O-(2-methoxyethyl) (2'-MOE), 2'-O-(N-methylacetamide), 2'-deoxy-2'-fluoronucleosides, 2'-fluoro-β-D-arabinonucleosides, locked nucleic acids (LNA), constrained ethyl 2'-4'-bridged nucleic acids (cEt), S-cEt, hexitol nucleic acids (HNA), and tricyclic analogs (e.g., tcDNA). In various embodiments, the gapmer oligonucleotide comprises 10 2'-O-(2-methoxyethyl) (2'-MOE) nucleosides. In various embodiments, five of the 2'-O-(2-methoxyethyl) (2'-MOE) nucleosides are positioned at the 3' end of the gapmer oligonucleotide, and five of the 2'-O-(2-methoxyethyl) (2'-MOE) nucleosides are positioned at the 5' end of the gapmer oligonucleotide. In various embodiments, at least one nucleoside having a modified sugar moiety or a nucleoside having a modified sugar moiety is a ribonucleoside. In various embodiments, the gapmer oligonucleotide comprises at least one deoxyribonucleoside. In various embodiments, the gapmer oligonucleotide comprises 2, 3, 4, 5, 6, 7, 8, 9, or 10 deoxyribonucleosides.

[0043] In various embodiments, the gapmer oligonucleotide comprises a gap segment comprising one or more of linked deoxyribonucleosides, 2'-fluoroarabinonucleic acid (FANA), and fluorocyclohexenyl nucleic acid (F-CeNA), a 5' wing region comprising linked nucleosides, and a 3' wing region comprising linked nucleosides, wherein the central region comprises at least eight nucleobases comprising at least four contiguous nucleobases disposed between the 5' wing segment and the 3' wing segment. wherein the region has at least 80% identity to an equal length portion of any one of SEQ ID NOS: 1-954, 1914-149354, 149362-158581, 167805-301566, SEQ ID NOS: 301692-301742, or the sequences in Tables 4A-4C, and the 5' wing region and the 3' wing region each comprise at least two linked nucleosides, and at least one nucleoside in each wing region comprises a modified sugar.

[0044] In various embodiments, at least two linked nucleosides in the 5' wing region are linked through phosphorothioate internucleoside linkages, and / or at least two linked nucleosides in the 3' wing region are independently linked through phosphorothioate internucleoside linkages. In various embodiments, all internucleoside linkages in the 5' wing region and / or all internucleoside linkages in the 3' wing region are independently phosphorothioate internucleoside linkages. In various embodiments, the 5' wing region further comprises at least one phosphodiester internucleoside linkage. In various embodiments, the 3' wing region further comprises at least one phosphodiester internucleoside linkage. In various embodiments, at least two linked nucleosides in the 5' wing region are linked via a phosphodiester internucleoside linkage, and / or at least two linked nucleosides in the 3' wing region are independently linked via a phosphodiester internucleoside linkage. In various embodiments, at least one of the internucleoside linkages in the central region is a phosphodiester linkage. In various embodiments, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, or at least nine of the internucleoside linkages in the central region are phosphodiester linkages. In various embodiments, at least one of the internucleoside linkages in the central region is a phosphorothioate internucleoside linkage. In various embodiments, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, or at least nine of the internucleoside linkages in the central region are phosphorothioate internucleoside linkages. In various embodiments, all of the internucleoside linkages of the gapmer oligonucleotide are phosphorothioate internucleoside linkages. In various embodiments, any one or all of the phosphorothioate internucleoside linkages are in the Rp conformation, the Sp conformation, or any combination of the Rp conformation and the Sp conformation.

[0045] In various embodiments, the gapmer oligonucleotide comprises at least one modified sugar moiety. In various embodiments, the 5'-wing region or the 3'-wing region comprises at least one modified sugar moiety. In various embodiments, the central region comprises at least one modified sugar moiety. In various embodiments, the at least one modified sugar moiety is any one of a 2'-OMe modified sugar moiety, a bicyclic sugar moiety, 2'-O-(2-methoxyethyl) (2'-MOE), 2'-O-(N-methylacetamide), a 2'-deoxy-2'-fluoronucleoside, a 2'-fluoro-β-D-arabinonucleoside, a locked nucleic acid (LNA), a constrained ethyl 2'-4'-bridged nucleic acid (cEt), an S-cEt, a tcDNA, a hexitol nucleic acid (HNA), and a tricyclic analog (e.g., a tcDNA). In various embodiments, the gapmer oligonucleotide comprises one or more 2'-MOE nucleosides. In various embodiments, the 5' or 3' wing region comprises one or more 2'-MOE nucleosides. In various embodiments, the 5' or 3' wing region comprises 2, 3, 4, 5, or 6 2'-MOE nucleosides. In various embodiments, all nucleosides in the 5' or 3' wing region are 2'-MOE nucleosides. In various embodiments, the central region comprises one or more 2'-MOE nucleosides. In various embodiments, the central region comprises 2, 3, 4, 5, 6, 7, 8, 9, or 10 2'-MOE nucleosides. In various embodiments, all nucleosides in the central region are 2'-MOE nucleosides. In various embodiments, one or more 2'-MOE nucleosides are linked via phosphorothioate internucleoside linkages.

[0046] In various embodiments, gapmer oligonucleotides comprise sugar modifications in any of the following patterns: eeeee-d10-eeeee, eeeee-d8-eeeee, eeeeee-d11-eeeeee, eee-d8-eee, eee-d10-eee, eeee-d10-eeee, and eeee-d8-eeee, where e=2'-MOE nucleoside and d=deoxyribonucleoside, and at least one "e" or at least one "d" is replaced with a spacer.In various embodiments, gapmer oligonucleotides have internucleoside linkages arranged in the following pattern: sssssooooooooosssss, ooooosssssssssooooo; oooooooooooooosssss; soosssssssssssssssss; sososssssssssssosos; sssssssssssssssss; sssssoooooooooooooo; ssssssssssssssssss; sssssoooooooooooooo; ssssssssssssssssss; sssssssssssss ssssssssssss;sssoooooooss;ooosssssssooo;ssssssssssss;sosssssssssos;sosssssssssss;ssssssssssssos;sssssssssss ooo;ooossssssssss;sssoooooooooss;ooosssssssssooo;ssssssssssssss;ssssssssssssooo;ooossssssssssss;sosssssssss ssos;sossssssssssss;sssssssssssssos;ssssooooooooosss;oooosssssssssssoo;ssssssssssssssss;ssssssssssssssssss ;ssssssssssssssss;sssssssssssssss;sssssssssssssssoos;oooosssssssssssss;ssssooooooosss;oooosssssssoooo;ssss wherein s=phosphorothioate linkage and o=phosphodiester linkage.

[0047] In various embodiments, gapmer oligonucleotides have a combination of sugar modifications and internucleoside linkages, respectively, in the following patterns: a) eeeee-d10-eeeee and sssssooooooooosssss; b) eeeee-d10-eeeee and ooooosssssssssooooo; c) eeeee-d10-eeeee and ssssssssssssssssssss; d) eee-d8-eee and sssooooooosss; e)eee-d8-eee and ooosssssssooo f) eee-d8-eee and sssssssssssss; g) eee-d10-eee and sssooooooooosss; h) eee-d10-eee and ooosssssssssooo; i) eee-d10-eee and ssssssssssssssss; j)eeee-d10-eeee and ssssooooooooossss; k)eeee-d10-eeee and oooosssssssssoooo; l)eeee-d10-eeee and ssssssssssssssssss; m)eeee-d8-eeee and ssssooooooossss, n)eeee-d8-eeee and oooosssssssoooo, o)eeee-d8-eeee and ssssssssssssssss, p)eeeeee-d11-eeeeee and sssssssssssssssssssss; q)eeeee-d10-eeeee and sosossssssssssssosos; r)eeeee-d10-eeeee and soooosssssssssssooos; s)eeeee-d10-eeeee and soooosssssssssssooss; t)eeeee-d8-eeeee and sossssssssssssssoss; u)eeeee-d10-eeeee and sosssssssssssssssosss; v)eeeeee-d10-eeee and ssssssssssssssssss; and e=2'-MOE nucleoside, d=deoxyribonucleoside, s=phosphorothioate linkage, o=phosphodiester linkage, and at least one "e" or at least one "d" of the gapmer oligonucleotide is replaced with a spacer.

[0048] In various embodiments, the gapmer oligonucleotide comprises at least one modified nucleobase. In various embodiments, the 5'-wing region or the 3'-wing region comprises at least one modified nucleobase. In various embodiments, the central region comprises at least one modified nucleobase. In various embodiments, the at least one modified nucleobase is 5-methylcytosine, pseudouridine, or 5-methoxyuridine. In various embodiments, all cytosines in the 5'-wing region or the 3'-wing region are 5-methylcytosines. In various embodiments, all cytosines in the central region are 5-methylcytosines. In various embodiments, the gapmer oligonucleotide comprises a combination of sugar modifications and internucleoside linkages of eeeee-d10-eeeee and sssssssssssssssssss, where e=2'-MOE nucleoside, d=deoxyribonucleoside, and s=phosphorothioate linkage, wherein at least one "e" or at least one "d" of the gapmer oligonucleotide is replaced by a spacer, and each cytosine of the 2'-MOE nucleoside is a 5-methylcytosine.

[0049] In various embodiments, the gapmer oligonucleotide further comprises a conjugate moiety, hi various embodiments, the conjugate moiety is a cholesterol conjugate positioned at the 3' end of the gapmer oligonucleotide.

[0050] Further disclosed herein is a pharmaceutical composition comprising a gapmer oligonucleotide disclosed herein, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0051] Further disclosed herein is a method of treating a neurological disease in a patient in need thereof, comprising administering to the patient a gapmer oligonucleotide disclosed herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition disclosed herein. In various embodiments, the neurological disease is 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's 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 neuropathy, e.g., chemotherapy. The present invention relates to a method-induced neuropathy, spinocerebellar ataxia (SCA), SCA type 2, spinal muscular atrophy (SMA), parkinsonism, Niemann-Pick disease type C (NPC), Charcot-Marie-Tooth disease (CMT), mucopolysaccharidosis type II (MPSIIA), mucolipidosis IV, GM1 gangliosidosis, sporadic inclusion body myositis (sIBM), Henoch-Schonlein purpura (HSP), limbic-predominant age-related TDP-43 encephalopathy (LATE), cerebral age-related TDP-43 with sclerosis (CARTS), Gaucher disease, and synaptic diseases such as facial-onset sensorimotor neuropathy, Guam Parkinsonism-Dementia Complex, multisystem proteinopathy, Perry disease, and autism.

[0052] In various embodiments, the gapmer oligonucleotide is administered topically, parenterally, intrathecally, orally, intrapulmonary, intratracheally, intranasally, transdermally, bucally, intrathalamic, intracerebroventricularly, intraocularly, sublingually, rectally, intravaginally, or intraduodenum. In various embodiments, the gapmer oligonucleotide is administered intrathecally. In various embodiments, a therapeutically effective amount of the gapmer oligonucleotide is administered. In various embodiments, the patient is a human. In various embodiments, the pharmaceutical composition is suitable for topical, parenteral, intrathecal, oral, intrapulmonary, intratracheally, intranasal, transdermal, buccal, intrathalamic, intracerebroventricularly, intraocularly, sublingually, rectally, intravaginally, or intraduodenum.

[0053] Further disclosed herein is the use of gapmer oligonucleotides in the manufacture of a medicament for the treatment of a neurological disease. 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's 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 neuropathy, e.g., chemotherapy. The gapmer oligonucleotide is selected from the group consisting of chemotherapy-induced neuropathy, spinocerebellar ataxia (SCA), SCA type 2, spinal muscular atrophy (SMA), parkinsonism, Niemann-Pick disease type C (NPC), Charcot-Marie-Tooth disease (CMT), mucopolysaccharidosis type II (MPSIIA), mucolipidosis IV, GM1 gangliosidosis, sporadic inclusion body myositis (sIBM), Henoch-Schönlein purpura (HSP), limbic-predominant age-related TDP-43 encephalopathy (LATE), cerebral age-related TDP-43 with sclerosis (CARTS), Gaucher disease, and synaptic diseases such as facial-onset sensorimotor neuropathy, Guam Parkinsonism-Dementia Complex, multisystem proteinopathy, Perry disease, and autism. In various embodiments, the gapmer oligonucleotide is a gapmer oligonucleotide disclosed herein.

[0054] Further disclosed herein is a method of treating a neurological disease in a patient in need thereof, comprising administering to the patient in need thereof a therapeutically effective amount of a pharmaceutical composition comprising a gapmer oligonucleotide and a pharmaceutically acceptable excipient. In various embodiments, the neurological disease is 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's 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 neuropathy, e.g., chemotherapy. The present invention relates to a method-induced neuropathy, spinocerebellar ataxia (SCA), SCA type 2, spinal muscular atrophy (SMA), parkinsonism, Niemann-Pick disease type C (NPC), Charcot-Marie-Tooth disease (CMT), mucopolysaccharidosis type II (MPSIIA), mucolipidosis IV, GM1 gangliosidosis, sporadic inclusion body myositis (sIBM), Henoch-Schonlein purpura (HSP), limbic-predominant age-related TDP-43 encephalopathy (LATE), cerebral age-related TDP-43 with sclerosis (CARTS), Gaucher disease, and synaptic diseases such as facial-onset sensorimotor neuropathy, Guam Parkinsonism-Dementia Complex, multisystem proteinopathy, Perry disease, and autism. In various embodiments, the gapmer oligonucleotide is a gapmer oligonucleotide disclosed herein, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition disclosed herein. In various embodiments, the pharmaceutical composition is administered topically, parenterally, orally, pulmonary, rectally, bucally, sublingually, intravaginally, intratracheally, intranasally, intrathecally, intracisternally, transdermally, or intraduodenum. In various embodiments, the pharmaceutical composition is administered intrathecally. In various embodiments, the patient is a human.

[0055] Further disclosed herein is a gapmer oligonucleotide, or a pharmaceutically acceptable salt thereof, for use as a pharmaceutical. Further disclosed herein is a gapmer oligonucleotide, or a pharmaceutically acceptable salt thereof, for use in the treatment of neurological diseases. In various embodiments, the neurological disease is 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, cerebral cortical Basal ganglionic degeneration (CBD) and / or neuropathy, for example, chemotherapy-induced neuropathy, spinocerebellar ataxia (SCA), SCA type 2, spinal muscular atrophy (SMA), parkinsonism, Niemann-Pick disease type C (NPC), Charcot-Marie-Tooth disease (CMT), mucopolysaccharidosis type II (MPSIIA), mucolipidosis IV, GM1 gangliosidosis, sporadic inclusion body myositis (sIBM), Henoch-Schönlein purpura (HSP), Gaucher disease, and facial onset sensorimotor neuropathy, Guam Parkinsonism-Dementia Complex, multisystem proteinopathy, Perry disease, and autism.

[0056] Further disclosed herein is a gapmer oligonucleotide comprising any one of the sequences of SEQ ID NOs: 301692-301742 or the sequences in Tables 4A-4C, or a pharmaceutically acceptable salt thereof, wherein the gapmer oligonucleotide further comprises a gap segment comprising one or more linked deoxyribonucleosides, 2'-fluoroarabinonucleic acid (FANA), and fluorocyclohexenyl nucleic acid (F-CeNA), a 5' wing region comprising linked nucleosides, and a 3' wing region comprising linked nucleosides, wherein the gapmer oligonucleotide comprises a sugar modification of eeeee-d10-eeeee, where e = 2'-MOE nucleoside and d = deoxyribonucleoside, and at least one "e" or at least one "d" is replaced with a spacer, and wherein each cytosine of the 2' MOE nucleoside is 5-methylcytosine.

[0057] In various embodiments, at least one internucleoside linkage of the gapmer oligonucleotide is a phosphorothioate linkage.In various embodiments, the phosphorothioate internucleoside linkage is in one of the Rp conformation or the Sp conformation.In various embodiments, all internucleoside linkages of the gapmer oligonucleotide are phosphorothioate linkages.In various embodiments, the gapmer oligonucleotide has a sugar modification and an internucleoside linkage combination in the following patterns, respectively: a) eeeee-d10-eeeee and sssssooooooooosssss; b) eeeee-d10-eeeee and ooooosssssssssooooo; c) eeeee-d10-eeeee and ssssssssssssssssssss; d) eee-d8-eee and sssooooooosss; e)eee-d8-eee and ooosssssssooo f) eee-d8-eee and sssssssssssss; g) eee-d10-eee and sssooooooooosss; h) eee-d10-eee and ooosssssssssooo; i) eee-d10-eee and ssssssssssssssss; j)eeee-d10-eeee and ssssooooooooossss; k)eeee-d10-eeee and oooosssssssssoooo; l)eeee-d10-eeee and ssssssssssssssssss; m)eeee-d8-eeee and ssssooooooossss, n)eeee-d8-eeee and oooosssssssoooo, o)eeee-d8-eeee and ssssssssssssssss, p)eeeeee-d11-eeeeee and sssssssssssssssssssss; q)eeeee-d10-eeeee and sosossssssssssssosos; r)eeeee-d10-eeeee and soooosssssssssssooos; s)eeeee-d10-eeeee and soooosssssssssssooss; t)eeeee-d8-eeeee and sossssssssssssssoss; u)eeeee-d10-eeeee and sossssssssssssssssosss, and At least one "e" or at least one "d" of the gapmer oligonucleotide is replaced with a spacer.

[0058] Further disclosed herein is a pharmaceutical composition comprising the antisense gapmer oligonucleotide disclosed herein or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient. In various embodiments, the spacer is a nucleoside substitute that includes a non-sugar surrogate, and the non-sugar surrogate does not include a ketone, aldehyde, ketal, hemiketal, acetal, hemiacetal, aminal, or hemiaminal moiety and cannot form a covalent bond with a nucleotide base. In various embodiments, the spacer is represented by formula (X):

[0059] [ka] wherein: Ring A is an optionally substituted 4- to 8-membered monocyclic cycloalkyl group or a 4- to 8-membered monocyclic heterocyclyl group, the heterocyclyl group containing 1 or 2 heteroatoms selected from O, S, and N, with the proviso that A cannot form a covalent bond with a nucleobase;

[0060] [ka] The symbol represents the point of attachment to the internucleoside linkage.

[0061] In various embodiments, the spacer has the formula (Xa):

[0062] [ka] is expressed by

[0063] In various embodiments, Ring A is an optionally substituted 4-8 membered monocyclic cycloalkyl group selected from cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl, or a 4-8 membered monocyclic heterocyclyl group selected from oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, 1,4-dioxanyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, and azepanyl. In various embodiments, Ring A is tetrahydrofuranyl. In various embodiments, Ring A is tetrahydropyranyl.

[0064] In various embodiments, the spacer has formula (I):

[0065] [ka] wherein: X is selected from —CH— and —O—; n is 0, 1, 2, or 3.

[0066] In various embodiments, the spacer has formula (I'):

[0067] [ka] is expressed by

[0068] In various embodiments, the spacer has formula (Ia):

[0069] [ka] is expressed by

[0070] In various embodiments, the spacer has formula (Ia'):

[0071] [ka] is expressed by

[0072] In various embodiments, the spacer has Formula II:

[0073] [ka] wherein: X is selected from -CH2- and -O-.

[0074] In various embodiments, the spacer has formula II':

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

[0076] In various embodiments, the spacer has formula (Iia):

[0077] [ka] is expressed by

[0078] In various embodiments, the spacer has the formula (Iia'):

[0079] [ka] is expressed by

[0080] In various embodiments, the spacer has Formula III:

[0081] [ka] wherein: X is selected from -CH2- and -O-.

[0082] In various embodiments, the spacer has formula III':

[0083] [ka] wherein: X is selected from -CH2- and -O-.

[0084] In various embodiments, the spacer has formula (IIIa):

[0085] [ka] is expressed by

[0086] In various embodiments, the spacer has formula (IIIa'):

[0087] [ka] is expressed by

[0088] Further provided are methods for treating neurological diseases and / or neuropathy in a patient in need thereof, comprising administering to the patient in need thereof a therapeutically effective amount of a gapmer oligonucleotide, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition, in combination with riluzole (Rilutek), troriluzole, edaravone (Radicava), rivastigmine, donepezil, QRL-101, QRL-201, galantamine, selective serotonin reuptake inhibitors, antipsychotics, cholinesterase inhibitors, memantine, benzodiazepine anxiolytics, AMX0035 (ELYBRIO®), ZILUCOPLAN (RA101495), dual intrathecal AON administration (e.g., BIIB067, BIIB078), BI Disclosed herein are methods comprising administering in combination with a second therapeutic agent selected from the group consisting of IB100, levodopa / carbidopa, dopaminergic agents (e.g., ropinirole, pramipexole, rotigotine), medroxyprogesterone, KCNQ2 / KCNQ3 openers, pridopidine, PrimeC (a combination of ciprofloxacin and Celebrex), olanzapine (Zyprexa), quetiapine (Seroquel), SSRIs, divalproex sodium (Depakote), carbamazepine (Tegretol), medroxyprogesterone, lithium, anticonvulsants and psychostimulants, respiratory care, physical therapy, occupational therapy, speech therapy, nutritional support, or any combination thereof. In various embodiments, the neurological disease is any one of amyotrophic lateral sclerosis (ALS), frontotemporal dementia (FTD), or ALS with FTD.

[0089] Further provided are methods of treating neurological diseases and / or neuropathy in a patient in need thereof, comprising administering to the patient in need thereof a therapeutically effective amount of a gapmer oligonucleotide, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, to a patient in need thereof, the method comprising administering to the patient ... Zadyne®, aducanumab, BAN2401, BIIB091 (goslanemab), BIIB076, BIIB080 (IONIS-MAPTRx), Elayta (CT1812), MK1942, allogeneic hMSCs, nilotinib, ABT-957, acitretin, ABT-354, GV1001, riluzole, CAD106, CNP520, AD-35, rilapladib, DHP1401, T-817 MA, TC-5619, TPI-287, RVT-101, LY450139, JNJ-54861911, dapagliflozin, GSK239512, PF-04360365, ASP0777, SB-742457 (5-HT6 receptor antagonist), PF-03654746 (H3 receptor antagonist), GSK933776 (Fc-inactivating anti-β amyloid (Aβ) monoclonal antibody (mAb)), Posiphen ((+) Disclosed herein are methods comprising administering in combination with a second therapeutic agent selected from the group including phenserine tartrate, AMX0035 (ELYBRIO®), coenzyme Q10, aducanamab (ADUHLEM), memantine (NAMENDA), namzeric, suvorexant (Belsomra), lecanemab, or any combination thereof. In various embodiments, the neurological disease is Alzheimer's disease.

[0090] Further provided is a method of treating a neurological disease and / or neuropathy in a patient in need thereof, comprising administering to a patient in need thereof a therapeutically effective amount of a gapmer oligonucleotide, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, including but not limited to levodopa, carbidopa-levidopa, pramipexole (MIRAPEX), ropinirole (REQUIP), rotigotine (NEUPRO), apomorphine (APOKYN, KYNMOBI), selegiline (EDLEPRYL, ZELAPAR), rasagiline, entacapone (COMTAN), tolcapone (TASMAR), amantadine (GOCOVRI, SYMMETREL, OSMOLEX), trihexyphenidyl (ARTANE), BIIB054 (cinepanemab), BIIB094, BIIB Disclosed herein are methods for treating Parkinson's disease, including administering in combination with a second therapeutic agent selected from the group consisting of 118, ABBV-0805, zonisamide, deep brain stimulation, brain-derived neurotrophic factor, stem cell transplantation, niacin, brainstem stimulation, nicotine, nabilone, PF-06649751, DNL201, LRRK2 inhibitors, CK1 inhibitors, isradipine, CLR4001, IRX4204, yohimbine, coenzyme Q10, OXB-102, duloxetine, pioglitazone, preladenant, istradefylline (NOURIANZ), safinamide (XADAGO), benztropine (COGENTIN), opicapone (ongentys), exenatide, reishi mushroom, caffeine, sarizotan, fetal dopamine cell transplantation, or any combination thereof. In various embodiments, the neurological disease is Parkinson's disease.

[0091] Further, there is provided a method of treating a neurological disease and / or neuropathy in a patient in need thereof, comprising administering to a patient in need thereof a therapeutically effective amount of a gapmer oligonucleotide, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, UCB0107, ABBV-8E12, F-18 AV1451, BIIB092, C2N-8E12, tideglusib, transcranial deep magnetic stimulation, lipoic acid, tolfenamic acid, lithium, AZP2006, glial cell line-derived neurotrophic factor, NBMI, suvorexant, zolpidem, TPI. 287, davunetide, pimavanserin, levodopa, carbidopa-levidopa, pramipexole, ropinirole, rotigotine, apomorphine, selegiline, rasagiline, entacapone, tolcapone, amantadine, trihexyphenidyl, BIIB054 (sinepanemab), BIIB094, BIIB118, ABBV-0805, zonisamide, deep brain stimulation, brain-derived neurotrophic factor, stem cell transplantation, niacin, brain Disclosed herein is a method comprising administering in combination with a second therapeutic agent selected from the group consisting of stem stimulants, nicotine, nabilone, PF-06649751, DNL201, LRRK2 inhibitors, CK1 inhibitors, isradipine, CLR4001, IRX4204, yohimbine, coenzyme Q10, OXB-102, duloxetine, pioglitazone, preladenant, or any combination thereof. In various embodiments, the neurological disease is progressive supranuclear palsy (PSP).

[0092] Further provided are methods of treating neurological diseases and / or neuropathy in a patient in need thereof, comprising administering to a patient in need thereof a therapeutically effective amount of a gapmer oligonucleotide, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, in combination with tetrabenazine, deutetrabenazine, physical therapy, risperidone, haloperidol, chlorpromazine, clonazepam, diazepam, benzodiazepines, selective serotonin reuptake inhibitors, quetiapine, carbatrol, valproate, lamotrigine, pridopidine, delta-9-tetrahydrocannabinol, cannabidiol, stem cell therapy, or the like. Disclosed herein are methods for treating a neurological disorder, the method comprising administering in combination with a second therapeutic agent selected from the group consisting of: , ISIS-443139, nilotinib, resveratrol, neflamapimod, fenofibrate, creatine, RO7234292, SAGE-718, WVE-120102, WVE-120101, dimebon, minocycline, deep brain stimulation, ursodiol, coenzyme Q10, OMS643762, VX15 / 2503, PF-02545920, BN82451B, SEN0014196, olanzapine, tiapridal (tiapride), or any combination thereof. In various embodiments, the neurological disorder is Huntington's disease.

[0093] Further disclosed herein is a method of treating a neurological disease and / or neuropathy in a patient in need thereof, comprising administering to the patient in need thereof a therapeutically effective amount of a gapmer oligonucleotide, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition in combination with a second therapeutic agent selected from the group consisting of anticoagulants, antidepressants, muscle relaxants, stimulants, anticonvulsants, anti-anxiety agents, erythropoietin, hyperbaric oxygen treatment, rehabilitation therapy (e.g., physical, occupational, speech, psychological, or vocational counseling), or any combination thereof. In various embodiments, the neurological disease is brain trauma.

[0094] Further disclosed herein is a method for treating a neurological disease and / or neuropathy in a patient in need thereof, comprising administering to the patient in need thereof a therapeutically effective amount of a gapmer oligonucleotide, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition in combination with a second therapeutic agent selected from the group consisting of AXER-204, glyburide, 5-hydroxytryptophan (5-HTP), L-3,4-dihydroxyphenylalanine (L-DOPA), or rehabilitation therapy (e.g., physical therapy, occupational therapy, recreational therapy, use of assistive devices, improved strategies for exercise and healthy eating), or any combination thereof. In various embodiments, the neurological disease is spinal cord injury.

[0095] Further disclosed herein is a method for treating a neurological disease and / or neuropathy in a patient in need thereof, comprising administering to the patient in need thereof a therapeutically effective amount of a gapmer oligonucleotide, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition in combination with a second therapeutic agent selected from the group consisting of TPI-287, lithium, occupational therapy, physical therapy, and speech therapy, or any combination thereof, which may be selected as an additional therapy. In various embodiments, the neurological disease is corticobasal degeneration.

[0096] Further disclosed herein is a method for treating a neurological disease and / or neuropathy in a patient in need thereof, comprising administering to the patient in need thereof a therapeutically effective amount of a gapmer oligonucleotide, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition in combination with a second therapeutic agent selected from the group consisting of gabapentin, pregabalin, lamotrigine, carbamazepine, duloxetine, gabapentinoids, tricyclic antidepressants, serotonin-norepinephrine reuptake inhibitors, opioids, neurotoxins, dextromethorphan, nicotinamide riboside, autoantibody-targeted neuroantigens (TS-HDS and FGFR3), or any combination thereof. In various embodiments, the neuropathy is chemotherapy-induced neuropathy.

[0097] Further provided is a method of treating a neurological disease and / or neuropathy in a patient in need thereof, comprising administering to a patient in need thereof a therapeutically effective amount of a gapmer oligonucleotide, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, or a combination thereof with or without amantadine, armodafinil, baclofen, buspirone, carbamazepine, citalopram, clonazepam, desvenlafaxine, diazepam, duloxetine, escitalopram, flunarizine, fluoxetine, or a combination thereof. Disclosed herein are methods comprising administering in combination with a second therapeutic agent selected from the group consisting of fluvoxamine, gabapentin, isoniazid, levetiracetam, levodopa, memantine, modafinil, ondansetron, paroxetine, pramipexole, primidone, riluzole, ropinirole, sertraline, tizanidine, topiramate, trihexyphenidyl, valproic acid, venlafaxine, BHV-4157, or a combination thereof. In various embodiments, the neurological disease is spinocerebellar ataxia.

[0098] Further provided are methods of treating neurological diseases and / or neuropathy in a patient in need thereof, comprising administering to the patient in need thereof a therapeutically effective amount of a gapmer oligonucleotide, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, in combination with brivaracetam (briviact), cannabidiol (epidiolex), carbamazepine (carbatrol, Tegretol), cenobamate (xcopri), diazepam (valium), lorazepam (Ativan), clonazepam (klonopin), eslicarbazepine (aptiom), ethosuximide (zarontin), felbamate (felbatol), fenfluramine (fintepla), lacosamide (VIMPAT), lamotrigine (Lamictal), levetiracetam (Keppra), oxcarbazepine (oxtellar). Disclosed herein are methods comprising administering in combination with a second therapeutic agent selected from the group including fluticasone (fluticasone), fluoxetine (fluoxetine ...

[0099] Further disclosed herein is a method of treating a neurological disease and / or neuropathy in a patient in need thereof, comprising administering to the patient in need thereof a therapeutically effective amount of a gapmer oligonucleotide, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition in combination with a second therapeutic agent selected from the group consisting of nusinersen (SPINRAZA), onasemnogene abeparvovec-xioi (ZOLGENSMA), risdiplam (EVRYSDI), or any combination thereof. In various embodiments, the neurological disease is spinal muscular atrophy.

[0100] Further disclosed herein is a method of treating a neurological disease and / or neuropathy in a patient in need thereof, comprising administering to the patient in need thereof a therapeutically effective amount of a gapmer oligonucleotide, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof in combination with a second therapeutic agent selected from the group consisting of an anti-seizure drug, speech therapy, physical therapy, occupational therapy, adrabetadex, arimoclomol, N-acetyl-L-leucine, or any combination thereof. In various embodiments, the neurological disease is Niemann-Pick disease type C.

[0101] Further disclosed herein is a method of treating a neurological disease and / or neuropathy in a patient in need thereof, comprising administering to the patient in need thereof a therapeutically effective amount of a gapmer oligonucleotide, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, in combination with a second therapeutic agent selected from the group consisting of physical and occupational therapy, orthopedic surgery, an orthopedic device, PXT3003, or any combination thereof. In various embodiments, the neurological disease is Charcot-Marie-Tooth disease (CMT).

[0102] Further disclosed herein is a method of treating a neurological disease and / or neuropathy in a patient in need thereof, comprising administering to the patient in need thereof a therapeutically effective amount of a gapmer oligonucleotide, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition in combination with a second therapeutic agent selected from the group consisting of enzyme replacement therapy, idursulfase (Elaprase), surgical intervention (tonsillectomy and / or adenoidectomy), RGX-121 gene therapy, adalimumab, MT2013-31, or any combination thereof. In various embodiments, the neurological disease is mucopolysaccharidosis type II (MPSIIA).

[0103] Further disclosed herein is a method of treating a neurological disease and / or neuropathy in a patient in need thereof, comprising administering to the patient in need thereof a therapeutically effective amount of a gapmer oligonucleotide, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition in combination with a second therapeutic agent selected from the group consisting of physical therapy, occupational therapy, and speech therapy, contact lenses and artificial tears, genetic counseling, or any combination thereof. In various embodiments, the neurological disease is mucolipidosis IV.

[0104] Further disclosed herein is a method of treating a neurological disease and / or neuropathy in a patient in need thereof, comprising administering to the patient in need thereof a therapeutically effective amount of a gapmer oligonucleotide, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition in combination with a second therapeutic agent selected from the group consisting of anticonvulsants, physical and occupational therapy, galactosidase, gene delivery of galactosidase, LYS-GM101 gene therapy, or any combination thereof. In various embodiments, the neurological disease is GM1 gangliosidosis.

[0105] Further disclosed herein is a method of treating a neurological disease and / or neuropathy in a patient in need thereof, comprising administering to the patient in need thereof a therapeutically effective amount of a gapmer oligonucleotide, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition in combination with a second therapeutic agent selected from the group consisting of physical and occupational therapy, the use of devices such as braces, walkers, wheelchairs, immunosuppressants, BYM338, or any combination thereof. In various embodiments, the neurological disease is sporadic inclusion body myositis (sIBM).

[0106] Further disclosed herein is a method of treating a neurological disease and / or neuropathy in a patient in need thereof, comprising administering to the patient in need thereof a therapeutically effective amount of a gapmer, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof in combination with a second therapeutic agent selected from the group consisting of corticosteroids, colchicine, dapsone, azathioprine, or any combination thereof. In various embodiments, the neurological disease is Henoch-Schönlein purpura (HSP).

[0107] Further disclosed herein is a method of treating a neurological disease and / or neuropathy in a patient in need thereof, comprising administering to the patient in need thereof a therapeutically effective amount of a gapmer oligonucleotide, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, in combination with a second therapeutic agent selected from the group consisting of enzyme replacement therapy, substrate reduction therapy, N-acetylcysteine, GZ / SAR402671, cerezyme, or any combination thereof. In various embodiments, the neurological disease is Gaucher disease. [Brief explanation of the drawings]

[0108] [Figure 1] Figure 1 shows an exemplary antisense oligonucleotide (AON), a portion of which is complementary to an mRNA or pre-mRNA transcript. Dashed lines indicate positions of the AON that may or may not be complementary to the corresponding position in the transcript. DETAILED DESCRIPTION OF THE INVENTION

[0109] The features and other details of the present disclosure will now be more particularly described. Before further describing the present invention, certain terms used in the specification, examples, and appended claims are collected here. These definitions should be read in light of the remainder of the disclosure and would be understood by one of ordinary skill 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.

[0110] definition The terms "treat," "treatment," "treating," and the like are used herein generally to mean obtaining a desired pharmacological and / or physiological effect. The effect may be therapeutic in terms of partial or complete cure of a disease and / or side effects attributable to a disease. The term "treatment," as used herein, encompasses any treatment of a disease in a mammal, particularly a human, including (a) inhibiting the disease, i.e., preventing the disease from increasing in severity or extent, (b) palliating the disease, i.e., causing partial or complete alleviation of the disease, or (c) preventing the recurrence of the disease, i.e., treating the symptoms of the disease or preventing the disease from returning to an active state after previous successful treatment of the disease.

[0111] "Preventing" includes delaying the onset of clinical symptoms, complications, or biochemical indicators of a condition, disorder, disease, or condition that develops in a subject who may be suffering from or predisposed to the condition, disorder, disease, or condition, but who has not yet experienced or displayed clinical or preclinical symptoms of the condition, disorder, disease, or condition. "Preventing" includes prophylactically treating a condition, disorder, disease, or condition that develops in or in a subject, including prophylactically treating clinical symptoms, complications, or biochemical indicators of a condition, disorder, disease, or condition that develops in or in a subject.

[0112] The terms "pharmaceutically acceptable carrier" or "pharmaceutically acceptable excipient" as used herein refer interchangeably to any and all solvents, dispersion media, coating agents, isotonic agents, absorption delaying agents, etc., 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 supplementary, additional, or enhanced therapeutic functions.

[0113] The term "pharmaceutical composition," as used herein, refers to a composition comprising at least one biologically active compound, e.g., an antisense oligonucleotide (AON) disclosed herein, formulated together with one or more pharmaceutically acceptable excipients.

[0114] The terms "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, most preferably humans. The compounds of the invention can be administered to mammals, such as humans, but also to other mammals, e.g., animals in need of veterinary treatment, such as farm animals (e.g., dogs, cats, etc.), livestock animals (e.g., cows, sheep, pigs, horses, etc.), and laboratory animals (e.g., rats, mice, guinea pigs, non-human primates, etc.). In some embodiments, the mammal treated in the methods of the invention is desirably a mammal in which modulation of target expression and / or activity is desired.

[0115] As used herein, "PPM1A" (protein phosphatase, Mg 2+ / Mn 2+Dependent 1A, protein phosphatase 1A (formerly 2C), magnesium-dependent, alpha isoform, protein phosphatase 1A, EC 3.1.3.16, protein phosphatase 2C isoform alpha, protein phosphatase IA, phosphatase 2C alpha, PP2C-alpha, PPPM1A, and PP2CA) refers to the gene or gene product (e.g., the protein or mRNA transcript (including pre-mRNA) encoded by the gene) identified by Entrez Gene Identification Number 5494 and allelic variants thereof, and orthologs found in non-human species (e.g., non-human primates or mice).

[0116] As used herein, "ATXN2" (also known as ataxin 2, ATX2, TNRC13, SCA2, trinucleotide repeat-containing gene 13 protein, spinocerebellar ataxia type 2 protein, ataxin-2, spinocerebellar ataxia 2 (olivopontocerebellar ataxia 2, autosomal dominant, ataxin 2), and trinucleotide repeat-containing 13) refers to the gene or gene product (e.g., the protein or mRNA transcript (including pre-mRNA) encoded by the gene) identified by Entrez Gene Identification Number 6311 and allelic variants thereof, and orthologs found in non-human species (e.g., non-human primates or mice).

[0117] As used herein, "SOD1" (also known as superoxide dismutase 1, IPOA, superoxide dismutase 1, soluble, superoxide dismutase [Cu-Zn], EC 1.15.1.1, HSod1, ALS1, ALS, amyotrophic lateral sclerosis 1 (adult), epididymal secretory protein Li 44, superoxide dismutase, systolic (cystolic), Cu / Zn superoxide dismutase, indophenol oxidase A, SOD, soluble, homodimeric, HEL-S-44, STAHP, and SOD) refers to the gene or gene product (e.g., the protein or mRNA transcript (including pre-mRNA) encoded by the gene) identified by Entrez Gene Identification Number 6647 and allelic variants thereof, and orthologs found in non-human species (e.g., non-human primates or mice).

[0118] As used herein, "MAPT" (also known as microtubule-associated protein Tau, MTBT1, PPP1R103, FTDP-17, MTBT2, MAPTL, PPND, MSTD, TAU, G-protein beta 1 / gamma 2, subunit interactor, protein phosphatase 1, regulatory subunit 103, microtubule-associated protein Tau, neurofibrillary tangle protein, paired helical filaments-Tau, MGC138549, FLJ31424, PHF-Tau, Tau-40, DDPAC, Tau, and microtubule-associated protein Tau, isoform 4) refers to the gene or gene product (e.g., the protein or mRNA transcript (including pre-mRNA) encoded by the gene) identified by Entrez Gene Identification Number 4137 and allelic variants thereof, and orthologs found in non-human species (e.g., non-human primates or mice).

[0119] As used herein, the term "therapeutically effective amount" refers to an amount of the subject AON that will elicit the biological or medical response in a tissue, system, animal, or human desired by a researcher, veterinarian, physician, or other clinician. The AONs of the present invention are administered in a therapeutically effective amount to treat and / or prevent a disease, condition, disorder, or condition, e.g., ALS, FTD, ALS with FTD, or another motor neuron disease or neurological disease or condition. Alternatively, a therapeutically effective amount of an AON is the amount required to achieve the desired therapeutic and / or prophylactic effect, e.g., an amount that results in the prevention or reduction of symptoms associated with a disease disclosed herein.

[0120] As used herein, the term "antisense oligonucleotide" or "AON" includes antisense oligonucleotides that target any of the genes or gene products of PPM1A, ATXN2, SOD1, or MAPT. "Antisense oligonucleotide" or "AON" includes parent oligonucleotides, oligonucleotide variants, oligonucleotides with one or more spacers, oligonucleotide variants with one or more spacers, gapmer antisense oligonucleotides (AONs), and gapmer AONs with one or more spacers. Examples of antisense oligonucleotides include antisense oligonucleotides comprising any one of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, or 167805-301566, or oligonucleotides comprising one or more spacers replacing one or more nucleosides of any one of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, or 167805-301566, e.g., any one of SEQ ID NOs: 301692-301742 or any one of the sequences in Tables 4A-4C.

[0121] The terms "parent oligonucleotide," "parent antisense oligonucleotide," or "parent AON" refer to an antisense oligonucleotide that is complementary to a portion of any one of the target gene products, e.g., PPM1A, ATXN2, SOD1, or MAPT mRNA or pre-mRNA transcripts. The parent oligonucleotide does not contain a spacer. In certain embodiments, the parent oligonucleotide contains 20 linked nucleosides. In such embodiments, the parent oligonucleotide is a 20-mer. Examples of parent oligonucleotides are AONs having the sequence of any one of SEQ ID NOS: 1-954, 1914-149354, 149362-158581, or 167805-301566. As described hereinafter, oligonucleotides with spacers, oligonucleotide variants, and gapmer AONs are described with reference to the corresponding parent oligonucleotides.

[0122] The term "oligonucleotide variant" refers to an antisense oligonucleotide that represents a modified version of a corresponding parent oligonucleotide. For example, an oligonucleotide variant represents a shortened version of a parent oligonucleotide. In various embodiments, an oligonucleotide variant is any one of a 15-mer, 16-mer, 17-mer, 18-mer, 19-mer, 20-mer, 21-mer, 22-mer, or 23-mer. For example, an AON variant may be a shorter or longer version of a corresponding parent oligonucleotide containing a nucleobase sequence selected from any one of SEQ ID NOS: 1-954, 1914-149354, 149362-158581, or 167805-301566. An example of an oligonucleotide variant is an oligonucleotide containing any one of SEQ ID NOS: 301590-301594. In various embodiments, an oligonucleotide variant includes one or more spacers.

[0123] 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, an oligonucleotide with one or more spacers can include 1 spacer, 2 spacers, 3 spacers, 4 spacers, 5 spacers, 6 spacers, 7 spacers, 8 spacers, 9 spacers, or 10 spacers. In various embodiments, an oligonucleotide comprising one or more spacers includes at least one segment having at most 10 linked nucleosides. For example, when written in the 5' to 3' direction, an oligonucleotide comprising a spacer can include a segment having 10 linked nucleosides, followed by a spacer, followed by a second segment of 9 linked nucleosides. In various embodiments, an oligonucleotide comprising one or more spacers includes at least one segment having at most 7 linked nucleosides. For example, when written in the 5' to 3' direction, an oligonucleotide containing a spacer can include 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.In this specification, the first segment of 7 linked nucleosides and the third segment of 7 linked nucleosides each represent a segment having 7 linked nucleosides.In various embodiments, an oligonucleotide containing one or more spacers includes at least one segment having at most 6 linked nucleosides.For example, when written in the 5' to 3' direction, an oligonucleotide containing a spacer can include a segment having 6 linked nucleosides, followed by a spacer, a second segment having 6 linked nucleosides, followed by a second spacer, and a third segment having 6 linked nucleosides.Herein, the first segment of 6 linked nucleosides, the second segment of 6 linked nucleosides, and the third segment of 6 linked nucleosides each represent a segment with 6 linked nucleosides.Herein, all segments of the oligonucleotide with one or more spacers have at most 6 linked nucleosides.In various embodiments, gapmer oligonucleotide comprises a segment with at most 10, 9, 8, 7, 6, 5, 4, 3 or 2 linked nucleosides.

[0124] As another example, an oligonucleotide containing a spacer may include a segment having 10 linked nucleosides, followed by a spacer, a second segment having 10 linked nucleosides, followed by a second spacer, and a third segment having 3 linked nucleosides. Herein, the third segment of 3 linked nucleosides refers to a segment having at most 6 or at most 7 linked nucleosides. In various embodiments, an oligonucleotide having one or more spacers includes multiple segments having at most 6 or at most 7 linked nucleosides. In various embodiments, all segments of an oligonucleotide having one or more spacers have at most 6 or at most 7 linked nucleosides. For example, an oligonucleotide may be a 20-mer and include two spacers that divide the 20-mer into three separate segments, each of which has 6 linked nucleosides. Thus, each segment of the oligonucleotide has at most 6 linked nucleosides. For example, an oligonucleotide may be a 23-mer and include two spacers that divide the 23-mer into three separate segments of seven linked nucleosides each. Thus, each segment of the oligonucleotide has at most seven linked nucleosides. Generally, oligonucleotides that include one or more spacers are described with reference to the corresponding parent oligonucleotide or corresponding oligonucleotide variant. Exemplary oligonucleotides that include one or more spacers include any of the sequences of SEQ ID NOs: 301692-301742 or Tables 4A-4C.

[0125] In various embodiments, one or more spacers can be located at one or more positions of oligonucleotide.Spacer can be located between the first position and the second position of oligonucleotide.As used herein, the spacer located between the first position and the second position includes the spacer that is located at the first position, that is located at the second position, or that is located at any position of the oligonucleotide sandwiched between the first position and the second position.

[0126] The term "gapmer antisense oligonucleotide," "gapmer AON," or "gapmer AON variant" refers to an AON with at least three distinct structural regions, including a 5' wing region, a central region, and a 3' wing region, in a "5 to 3" orientation. The central region contains a stretch of nucleosides that allows for the recruitment and activation of RNAse H. For example, the central region contains linked DNA nucleosides, 2'-fluoroarabinonucleic acid (FANA), and fluorocyclohexenyl nucleic acid (F-CeNA). Exemplary gapmer AONs or gapmer oligonucleotide variants include any of SEQ ID NOs: 301595-301607. In various embodiments, a gapmer AON comprises a sequence that shares at least 85%, at least 90%, at least 95%, at least 98%, or 100% identity with an equal-length portion of any one of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, or 167805-301566. In various embodiments, a gapmer AON can comprise one or more spacers. Exemplary gapmer AONs comprising one or more spacers comprise a sequence that shares at least 85%, at least 90%, at least 95%, at least 98%, or 100% identity with an equal-length portion of any one of SEQ ID NOs: 301692-301742 or the sequences in Tables 4A-4C.

[0127] The term " pharmaceutically acceptable salt " as used herein refers to the salt of acidic or basic groups that may be present in the antisense oligonucleotide used in the present composition.The antisense oligonucleotide contained in the present composition is basic in nature and can form a wide range 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, e.g., salts with 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, oleic acid, tannic acid, pantothenic acid, bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucuronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, and pamoate salts (e.g., 1,1'-methylene-bis-(2-hydroxy-3-naphthoate)). Antisense oligonucleotides contained in the present compositions that contain amino moieties can form pharmaceutically acceptable salts with various amino acids in addition to the acids mentioned above. Compounds contained in the present compositions that are acidic in nature can form base salts with various pharmacologically acceptable cations. Examples of such salts include alkali metal or alkaline earth metal salts, specifically calcium, magnesium, sodium, lithium, zinc, potassium, and iron salts. Pharmaceutically acceptable salts of the present disclosure include, for example, pharmaceutically acceptable salts of AONs containing any of the nucleotide sequences set forth in SEQ ID NOS: 1-954, 1914-149354, 149362-158581, 167805-301566, SEQ ID NOS: 301692-301742, or the sequences in Tables 4A-4C.

[0128] The antisense oligonucleotides of the present disclosure may contain one or more chiral centers, groups, linkages, and / or double bonds, and therefore may exist as stereoisomers, for example, geometric isomers, enantiomers, or diastereomers. The term "stereoisomer" as used herein refers to all geometric isomers, enantiomers, or diastereomers. These compounds may be designated by the symbols "R" or "S" (or "Rp" or "Sp") depending on the conformation of the substituents around the stereogenic atom, for example, the stereogenic carbon, phosphorus, or sulfur atom. In some embodiments, one or more linkages in a compound may have an Rp or Sp conformation (e.g., one or more phosphorothioate linkages have either an Rp or Sp conformation). The conformation of each phosphorothioate linkage may be independent of other phosphorothioate linkages (e.g., one phosphorothioate linkage has an Rp conformation, and a second phosphorothioate linkage has an Sp conformation). The present invention encompasses various stereoisomers of these compounds or mixtures thereof. Stereoisomers include enantiomers and diastereomers. Mixtures of enantiomers or diastereomers may be designated by "(±)" in nomenclature, but those skilled in the art will recognize that certain structures may implicitly exhibit chiral centers. Individual stereoisomers of the antisense oligonucleotides of the present invention can be prepared by synthesis from commercially available starting materials containing asymmetric or stereogenic centers, or by preparation of a racemic mixture followed by resolution methods well known to those skilled in the art. These resolution methods are exemplified by (1) binding the mixture of enantiomers to a chiral auxiliary, recrystallizing or chromatographically separating the resulting mixture of diastereomers, and liberating the optically pure product from the auxiliary; (2) forming a salt using an optically active resolving agent; or (3) directly separating the mixture of optical enantiomers on a chiral chromatography column.Stereoisomeric mixtures can also be resolved into their component stereoisomers by well-known methods, such as chiral-phase gas chromatography, chiral-phase supercritical fluid chromatography, chiral-phase simulated moving bed chromatography, chiral-phase high performance liquid chromatography, crystallizing the compound as a chiral salt complex, or crystallizing the compound in a chiral solvent. Stereoisomers can also be obtained from stereomerically pure intermediates, reagents, and catalysts by well-known asymmetric synthetic methods.

[0129] Individual stereoisomers of the antisense oligonucleotides of the present invention can be prepared by synthesis from commercially available starting materials containing asymmetric or stereogenic centers, or by preparing racemic mixtures followed by resolution methods well known to those skilled in the art. These resolution methods are exemplified by (1) binding the mixture of enantiomers to a chiral auxiliary, recrystallizing or chromatographically separating the resulting diastereomeric mixture, and liberating the optically pure product from the auxiliary; (2) forming a salt using an optically active resolving agent; or (3) directly separating the mixture of optical enantiomers on a chiral chromatography column. Stereoisomeric mixtures can also be resolved into their component stereoisomers by well-known methods, such as chiral-phase supercritical fluid chromatography, chiral-phase simulated moving bed chromatography, chiral-phase gas chromatography, chiral-phase high-performance liquid chromatography, crystallizing the compound as a chiral salt complex, or crystallizing the compound in a chiral solvent. Stereoisomers can also be obtained from stereopure intermediates, reagents, and catalysts by well-known asymmetric synthesis methods.

[0130] The antisense oligonucleotides disclosed herein may exist in solvated as well as unsolvated forms with pharmaceutically acceptable solvents such as water, ethanol, and the like, and the present invention is intended to encompass both solvated and unsolvated forms.

[0131] The present invention also encompasses isotopically labeled compounds of the present invention (e.g., isotopically labeled antisense oligonucleotides) that are identical to those listed herein, except that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number most abundantly found in nature. Examples of isotopes that can be incorporated into compounds of the present invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, and chlorine, such as: 2 H, 3 H, 11 C. 13 C. 14 C. 15 N, 18 O. 17 O. 31 P, 32 P, 35 S, 18 F, and 36 Examples include Cl.

[0132] Certain isotopically labeled disclosed compounds (e.g., 3 H and 14 C) are useful in compound and / or substrate tissue distribution assays. 3 H), carbon-14 (i.e. 14 C), or 35 S phosphorothioate isotopes are particularly preferred due to their ease of preparation and detectability. Additionally, heavier isotopes, such as deuterium (i.e. 2 H) may confer 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.

[0133] As used herein, "2'-O-(2-methoxyethyl)" (also 2'-MOE and 2'-O(CH2)2OCH3 and MOE) refers to an O-methoxyethyl modification at the 2' position of the furanose ring. 2'-O-(2-methoxyethyl) is used interchangeably with "2'-O-methoxyethyl" in this disclosure. The sugar moiety in a 2'-MOE modified nucleoside is the modified sugar.

[0134] As used herein, "2'-MOE nucleoside" (also 2'-O-(2-methoxyethyl) nucleoside) means a nucleoside that includes a 2'-MOE modified sugar moiety.

[0135] As used herein, "2'-substituted nucleoside" means a nucleoside that includes 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.

[0136] As used herein, "5-methylcytosine" (5-MeC) means a cytosine modified with a methyl group attached to position 5. 5-Methylcytosine (5-MeC) is a modified nucleobase.

[0137] As used herein, "bicyclic sugar" means a furanose ring modified by bridging two atoms. A bicyclic sugar is a modified sugar.

[0138] As used herein, "bicyclic nucleoside" (also BNA) refers to a nucleoside having a sugar moiety that includes a bridge connecting two carbon atoms of the sugar ring, thereby forming a bicyclic ring system. In certain embodiments, the bridge connects the 4' and 2' carbons of the sugar ring.

[0139] As used herein, "cEt" or "constrained ethyl" means a bicyclic nucleoside having a sugar moiety containing a bridge connecting the 4' and 2' carbons, where the bridge has the formula: 4'-CH(CH3)-O-2'.

[0140] As used herein, "constrained ethyl nucleoside" (also cEt nucleoside) refers to a nucleoside containing a bicyclic sugar moiety containing a 4'-CH(CH3)-O-2' bridge. In some embodiments, the cEt may be modified. In some embodiments, the cEt may be an S-cEt (S-constrained ethyl 2'-4' bridged nucleoside). In some other embodiments, the cEt may be an R-cEt.

[0141] As used herein, "internucleoside linkage" refers to the atoms or groups connecting the 3' and 5' positions of a sugar or the corresponding positions of a sugar mimetic. In some embodiments, as used herein, "non-natural linkage" refers to an "modified internucleoside linkage."

[0142] As used herein, " contiguous " in the context of oligonucleotide refers to nucleosides, nucleic acid bases, sugar moieties, or internucleoside linkages that are directly adjacent to each other.For example, " contiguous nucleic acid bases " refers to the nucleic acid bases that are directly adjacent to each other in sequence.In contrast, two nucleosides that are separated by a spacer are not contiguous.

[0143] As used herein, "modified nucleobase" refers to any nucleobase other than adenine, cytosine, guanine, thymine, or uracil. Examples of modified nucleobases include 5-methylcytosine, pseudouridine, or 5-methoxyuridine. "Unmodified nucleobase" refers to the purine bases adenine (A) and guanine (G), and the pyrimidine bases thymine (T), cytosine (C), and uracil (U).

[0144] As used herein, "spacer" refers to a nucleoside substitute group (for example, a non-nucleoside group that replaces the nucleoside present in parent oligonucleotide).A spacer is characterized by the absence of a nucleotide base and the replacement of the sugar moiety of the nucleoside with a non-sugar substitute.The non-sugar substitute group of the spacer lacks an aldehyde, ketone, acetal, ketal, hemiacetal, or hemiketal group.Thus, the non-sugar substitute group of the spacer can be connected to the 3' and 5' positions of the nucleoside adjacent to the spacer through the internucleoside linker described herein, but cannot form a covalent bond with the nucleotide base (i.e., cannot link the nucleobase to another group, such as an internucleoside linkage, a conjugate group, or a terminal group in an oligonucleotide).Generally, antisense oligonucleotides with spacers are described in relation to parent antisense oligonucleotides, where the spacer replaces the nucleoside of the parent antisense oligonucleotide. In all embodiments of the present disclosure, the spacer cannot hybridize to the nucleoside containing the nucleic acid base at the corresponding position of the transcript (e.g., pre-mRNA or mRNA transcript) within the numerical order of the length of the AON (i.e., if the spacer is placed after nucleoside 4 of the AON (i.e., 5 positions from the 5' end), the spacer is not complementary to the nucleoside (A, C, G, or U) at the same corresponding position of the target transcript).

[0145] As used herein, "modified nucleoside" refers to a nucleoside that has independently modified sugar moieties and / or modified nucleobases.A universal base is a modified nucleobase that can pair with any one of five unmodified nucleobases.Modified nucleosides include abasic nucleosides that lack nucleobases.However, modified nucleosides do not contain spacers or other groups that cannot be linked to nucleobases.

[0146] As used herein, "linked nucleosides" are nucleosides that are connected in a contiguous sequence (i.e., no additional nucleosides are present between the linked ones). In various embodiments, an oligonucleotide can have different segments of linked nucleosides that are connected through a spacer. As used herein, a spacer (i.e., a nucleoside substitute) is not considered a nucleoside and thus divides the oligonucleotide into two segments of linked nucleosides. An oligonucleotide can have a first segment of Y linked nucleosides (e.g., Y nucleosides connected in a contiguous sequence), followed by a spacer, and then a second segment of Z linked nucleosides. As used herein, the Y and Z linked nucleosides are described in either the 5' to 3' direction or the 3' to 5' direction. In various embodiments, the first segment consists of seven or fewer linked nucleosides (e.g., Y=7 or less), while the second segment comprises eight or more linked nucleosides (e.g., Z=8 or more).

[0147] 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, thereby forming 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) ethyleneoxy (4'-(CH2)2-O-2') LNA, (D) aminooxy (4'-CH2-ON(R)-2') LNA, and (E) oxyamino (4'-CH2-N(R)-O-2') LNA, where R is H, C1-C 12alkyl, or a protecting group (see U.S. Patent No. 7,427,672, issued September 23, 2008).

[0148] As used herein, LNA compounds include, but are not limited to, compounds with at least one bridge between the 4' and 2' sugar positions, where each of the bridges is independently -[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)-, where x is 0, 1, or 2, and n is 1, 2, 3, or 4; and each R1 and R2 is independently H, a protecting group, hydroxyl, 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 Aryl, substituted C5-C 20 aryl, heterocyclic radical, substituted heterocyclic radical, heteroaryl, substituted heteroaryl, C5-C7 cycloaliphatic radical, substituted C5-C7 cycloaliphatic radical, halogen, OJ1, NJ1J2, SJ1, N3, COOJ1, acyl (C(=O)-H), substituted acyl, CN, sulfonyl (S(=O)2-J1), or sulfoxyl (S(=O)-J1), wherein each J1 and J2 is 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 Aryl, substituted C5-C 20 Aryl, acyl (C(=O)-H), substituted acyl, heterocyclic radical, substituted heterocyclic radical, C1-C12 Aminoalkyl, substituted C1-C 12 aminoalkyl, or a protecting group.

[0149] Examples of 4'-2' bridging groups encompassed within the definition of LNA include groups of the formula: -[C(R1)(R2)] n -, -[C(R1)(R2)] n Additionally, other bridging groups encompassed within the definition of LNA include, but are not limited to, one of: -O-, -C(R1R2)-N(R1)-O-, or -C(R1R2)-ON(R1)-. Additionally, other bridging groups encompassed within 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, where each R1 and R2 is independently H, a protecting group, or C1-C 12 It is alkyl.

[0150] Also included within the definition of LNA according to the present invention are LNAs in which the 2'-hydroxyl group of the ribosyl sugar ring is connected to the 4' carbon atom of the sugar ring, thereby forming a bridge to form a bicyclic sugar moiety. The bridge can be a methylene (-CH2-) group connecting the 2' oxygen atom and the 4' carbon atom, for which the term methyleneoxy(4'-CH2-O-2')LNA is used. Furthermore, in the case of a bicyclic sugar moiety having an ethylene bridging group at this position, the term ethyleneoxy(4'-CH2CH2-O-2')LNA is used. The isomer of methyleneoxy(4'-CH2-O-2')LNA, AL-methyleneoxy(4'-CH2-O-2'), is also included within the definition of LNA used herein.

[0151] As used herein, "hybridization" refers to the pairing or annealing of complementary oligonucleotides and / or nucleic acids.Although not limited to a specific mechanism, the most common mechanism of hybridization involves hydrogen bonding, which can be Watson-Crick, Hoosteen, or reversed Hoosteen hydrogen bonding between complementary nucleic acid bases.

[0152] As used herein, "increasing the amount of activity" refers to more transcriptional expression, more accurate splicing resulting in full-length mature mRNA and / or protein expression, and / or more activity compared to transcriptional expression or activity in an untreated or control sample.

[0153] As used herein, a "mismatch" or "non-complementary nucleobase" refers to an instance where a group (e.g., nucleobase) of a first nucleic acid cannot pair with a corresponding group (e.g., nucleobase) of a second or target nucleic acid.

[0154] As used herein, a "modified internucleoside linkage" refers to a substitution or any change from a naturally occurring internucleoside linkage (e.g., a phosphodiester internucleoside bond). A "phosphorothioate linkage" is a modified internucleoside linkage in which one of the non-bridging oxygen atoms of a phosphodiester internucleoside linkage is replaced with a sulfur atom.

[0155] As used herein, "modified oligonucleotide" means an oligonucleotide that contains at least one (i.e., one or more) modified internucleoside linkage, modified sugar, and / or modified nucleobase.

[0156] As used herein, "modified sugar" or "modified sugar moiety" means a modified furanosyl sugar moiety or a modified sugar moiety having other than a furanosyl moiety that can link a nucleobase to another group, e.g., an internucleoside linkage in an oligonucleotide, a conjugate group, or a terminal group.

[0157] 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.

[0158] As used herein, "motif" means a pattern of unmodified and modified nucleosides in an antisense compound.

[0159] As used herein, "natural sugar moiety" means a sugar moiety found in DNA (2'-H) or RNA (2'-OH).

[0160] As used herein, a "naturally occurring internucleoside linkage" means a 3' to 5' phosphodiester linkage.

[0161] As used herein, "nucleobase" means a heterocyclic moiety capable of pairing with a base of another nucleic acid.

[0162] As used herein, " nucleobase complementarity " refers to the nucleobase that can base pair with another nucleobase.For example, in the case of DNA, adenine (A) is complementary to thymine (T).For example, in the case of RNA, adenine (A) is complementary to uracil (U).In certain embodiments, complementary nucleobase refers to the nucleobase of antisense compound that can base pair with the nucleobase of its target nucleic acid.For example, if the nucleobase at a certain position of antisense compound can hydrogen bond with the nucleobase at a certain position of target nucleic acid, the hydrogen bond position between oligonucleotide and target nucleic acid is considered to be complementary in this nucleobase pair.

[0163] As used herein, "non-complementary nucleobases" refers to a pair of nucleobases that do not form hydrogen bonds with each other or otherwise support hybridization.

[0164] As used herein, " nucleic acid " refers to the molecule that is composed of monomeric nucleotide.Nucleic acid includes but is not limited to ribonucleic acid (RNA), deoxyribonucleic acid (DNA), single-stranded nucleic acid, double-stranded nucleic acid, non-coding RNA, small interfering ribonucleic acid (siRNA), short hairpin RNA (shRNA), circular RNA, circular DNA and microRNA (miRNA).

[0165] As used herein, "nucleobase sequence" means the order of nucleobases independent of any sugar, linkage, and / or nucleobase modifications.

[0166] As used herein, "nucleoside" refers to a nucleobase linked to a sugar. The term "nucleoside" also includes "modified nucleosides" having, independently, modified sugar moieties and / or modified nucleobases.

[0167] As used herein, "oligonucleotide unit" refers to either a nucleoside (e.g., a nucleoside containing a sugar and / or a nucleic acid base) or a nucleoside substitute (e.g., a spacer) of an oligonucleotide. Oligonucleotide units include nucleosides, modified nucleosides, and spacers. For example, an oligonucleotide may have 20 oligonucleotide units, where one of the oligonucleotide units is a spacer, and the other 19 oligonucleotide units are nucleosides or modified nucleosides. As another example, an oligonucleotide may have 20 oligonucleotide units, where two of the oligonucleotide units are spacers, and the other 18 oligonucleotide units are nucleosides or modified nucleosides.

[0168] As used herein, "nucleoside mimic" includes structures used to replace the sugar, or sugar and base, and optionally the linkage at one or more positions of an oligomeric compound, such as, for example, morpholino, cyclohexenyl, cyclohexyl, tetrahydropyranyl, bicyclo, or tricyclo sugar mimics, e.g., nucleoside mimics with non-furanose sugar units. Nucleotide mimics include structures used to replace the nucleoside and linkage at one or more positions of an oligomeric compound, such as, for example, peptide nucleic acids or morpholinos (morpholinos linked by phosphorodiamidate or other non-phosphodiester linkages). Sugar surrogate overlaps slightly with the broader term nucleoside mimic, but is intended to refer to the replacement of only the sugar unit (furanose ring). The tetrahydropyranyl ring provided herein represents an example of a sugar surrogate in which the furanose sugar group is replaced with a tetrahydropyranyl ring system. "Mimetic" refers to groups that substitute for the sugar, nucleobase, and / or internucleoside linkage. Generally, a mimetic is used in place of the sugar or sugar-internucleoside linkage combination, while maintaining the nucleobase for hybridization to a selected target.

[0169] As used herein, "nucleotide" means a nucleoside having a phosphate group covalently linked to the sugar portion of the nucleoside.

[0170] As used herein, "oligomeric compound" or "oligomer" means a polymer of linked monomeric subunits that is capable of hybridizing to at least some region of a nucleic acid molecule.

[0171] As used herein, "oligonucleotide" means a polymer of one or more segments of linked nucleosides, each of which may or may not be modified independently of the other.

[0172] The present disclosure relates to the treatment 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 neuropathy, e.g., chemotherapy-induced neuropathy, spinocerebellar neuropathy, Provided is a method for treating, alleviating, or preventing diseases such as, but not limited to, spinal muscular atrophy (SCA), SCA type 2, spinal muscular atrophy (SMA), parkinsonism, Niemann-Pick disease type C (NPC), Charcot-Marie-Tooth disease (CMT), mucopolysaccharidosis type II (MPSIIA), mucolipidosis IV, GM1 gangliosidosis, sporadic inclusion body myositis (sIBM), Henoch-Schonlein purpura (HSP), Gaucher disease, and facial onset sensorimotor neuropathy, Guam Parkinsonism-Dementia Complex, multisystem proteinopathy, Perry disease, and autism, comprising the step of administering to a patient a composition comprising a therapeutically effective amount of an antisense oligonucleotide and a pharmaceutically acceptable excipient.

[0173] Also provided herein is a method for treating, alleviating, or preventing a disease, comprising administering to a patient a composition comprising a therapeutically effective amount of an antisense oligonucleotide and a pharmaceutically acceptable excipient.For example, in some embodiments, the method for treating, alleviating, or preventing a disease comprises administering to a patient a pharmaceutically acceptable composition, such as a pharmaceutically acceptable formulation, comprising one or more antisense oligonucleotides.The antisense oligonucleotide disclosed herein can target any one of the transcripts (e.g., mRNA or pre-mRNA transcripts) of PPM1A, ATXN2, SOD1, or MAPT.The antisense oligonucleotide can modulate the expression of the transcript to treat the disease disclosed herein.

[0174] The present disclosure also provides pharmaceutical compositions comprising the antisense oligonucleotide disclosed herein, formulated with one or more pharmaceutically or cosmetically acceptable excipients.These formulations include those suitable for oral, sublingual, intratracheal, intranasal, intravaginal, rectal, topical, transdermal, intrapulmonary, intrathecal, intracisternal, intrathecal, intrathalamic, intraventricular, intraocular, buccal, and parenteral (for example, subcutaneous, intramuscular, intradermal, intraduodenal, or intravenous) administration, or those suitable for topical use, for example, as part of a composition suitable for topical application to skin and / or mucous membrane, for example, in the form of gel, paste, wax, cream, spray, liquid, foam, lotion, ointment, topical solution, transdermal patch, powder, vapor, or tincture.However, the most suitable form of administration in any given case will depend on the degree and severity of the condition being treated and the properties of the specific antisense oligonucleotide being used.

[0175] The present invention also provides pharmaceutical compositions comprising antisense oligonucleotides or pharmaceutically acceptable salts thereof. The present disclosure also provides methods that involve the use of pharmaceutical compositions comprising the antisense oligonucleotides disclosed herein (e.g., AONs comprising any one of SEQ ID NOS: 1-954, 1914-149354, 149362-158581, or 167805-301566, or AONs comprising one or more spacers replacing one or more nucleosides of any one of SEQ ID NOS: 1-954, 1914-149354, 149362-158581, or 167805-301566, e.g., AONs comprising SEQ ID NOS: 301692-301742 or any one of the sequences in Tables 4A-4C) formulated with one or more pharmaceutically acceptable excipients. Exemplary compositions provided herein include compositions comprising the above-mentioned antisense oligonucleotide and one or more pharmaceutically acceptable excipients.Preparations include those suitable for oral, sublingual, intratracheal, intranasal, rectal, intravaginal, topical, transdermal, intrapulmonary, intrathecal, intracisternal, intrathecal, intrathalamic, intraventricular, intraocular, buccal, and parenteral (for example, subcutaneous, intramuscular, intradermal, intraduodenal, or intravenous) administration or topical use.The most suitable form of administration in any given case will depend on the clinical symptoms, comorbidities, or biochemical indicators of the condition, disorder, disease, or condition that is to be prevented in the subject, the condition, disorder, disease, or condition that is to be prevented in the subject, and / or the nature of the specific compound and / or composition that is used.

[0176] Antisense therapeutic drugs Antisense therapeutic agents are a class of nucleic acid-based compounds that can be used to inhibit gene expression.Antisense therapeutic agents can be single-stranded or double-stranded deoxyribonucleic acid (DNA), ribonucleic acid (RNA), or DNA / RNA chemical analog compounds.Generally, antisense therapeutic agents are designed to contain nucleotide sequences that are complementary or nearly complementary to the mRNA or pre-mRNA sequence of a given gene, so as to promote the binding between the antisense therapeutic agent and pre-mRNA or mRNA.Examples of given genes disclosed herein include, but are not limited to, PPM1A, ATXN2, SOD1, or MAPT.

[0177] Without being bound by theory, in most cases, antisense therapeutic agents are believed to act by binding to mRNA or pre-mRNA, thereby inhibiting protein translation, altering the splicing of pre-mRNA into mature mRNA, and / or causing mRNA destruction. In most cases, the nucleotide sequence of the antisense therapeutic agent is complementary to a portion of the sense sequence of the targeted gene, mRNA, or pre-mRNA. The antisense therapeutic agents described herein are oligonucleotide-based compounds that include an oligonucleotide sequence complementary to the sense sequence of a gene, the sense sequence of a pre-mRNA, and / or the sense sequence of an mRNA, or a portion thereof. The antisense therapeutic agents described herein may also be compounds based on nucleotide chemical analogs that can bind to the sense sequence of a gene, the sense sequence of a pre-mRNA, and / or the sense sequence of an mRNA, or a portion thereof. Antisense therapeutic agents include antisense oligonucleotides, shRNAs, siRNAs, PNAs, LNAs, and morpholino oligomers.

[0178] Antisense oligonucleotides (AONs) are short oligonucleotide-based sequences that contain an oligonucleotide sequence complementary to a target RNA sequence. AONs are typically 8 to 50 nucleotides in length, e.g., 18 nucleotides, 20 nucleotides, or 23 nucleotides in length. AONs may contain chemically modified nucleosides (e.g., 2'-O-methylated nucleosides or 2'-O-(2-methoxyethyl) nucleosides) and modified internucleoside linkages (e.g., phosphorothioate linkages). AONs described herein include oligonucleotide sequences that are complementary to an RNA sequence, e.g., an mRNA or pre-mRNA transcript. AONs described herein may contain chemically modified nucleosides and modified internucleoside linkages (e.g., phosphorothioate linkages).

[0179] Peptide nucleic acids (PNAs) are short, artificially synthesized polymers with structures that mimic DNA or RNA. PNAs contain a backbone composed of repeating N-(2-aminoethyl)-glycine units linked by peptide bonds. The PNAs described herein bind to mRNA or pre-mRNA sequences with high specificity and can be used as antisense therapeutic agents to inhibit target gene expression.

[0180] Locked nucleic acid (LNA) is an oligonucleotide sequence that contains one or more modified RNA nucleotides, in which the ribose moiety is modified with an extra bridge connecting the 2' oxygen and 4' carbon.LNA is believed to have a higher Tm than similar oligonucleotide sequences.The LNA described herein binds to RNA sequences with high specificity and can be used as an antisense therapeutic agent to inhibit target gene expression.

[0181] Morpholino oligomers are oligonucleotide compounds containing DNA bases attached to a backbone of methylene morpholine rings linked through phosphorodiamidate groups. The morpholino oligomers of the present invention can be designed to bind to a specific RNA sequence of interest (e.g., a specific mRNA or pre-mRNA sequence) and thereby inhibit gene expression. The morpholino oligomers described herein can be used as antisense therapeutic agents that bind to mRNA sequences with high specificity and inhibit gene expression. The morpholino oligomers described herein can also be used to bind to pre-mRNA sequences and alter pre-mRNA splicing and gene expression.

[0182] Small hairpin RNA (shRNA) is generally an RNA molecule with a hairpin-like structure that can be used to silence gene expression. shRNA is generally expressed from a plasmid encoding the shRNA sequence, and can be expressed from a viral vector that allows lentivirus, adenovirus, or adeno-associated virus expression. Without being bound by theory, shRNA is thought to inhibit gene expression by utilizing the RNA interference (RNAi) process. Briefly, shRNA transcripts are processed by Drosha and Dicer, and then loaded into an RNA-induced silencing complex (RISC), allowing for the targeting of specific mRNA and either mRNA degradation or suppression of protein translation. The shRNA described herein can inhibit target gene expression.

[0183] Small interfering RNA (siRNA) is a double-stranded RNA molecule, approximately 20-25 base pairs in length, that utilizes RNAi mechanisms (e.g., Drosha and RISC) to bind to and target mRNA for degradation. siRNA does not rely on a plasmid or vector for expression and can generally be delivered directly to target cells, for example, by transfection. siRNA is a double-stranded RNA sequence that contains an RNA sequence complementary to an mRNA sequence and prevents protein translation.

[0184] The number of nucleotides contained in antisense therapeutic agents, e.g., the antisense oligonucleotides described herein, can vary. For example, in some embodiments, the antisense oligonucleotide is 12 to 15 oligonucleotide units in length. In some embodiments, the antisense oligonucleotide is 15 to 20 oligonucleotide units in length. In some embodiments, the antisense oligonucleotide is 20 to 40 oligonucleotide units in length. In some embodiments, the antisense oligonucleotide is 20 to 22 oligonucleotide units in length. In some embodiments, the antisense oligonucleotide is 22 to 40 oligonucleotide units in length. In some embodiments, the antisense oligonucleotide is 20 to 30, 25 to 35, or 30 to 40 oligonucleotide units in length. In certain embodiments, the antisense oligonucleotide is 18 oligonucleotide units in length. In certain embodiments, the antisense oligonucleotide is 20 oligonucleotide units in length. In certain embodiments, the antisense oligonucleotide is 23 oligonucleotide units in length.

[0185] antisense oligonucleotides Antisense oligonucleotides (AONs) described herein are short synthetic oligonucleotide sequences complementary to any portion of a gene product, e.g., a PPM1A, ATXN2, SOD1, or MAPT transcript (e.g., a PPM1A, ATXN2, SOD1, or MAPT mRNA transcript, or a PPM1A, ATXN2, SOD1, or MAPT pre-mRNA transcript).

[0186] In various embodiments, AONs comprise linked nucleosides having a nucleobase sequence that is 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 100% complementary to a portion of a target gene product, e.g., mRNA or pre-mRNA sequence. In some embodiments, AONs may comprise non-duplex oligonucleotides. In some embodiments, AONs may comprise a duplex of two oligonucleotides, where the first oligonucleotide comprises a nucleotide sequence that is fully or nearly fully complementary to the target mRNA sequence, and the second oligonucleotide comprises a nucleotide sequence that is complementary to the nucleotide sequence of the first oligonucleotide. The binding specificity of AONs can be assessed by measuring parameters such as dissociation constants, melting temperatures (Tm), or other criteria, e.g., changes in protein or RNA expression levels, or other assays that measure target activity or expression.

[0187] AONs, such as those disclosed herein, can be oligonucleotide sequences that are 5 to 100 oligonucleotide units in length, e.g., 10 to 40 oligonucleotide units in length, e.g., 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 oligonucleotide units in length, 15 to 22 oligonucleotide units in length, 18 to 21 oligonucleotide units in length, or 18, 19, 20, 21, 22, 23, 24, or 25 oligonucleotide units in length. In certain embodiments, the AON is 18 oligonucleotide units in length. In certain embodiments, the AON is 20 oligonucleotide units in length. In certain embodiments, the AON is 23 oligonucleotide units in length. In certain embodiments, the AON comprises 20 linked nucleosides. As used herein, "parent oligonucleotide" refers to an AON comprising 20 linked nucleosides.

[0188] AONs described herein also include antisense oligonucleotides comprising the oligonucleotide sequences listed in Tables 1A-1D. Specifically, Table 1A shows exemplary PPM1A AONs. In various embodiments, the PPM1A AON comprises the sequence of any one of SEQ ID NOs: 1-954. Table 1B shows exemplary ATXN2 AONs. In various embodiments, the ATXN2 AON comprises the sequence of any one of SEQ ID NOs: 1914-149354. Table 1C shows exemplary SOD1 AONs. In various embodiments, the SOD1 AON comprises the sequence of any one of SEQ ID NOs: 149362-158581. Table 1D shows exemplary MAPT AONs. In various embodiments, the MAPT AON comprises the sequence of any one of SEQ ID NOs: 167805-301566.

[0189] In various embodiments, the AONs shown in Tables 1A-1D comprise at least one nucleoside linkage selected from a phosphorothioate linkage, an alkylphosphate linkage, an alkylphosphonate linkage, a 3-methoxypropylphosphonate linkage, a phosphorodithioate linkage, a phosphotriester linkage, a methylphosphonate linkage, an aminoalkylphosphotriester linkage, an alkylenephosphonate linkage, a phosphinate linkage, a phosphoramidate linkage, a phosphoramidothioate linkage, a phosphorodiamidate (e.g., comprising a phosphorodiamidate morpholino (PMO), a 3' aminoribose, or a 5' aminoribose) linkage, an aminoalkylphosphoramidate linkage, a thiophosphoramidate linkage, a thionoalkylphosphonate linkage, a thionoalkylphosphotriester linkage, a thiophosphate linkage, a selenophosphate linkage, and a boranophosphate linkage.

[0190] In certain embodiments, disclosed herein are gapmer oligonucleotides comprising a sequence that shares at least 85% identity with an equal length portion of any one of SEQ ID NOS: 1-954, 1914-149354, 149362-158581, 167805-301566, SEQ ID NOS: 301692-301742, or the sequences in Tables 4A-4C. In certain embodiments, disclosed herein are gapmer oligonucleotides comprising a sequence that shares at least 90% identity with an equal length portion of any one of SEQ ID NOS: 1-954, 1914-149354, 149362-158581, 167805-301566, SEQ ID NOS: 301692-301742, or the sequences in Tables 4A-4C. In certain embodiments, disclosed herein are gapmer oligonucleotides comprising a sequence that shares at least 95% identity with an equal length portion of any one of SEQ ID NOS: 1-954, 1914-149354, 149362-158581, 167805-301566, SEQ ID NOS: 301692-301742, or the sequences in Tables 4A-4C. In certain embodiments, disclosed herein are gapmer oligonucleotides comprising a sequence that shares at least 98% identity with an equal length portion of any one of SEQ ID NOS: 1-954, 1914-149354, 149362-158581, 167805-301566, SEQ ID NOS: 301692-301742, or the sequences in Tables 4A-4C. In certain embodiments, disclosed herein are gapmer oligonucleotides comprising a sequence that shares 100% identity with an equal length portion of any one of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, 167805-301566, SEQ ID NOs: 301692-301742, or the sequences in Tables 4A-4C.

[0191] Oligonucleotide Variants In various embodiments, AONs include various variants (e.g., AONs of different lengths), hereinafter referred to as AON variants. AON variants can be oligonucleotide sequences that are 5 to 100 nucleobases in length, e.g., 10 to 40 nucleobases in length, e.g., 14 to 40 nucleobases in length, 10 to 30 nucleobases in length, e.g., 14 to 30 nucleobases in length, e.g., 16 to 28 nucleobases in length, e.g., 19 to 23 nucleobases in length, e.g., 18 to 21 nucleobases in length, or e.g., 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleobases in length. AON variants can be oligonucleotide sequences that are complementary to a portion of a target mRNA sequence or a target pre-mRNA sequence.

[0192] In various embodiments, an AON variant represents a modified version of a corresponding parent oligonucleotide comprising a nucleobase sequence selected from any one of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, or 167805-301566. In some embodiments, an AON variant comprises a nucleobase sequence that represents a truncated version of the nucleobase sequence of an AON selected from any one of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, or 167805-301566. As an example, if a parent oligonucleotide comprises a 20-mer (e.g., 20 nucleotide bases in length), the variant (e.g., AON variant) can comprise a shorter version of the 20-mer parent oligonucleotide (e.g., a 15-mer, 16-mer, 17-mer, 18-mer, or 19-mer) or a longer version of the 20-mer parent oligonucleotide (e.g., a 21-mer, 22-mer, 23-mer, 24-mer, 25-mer, 26-mer, 27-mer, 28-mer, 29-mer, or 30-mer). In one embodiment, the nucleobase sequence of the AON variant differs from the corresponding nucleobase sequence of the parent oligonucleotide in that 1, 2, 3, 4, 5, or 6 nucleotide bases have been removed from or added to one or both of the 3' and 5' ends of the nucleobase sequence of the parent oligonucleotide.

[0193] In one embodiment, the corresponding AON variant may comprise an 18-mer in which one nucleotide base has been removed from each of the 3' and 5' ends of the 20-mer in the parent oligonucleotide. For example, the 18-mer AON variant may comprise a sequence in which one nucleotide base has been removed from each of the 3' and 5' ends of any one of SEQ ID NOS: 1-954, 1914-149354, 149362-158581, or 167805-301566.

[0194] In one embodiment, the corresponding AON variant may comprise an 18-mer in which two nucleotide bases have been removed from the 3' end of a 20-mer contained in a parent oligonucleotide. For example, an 18-mer AON variant may comprise a sequence in which two nucleotide bases have been removed from the 3' end of any one of SEQ ID NOS: 1-954, 1914-149354, 149362-158581, or 167805-301566. In one embodiment, the corresponding AON variant may comprise an 18-mer in which two nucleotide bases have been removed from the 5' end of a 20-mer contained in a parent oligonucleotide. For example, an 18-mer AON variant may comprise a sequence in which two nucleotide bases have been removed from the 5' end of any one of SEQ ID NOS: 1-954, 1914-149354, 149362-158581, or 167805-301566.

[0195] In one embodiment, the corresponding AON variant may comprise a 19-mer in which one nucleotide base has been removed from either the 3' or 5' end of the 20-mer contained in the parent oligonucleotide. For example, a 19-mer AON variant may comprise a sequence in which one nucleotide base has been removed from the 3' end of any one of SEQ ID NOS: 1-954, 1914-149354, 149362-158581, or 167805-301566. For example, a 19-mer AON variant may comprise a sequence in which one nucleotide base has been removed from the 5' end of any one of SEQ ID NOS: 1-954, 1914-149354, 149362-158581, or 167805-301566.

[0196] In one embodiment, the corresponding AON variant may comprise a 23-mer in which three nucleotide bases have been added to either the 3' or 5' end of a 20-mer contained in a parent oligonucleotide. For example, a 19-mer AON variant may comprise a sequence in which three nucleotide bases have been added to either the 3' or 5' end of any one of SEQ ID NOS: 1-954, 1914-149354, 149362-158581, or 167805-301566.

[0197] Exemplary sequences of AON variants are shown below in Tables 2A-2B.

[0198] [Table 1]

[0199] [Table 2] In Tables 2A and 2B, an "*" indicates that at least one nucleoside linkage of the nucleobase sequence is selected from 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 phosphoramidate linkage, a phosphoramidothioate linkage, a thiophosphorodiamidate linkage, a phosphorodiamidate (e.g., comprising a phosphorodiamidate morpholino (PMO), a 3' aminoribose, or a 5' aminoribose) linkage, an aminoalkylphosphoramidate linkage, a thiophosphoramidate linkage, a thionoalkylphosphonate linkage, a thionoalkylphosphotriester linkage, a thiophosphate linkage, a selenophosphate linkage, and a boranophosphate linkage.

[0200] Gapmar AON In certain embodiments, the AONs disclosed herein have a gapmer design or structure, also referred to herein as "gapmers" or "gapmer AONs." In a gapmer structure, the AON comprises at least three distinct structural regions in a "5 to 3" orientation, including a 5' wing region, a central region, and a 3' wing region.

[0201] In various embodiments, the 5' wing region comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 oligonucleotide units. In various embodiments, at least one of the oligonucleotide units in the 5' wing region comprises a spacer. In various embodiments, the 5' wing region comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 linked nucleosides. In various embodiments, the 3' wing region comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 oligonucleotide units. In various embodiments, at least one of the oligonucleotide units in the 3' wing region comprises a spacer. In various embodiments, the 3' wing region comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 linked nucleosides. The 5' and 3' wing regions (also referred to as flanking regions) comprise at least one oligonucleotide unit adjacent to the central region, which in some embodiments comprises a stretch of consecutive nucleosides. The 5' and 3' wing regions may be symmetric or asymmetric with respect to the number of oligonucleotide units or linked nucleosides they contain.

[0202] In various embodiments, the 5' wing region comprises one or more RNA nucleosides (e.g., ribonucleosides). In various embodiments, the 5' wing region comprises one or more DNA nucleosides (e.g., deoxyribonucleosides). In various embodiments, the 5' wing region comprises both RNA and DNA nucleosides. In various embodiments, the 3' wing region comprises one or more RNA nucleosides. In various embodiments, the 3' wing region comprises one or more DNA nucleosides. In various embodiments, the 3' wing region comprises both RNA and DNA nucleosides.

[0203] In various embodiments, the central region comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 oligonucleotide units. In various embodiments, the central region comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 linked nucleosides. In some embodiments, the central region comprises a stretch of nucleosides that allows for the recruitment and activation of RNAse H. In some embodiments, the central region comprises one or more of linked DNA nucleosides, 2'-fluoroarabinonucleic acid (FANA), and fluorocyclohexenyl nucleic acid (F-CeNA). In some embodiments, all nucleosides in the central region are DNA nucleosides. In some embodiments, the central region comprises a contiguous stretch of 5-16 DNA nucleosides. In some embodiments, the central region comprises a contiguous stretch of 6-15, 7-14, 8-13, or 9-11 DNA nucleosides. In various embodiments, the central region comprises a mix of DNA and RNA nucleosides. In various embodiments, at least one oligonucleotide unit in the central region is a spacer. In some embodiments, one oligonucleotide unit in the central region is a spacer. In some embodiments, two oligonucleotide units in the central region are spacers.

[0204] In some embodiments, all of the nucleosides in the central region are DNA nucleosides. In further embodiments, the central region can be composed of a mixture of DNA nucleosides and other nucleosides capable of mediating RNase H cleavage, such as 2'-fluoroarabinonucleic acid (FANA) and fluorocyclohexenyl nucleic acid (F-CeNA). In some embodiments, at least 50% of the nucleosides in the central region are DNA nucleosides, e.g., at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% are DNA nucleosides.

[0205] In certain embodiments, the AON comprises a 5' wing region of five linked nucleosides, a central region of ten linked nucleosides, and a 3' wing region of five linked nucleosides, also referred to as a 5-10-5 gapmer. In certain embodiments, the AON comprises a 5' wing region of three linked nucleosides, a central region of eight linked nucleosides, and a 3' wing region of three linked nucleosides, also referred to as a 3-8-3 gapmer. In certain embodiments, the AON comprises a 5' wing region of three linked nucleosides, a central region of ten linked nucleosides, and a 3' wing region of three linked nucleosides, also referred to as a 3-10-3 gapmer. In certain embodiments, the AON comprises a 5' wing region of four linked nucleosides, a central region of ten linked nucleosides, and a 3' wing region of four linked nucleosides, also referred to as a 4-10-4 gapmer. In certain embodiments, the AON comprises a 5' wing region of four linked nucleosides, a central region of eight linked nucleosides, and a 3' wing region of four linked nucleosides, also referred to as a 4-8-4 gapmer. In various embodiments, at least one of the nucleosides in either the 3' wing region, the 5' wing region, or the central region is replaced with a spacer. Embodiments of gapmer AONs having one or more spacers are described in further detail herein.

[0206] In certain embodiments, a gapmer AON disclosed herein comprises any one of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, or 167805-301566, wherein the gapmer AON comprises a 5' wing region comprising one or more RNA nucleosides, a central region comprising one or more DNA nucleosides, and a 3' wing region comprising one or more RNA nucleosides. In certain embodiments, a gapmer AON disclosed herein comprises any one of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, or 167805-301566, and is a 5-10-5 gapmer (e.g., a 5' wing region of 5 linked nucleosides, a central region of 10 linked nucleosides, and a 3' wing region of 5 linked nucleosides). In certain embodiments, a gapmer AON disclosed herein comprises any one of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, or 167805-301566, and is a 6-11-6 gapmer (e.g., a 5' wing region of 6 linked nucleosides, a central region of 11 linked nucleosides, and a 3' wing region of 6 linked nucleosides). In certain embodiments, a gapmer AON disclosed herein comprises any one of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, or 167805-301566, and is a 5-8-5 gapmer (e.g., a 5' wing region of 5 linked nucleosides, a central region of 8 linked nucleosides, and a 3' wing region of 5 linked nucleosides).

[0207] Further exemplary gapmer AONs described herein can include those identified in Tables 3A-3D below. Specifically, Table 3A shows exemplary PPM1A gapmer AONs, Table 3B shows exemplary ATXN2 gapmer AONs, Table 3C shows exemplary SOD1 gapmer AONs, and Table 3D shows exemplary MAPT gapmer AONs. In various embodiments, all cytosines in the gapmer AONs shown in Tables 3A-3D are 5-methylcytosines (5-MeC).

[0208] [Table 3]

[0209] [Table 4]

[0210] [Table 5]

[0211] [Table 6]

[0212] In various embodiments, gapmer AONs, e.g., gapmer AONs comprising any one of SEQ ID NOS: 1-954, 1914-149354, 149362-158581, or 167805-301566, or any of the gapmer AON sequences set forth in Tables 3A-3D (e.g., SEQ ID NOS: 301595-301607), comprise a mixture of ribonucleosides and deoxyribonucleosides (including modified ribonucleosides and / or modified deoxyribonucleosides) in the 5' wing region and / or the 3' wing region.

[0213] In certain embodiments, gapmer AONs, such as those comprising any one of SEQ ID NOS: 1-954, 1914-149354, 149362-158581, or 167805-301566, or any of the gapmer AON sequences set forth in Tables 3A-3D (e.g., SEQ ID NOS: 301595-301607), contain modified ribonucleosides in the 5' and / or 3' wing regions. In certain embodiments, each nucleoside in the 5' and 3' wing regions is a modified nucleoside, such as a 2'-O-(2-methoxyethyl) (2'-MOE) nucleoside. For example, each guanosine in the 5' or 3' wing region can be a modified 2'-O-(2-methoxyethyl) guanosine. For example, each adenosine in the 5' or 3' wing region can be a modified 2'-O-(2-methoxyethyl) adenosine. For example, each cytosine in the 5' and 3' wing regions can be a 2'-O-(2-methoxyethyl)-5-methylcytosine. For example, each thymidine in the 5' and 3' wing regions can be a 2'-O-(2-methoxyethyl) thymidine.

[0214] In various embodiments, exemplary gapmer AONs have one or more modified internucleoside linkages, such as any of a phosphorothioate linkage, alkylphosphate linkage, alkylphosphonate linkage, 3-methoxypropylphosphonate linkage, phosphorodithioate linkage, phosphotriester linkage, methylphosphonate linkage, aminoalkylphosphotriester linkage, alkylenephosphonate linkage, phosphinate linkage, phosphoramidate linkage, phosphoramidothioate linkage, phosphorodiamidate (e.g., including phosphorodiamidate morpholino (PMO), 3' aminoribose, or 5' aminoribose) linkage, aminoalkylphosphoramidate linkage, thiophosphoramidate linkage, thionoalkylphosphonate linkage, thionoalkylphosphotriester linkage, thiophosphate linkage, selenophosphate linkage, and boranophosphate linkage.

[0215] Antisense oligonucleotides with one or more spacers The embodiments disclosed herein include antisense oligonucleotides (AONs) containing one or more spacers. In certain embodiments, the AON contains one spacer. In certain embodiments, the AON contains two spacers. In certain embodiments, the AON contains three spacers. Generally, a spacer refers to a nucleoside substitute group lacking a nucleobase, and the nucleoside sugar moiety is replaced by a non-sugar surrogate group. The non-sugar surrogate group cannot be linked to a nucleobase, but can be linked to the 3' and 5' positions of the nucleoside adjacent to the spacer through an internucleoside linking group.

[0216] In various embodiments, an AON having one or more spacers comprises a parent AON (e.g., an antisense oligonucleotide complementary to a portion of any one of a target gene product, e.g., a PPM1A, ATXN2, SOD1, or MAPT mRNA transcript) in which one or more of the nucleosides of the parent oligonucleotide (e.g., any of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, or 167805-301566) have been replaced with a spacer.

[0217] In various embodiments, AONs having one or more spacers include oligonucleotide variants (e.g., antisense oligonucleotides representing modified versions (e.g., shorter or longer) of the corresponding parent oligonucleotides) in which one or more of the nucleosides of the oligonucleotide variants (e.g., shorter or longer versions of the corresponding parent oligonucleotides comprising a nucleobase sequence selected from any one of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, or 167805-301566) have been replaced with a spacer.

[0218] In various embodiments, AONs having one or more spacers include gapmer AONs (e.g., gapmer AONs comprising any one of SEQ ID NOS: 1-954, 1914-149354, 149362-158581, or 167805-301566, or any of the gapmer AON sequences set forth in Tables 3A-3D (e.g., SEQ ID NOS: 301595-301607)) that have one or more of their nucleosides replaced with a spacer. In certain embodiments, for gapmer AONs having one or more spacers, a nucleoside in the 5' wing region is replaced with a spacer. In certain embodiments, for gapmer AONs having one or more spacers, a nucleoside in the 3' wing region is replaced with a spacer. In certain embodiments, for gapmer AONs having one or more spacers, nucleosides in the central region are replaced with spacers. In certain embodiments, for gapmer AONs having one or more spacers, nucleosides in the 5' wing region are replaced with spacers and nucleosides in the central region are replaced with spacers. In certain embodiments, for gapmer AONs having one or more spacers, nucleosides in the 5' wing region are replaced with spacers and nucleosides in the 3' wing region are replaced with spacers. In certain embodiments, for gapmer AONs having one or more spacers, nucleosides in the central region are replaced with spacers and nucleosides in the 3' wing region are replaced with spacers.

[0219] In certain embodiments, gapmer oligonucleotides comprise sugar modifications in any of the following patterns: eeeee-d10-eeeee, eeeee-d8-eeeee, eeeeee-d11-eeeeee, eee-d8-eee, eee-d10-eee, eeee-d10-eeee, and eeee-d8-eeee, where e=2'-MOE nucleoside and d=deoxyribonucleoside. In such embodiments, the spacer may replace at least one "e" (e.g., a 2'-MOE nucleoside) or at least one "d" (e.g., a deoxyribonucleoside) in the gapmer oligonucleotide.

[0220] As used herein, "AONs with one or more spacers" refers to parent AONs with one or more spacers (e.g., any of SEQ ID NOS: 1-954, 1914-149354, 149362-158581, or 167805-301566), AON variants with one or more spacers (e.g., AON variants selected from any one of SEQ ID NOS: 1-954, 1914-149354, 149362-158581, or 167805-301566), and AON variants with one or more spacers (e.g., AON variants selected from any one of SEQ ID NOS: 1-954, 1914-149354, 149362-158581, or 167805-301566). AONs that have one or more spacers (e.g., a gapmer AON comprising any one of SEQ ID NOS: 1-954, 1914-149354, 149362-158581, or 167805-301566, or any of the gapmer AON sequences shown in Tables 3A-3D (e.g., SEQ ID NOS: 301595-301607)). In certain embodiments, an AON having one or more spacers can be an oligonucleotide having a length of 5 to 100 oligonucleotide units, e.g., a length of 10 to 60 oligonucleotide units, e.g., a length of 12 to 50 oligonucleotide units, a length of 14 to 40 oligonucleotide units, a length of 10 to 30 oligonucleotide units, e.g., a length of 14 to 30 oligonucleotide units, e.g., a length of 14 to 25 or 15 to 22 oligonucleotide units, or a length of 18, 19, 20, 21, 22, 23, 24, or 25 oligonucleotide units. As used herein, "oligonucleotide unit" refers to either a nucleoside (e.g., a nucleoside comprising a sugar and / or nucleobase) or a nucleoside-substituting group (e.g., a spacer) of an oligonucleotide.

[0221] In certain embodiments, the AONs with one or more spacers are 25 oligonucleotide units in length. In certain embodiments, the AONs with one or more spacers are 23 oligonucleotide units in length. In certain embodiments, the AONs with one or more spacers are 21 oligonucleotide units in length. In certain embodiments, the AONs with one or more spacers are 20 oligonucleotide units in length. In certain embodiments, the AONs with one or more spacers are 19 oligonucleotide units in length. In certain embodiments, the AONs with one or more spacers are 18 oligonucleotide units in length. In various embodiments, the AONs with one or more spacers are at least 15, at least 16, at least 17, 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 AONs with one or more spacers are at least 15 oligonucleotide units in length. In various embodiments, the AONs having one or more spacers are at least 16 oligonucleotide units in length. In various embodiments, the AONs having one or more spacers are at least 17 oligonucleotide units in length. In various embodiments, the AONs having one or more spacers are at least 18 oligonucleotide units in length. In various embodiments, the AONs having one or more spacers are at least 19 oligonucleotide units in length. In various embodiments, the AONs having one or more spacers are at least 20 oligonucleotide units in length. In various embodiments, the AONs having one or more spacers are at least 21 oligonucleotide units in length.In various embodiments, the AONs having one or more spacers are at least 22 oligonucleotide units in length. In various embodiments, the AONs having one or more spacers are at least 23 oligonucleotide units in length. In various embodiments, the AONs having one or more spacers are at least 24 oligonucleotide units in length. In various embodiments, the AONs having one or more spacers are at least 25 oligonucleotide units in length.

[0222] In various embodiments, the AON with one or more spacers comprises a sequence that shares at least 80% identity with an equal length portion of any one of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, or 167805-301566. In various embodiments, the AON with one or more spacers comprises a sequence that shares at least 85% identity with an equal length portion of any one of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, or 167805-301566. In various embodiments, the AON with one or more spacers comprises a sequence that shares at least 90% identity with an equal length portion of any one of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, or 167805-301566. In various embodiments, the AON having one or more spacers comprises a sequence that shares at least 95% identity with an equal length portion of any one of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, or 167805-301566. In various embodiments, the AON comprises a sequence that shares 98% identity with an equal length portion of any one of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, or 167805-301566.

[0223] In some embodiments, the spacer has the formula (X):

[0224] [ka] It is of wherein ring A is as defined herein.

[0225] In some embodiments, the spacer has the formula (Xa):

[0226] [ka] It is of wherein ring A is as defined herein and the -CH2-O- group is on the ring A atom adjacent to the -O- group.

[0227] As generally defined herein, ring A of formula (X) and (Xa) is an optionally substituted 4- to 8-membered monocyclic cycloalkyl group (e.g., ring A is cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl), or a 4- to 8-membered monocyclic heterocyclyl group, wherein the heterocyclyl group 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 halo (e.g., -F, -Cl), -OMe, -OEt -O(CH)OMe, -O(CH)OMe, and CN. In some embodiments, the spacer is represented by formula (I):

[0228] [ka] wherein: X is selected from —CH— and —O—; n is 0, 1, 2, or 3.

[0229] In some embodiments, the spacer has formula (I'):

[0230] [ka] wherein: X is selected from —CH— and —O—; n is 0, 1, 2, or 3.

[0231] In some embodiments, the spacer has formula (Ia):

[0232] [ka] wherein: n is 0, 1, 2, or 3.

[0233] In some embodiments, the spacer has formula (Ia'):

[0234] [ka] wherein: n is 0, 1, 2, or 3.

[0235] As generally defined herein, X is selected from -CH2- and -O-. In some embodiments, X is -CH2-. In other embodiments, X is -O-.

[0236] 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.

[0237] In some embodiments, the spacer has formula (II):

[0238] [ka] wherein: X is selected from -CH2- and -O-.

[0239] In some embodiments, the spacer has formula (II'):

[0240] [ka] wherein: X is selected from -CH2- and -O.

[0241] In some embodiments, the spacer has formula (Iia):

[0242] [ka] is expressed by

[0243] In some embodiments, the spacer has the formula (Iia'):

[0244] [ka] is expressed by

[0245] In some embodiments, the spacer has formula (III):

[0246] [ka] wherein: X is selected from -CH2- and -O-.

[0247] In some embodiments, the spacer has formula (III'):

[0248] [ka] wherein: X is selected from -CH2- and -O.

[0249] In some embodiments, the spacer has formula (IIIa):

[0250] [ka] is expressed by

[0251] In some embodiments, the spacer has formula (IIIa'):

[0252] [ka] is expressed by

[0253] In some embodiments, vacant positions (i.e., positions not specifically shown as having exclusively hydrogen atoms, including the -CH- group of 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 halo (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.

[0254] As further described below, AONs having one or more spacers are described with reference to the corresponding parent oligonucleotide, corresponding oligonucleotide variant, or corresponding gapmer oligonucleotide. In various embodiments, oligonucleotides having one or more spacers differ from parent oligonucleotides, oligonucleotide variants, or gapmer oligonucleotides in that each of the one or more spacers replaces a nucleoside in the parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide. As used hereinafter, the "position" of an oligonucleotide refers to a specific position counted from the 5' end of the 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 parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide. In certain embodiments, a spacer replaces the nucleoside at position 7 of a parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide. In certain embodiments, a spacer replaces the nucleoside at position 8 of a parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide. In certain embodiments, a spacer replaces the nucleoside at position 11 of a parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide. In certain embodiments, a spacer replaces the nucleoside at position 14 of a parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide. In certain embodiments, a spacer replaces the nucleoside at position 16 of a parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide.In certain embodiments, the spacer replaces the nucleoside at position 19 of a parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide. In certain embodiments, the spacer replaces the nucleoside at position 22 of an oligonucleotide variant or gapmer oligonucleotide.

[0255] In various embodiments, the oligonucleotide comprises a spacer replacing a nucleoside in the parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide (e.g., a spacer replaces a nucleoside in the parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide). Exemplary oligonucleotides having a spacer are shown in Table 4A below. In certain embodiments, the spacer replaces a nucleoside between positions 9 and 15 of the parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide. In certain embodiments, the spacer replaces a nucleoside between positions 9 and 12 of the parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide. In certain embodiments, the spacer replaces a nucleoside at position 9 of the parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide. In certain embodiments, the spacer replaces a nucleoside at position 10 of the parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide. In certain embodiments, a spacer replaces the nucleoside at position 11 of a parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide. In certain embodiments, a spacer replaces the nucleoside at position 12 of a parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide. In certain embodiments, a spacer replaces a nucleoside between positions 12 and 16 of a parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide. In certain embodiments, a spacer replaces the nucleoside at position 15 of a parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide.

[0256] In various embodiments, an oligonucleotide comprising one spacer has two segments, and at least one of the two segments has at most 11 linked nucleosides. For example, the oligonucleotide can be 23 oligonucleotide units in length, and the spacer can be located at position 12. Thus, the oligonucleotide has two segments separated by a spacer, and both of the two segments are 11 nucleosides in length. In various embodiments, an oligonucleotide comprising one spacer has two segments, and at least one of the two segments has at most 10 linked nucleosides. For example, the oligonucleotide can be 21 oligonucleotide units in length, and the spacer can be located at position 11. Thus, the oligonucleotide has two segments separated by a spacer, and both of the two segments are 10 nucleosides in length. As another example, the oligonucleotide can be 25 oligonucleotide units in length, and the spacer can be located at position 15. Thus, the oligonucleotide has two segments separated by a spacer, one of which is 14 nucleobases in length, and the second of which is 10 nucleobases in length.As another example, the oligonucleotide can be 20 oligonucleotide units in length, and the spacer can be located at position 11.Therefore, the oligonucleotide has two segments separated by a spacer, one of which is 10 nucleobases in length, and the second of which is 9 nucleobases in length.As another example, the oligonucleotide can be 18 oligonucleotide units in length, and the spacer can be located at position 11.Therefore, the oligonucleotide has two segments separated by a spacer, one of which is 10 nucleobases in length, and the second of which is 7 nucleobases in length.As another example, the oligonucleotide may be 18 oligonucleotide units in length and the spacer may be located at position 9. Thus, the oligonucleotide has two segments separated by the spacer, one of the two segments being 8 nucleobases in length and the second of the two segments being 9 nucleobases in length.

[0257] In various embodiments, the oligonucleotide comprises two spacers, each replacing a nucleoside in the parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide (e.g., two spacers replacing two separate nucleosides in the parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide). Exemplary oligonucleotides having two spacers are shown in Table 4B and Table 4C below. In various embodiments, the first spacer and the second spacer are spaced apart 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 certain embodiments, the first spacer and the second spacer are spaced apart by at least 5 nucleobases, at least 6 nucleobases, or at least 7 nucleobases. In certain embodiments, the first spacer and the second spacer are not adjacent to each other in the oligonucleotide.

[0258] In certain embodiments, a first spacer replaces a nucleoside between positions 7 and 11 of a parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide. In various embodiments, a first spacer replaces a nucleoside between positions 8 and 11, 9 and 11, 10 and 11, 7 and 10, 7 and 9, 7 and 8, 8 and 10, 8 and 9, or 9 and 10 of a parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide. In certain embodiments, a second spacer replaces a nucleoside between positions 14 and 22 of a parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide. In various embodiments, the second spacer is located between positions 15 and 22, between positions 16 and 22, between positions 17 and 22, between positions 18 and 22, between positions 19 and 22, between positions 20 and 22, between positions 21 and 22, between positions 15 and 21, between positions 16 and 21, between positions 17 and 21, between positions 18 and 21, between positions 19 and 21, and replacing a nucleoside between positions 1 and 2, between positions 20 and 21, between positions 15 and 20, between positions 16 and 20, between positions 17 and 20, between positions 18 and 20, between positions 19 and 20, between positions 15 and 19, between positions 16 and 19, between positions 17 and 19, between positions 18 and 19, between positions 15 and 18, between positions 16 and 18, between positions 17 and 18, between positions 15 and 17, between positions 16 and 17, or between positions 15 and 16.

[0259] In a preferred embodiment, a first spacer replaces the nucleoside at position 7 of a parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide, and a second spacer replaces the nucleoside at position 14 of a parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide. In a preferred embodiment, a first spacer replaces the nucleoside at position 7 of a parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide, and a second spacer replaces the nucleoside at position 15 of a parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide. In a preferred embodiment, a first spacer replaces the nucleoside at position 7 of a parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide, and a second spacer replaces the nucleoside at position 19 of a parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide. In a preferred embodiment, a first spacer replaces the nucleoside at position 8 of a parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide, and a second spacer replaces the nucleoside at position 16 of a parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide. In a preferred embodiment, a first spacer replaces the nucleoside at position 11 of a parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide, and a second spacer replaces the nucleoside at position 19 of a parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide. In a preferred embodiment, a first spacer replaces the nucleoside at position 11 of a parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide, and a second spacer replaces the nucleoside at position 22 of a parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide.In a preferred embodiment, a first spacer replaces the nucleoside at position 9 of a parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide, and a second spacer replaces the nucleoside at position 19 of a parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide. In a preferred embodiment, a first spacer replaces the nucleoside at position 5 of a parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide, and a second spacer replaces the nucleoside at position 17 of a parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide.

[0260] In various embodiments, the oligonucleotide comprises three spacers, each replacing a nucleoside in the parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide (e.g., three spacers replacing three separate nucleosides in the parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide). In certain embodiments, the first spacer replaces a nucleoside between positions 7 and 11 of the parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide. In certain embodiments, the second spacer replaces a nucleoside between positions 14 and 22 of the parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide. In certain embodiments, the third spacer replaces a nucleoside between positions 21 and 24 of the parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide. In some embodiments, the first spacer replaces a nucleoside between positions 2 and 5 of the parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide. In certain embodiments, the second spacer replaces a nucleoside between positions 8 and 12 of a parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide. In certain embodiments, the third spacer replaces a nucleoside between positions 18 and 22 of a parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide.

[0261] In various embodiments, the three spacers in the oligonucleotide are arranged so that each of the four segments of the oligonucleotide is at most seven linked nucleosides in length.For example, the oligonucleotide may have a first segment with seven linked nucleosides connected to a first spacer, then a second segment with seven linked nucleosides connected to the first spacer at one end and to a second spacer at the other end, then a third segment with six linked nucleosides connected to the second spacer at one end and to a third spacer at the other end, and then a fourth segment with six linked nucleosides connected to the third spacer.

[0262] In various embodiments, one or more spacers are arranged in the oligonucleotide to replace one or more adenosine or thymine nucleosides (as opposed to guanine or cytosine nucleosides).For example, one or more spacers can replace 1, 2, 3, 4, 5, 6, 7, 8, or 9 adenosine or thymine nucleosides in the oligonucleotide.In various embodiments, one or more spacers are arranged in the oligonucleotide to replace one or more guanine or cytosine nucleosides (as opposed to adenosine or thymine nucleosides).For example, one or more spacers can replace 1, 2, 3, 4, 5, 6, 7, 8, or 9 guanine or cytosine nucleosides in the oligonucleotide.In various embodiments, spacers are arranged in the oligonucleotide to replace an equal number of adenosine / thymine nucleosides and guanine / cytosine nucleosides. For example, a first spacer in an oligonucleotide can replace an adenosine / thymine nucleoside and a second spacer in an oligonucleotide can replace a guanine / cytosine nucleoside.

[0263] In various embodiments, one or more spacers are arranged in the oligonucleotide to control the sequence content in the oligonucleotide.For example, one or more spacers are arranged so that at least one of the spacers is located adjacent to a guanine group.In various embodiments, the oligonucleotide with spacers can include one spacer adjacent to a guanine group, two spacers adjacent to a guanine group, three spacers adjacent to a guanine group, four spacers adjacent to a guanine group, or five spacers adjacent to a guanine group.In one embodiment, when counting from the 5' end of the oligonucleotide, the spacer is located immediately before the guanine group in the sequence.Therefore, in various embodiments, the oligonucleotide with spacers can include one spacer immediately before the guanine group, two spacers each immediately before the guanine group, three spacers each immediately before the guanine group, four spacers each immediately before the guanine group, or five spacers each immediately before the guanine group.In one embodiment, when counting from the 5' end of the oligonucleotide, the guanine group is immediately followed by a spacer. Therefore, in various embodiments, the oligonucleotide having spacers 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 arranged to maximize the number of spacers adjacent to the guanine group.

[0264] In various embodiments, one or more spacers are arranged in the oligonucleotide to replace one or more adenosine or thymine nucleosides, such that one or more spacers are positioned adjacent to a guanine group.For example, two spacers can replace adenosine or thymine nucleosides in the oligonucleotide, and each of the two spacers is positioned adjacent to a guanine group.

[0265] In various embodiments, the oligonucleotide with one or more spacers has a specific GC content.As used herein, GC content (or guanine-cytosine content) refers to the percentage of nitrogenous bases in an oligonucleotide that are either guanine (G) or cytosine (C).In various embodiments, the oligonucleotide with one or more spacers has 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 certain embodiments, the oligonucleotide with one or more spacers has at least 30% GC content.In certain embodiments, the oligonucleotide with one or more spacers has at least 40% GC content. In various embodiments, one or more spacers are positioned in the oligonucleotide to maximize GC content. For example, instead of selecting guanine or cytosine for replacement by a spacer in the oligonucleotide, thymine or adenine can be selected for replacement by the spacer.

[0266] In various embodiments, oligonucleotides having spacers are designed such that 1) each segment of the oligonucleotide has at most seven linked nucleosides, and 2) at least two, three, or four spacers are positioned adjacent to a guanine group. In some embodiments, oligonucleotides having spacers are designed such that 1) each segment of the oligonucleotide has at most seven linked nucleosides, and 2) each of two spacers precedes a guanine group.

[0267] In various embodiments, the inclusion of one or more spacers in the oligonucleotide does not reduce the effectiveness of the oligonucleotide with spacers in reducing the frequency of target mRNA or pre-mRNA transcripts compared to the effect of corresponding parent oligonucleotide, oligonucleotide variant or gapmer oligonucleotide.Exemplary mRNA or pre-mRNA transcripts include any of the gene products of PPM1A, ATXN2, SOD1 or MAPT.In various embodiments, the inclusion of one or more spacers in the oligonucleotide increases the effectiveness of the oligonucleotide with spacers in reducing the frequency of target mRNA or pre-mRNA transcripts compared to the effect of corresponding parent oligonucleotide, oligonucleotide variant or gapmer oligonucleotide.

[0268] Tables 4A, 4B, and 4C document exemplary AONs with one or more spacers, as well as their association with the corresponding parent oligonucleotides, corresponding oligonucleotide variants, and / or corresponding gapmer oligonucleotides. Each oligonucleotide is assigned a sequence name. As used hereinafter, the nomenclature of the sequence names is represented as "X_spA" (AON with one spacer), "X_spA_spB" (AON with two spacers), or "X_spA_spB_spC" (AON with three spacers). In this specification, "X" refers to the length of the AON, "A" refers to the position in the AON where the first spacer is located, "B" refers to the position in the AON where the second spacer is located, and "C" refers to the position in the AON where the third spacer is located, if present.

[0269] Exemplary AONs with one spacer are documented below in Table 4A.

[0270] [Table 7-1]

[0271] [Table 7-2]

[0272] [Table 7-3]

[0273] [Table 7-4]

[0274] In various embodiments, the oligonucleotides disclosed herein comprise two spacers, hi various embodiments, the inclusion of the two spacers divides the oligonucleotide into three separate segments, at least one of which is at most seven linked nucleosides in length.

[0275] In certain embodiments, the first spacer is positioned between positions 5 and 11 of the oligonucleotide. In various embodiments, the first spacer is positioned between positions 7 and 11 of the oligonucleotide. In various embodiments, the second spacer is positioned between positions 15 and 19 of the oligonucleotide. In various embodiments, the first spacer and the second spacer are spaced apart in the oligonucleotide by at least 5 nucleobases, at least 6 nucleobases, or at least 7 nucleobases.

[0276] In various embodiments, the first spacer is positioned between positions 5 and 11 of the oligonucleotide, and the second spacer is positioned between positions 15 and 19 of the oligonucleotide. In various embodiments, the first spacer is positioned at position 8 of the oligonucleotide, and the second spacer is positioned at position 16 of the oligonucleotide. In various embodiments, the first spacer is positioned at position 5 of the oligonucleotide, and the second spacer is positioned at position 17 of the oligonucleotide. In various embodiments, the first spacer is positioned at position 7 of the oligonucleotide, and the second spacer is positioned at position 14 of the oligonucleotide. In various embodiments, the first spacer is positioned at position 7 of the oligonucleotide, and the second spacer is positioned at position 15 of the oligonucleotide. In various embodiments, the first spacer is positioned at position 11 of the oligonucleotide, and the second spacer is positioned at position 19 of the oligonucleotide. Exemplary AONs with two spacers are documented below in Table 4B.

[0277] [Table 8-1]

[0278] [Table 8-2]

[0279] [Table 8-3]

[0280] [Table 8-4]

[0281] [Table 8-5]

[0282] In various embodiments, the AONs with one or more spacers are of reduced or increased length compared to the AONs described above in Tables 4A and 4B. For example, such AONs can be oligonucleotide variants with one or more spacers. In various embodiments, the oligonucleotide variants with one or more spacers are 23-mers, 21-mers, 19-mers, or 18-mers. In various embodiments, the oligonucleotide variants include two spacers such that the oligonucleotide variants include three segments separated by two spacers. In various embodiments, at least one of the three segments has at most seven linked nucleosides. In various embodiments, each of the three segments has at most seven linked nucleosides. Exemplary oligonucleotide variants with one or more spacers are shown in Table 4C below.

[0283] [Table 9-1]

[0284] [Table 9-2]

[0285] [Table 9-3]

[0286] In various embodiments, the antisense oligonucleotides disclosed herein (e.g., PPM1A AONs, ATXN2 AONs, SOD1 AONs, or MAPT AONs) comprise one or more locked nucleic acids (LNAs). In certain embodiments, the antisense oligonucleotides comprise one LNA. In certain embodiments, the antisense oligonucleotides comprise two LNAs. In certain embodiments, the antisense oligonucleotides comprise three LNAs. Generally, an LNA 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, thereby forming a bicyclic sugar.

[0287] In some embodiments, the antisense oligonucleotides disclosed herein (e.g., PPM1A AONs, ATXN2 AONs, SOD1 AONs, or MAPT AONs) comprise one or more spacers and one or more locked nucleic acids (LNAs). In some embodiments, the antisense oligonucleotides disclosed herein (e.g., PPM1A AONs, ATXN2 AONs, SOD1 AONs, or MAPT AONs) comprise two spacers and two LNAs. In some embodiments, the antisense oligonucleotides disclosed herein comprise two spacers and three LNAs.

[0288] In various embodiments, spacer and LNA are positioned adjacent to each other in antisense oligonucleotide.For example, spacer can be positioned at M position of antisense oligonucleotide.In addition, LNA can be positioned at M+1 position or M-1 position of antisense oligonucleotide.In various embodiments, the first spacer is positioned adjacent to the first LNA in antisense oligonucleotide, and the second spacer is positioned adjacent to the second LNA.For example, the first spacer is positioned at M position of antisense oligonucleotide, and the second spacer is positioned at N position of antisense oligonucleotide.The first LNA is positioned at M+1 position or M-1 position of antisense oligonucleotide, and the second LNA is positioned at N+1 position or N-1 position of antisense oligonucleotide.In other embodiments, one or more spacers and one or more LNA are not positioned adjacent to each other in antisense oligonucleotide. For example, there may be 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 oligonucleotide units between the spacer and the LNA in the antisense oligonucleotide.

[0289] Chemical modifications to AONs As described herein, AONs, such as those having any one of the sequences set forth in SEQ ID NOS: 1-954, 1914-149354, 149362-158581, or 167805-301566, or one or more spacers replacing one or more nucleosides in any one of the sequences set forth in SEQ ID NOS: 1-954, 1914-149354, 149362-158581, or 167805-301566, such as those having any one of SEQ ID NOS: 301692-301742 or any one of the sequences in Tables 4A-4C, can contain one or more chemical modifications to one or more nucleosides and / or one or more internucleoside linkages. A nucleoside is a base-sugar combination. The nucleobase (also known as the base) portion of a nucleoside is typically a heterocyclic base moiety. Nucleotides are nucleosides that further comprise a phosphate group covalently linked to the sugar moiety of the nucleoside.For nucleosides that comprise a pentofuranosyl sugar, the phosphate group can be linked to the 2', 3', or 5' hydroxyl moiety of the sugar.Oligonucleotides are formed by covalently linking adjacent nucleosides to each other to form linear polymeric oligonucleotides.Within the oligonucleotide structure, the phosphate group is generally referred to as forming the internucleoside linkage of the oligonucleotide.

[0290] Modifications to AONs include substitutions or changes to internucleoside linkages and / or nucleosides (e.g., sugar moieties or nucleobases of nucleosides). Modified AONs may be preferred over native forms due to desirable properties, such as enhanced cellular uptake, enhanced affinity for nucleic acid targets, increased stability in the presence of nucleases, or increased inhibitory activity. Chemically modified nucleosides, nucleobases, and internucleoside linkages are described in Agrawal and Gait, History and Development of Nucleotide Analogues in Nucleic Acids Drugs, in Drug Discovery Series No. 68, Advances in Nucleic Acid Therapeutics, 1-21 (Agrawal and Gait eds., 2019), the contents of which are incorporated herein by reference.

[0291] Modified Internucleoside Linkages In various embodiments, AONs, such as those comprising any one of SEQ ID NOS: 1-954, 1914-149354, 149362-158581, or 167805-301566, or AONs having one or more spacers replacing one or more nucleosides in any one of SEQ ID NOS: 1-954, 1914-149354, 149362-158581, or 167805-301566, such as those having SEQ ID NOS: 301692-301742 or any one of the sequences in Tables 4A-4C, comprise one or more modified internucleoside linkages. The naturally occurring internucleoside linkage in RNA and DNA is a 3' to 5' phosphodiester linkage. AONs having one or more modified, i.e., non-naturally occurring, internucleoside linkages may be selected over antisense compounds having naturally occurring internucleoside linkages due to desirable properties, such as enhanced cellular uptake, enhanced affinity for target nucleic acids, and increased stability in the presence of nucleases.

[0292] Modified internucleoside linkages include internucleoside linkages that retain a phosphorus atom and internucleoside linkages that do not have a phosphorus atom.Representative phosphorus-containing internucleoside linkages include, but are not limited to, phosphodiesters, phosphotriesters, methylphosphonates, phosphoramidates, and phosphorothioates.Methods for preparing phosphorus-containing and non-phosphorus-containing linkages are well known. Examples of modified internucleoside linkages include any one of a phosphorothioate linkage, an alkylphosphate linkage, an alkylphosphonate linkage, a 3-methoxypropylphosphonate linkage, a phosphorodithioate linkage, a phosphotriester linkage, a methylphosphonate linkage, an aminoalkylphosphotriester linkage, an alkylenephosphonate linkage, a phosphinate linkage, a phosphoramidate linkage, a phosphoramidothioate linkage, a phosphorodiamidate (e.g., including a phosphorodiamidate morpholino (PMO), a 3' aminoribose, or a 5' aminoribose) linkage, an aminoalkylphosphoramidate linkage, a thiophosphoramidate linkage, a thionoalkylphosphonate linkage, a thionoalkylphosphotriester linkage, a thiophosphate linkage, a selenophosphate linkage, and a boranophosphate linkage.

[0293] In various embodiments, the AON comprises one or more modified internucleoside linkages linking the oligonucleotide units, hi various embodiments, the AON comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 modified internucleoside linkages linking the oligonucleotide units.

[0294] In various embodiments, each modified internucleoside linkage of an AON can be designed independently of other modified internucleoside linkages of the AON. In other words, the modified internucleoside linkages of an AON do not all have to be the same type of modified internucleoside linkage. In various embodiments, modified internucleoside linkages are interspersed throughout the antisense compound.

[0295] In various embodiments, the AON comprises at least one phosphorothioate linkage. In various embodiments, the AON comprises at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, or at least 19 phosphorothioate linkages. In certain embodiments, the AON comprises 13, 14, 15, 16, 17, 18, 19, 20, 21, or 22 phosphorothioate linkages. In certain embodiments, the AON comprises 17 phosphorothioate linkages. In certain embodiments, the AON comprises 19 phosphorothioate linkages. In certain embodiments, the AON comprises 22 phosphorothioate linkages. In certain embodiments, all internucleoside linkages of the AON are phosphorothioate linkages.

[0296] In various embodiments, AONs comprise a mixture of modified internucleoside linkages and naturally occurring phosphodiester linkages. For example, AONs comprise at least one phosphodiester linkage and at least one phosphorothioate linkage. In various embodiments, AONs comprise 6-10, 6-9, 6-8, 7-10, 7-9, or 6, 7, or 8 phosphorothioate linkages. In some embodiments, AONs comprise 6, 7, 8, 9, or 10 phosphorothioate linkages. In some embodiments, AONs comprise 6-10, 6-9, 6-8, 7-10, 7-9, or 6, 7, or 8 phosphodiester linkages. In some embodiments, AONs comprise 6, 7, 8, 9, or 10 phosphodiester linkages.

[0297] In certain embodiments, the AON comprises 10 phosphorothioate linkages and 9 phosphodiester linkages. In certain embodiments, the AON comprises 6 phosphorothioate linkages and 7 phosphodiester linkages. In certain embodiments, the AON comprises 6 phosphorothioate linkages and 9 phosphodiester linkages. In certain embodiments, the AON comprises 8 phosphorothioate linkages and 9 phosphodiester linkages. In certain embodiments, the AON comprises 8 phosphorothioate linkages and 7 phosphodiester linkages. In certain embodiments, the AON comprises 12 phosphorothioate linkages and 7 phosphodiester linkages. In certain embodiments, the AON comprises 15 phosphorothioate linkages and 4 phosphodiester linkages. In certain embodiments, the AON comprises 15 phosphorothioate linkages and 2 phosphodiester linkages. In certain embodiments, the AON comprises 17 phosphorothioate linkages and 2 phosphodiester linkages.

[0298] In some embodiments, the AON comprises internucleoside linkages designed according to the gapmer design of the AON. In some embodiments, the 5' wing region comprises at least one modified internucleoside linkage (e.g., modified from the naturally occurring internucleoside linkage of a 3' to 5' phosphodiester linkage). In some embodiments, the 5' wing region comprises at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten modified internucleoside linkages. In some embodiments, the 3' wing region comprises at least one modified internucleoside linkage. In some embodiments, the 3' wing region comprises at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten modified internucleoside linkages. In some embodiments, the central region comprises at least one modified internucleoside linkage. In some embodiments, the central region comprises at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 modified internucleoside linkages.

[0299] In certain embodiments, all internucleoside linkages in the 5' wing region are modified internucleoside linkages, e.g., phosphorothioate linkages. In certain embodiments, all internucleoside linkages in the 3' wing region are modified internucleoside linkages, e.g., phosphorothioate linkages. In certain embodiments, all internucleoside linkages in the central region are modified internucleoside linkages, e.g., phosphorothioate linkages. In certain embodiments, all internucleoside linkages in each of the 5' wing region, 3' wing region, and central region are modified internucleoside linkages, e.g., phosphorothioate linkages.

[0300] In various embodiments, two internucleoside linkages in the 5' wing region are phosphorothioate linkages. In various embodiments, two internucleoside linkages in the 5' wing region are phosphodiester linkages. In various embodiments, two internucleoside linkages in the 5' wing region are phosphorothioate linkages and two other internucleoside linkages in the 5' wing region are phosphodiester linkages. In various embodiments, two internucleoside linkages in the 3' wing region are phosphorothioate linkages. In various embodiments, two internucleoside linkages in the 3' wing region are phosphodiester linkages. In various embodiments, two internucleoside linkages in the 3' wing region are phosphorothioate linkages and two other internucleoside linkages in the 3' wing region are phosphodiester linkages.

[0301] In various embodiments, four internucleoside linkages in the 5' wing region are phosphorothioate linkages. In various embodiments, four internucleoside linkages in the 5' wing region are phosphorothioate linkages and two internucleoside linkages in the 3' wing region are phosphorothioate linkages. In various embodiments, one internucleoside linkage in the 5' wing region is phosphorothioate linkage. In various embodiments, one internucleoside linkage in the 3' wing region is phosphorothioate linkage. In various embodiments, one internucleoside linkage in the 5' wing region is phosphorothioate linkage and one internucleoside linkage in the 3' wing region is phosphorothioate linkage.

[0302] In some embodiments, one or more modified internucleoside linkages in the 5' wing region, 3' wing region, or central region are phosphorothioate internucleoside linkages. In some embodiments, the phosphorothioate linkages are stereochemically pure phosphorothioate linkages. In some embodiments, the phosphorothioate linkages are Sp phosphorothioate linkages. In other embodiments, the phosphorothioate linkages are Rp phosphorothioate linkages.

[0303] In some embodiments, the one or more modified internucleoside linkages in the 5' wing region, 3' wing region, or central region can be any of an alkyl phosphate linkage, alkyl phosphonate linkage, 3-methoxypropyl phosphonate linkage, phosphorodithioate linkage, phosphotriester linkage, methyl phosphonate linkage, aminoalkyl phosphotriester linkage, alkylene phosphonate linkage, phosphinate linkage, phosphoramidate linkage, phosphoramidothioate linkage, phosphorodiamidate (e.g., including phosphorodiamidate morpholino (PMO), 3' aminoribose, or 5' aminoribose) linkage, aminoalkyl phosphoramidate linkage, thiophosphoramidate linkage, thionoalkylphosphonate linkage, thionoalkylphosphotriester linkage, thiophosphate linkage, selenophosphate linkage, and boranophosphate linkage. In various embodiments, each modified internucleoside linkage in the 5' wing region, 3' wing region, or central region can be designed independently of other modified internucleoside linkages. In other words, the modified internucleoside linkages in the 5' wing region, 3' wing region, and central region do not all have to be the same type of modified internucleoside linkage. In various embodiments, modified internucleoside linkages are interspersed throughout the antisense compound.

[0304] In various embodiments, one or more internucleoside linkages in the 5' wing region, 3' wing region, or central region are naturally occurring linkages (e.g., phosphodiester linkages). In various embodiments, all internucleoside linkages in the central region are unmodified internucleoside linkages (e.g., phosphodiester linkages).

[0305] In various embodiments, the internucleoside linkages in one region (e.g., the 5' wing region, the 3' wing region, or the central region) can be different from the internucleoside linkages in another region. In certain embodiments, the 5' wing region comprises at least one modified internucleoside linkage, the 3' wing region comprises at least one modified internucleoside linkage, and all internucleoside linkages in the central region are unmodified internucleoside linkages (e.g., phosphodiester linkages). In some embodiments, the central region of the oligonucleotide comprises phosphodiester linkages, and the 5' wing region and the 3' wing region each comprise one or more phosphorothioate linkages. In certain embodiments, all of the internucleoside linkages in the 5' wing region are modified internucleoside linkages, all of the internucleoside linkages in the 3' wing region are modified internucleoside linkages, and all of the internucleoside linkages in the central region are unmodified internucleoside linkages (e.g., phosphodiester linkages).

[0306] In certain embodiments, the gapmer AON is a 5-10-5 gapmer and the internucleoside linkages of the gapmer AON are designated as ssssssssssssssssssss (where "s" refers to a phosphorothioate linkage). In certain embodiments, the gapmer AON is a 5-10-5 gapmer and the internucleoside linkages of the gapmer AON are designated as sssssssssssssssss, sosssssssssssssss, sssssooooooooossss, sosossssssssssssosos, sooooosssssssssooos, sooooossssssssss , ooooossssssssooooo, oooooooooooooosssss, sooossssssssssssssss, sooosssssssssssssss, ssssssssssssssssss, and sssssoooooooooooooo (where "s" refers to a phosphorothioate linkage and "o" refers to a phosphodiester linkage). In certain embodiments, the gapmer AON is a 3-8-3 gapmer, and the internucleoside linkages of the gapmer AON are shown as sssssssssssss (where "s" refers to a phosphorothioate linkage).

[0307] In certain embodiments, the gapmer AON is a 3-8-3 gapmer, and the internucleoside linkages of the gapmer AON are designated as any of sssooooooosss, ooosssssssooo, sssssssssooo, sossssssssss, sossssssssss, sssssssssss, and ooossssssssss (where "s" refers to a phosphorothioate linkage and "o" refers to a phosphodiester linkage). In certain embodiments, the gapmer AON is a 3-10-3 gapmer, and the internucleoside linkages of the gapmer AON are designated as sssssssssssss (where "s" refers to a phosphorothioate linkage).

[0308] In certain embodiments, the gapmer AON is a 3-10-3 gapmer, and the internucleoside linkages of the gapmer AON are designated as any of sssoooooooosss, ooosssssssssooo, sssssssssssooo, sosssssssssssss, sosssssssssssss, sssssssssssss, and ooossssssssssss (where "s" refers to a phosphorothioate linkage and "o" refers to a phosphodiester linkage). In certain embodiments, the gapmer AON is a 4-10-4 gapmer, and the internucleoside linkages of the gapmer AON are designated as sssssssssssssss (where "s" refers to a phosphorothioate linkage).

[0309] In certain embodiments, the gapmer AON is a 4-10-4 gapmer, and the internucleoside linkages of the gapmer AON are designated as any of ssssooooooooossss, oooosssssssssoooo, sssssssssssssoooo, sooossssssssssss, sooosssssssssssss, ssssssssssssssss, and oooossssssssssss (where "s" refers to a phosphorothioate linkage and "o" refers to a phosphodiester linkage). In certain embodiments, the gapmer AON is a 4-8-4 gapmer, and the internucleoside linkages of the gapmer AON are designated as sssssssssssss (where "s" refers to a phosphorothioate linkage).

[0310] In certain embodiments, the gapmer AON is a 4-8-4 gapmer, and the internucleoside linkages of the gapmer AON are designated as any of ssssooooooossss, oooosssssssoooo, sssssssssssoooo, soossssssssssoos, sooosssssssssss, ssssssssssssoos, and oooosssssssssss (where "s" refers to a phosphorothioate linkage and "o" refers to a phosphodiester linkage).

[0311] In certain embodiments, the gapmer AON is a 5-8-5 gapmer, and the internucleoside linkages of the gapmer AON are designated as either ssssssssssssssssss or sosssssssssssssoss ("s" refers to a phosphorothioate linkage and "o" refers to a phosphodiester linkage).

[0312] Modified sugar moieties AONs, such as those having any one of SEQ ID NOS: 1-954, 1914-149354, 149362-158581, or 167805-301566, or one or more spacers replacing one or more nucleosides in any one of SEQ ID NOS: 1-954, 1914-149354, 149362-158581, or 167805-301566, such as those having SEQ ID NOS: 301692-301742 or any one of the sequences in Tables 4A-4C, can contain one or more nucleosides with modified sugar groups. Such sugar-modified nucleosides can confer enhanced nuclease stability, increased binding affinity, or some other beneficial biological property to the antisense compound.

[0313] In various embodiments, nucleosides having modified sugar moieties include ribose (wherein R is alkyl or aryl and R is alkylene) in which the 2'-OH group may be replaced by any one selected from the group consisting of OR, R, R'OR, SH, SR, NH, NR, N, CN, F, Cl, Br, and I, 2'-O-methyl (2'-OMe) nucleosides, 2'-O-(2-methoxyethyl) ... OE) nucleosides, peptide nucleic acids (PNAs), bicyclic nucleic acids (BNAs), 2'-deoxy-2'-fluoronucleosides, 2'-fluoro-β-D-arabinonucleosides, locked nucleic acids (LNAs), constrained ethyl 2'-4'-bridged nucleic acids (cEts), S-cEts, morpholino oligomers, tcDNA, 2'-O,4'-C-ethylene-linked nucleic acids (ENAs), hexitol nucleic acids (HNAs), and tricyclic analogs (e.g., tcDNA).

[0314] In certain embodiments, a nucleoside comprises a chemically modified ribofuranose ring moiety. Examples of chemically modified ribofuranose rings include, but are not limited to, the addition of substituents (including 5' and 2' substituents), bridging of non-geminal ring atoms to form bicyclic nucleic acids (BNAs), and replacement of a ribosyl ring oxygen atom with S, N(R), or C(R1)(R2), where R, R1, and R2 are each independently H, C1-C 12 alkyl, or protecting groups), as well as combinations thereof. Examples of chemically modified sugars include 2'-F-5'-methyl substituted nucleosides (see PCT International Application WO2008 / 101157, published August 21, 2008, for other disclosed 5',2'-bissubstituted nucleosides), or those with further substitution at the 2' position in addition to replacing the ribosyl ring oxygen atom with S or CF2 (see published U.S. Patent Application No. US2005-0130923, published June 16, 2005), or alternatively, 5' substitution of BNA (see PCT International Application WO2007 / 134181, published November 22, 2007, where LNA is substituted, for example, with a 5'-methyl or 5'-vinyl group).

[0315] Examples of nucleosides having modified sugar moieties include, but are not limited to, nucleosides containing 5'-vinyl, 5'-methyl (R or 5), 4'-S, 2'-F, 2'-OCH, 2'-OCHCH, 2'-OCHCHF, and 2'-O(CH)OCH. Substituents at the 2' position also include allyl, amino, azido, thio, O-allyl, O-C-C 10 Alkyl, OCF3, OCH2F, O(CH2)2S CH3, O(CH2)2-ON(R m )(R n ), O-CH2-C(=O)-N(R m )(R n ), and O-CH2-C(=O)-N(R1)-(CH2)2-N(R m )(R n )-, where each R l , R m , and R n are independently H or substituted or unsubstituted C-C 10 It is alkyl.

[0316] Additional examples of modified sugar moieties include 2'-OMe modified sugar moieties, bicyclic sugar moieties, 2'-O-(2-methoxyethyl) (2'-MOE), 2'-O-(N-methylacetamide), 2'-deoxy-2'-fluoronucleosides, 2'-fluoro-β-D-arabinonucleosides, locked nucleic acids (LNA), constrained ethyl 2'-4'-bridged nucleic acids (cEt) (4'-CH(CH)-O-2'), S-constrained ethyl (S-cEt) 2'-4'-bridged nucleic acids, 4'-CH-O-CH-2', 4'-CH-N(R)-2', 4'-CH(CHOCH)-O-2' ("constrained MOE" or "cMOE"), hexitol nucleic acids (HNA), and tricyclic analogs (e.g., tcDNA).

[0317] In some embodiments, the AONs are 2'-O-methylnucleosides (2'OMe) (e.g., AONs containing one or more 2'OMe-modified sugars), 2'-O-(2-methoxyethyl) (2'-MOE) (e.g., AONs containing one or more 2'-MOE-modified sugars (e.g., 2'-MOE)), peptide nucleic acids (PNAs) (e.g., AONs containing one or more N-(2-aminoethyl)-glycine units linked by amide bonds or carbonyl methylene linkages as repeat units instead of a sugar-phosphate backbone), locked nucleic acids (LNAs) (e.g., AONs containing one or more locked riboses, which may be a mixture of 2'-deoxynucleotides or 2'OMe nucleotides), or conjugated nucleic acids (CNAs). These include ethyl 2'-4'-bridged nucleic acids (c-ET) (e.g., AONs comprising one or more cET sugars), cMOE (e.g., AONs comprising one or more cMOE sugars), morpholino oligomers (e.g., AONs comprising a backbone comprising one or more PMOs), deoxy-2'-fluoronucleosides (e.g., AONs comprising one or more 2'-fluoro-β-D-arabinonucleosides), 2'-O,4'-C-ethylene-linked nucleic acids (ENA) (e.g., AONs comprising one or more ENA-modified sugars), hexitol nucleic acids (HNA) (e.g., AONs comprising one or more HNA-modified sugars), or tricyclic analogs (tcDNA) (e.g., AONs comprising one or more tcDNA-modified sugars).

[0318] As used herein, " bicyclic nucleoside " refers to a modified nucleoside that comprises a bicyclic sugar moiety. Examples of bicyclic nucleosides include, but are not limited to, nucleosides that comprise a bridge between 4' and 2' ribosyl ring atoms.In certain embodiments, the antisense compounds provided herein comprise one or more bicyclic nucleosides that comprise a bridge between 4' and 2'. Examples of such 4' to 2' bridged bicyclic nucleosides include those of the formula: 4'-(CH2)-O-2' (LNA), 4'-(CH2)-S-2', 4'-(CH2)2-O-2' (ENA), 4'-CH(CH3)-O-2', and 4'-CH(CHOCH3)-O-2' (and analogs thereof (see U.S. Patent No. 7,399,845 issued July 15, 2008)), 4'-C(CH3)(CH3)-O-2' (and analogs thereof (see U.S. Patent No. 7,399,845 issued January 8, 2009)).

[0013] Examples of suitable amines include 4'-CH-N(OCH)-2' (and analogs thereof (see published International Application No. WO / 2009 / 006478, published on Dec. 11, 2008)), 4'-CH-N(OCH)-2' (and analogs thereof (see published International Application No. WO / 2008 / 150729, published on Dec. 11, 2008)), 4'-CH-ON(CH)-2' (see published U.S. Patent Application No. US2004-0171570, published on Sep. 2, 2004), 4'-CH-N(R)-O-2' (where R is H, C-C 12 alkyl, or a protecting group) (see U.S. Pat. No. 7,427,672 issued Sep. 23, 2008), 4'-CH2-C(H)(CH3)-2' (see Chattopadhyaya et al., J. Org. Chem., 2009, 74, 118-134), and 4'-CH2-C-(=CH2)-2' (and analogs thereof (see published International Publication WO2008 / 154401 published Dec. 8, 2008)).

[0319] Further reports relating to bicyclic nucleosides can also be found in the published literature (e.g., Singh et al., Chem. Commun., 1998, 4, 455-456; Koshkin et al., Tetrahedron, 1998, 54, 3607-3630; Wahlestedt et al., Proc. Natl. Acad. Sci. USA, 2000, 97, 5633-5638; Kumar et al., Bioorg. Med. Chem. Lett., 1998, 8, 2219-2222; Singh et al., J. Org. Chem., 1998, 63, 10035-10039; Srivastava et al., J. Am. Chem. Soc., 2007, 129(26) 8362-8379, Elayadi et al., Curr. Opinion Invest. Drugs, 2001, 2, 558-561, Braasch et al., Chem. Biol., 2001, 8, 1-7, and Orum et al., Curr. Opinion Mol. Ther., 2001, 3, 239-243, U.S. Patent Nos. 6,268,490, 6,525,191, 6,670,461, 6,770,748, 6,794,499, 7,034,133, 7,053,207, 7,399,845, 7,547,684, and 7,696,345, U.S. Patent Publication Nos. US2008-0039618, US2009-0012281, U.S. Patent Application Nos. 60 / 989,574, 61 / 026, 995, 61 / 026,998, 61 / 056,564, 61 / 086,231, 61 / 097,787, and 61 / 099,844, and published PCT International Applications WO1994 / 014226, WO2004 / 106356, WO2005 / 021570, WO2007 / 134181, WO2008 / 150729, WO2008 / 154401, and WO2009 / 006478.Each of the foregoing bicyclic nucleosides can be prepared with one or more stereochemical sugar configurations, including, for example, α-L-ribofuranose and β-D-ribofuranose (see PCT International Application No. PCT / DK98 / 00393, published as International Publication No. WO 99 / 14226 on March 25, 1999).

[0320] In certain embodiments, the bicyclic sugar moiety of a BNA nucleoside includes, but is not limited to, compounds having at least one bridge between the 4' and 2' positions of the pentofuranosyl sugar moiety, where such bridge is independently selected from the group consisting of -[C(R a )(R b )] n -, -C(R a )=C(R b )-, -C(R a )=N-, -C(=O)-, -C(=NR a )-, -C(=S)-, -O-, -Si(R a )2-, -S(=O) x - and -N(R a )-, and During the ceremony, x is 0, 1, or 2; n is 1, 2, 3, or 4; Each R a and R b are independently H, a protecting group, hydroxyl, 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 Aryl, substituted C5-C 20aryl, heterocyclic radical, substituted heterocyclic radical, heteroaryl, substituted heteroaryl, C5-C7 cycloaliphatic radical, substituted C5-C7 cycloaliphatic radical, halogen, OJ1, NJ1J2, SJ1, N3, COOJ1, acyl (C(=O)-H), substituted acyl, CN, sulfonyl (S(=O)2-J1), or sulfoxyl (S(=O)-J1); Each J1 and J2 is 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 Aryl, substituted C5-C 20 Aryl, acyl (C(=O)-H), substituted acyl, heterocyclic radical, substituted heterocyclic radical, C1-C 12 Aminoalkyl, substituted C1-C 12 aminoalkyl, or a protecting group.

[0321] In certain embodiments, the bridge of the bicyclic sugar moiety is —[C(R a )(R b )] n -,-[-[C(R a )(R b )] n -O-, -C(R a R b )-N(R)-O-, or -C(R a R b )-ON(R)-. In certain embodiments, the bridges are 4'-CH2-2', 4'-(CH2)2-2', 4'-(CH2)3-2', 4'-CH2-O-2', 4'-(CH2)2-O-2', 4'-CH2-ON(R)-2', and 4'-CH2-N(R)-O-2'-, where each R is independently H, a protecting group, or C1-C 12 alkyl, and each R a and R b are independently H, a protecting group, hydroxyl, 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 It is aryl, heterocyclic radical, substituted heterocyclic radical, heteroaryl, substituted heteroaryl, C5-C7 cycloaliphatic radical, substituted C5-C7 cycloaliphatic radical, halogen, OJ1, NJ1J2, SJ1, N3, COOJ1, acyl (C(=O)--H), substituted acyl, CN, sulfonyl (S(=O)2-J1), or sulfoxyl (S(=O)-J1).

[0322] In certain embodiments, bicyclic nucleosides are further defined by their isomeric conformation.For example, nucleosides comprising 4'-2' methylene-oxy bridges can be in α-L conformation or β-D conformation.Previously, α-L-methyleneoxy (4'-CH2-O-2') BNA has been incorporated into antisense oligonucleotides that exhibit antisense activity (Frieden et al., Nucleic Acids Research, 2003, 21, 6365-6372).

[0323] In certain embodiments, bicyclic nucleosides include, but are not limited to, α-L-methyleneoxy (4'-CH2-O-2') BNAs, β-D-methyleneoxy (4'-CH2-O-2') BNAs, ethyleneoxy (4'-(CH2)2-O-2') BNAs, aminooxy (4'-CH2-ON(R)-2') BNAs, oxyamino (4'-CH2-N(R)-O-2') BNAs, methyl(methyleneoxy) (4'-CH(CH3)-O-2') BNAs, methylene-thio (4'-CH2-S-2') BNAs, methylene-amino (4'-CH2-N(R)-2') BNAs, methyl carbocyclic (4'-CH2-CH(CH3)-2') BNAs, and propylene carbocyclic (4'-(CH2)3-2') BNAs.

[0324] As used herein, "locked nucleic acid" or "LNA" or "LNA nucleoside" refers to a modified nucleoside having a bridge (e.g., a methylene, ethylene, aminooxy, or oximino bridge) connecting the two carbon atoms between the 4' and 2' positions of the nucleoside sugar unit, thereby forming 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) ethyleneoxy (4'-(CH2)2-O-2') LNA, (D) aminooxy (4'-CH2-ON(R)-2') LNA, and (E) oxyamino (4'-CH2-N(R)-O-2') LNA, where R is H, C1-C 12 alkyl, or a protecting group (see U.S. Patent No. 7,427,672, issued September 23, 2008).

[0325] As used herein, LNA nucleosides include, but are not limited to, nucleosides having at least one bridge between the 4' and 2' sugar positions, where each bridge is independently -[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)-, where x is 0, 1, or 2, and n is 1, 2, 3, or 4; and each R1 and R2 is independently H, a protecting group, hydroxyl, 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 Aryl, substituted C5-C 20aryl, heterocyclic radical, substituted heterocyclic radical, heteroaryl, substituted heteroaryl, C5-C7 cycloaliphatic radical, substituted C5-C7 cycloaliphatic radical, halogen, OJ1, NJ1J2, SJ1, N3, COOJ1, acyl (C(=O)-H), substituted acyl, CN, sulfonyl (S(=O)2-J1), or sulfoxyl (S(=O)-J1), wherein each J1 and J2 is 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 Aryl, substituted C5-C 20 Aryl, acyl (C(=O)-H), substituted acyl, heterocyclic radical, substituted heterocyclic radical, C1-C 12 Aminoalkyl, substituted C1-C 12 aminoalkyl, or a protecting group.

[0326] Examples of 4'-2' bridging groups encompassed within the definition of LNA include groups of the formula: -[C(R1)(R2)] n -, -[C(R1)(R2)] n Additionally, other bridging groups encompassed within the definition of LNA include, but are not limited to, one of: -O-, -C(R1R2)-N(R1)-O-, or -C(R1R2)-ON(R1)-. Additionally, other bridging groups encompassed within 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, where each R1 and R2 is independently H, a protecting group, or C1-C 12 It is alkyl.

[0327] Also included within the definition of LNA according to the present invention are LNAs in which the 2'-hydroxyl group of the ribosyl sugar ring is connected to the 4' carbon atom of the sugar ring, thereby forming a bridge to form a bicyclic sugar moiety. The bridge can be a methylene (-CH2-) group connecting the 2' oxygen atom and the 4' carbon atom, for which the term methyleneoxy(4'-CH2-O-2')LNA is used. Furthermore, in the case of a bicyclic sugar moiety having an ethylene bridging group at this position, the term ethyleneoxy(4'-CH2CH2-O-2')LNA is used. The isomer of methyleneoxy(4'-CH2-O-2')LNA, α-L-methyleneoxy(4'-CH2-O-2'), is also included within the definition of LNA used herein.

[0328] In various embodiments, the AONs disclosed herein comprise one or more 2'-O-(2-methoxyethyl) (2'-MOE) nucleosides. In various embodiments, the AONs disclosed herein comprise 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 2'-O-(2-methoxyethyl) (2'-MOE) nucleosides. In various embodiments, the AONs disclosed herein comprise more than 25 2'-O-(2-methoxyethyl) (2'-MOE) nucleosides.

[0329] In some embodiments, the AON comprises modified sugar moieties designed according to the gapmer design of the gapmer AON. In various embodiments, the gapmer AON comprises one or more modified sugar moieties. In various embodiments, the 5' wing region comprises at least one modified sugar moiety. In various embodiments, the 3' wing region comprises at least one modified sugar moiety. In various embodiments, the 5' wing region comprises at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten modified sugar moieties. In various embodiments, the 3' wing region comprises at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten modified sugar moieties. In some embodiments, the 5' wing region and / or the 3' wing region each comprise 1 to 7 modified sugar moieties, e.g., 2 to 6 modified sugar moieties, 2 to 5 modified sugar moieties, 2 to 4 modified sugar moieties, or 1 to 3 modified sugar moieties. In certain embodiments, the 5' wing region comprises 3 modified sugar moieties and the 3' wing region comprises 3 modified sugar moieties. In certain embodiments, the 5' wing region comprises 4 modified sugar moieties and the 3' wing region comprises 4 modified sugar moieties. In certain embodiments, the 5' wing region comprises 5 modified sugar moieties and the 3' wing region comprises 5 modified sugar moieties.

[0330] In various embodiments, nucleosides having modified sugar moieties in the 5' and 3' wing regions include ribose (wherein R is alkyl or aryl and R' is alkylene) in which the 2'-OH group can be replaced by any one selected from the group consisting of OR, R, R'OR, SH, SR, NH, NR, N, CN, F, Cl, Br, and I, 2'-O-methyl (2'-OMe) nucleosides, 2'-O-(2-methoxyethyl) (2'-O-methyl) ... 2'-MOE) nucleosides, peptide nucleic acids (PNAs), bicyclic nucleic acids (BNAs), 2'-deoxy-2'-fluoronucleosides, 2'-fluoro-β-D-arabinonucleosides, locked nucleic acids (LNAs), constrained ethyl 2'-4'-bridged nucleic acids (cEts), S-cEts, morpholino oligomers, tcDNAs, 2'-O,4'-C-ethylene-linked nucleic acids (ENAs), hexitol nucleic acids (HNAs), and tricyclic analogs (e.g., tcDNAs).

[0331] In some embodiments, the 5' wing region and / or the 3' wing region comprises at least one 2'-MOE nucleoside. In some embodiments, both the 5' and 3' wing regions comprise at least one 2'-MOE nucleoside. In some embodiments, the 5' wing region and the 3' wing region each comprise 2, 3, 4, 5, 6, 7, 8, 9, or 10 2'-MOE nucleosides. In certain embodiments, the 5' wing region and the 3' wing region each comprise 4 2'-MOE nucleosides. In certain embodiments, the 5' wing region and the 3' wing region each comprise 5 2'-MOE nucleosides. In some embodiments, all nucleosides in each of the 5' wing region and the 3' wing region are 2'-MOE nucleosides.

[0332] In other embodiments, the wing regions may include both 2'-MOE nucleosides and other nucleosides (mixed wings), such as DNA nucleosides and / or non-MOE modified nucleosides, such as bicyclic nucleosides (BNAs) (e.g., locked nucleic acid (LNA) nucleosides or constrained ethyl 2'-4'-bridged nucleic acid (cEt) nucleosides), 2'-O-methyl nucleosides, tricyclic DNA, S-cEt, morpholino, or other 2'-substituted nucleosides.

[0333] In some embodiments, the 5' or 3' wing region comprises at least one BNA (e.g., at least one LNA nucleoside or cET nucleoside). In some embodiments, each of the 5' and 3' wing regions comprises a BNA. In some embodiments, all nucleosides in the 5' and 3' wing regions are BNAs. In further embodiments, the BNAs in the 5' and / or 3' wing regions are independently selected from the group comprising oxy-LNA, thio-LNA, amino-LNA, cET, and / or ENA, in either beta-D or alpha-L conformation or combinations thereof.

[0334] In some embodiments, the 5' and / or 3' wing comprises at least one 2'-O-methyl nucleoside. In some embodiments, the 5' wing comprises at least one 2'-O-methyl nucleoside. In some embodiments, both the 5' and 3' wing regions comprise 2'-O-methyl nucleosides. In some embodiments, all nucleosides in the wing regions are 2'-O-methyl nucleosides.

[0335] Modified nucleobases AONs, such as those having any one of SEQ ID NOS: 1-954, 1914-149354, 149362-158581, or 167805-301566, or one or more spacers replacing one or more nucleosides in any one of SEQ ID NOS: 1-954, 1914-149354, 149362-158581, or 167805-301566, such as those having any one of SEQ ID NOS: 301692-301742 or any one of the sequences in Tables 4A-4C, include one or more modified nucleobases. Examples of modified nucleobases include 5-methylpyrimidines, such as 5-methylcytosine or 5-methoxyuridine, 5-methylpurines, such as 5-methylguanine, or pseudouridine.

[0336] In various embodiments, the AON comprises at least one modified nucleobase. In various embodiments, the AON comprises 2, 3, 4, 5, 6, 7, 8, 9, or 10 modified nucleobases. In various embodiments, the AON comprises at least one 5-methylcytosine nucleobase. In various embodiments, the AON comprises 2, 3, 4, 5, 6, 7, 8, 9, or 10 5-methylcytosine nucleobases.

[0337] In various embodiments, AON comprises both modified and unmodified nucleobases.For example, AON can comprise both cytosine and 5-methylcytosine.In some embodiments, AON can comprise 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 cytosines, and can also comprise 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 5-methylcytosines.

[0338] In various embodiments, each specific type of nucleobase in AON is replaced with corresponding modified nucleobase.For example, all guanine in AON is replaced with 5-methylguanine.As another example, all cytosine in AON is replaced with 5-methylcytosine.

[0339] In some embodiments, the AON comprises modified nucleobases designed according to the gapmer design of gapmer AONs. In various embodiments, the linked nucleosides of the 5'-wing region, the linked nucleosides of the 3'-wing region, or the linked nucleosides of the central region comprise one or more modified nucleobases. In some embodiments, the 5'-wing region and / or the 3'-wing region comprise 1 to 10 modified nucleobases, e.g., 2 to 8 modified nucleobases, 3 to 6 modified nucleobases, or 4 to 5 modified nucleobases. In some embodiments, the 5'-wing region and / or the 3'-wing region comprise 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 modified nucleobases. In some embodiments, the central region comprises 1 to 10 modified nucleobases, e.g., 2 to 8 modified nucleobases, 3 to 6 modified nucleobases, or 4 to 5 modified nucleobases. In some embodiments, the central region comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 modified nucleobases. Examples of modified nucleobases include 5-methylpyrimidines, such as pseudouridine, 5-methylcytosine, or 5-methoxyuridine, 5-methylpurines, such as 5-methylguanine.

[0340] In various embodiments, at least one cytosine in the 5'-wing region and / or 3'-wing region of the AON is replaced with a modified nucleobase, for example, 5-methylcytosine. In various embodiments, at least one cytosine in the 5'-wing region is replaced with a modified nucleobase, for example, 5-methylcytosine. In various embodiments, at least one cytosine in the 3'-wing region is replaced with a modified nucleobase, for example, 5-methylcytosine. In various embodiments, at least one cytosine in the central region is replaced with a modified nucleobase, for example, 5-methylcytosine. In various embodiments, all cytosines in the 5'-wing region are replaced with modified nucleobases, for example, 5-methylcytosine. In various embodiments, all cytosines in the 3'-wing region are replaced with modified nucleobases, for example, 5-methylcytosine. In various embodiments, all cytosines in the central region are replaced with modified nucleobases, for example, 5-methylcytosine.

[0341] In certain embodiments, all cytosines in the 5' wing region, all cytosines in the 3' wing region, and all cytosines in the central region are replaced with modified nucleobases, e.g., 5-methylcytosine. In certain embodiments, all cytosines in the 5' wing region, all cytosines in the 3' wing region are replaced with modified nucleobases, e.g., 5-methylcytosine, while all cytosines in the central region are unmodified nucleobases.

[0342] Modified oligonucleotides Further embodiments of modified oligonucleotides are described herein, which may include any of the above-described modified internucleoside linkages and / or modified nucleosides (e.g., modified sugar moieties and / or modified nucleobases).

[0343] In some embodiments, the AON or pharmaceutically acceptable salt thereof comprises any one of SEQ ID NOS: 1-954, 1914-149354, 149362-158581, or 167805-301566, or one or more spacers replacing one or more nucleosides in any one of SEQ ID NOS: 1-954, 1914-149354, 149362-158581, or 167805-301566, e.g., SEQ ID NOS: 301692-301742 or any of the sequences in Tables 4A-4C. and (AONs having any one of the following structures): wherein at least one nucleoside of the sequence is substituted with a 2'-O-(2-methoxyethyl)nucleoside, a 2'-O-methylnucleoside, a 2'-O-(N-methylacetamido)nucleoside, a 2'-deoxy-2'-fluoronucleoside, a 2'-fluoro-β-D-arabinonucleoside, a bicyclic nucleic acid, a bridged nucleic acid, a locked nucleic acid (LNA), a constrained ethyl (cET) nucleic acid, a tricyclo-DNA (tcDNA), a 2'-O,4'-C-ethylene-linked nucleic acid (ENA), or a peptide nucleic acid (PNA). In certain embodiments, at least one internucleoside linkage of the AON is a phosphorothioate linkage. In some embodiments, all internucleoside linkages of the AON are phosphorothioate linkages. Also described herein are pharmaceutical compositions comprising any of the foregoing antisense oligonucleotides, or pharmaceutically acceptable salts thereof, and a pharmaceutically acceptable excipient.

[0344] The AONs described herein can include chemically modified nucleosides, including modified ribonucleosides and / or modified deoxyribonucleosides. Chemically modified nucleosides include 2'-substituted nucleosides, in which the 2'-position of the sugar ring contains a moiety other than -H or -OH (e.g., -F or an O-alkyl group). For example, chemically modified nucleosides include, but are not limited to, 2'-O-(2-methoxyethyl) modifications, such as 2'-O-(2-methoxyethyl) guanosine, 2'-O-(2-methoxyethyl) adenosine, 2'-O-(2-methoxyethyl) cytosine, 2'-O-(2-methoxyethyl) thymidine, and 2'-O-(2-methoxyethyl)-5-methylcytosine.

[0345] In some embodiments, AONs may include chemically modified nucleosides, such as 2'O-methylribonucleosides, e.g., 2'O-methylcytidine, 2'O-methylguanosine, 2'O-methyluridine, and / or 2'O-methyladenosine. The AONs described herein may also include one or more chemically modified bases, including 5-methylpyrimidines, e.g., 5-methylcytosine, and / or 5-methylpurines, e.g., 5-methylguanine. The AONs described herein may also include any of the following chemically modified nucleosides: 5-methyl-2'-O-methylcytidine, 5-methyl-2'-O-methylthymidine, 5-methylcytidine, 5-methyluridine, and / or 5-methyl-2'-deoxycytidine.

[0346] In some embodiments, the disclosed AONs may optionally have at least one modified nucleobase, e.g., 5-methylcytosine, and / or at least one methylphosphonate nucleotide, located, for example, at either only the 5' or 3' end or both the 5' and 3' ends, or along the oligonucleotide sequence.

[0347] In certain embodiments, the present disclosure provides mixed modalities of AONs with combinations of modified nucleosides, such as combinations of peptide nucleic acids (PNAs) and locked nucleic acids (LNAs). Chemically modified nucleosides also include, but are not limited to, locked nucleic acid (LNA), 2'-O-methyl, 2'-fluoro, and 2'-fluoro-β-D-arabinonucleotide (FANA) modifications. Chemically modified nucleosides that can be included in the AONs described herein are described in Johannes and Lucchino, (2018) "Current Challenges in Delivery and Cytosolic Translocation of Therapeutic RNAs" Nucleic Acid Ther. 28(3): 178-93, Rettig and Behlke, (2012) "Progress toward in vivo use of siRNAs-II" Mol Ther 20:483-512, and Khvorova and Watts, (2017) "The chemical evolution of oligonucleotide therapies of clinical utility" Nat Biotechnol., 35(3):238-48, the contents of each of which are incorporated herein by reference.

[0348] The AONs described herein may include chemical modifications and phosphatase-resistant analogs of 5'-phosphate that promote the stabilization of the terminal 5'-phosphate of oligonucleotides.Chemical modifications or phosphatase-resistant analogs of 5'-phosphate that promote the stabilization of the terminal 5'-phosphate of oligonucleotides include, but are not limited to, 5'-methyl phosphonate, 5'-methylene phosphonate, 5'-methylene phosphonate analogs, 5'-E-vinyl phosphonate (5'-E-VP), 5'-phosphorothioate, and 5'-C-methyl analogs.Chemical modifications and phosphatase-resistant analogs of 5'-phosphate that promote the stabilization of the terminal 5'-phosphate of AONs are described in Khvorova and Watts, (2017) "The chemical evolution of oligonucleotide therapies of clinical utility" Nat Biotechnol., 35(3):238-48, the contents of which are incorporated herein by reference.

[0349] In some embodiments described herein, the AON or pharmaceutically acceptable salt thereof is a modified oligonucleotide comprising the sequence of any one of SEQ ID NOS: 1-954, 1914-149354, 149362-158581, or 167805-301566, or one or more spacers replacing one or more nucleosides of the sequence of any one of SEQ ID NOS: 1-954, 1914-149354, 149362-158581, or 167805-301566, e.g., an AON having SEQ ID NOS: 301692-301742 or any one of the sequences in Tables 4A-4C), wherein at least one nucleoside linkage of the nucleotide sequence is a phosphorothioate. The internucleoside linkages include, for example, alkyl phosphate linkages, alkyl phosphonate linkages, 3-methoxypropyl phosphonate linkages, phosphorodithioate linkages, phosphotriester linkages, methyl phosphonate linkages, aminoalkyl phosphotriester linkages, alkylene phosphonate linkages, phosphinate linkages, phosphoramidate linkages, phosphoramidothioate linkages, phosphorodiamidate (e.g., including phosphorodiamidate morpholino (PMO), 3' aminoribose, or 5' aminoribose) linkages, aminoalkyl phosphoramidate linkages, thiophosphoramidate linkages, thionoalkyl phosphonate linkages, thionoalkyl phosphotriester linkages, thiophosphate linkages, selenophosphate linkages, and boranophosphate linkages. In certain embodiments, at least one internucleoside linkage of the nucleotide sequence is a phosphorothioate linkage. In some embodiments of the AONs described herein, one, two, three, or more internucleoside linkages of the nucleotide sequence are phosphorothioate linkages. In various embodiments of the AONs described herein, all internucleoside linkages of the nucleotide sequence are phosphorothioate linkages.Thus, in some embodiments, all of the nucleotide linkages of an AON sequence of any one of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, or 167805-301566, or an AON having one or more spacers replacing one or more nucleosides of any one of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, or 167805-301566, e.g., SEQ ID NOs: 301692-301742 or any one of the sequences in Tables 4A-4C, are phosphorothioate linkages.

[0350] Contemplated AONs may optionally include at least one modified sugar, for example, the sugar moiety of at least one nucleotide constituting the oligonucleotide is ribose in which the 2'-OH group may be replaced by any one selected from the group consisting of OR, R, R'OR, SH, SR, NH, NR, N, CN, F, Cl, Br, and I (wherein R is alkyl or aryl and R' is alkylene).

[0351] In certain embodiments, the AON has the structure eeeee-d10-eeeee ("e" represents a 2'-MOE modified nucleoside and "d10" represents a sequence of 10 consecutive DNA nucleobases). In this embodiment, the 5' wing region comprises five 2'-MOE modified nucleosides, the gap region comprises 10 consecutive DNA nucleobases, and the 3' wing region comprises five 2'-MOE modified nucleosides. In various embodiments, the internucleoside linkages of a 5-10-5 gapmer AON can have the sequence sssssooooooooosssss ("s" refers to phosphorothioate linkages and "o" refers to phosphodiester linkages), with all phosphorothioate linkages in the 5' or 3' wing regions of the AON and all phosphodiester linkages in the central region. In various embodiments, the AON contains unmodified cytosines. In various embodiments, the AON comprises modified cytosines (e.g., 5-methylcytosine). In various embodiments, all cytosines in the 5' and 3' wing regions are modified cytosines (e.g., 5-methylcytosine). In various embodiments, all cytosines in the central region are modified cytosines (e.g., 5-methylcytosine).

[0352] In certain embodiments, the AON has the structure eeeee-d10-eeeee ("e" represents a 2'-MOE modified nucleoside and "d10" represents a sequence of 10 consecutive DNA nucleobases). In this embodiment, the 5' wing region comprises five 2'-MOE modified nucleosides, the gap region comprises 10 consecutive DNA nucleobases, and the 3' wing region comprises five 2'-MOE modified nucleosides. The internucleoside linkages of the AON may have the sequence ooooosssssssssooooo ("s" refers to a phosphorothioate linkage and "o" refers to a phosphodiester linkage). In various embodiments, the AON comprises an unmodified cytosine. In various embodiments, the AON comprises a modified cytosine (e.g., 5-methylcytosine). In various embodiments, all cytosines in the 5' and 3' wing regions are modified cytosines (e.g., 5-methylcytosines). In various embodiments, all cytosines in the central region are modified cytosines (e.g., 5-methylcytosines).

[0353] In certain embodiments, the AON has the structure eeeee-d10-eeeee ("e" represents a 2'-MOE modified nucleoside and "d10" represents a sequence of 10 consecutive DNA nucleobases). In this embodiment, the 5' wing region comprises five 2'-MOE modified nucleosides, the gap region comprises 10 consecutive DNA nucleobases, and the 3' wing region comprises five 2'-MOE modified nucleosides. The internucleoside linkages of the AON may have the sequence sssssssssssssssssss ("s" refers to a phosphorothioate linkage), where all internucleoside linkages of the AON are phosphorothioate linkages. In various embodiments, the AON comprises an unmodified cytosine. In various embodiments, the AON comprises a modified cytosine (e.g., 5-methylcytosine). In various embodiments, all cytosines in the 5' and 3' wing regions are modified cytosines (e.g., 5-methylcytosines). In various embodiments, all cytosines in the central region are modified cytosines (e.g., 5-methylcytosines).

[0354] In certain embodiments, the AON has the structure eee-d8-eee ("e" represents a 2'-MOE modified nucleoside and "d8" represents a sequence of eight consecutive DNA nucleobases). In this embodiment, the 5' wing region comprises three 2'-MOE modified nucleosides, the gap region comprises eight oligonucleotide units comprising at least four consecutive nucleobases, and the 3' wing region comprises three 2'-MOE modified nucleosides. The internucleoside linkages of the AON may have the sequence sssooooooosss ("s" refers to phosphorothioate linkages and "o" refers to phosphodiester linkages), with all phosphorothioate linkages in the 5' or 3' wing region of the AON and all phosphodiester linkages in the central region. In various embodiments, the AON contains unmodified cytosines. In various embodiments, the AON comprises modified cytosines (e.g., 5-methylcytosine). In various embodiments, all cytosines in the 5' and 3' wing regions are modified cytosines (e.g., 5-methylcytosine). In various embodiments, all cytosines in the central region are modified cytosines (e.g., 5-methylcytosine).

[0355] In certain embodiments, the AON has the structure eee-d8-eee ("e" represents a 2'-MOE modified nucleoside and "d8" represents a sequence of eight consecutive DNA nucleobases). In this embodiment, the 5' wing region comprises three 2'-MOE modified nucleosides, the gap region comprises eight oligonucleotide units comprising four consecutive DNA nucleobases, and the 3' wing region comprises three 2'-MOE modified nucleosides. The internucleoside linkages of the AON may have the sequence ooosssssssooo ("s" refers to phosphorothioate linkages and "o" refers to phosphodiester linkages), with all phosphodiester linkages in the 5' or 3' wing region of the AON and all phosphorothioate linkages in the central region. In various embodiments, the AON contains unmodified cytosines. In various embodiments, the AON comprises modified cytosines (e.g., 5-methylcytosine). In various embodiments, all cytosines in the 5' and 3' wing regions are modified cytosines (e.g., 5-methylcytosine). In various embodiments, all cytosines in the central region are modified cytosines (e.g., 5-methylcytosine).

[0356] In certain embodiments, the AON has the structure eee-d8-eee ("e" represents a 2'-MOE modified nucleoside and "d8" represents a sequence of eight consecutive DNA nucleobases). In this embodiment, the 5' wing region comprises three 2'-MOE modified nucleosides, the gap region comprises eight oligonucleotide units comprising four consecutive DNA nucleobases, and the 3' wing region comprises three 2'-MOE modified nucleosides. The internucleoside linkages of the AON may have the sequence sssssssssssss ("s" refers to a phosphorothioate linkage), where all internucleoside linkages of the AON are phosphorothioate linkages. In various embodiments, the AON comprises an unmodified cytosine. In various embodiments, the AON comprises a modified cytosine (e.g., 5-methylcytosine). In various embodiments, all cytosines in the 5' and 3' wing regions are modified cytosines (e.g., 5-methylcytosines). In various embodiments, all cytosines in the central region are modified cytosines (e.g., 5-methylcytosines).

[0357] In certain embodiments, the AON has the structure eee-d10-eee ("e" represents a 2'-MOE modified nucleoside and "d10" represents a sequence of 10 consecutive DNA nucleobases). In this embodiment, the 5' wing region comprises three 2'-MOE modified nucleosides, the gap region comprises 10 consecutive DNA nucleobases, and the 3' wing region comprises three 2'-MOE modified nucleosides. The internucleoside linkages of the AON may have the sequence sssooooooooosss ("s" refers to phosphorothioate linkages and "o" refers to phosphodiester linkages), with all phosphorothioate linkages in the 5' or 3' wing region of the AON and all phosphodiester linkages in the central region. In various embodiments, the AON contains unmodified cytosines. In various embodiments, the AON comprises modified cytosines (e.g., 5-methylcytosine). In various embodiments, all cytosines in the 5' and 3' wing regions are modified cytosines (e.g., 5-methylcytosine). In various embodiments, all cytosines in the central region are modified cytosines (e.g., 5-methylcytosine).

[0358] In certain embodiments, the AON has the structure eee-d10-eee ("e" represents a 2'-MOE modified nucleoside and "d10" represents a sequence of 10 consecutive DNA nucleobases). In this embodiment, the 5' wing region comprises three 2'-MOE modified nucleosides, the gap region comprises 10 consecutive DNA nucleobases, and the 3' wing region comprises three 2'-MOE modified nucleosides. The internucleoside linkages of the AON may have the sequence ooossssssssooo ("s" refers to a phosphorothioate linkage and "o" refers to a phosphodiester linkage). In various embodiments, the AON comprises an unmodified cytosine. In various embodiments, the AON comprises a modified cytosine (e.g., 5-methylcytosine). In various embodiments, all cytosines in the 5' and 3' wing regions are modified cytosines (eg, 5-methylcytosines).

[0359] In certain embodiments, the AON has the structure eee-d10-eee ("e" represents a 2'-MOE modified nucleoside and "d10" represents a sequence of 10 consecutive DNA nucleobases). In this embodiment, the 5' wing region comprises three 2'-MOE modified nucleosides, the gap region comprises 10 consecutive DNA nucleobases, and the 3' wing region comprises three 2'-MOE modified nucleosides. The internucleoside linkages of the AON may have the sequence sssssssssssssss ("s" refers to a phosphorothioate linkage), where all internucleoside linkages of the AON are phosphorothioate linkages. In various embodiments, the AON comprises an unmodified cytosine. In various embodiments, the AON comprises a modified cytosine (e.g., 5-methylcytosine). In various embodiments, all cytosines in the 5' and 3' wing regions are modified cytosines (e.g., 5-methylcytosines). In various embodiments, all cytosines in the central region are modified cytosines (e.g., 5-methylcytosines).

[0360] In certain embodiments, the AON has the structure eeee-d10-eeee ("e" represents a 2'-MOE modified nucleoside and "d10" represents a sequence of 10 consecutive DNA nucleobases). In this embodiment, the 5' wing region comprises four 2'-MOE modified nucleosides, the gap region comprises 10 consecutive DNA nucleobases, and the 3' wing region comprises four 2'-MOE modified nucleosides. The internucleoside linkages of the AON may have the sequence ssssooooooooossss ("s" refers to phosphorothioate linkages and "o" refers to phosphodiester linkages), with all phosphorothioate linkages in the 5' or 3' wing region of the AON and all phosphodiester linkages in the central region. In various embodiments, the AON contains unmodified cytosines. In various embodiments, the AON comprises modified cytosines (e.g., 5-methylcytosine). In various embodiments, all cytosines in the 5' and 3' wing regions are modified cytosines (e.g., 5-methylcytosine). In various embodiments, all cytosines in the central region are modified cytosines (e.g., 5-methylcytosine).

[0361] In certain embodiments, the AON has the structure eeee-d10-eeee ("e" represents a 2'-MOE modified nucleoside and "d10" represents a sequence of 10 consecutive DNA nucleobases). In this embodiment, the 5' wing region comprises four 2'-MOE modified nucleosides, the gap region comprises 10 consecutive DNA nucleobases, and the 3' wing region comprises four 2'-MOE modified nucleosides. The internucleoside linkages of the AON may have the sequence oooosssssssssoooo ("s" refers to a phosphorothioate linkage and "o" refers to a phosphodiester linkage). In various embodiments, the AON comprises an unmodified cytosine. In various embodiments, the AON comprises a modified cytosine (e.g., 5-methylcytosine). In various embodiments, all cytosines in the 5' and 3' wing regions are modified cytosines (e.g., 5-methylcytosines). In various embodiments, all cytosines in the central region are modified cytosines (e.g., 5-methylcytosines).

[0362] In certain embodiments, the AON has the structure eeee-d10-eeee ("e" represents a 2'-MOE modified nucleoside and "d10" represents a sequence of 10 consecutive DNA nucleobases). In this embodiment, the 5' wing region comprises four 2'-MOE modified nucleosides, the gap region comprises 10 consecutive DNA nucleobases, and the 3' wing region comprises four 2'-MOE modified nucleosides. The internucleoside linkages of the AON may have the sequence sssssssssssssssss ("s" refers to a phosphorothioate linkage), where all internucleoside linkages of the AON are phosphorothioate linkages. In various embodiments, the AON comprises an unmodified cytosine. In various embodiments, the AON comprises a modified cytosine (e.g., 5-methylcytosine). In various embodiments, all cytosines in the 5' and 3' wing regions are modified cytosines (e.g., 5-methylcytosines). In various embodiments, all cytosines in the central region are modified cytosines (e.g., 5-methylcytosines).

[0363] In certain embodiments, the AON has the structure eeee-d8-eeee ("e" represents a 2'-MOE modified nucleoside and "d8" represents a sequence of eight consecutive DNA nucleobases). In this embodiment, the 5' wing region comprises four 2'-MOE modified nucleosides, the gap region comprises eight oligonucleotide units comprising four consecutive DNA nucleobases, and the 3' wing region comprises four 2'-MOE modified nucleosides. The internucleoside linkages of the AON may have the sequence ssssooooooossss ("s" refers to phosphorothioate linkages and "o" refers to phosphodiester linkages), with all phosphorothioate linkages in the 5' or 3' wing region of the AON and all phosphodiester linkages in the central region. In various embodiments, the AON contains unmodified cytosines. In various embodiments, the AON comprises modified cytosines (e.g., 5-methylcytosine). In various embodiments, all cytosines in the 5' and 3' wing regions are modified cytosines (e.g., 5-methylcytosine). In various embodiments, all cytosines in the central region are modified cytosines (e.g., 5-methylcytosine).

[0364] In certain embodiments, the AON has the structure eeee-d8-eeee ("e" represents a 2'-MOE modified nucleoside and "d8" represents a sequence of eight consecutive DNA nucleobases). In this embodiment, the 5' wing region comprises four 2'-MOE modified nucleosides, the gap region comprises eight oligonucleotide units comprising four consecutive DNA nucleobases, and the 3' wing region comprises four 2'-MOE modified nucleosides. The internucleoside linkages of the AON may have the sequence oooosssssssoooo ("s" refers to a phosphorothioate linkage and "o" refers to a phosphodiester linkage). In various embodiments, the AON comprises an unmodified cytosine. In various embodiments, the AON comprises a modified cytosine (e.g., 5-methylcytosine). In various embodiments, all cytosines in the 5' and 3' wing regions are modified cytosines (e.g., 5-methylcytosines). In various embodiments, all cytosines in the central region are modified cytosines (e.g., 5-methylcytosines).

[0365] In certain embodiments, the AON has the structure eeee-d8-eeee ("e" represents a 2'-MOE modified nucleoside and "d8" represents a sequence of eight consecutive DNA nucleobases). In this embodiment, the 5' wing region comprises four 2'-MOE modified nucleosides, the gap region comprises eight oligonucleotide units comprising four consecutive DNA nucleobases, and the 3' wing region comprises four 2'-MOE modified nucleosides. The internucleoside linkages of the AON may have the sequence sssssssssssssss ("s" refers to a phosphorothioate linkage), where all internucleoside linkages of the AON are phosphorothioate linkages. In various embodiments, the AON comprises an unmodified cytosine. In various embodiments, the AON comprises a modified cytosine (e.g., 5-methylcytosine). In various embodiments, all cytosines in the 5' and 3' wing regions are modified cytosines (eg, 5-methylcytosines).

[0366] In certain embodiments, the AON has the structure eeeeee-d11-eeeeee ("e" represents a 2'-MOE modified nucleoside and "d11" represents a sequence of 11 consecutive DNA nucleobases). In this embodiment, the 5' wing region comprises six 2'-MOE modified nucleosides, the gap region comprises 11 consecutive DNA nucleobases, and the 3' wing region comprises six 2'-MOE modified nucleosides. In various embodiments, the internucleoside linkages of a 6-11-6 gapmer AON can have the sequence ssssssssssssssssssssss ("s" refers to a phosphorothioate linkage). In various embodiments, the AON comprises an unmodified cytosine. In various embodiments, the AON comprises a modified cytosine (e.g., 5-methylcytosine). In various embodiments, all cytosines in the 5' and 3' wing regions are modified cytosines (e.g., 5-methylcytosines). In various embodiments, all cytosines in the central region are modified cytosines (e.g., 5-methylcytosines).

[0367] In certain embodiments, the AON has the structure eeeee-d10-eeeee ("e" represents a 2'-MOE modified nucleoside and "d10" represents a sequence of 10 consecutive DNA nucleobases). In this embodiment, the 5' wing region comprises five 2'-MOE modified nucleosides, the gap region comprises 10 consecutive DNA nucleobases, and the 3' wing region comprises five 2'-MOE modified nucleosides. In various embodiments, the internucleoside linkages of a 5-10-5 gapmer AON can have the sequence sosossssssssssssosos ("s" refers to a phosphorothioate linkage and "o" refers to a phosphodiester linkage). In various embodiments, the AON comprises an unmodified cytosine. In various embodiments, the AON comprises a modified cytosine (e.g., 5-methylcytosine). In various embodiments, all cytosines in the 5' and 3' wing regions are modified cytosines (e.g., 5-methylcytosines). In various embodiments, all cytosines in the central region are modified cytosines (e.g., 5-methylcytosines).

[0368] In certain embodiments, the AON has the structure eeeee-d10-eeeee ("e" represents a 2'-MOE modified nucleoside and "d10" represents a sequence of 10 consecutive DNA nucleobases). In this embodiment, the 5' wing region comprises five 2'-MOE modified nucleosides, the gap region comprises 10 consecutive DNA nucleobases, and the 3' wing region comprises five 2'-MOE modified nucleosides. In various embodiments, the internucleoside linkages of a 5-10-5 gapmer AON can have the sequence soooossssssssssooos ("s" refers to a phosphorothioate linkage and "o" refers to a phosphodiester linkage). In various embodiments, the AON comprises an unmodified cytosine. In various embodiments, the AON comprises a modified cytosine (e.g., 5-methylcytosine). In various embodiments, all cytosines in the 5' and 3' wing regions are modified cytosines (e.g., 5-methylcytosines). In various embodiments, all cytosines in the central region are modified cytosines (e.g., 5-methylcytosines).

[0369] In certain embodiments, the AON has the structure eeeee-d10-eeeee ("e" represents a 2'-MOE modified nucleoside and "d10" represents a sequence of 10 consecutive DNA nucleobases). In this embodiment, the 5' wing region comprises five 2'-MOE modified nucleosides, the gap region comprises 10 consecutive DNA nucleobases, and the 3' wing region comprises five 2'-MOE modified nucleosides. In various embodiments, the internucleoside linkages of a 5-10-5 gapmer AON can have the sequence soooosssssssssssooss ("s" refers to a phosphorothioate linkage and "o" refers to a phosphodiester linkage). In various embodiments, the AON comprises an unmodified cytosine. In various embodiments, the AON comprises a modified cytosine (e.g., 5-methylcytosine). In various embodiments, all cytosines in the 5' and 3' wing regions are modified cytosines (e.g., 5-methylcytosines). In various embodiments, all cytosines in the central region are modified cytosines (e.g., 5-methylcytosines).

[0370] In certain embodiments, the AON has the structure eeeee-d8-eeeee ("e" represents a 2'-MOE modified nucleoside and "d8" represents a sequence of eight consecutive DNA nucleobases). In this embodiment, the 5' wing region comprises five 2'-MOE modified nucleosides, the gap region comprises eight oligonucleotide units comprising four consecutive DNA nucleobases, and the 3' wing region comprises five 2'-MOE modified nucleosides. In various embodiments, the internucleoside linkages of a 5-8-5 gapmer AON can have the sequence sossssssssssssss ("s" refers to a phosphorothioate linkage and "o" refers to a phosphodiester linkage). In various embodiments, the AON comprises an unmodified cytosine. In various embodiments, the AON comprises a modified cytosine (e.g., 5-methylcytosine). In various embodiments, all cytosines in the 5' and 3' wing regions are modified cytosines (e.g., 5-methylcytosines). In various embodiments, all cytosines in the central region are modified cytosines (e.g., 5-methylcytosines).

[0371] In certain embodiments, the AON has the structure eeeee-d10-eeeee ("e" represents a 2'-MOE modified nucleoside and "d10" represents a sequence of 10 consecutive DNA nucleobases). In this embodiment, the 5' wing region comprises five 2'-MOE modified nucleosides, the gap region comprises 10 consecutive DNA nucleobases, and the 3' wing region comprises five 2'-MOE modified nucleosides. In various embodiments, the internucleoside linkages of a 5-10-5 gapmer AON can have the sequence sossssssssssssssss ("s" refers to a phosphorothioate linkage and "o" refers to a phosphodiester linkage). In various embodiments, the AON comprises an unmodified cytosine. In various embodiments, the AON comprises a modified cytosine (e.g., 5-methylcytosine). In various embodiments, all cytosines in the 5' and 3' wing regions are modified cytosines (e.g., 5-methylcytosines). In various embodiments, all cytosines in the central region are modified cytosines (e.g., 5-methylcytosines).

[0372] In certain embodiments, the AON has the structure eeeeee-d10-eeee ("e" represents a 2'-MOE modified nucleoside and "d10" represents a sequence of 10 consecutive DNA nucleobases). In this embodiment, the 5' wing region comprises six 2'-MOE modified nucleosides, the gap region comprises 10 consecutive DNA nucleobases, and the 3' wing region comprises four 2'-MOE modified nucleosides. The internucleoside linkages of the AON may have the sequence sssssssssssssss ("s" refers to a phosphorothioate linkage), where all internucleoside linkages of the AON are phosphorothioate linkages. In various embodiments, the AON comprises an unmodified cytosine. In various embodiments, the AON comprises a modified cytosine (e.g., 5-methylcytosine). In various embodiments, all cytosines in the 5' and 3' wing regions are modified cytosines (e.g., 5-methylcytosines). In various embodiments, all cytosines in the central region are modified cytosines (e.g., 5-methylcytosines).

[0373] Target gene product Generally, the AONs disclosed herein comprise linked nucleosides having a nucleobase sequence that is 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 100% complementary to a portion of a target gene product. Exemplary target gene products include any of the PPM1A, ATXN2, SOD1, or MAPT gene products. In certain embodiments, the AONs disclosed herein comprise linked nucleosides having a nucleobase sequence that is 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 100% complementary to a portion of a gene product. In certain embodiments, the AONs disclosed herein comprise linked nucleosides having a nucleobase sequence that is 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 100% complementary to a portion of the ATXN2 gene product. In certain embodiments, the AONs disclosed herein comprise linked nucleosides having a nucleobase sequence that is 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 100% complementary to a portion of the SOD1 gene product. In certain embodiments, the AONs disclosed herein comprise linked nucleosides having a nucleobase sequence that is 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 100% complementary to a portion of a MAPT gene product.

[0374] In embodiments of the invention described herein, AONs may target gene products of genes of one or more species. For example, AONs may target gene products of mammalian genes, such as human (i.e., Homo sapiens) genes, rodent genes (e.g., mouse (Mus musculus) genes), and / or primate genes (e.g., cynomolgus monkey (Macaca fascicularis) genes or rhesus monkey (Macaca mulatta) genes). In certain embodiments, AONs target human gene products.

[0375] The gene product can be, for example, an RNA gene product, e.g., an mRNA gene product, or a protein product of the target gene. In some embodiments, the AON comprises a sequence that is 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 100% complementary to the sequence of a gene or RNA, e.g., an mRNA or pre-mRNA, or a portion thereof. In some embodiments, the AON comprises a nucleobase sequence that is complementary to a portion of a sequence shared between multiple species of genes or RNA (e.g., mRNA). For example, in some embodiments, the AON is an antisense therapeutic, e.g., an AON that is complementary to a sequence shared by human, mouse, and / or primate genes or mRNAs.

[0376] PPM1A In some embodiments of the present disclosure, the PPM1A gene product is a PPM1A mRNA or PPM1A pre-mRNA that includes a sequence from nucleotide 41,932 to nucleotide 42,787 and nucleotide 44,874 to nucleotide 44,990 of the PPM1A gene sequence (e.g., the PPM1A gene sequence or PPM1A coding sequence of NCBI Reference Sequence NG_029698.1 (SEQ ID NO: 1909)), or a portion thereof. In some embodiments of the present disclosure, the PPM1A gene product comprises a sequence that shares at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, 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 100% identity with a PPM1A mRNA or PPM1A pre-mRNA, including a sequence derived from nucleotides 41,932 to 42,787 and nucleotides 44,874 to 44,990, or a portion thereof, of the PPM1A gene sequence (e.g., the PPM1A gene sequence or PPM1A coding sequence of NCBI Reference Sequence NG_029698.1 (SEQ ID NO: 1909)).

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[0394] In some embodiments of the present disclosure, the PPM1A gene product is a PPM1A gene product comprising a sequence derived from one or more of nucleotides 8470-8926, 41933-42787, 44874-45990, 49055-49164, 50647-50704, and 51703-58336 of the PPM1A gene sequence of NCBI Reference Sequence NG_029698.1 (SEQ ID NO: 1909). mRNA or PPM1A pre-mRNA. In some embodiments of the present disclosure, the PPM1A gene product is a PPM1A mRNA or PPM1A pre-mRNA comprising a sequence derived from the coding region of the PPM1A gene sequence, e.g., the coding region comprising nucleotides 8470-8926, 41933-42787, 44874-45990, 49055-49164, 50647-50704, and 51703-58336 of the PPM1A gene sequence (e.g., NCBI Reference Sequence NG_029698.1 (SEQ ID NO: 1909)). In various embodiments, the PPM1A mRNA is PPM1A mRNA transcript variant 1, which corresponds to NCBI Reference Sequence NM_021003.5 (SEQ ID NO: 1910).

[0395] In some embodiments of the present disclosure, the PPM1A gene product is a PPM1A mRNA or PPM1A pre-mRNA that includes sequence derived from a coding region of a PPM1A gene sequence, e.g., a coding region comprising one or more of nucleotides 8470-8926, 9629-9730, 41933-42787, and 44874-47804 of the PPM1A gene sequence (e.g., NCBI Reference Sequence NG_029698.1 (SEQ ID NO: 1909)). In some embodiments of the present disclosure, the PPM1A gene product is a PPM1A mRNA or PPM1A pre-mRNA that includes sequence derived from a coding region of a PPM1A gene sequence (e.g., a coding region comprising one or more of nucleotides 8470-8926, 9629-9730, 41933-42787, and 44874-47804 of the PPM1A gene sequence of NCBI Reference Sequence NG_029698.1 (SEQ ID NO: 1909). mRNA or PPM1A pre-mRNA. In various embodiments, the PPM1A mRNA is PPM1A mRNA transcript variant 2, which corresponds to NCBI reference sequence NM_177951.3 (SEQ ID NO: 1911).

[0396] In some embodiments of the present disclosure, the PPM1A gene product comprises a PPM1A gene sequence (e.g., a PPM1A gene comprising a sequence derived from one or more of nucleotides 4999-5295, 41933-42787, 44874-44990, 49055-49164, 50647-50704, 51703-58336 of the PPM1A gene sequence in NCBI Reference Sequence NG_029698.1 (SEQ ID NO: 1909)). mRNA or PPM1A pre-mRNA. In some embodiments of the present disclosure, the PPM1A gene product is a PPM1A mRNA or PPM1A pre-mRNA that includes a sequence derived from one or more coding regions of the PPM1A gene sequence, e.g., a coding region including nucleotides 4999-5295, 41933-42787, 44874-44990, 49055-49164, 50647-50704, or 51703-58336 of the PPM1A gene sequence of NCBI Reference Sequence NG_029698.1 (SEQ ID NO: 1909). In various embodiments, the PPM1A mRNA is PPM1A mRNA transcript variant 3, which corresponds to NCBI Reference Sequence NM_177952.3 (SEQ ID NO: 1912).

[0397] In some embodiments of the present disclosure, the PPM1A gene product comprises a sequence including nucleotides 457 to 1429 of PPM1A mRNA transcript variant 1 (i.e., nucleotides 457 to 1429 of PPM1A mRNA transcript variant 1, which correspond to, for example, NCBI reference sequence NM_021003.5 (SEQ ID NO: 1910)), or a portion thereof. In some embodiments of the present disclosure, the PPM1A gene product comprises a sequence sharing at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, 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 100% identity to nucleotides 457-1429 of PPM1A mRNA transcript variant 1 (i.e., nucleotides 457-1429 of PPM1A mRNA transcript variant 1 corresponding to, e.g., NCBI reference sequence NM_021003.5 (SEQ ID NO: 1910)), or a portion thereof.

[0398] In some embodiments described herein, the PPM1A gene product is a PPM1A mRNA isoform transcript (e.g., PPM1A mRNA transcript variant 1 corresponding to NCBI Reference Sequence NM_021003.5 (SEQ ID NO: 1910)) or a portion thereof. In some embodiments described herein, the PPM1A gene product comprises a sequence that shares at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, 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 100% identity with a PPM1A mRNA isoform transcript (e.g., PPM1A mRNA transcript variant 1 corresponding to NCBI Reference Sequence NM_021003.5 (SEQ ID NO: 1910)) or a portion thereof. PPM1A mRNA transcript variant 1 corresponding to NCBI reference sequence NM_021003.5 (SEQ ID NO: 1910)

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[0404] In some embodiments described herein, the PPM1A gene product is a PPM1A mRNA isoform transcript (e.g., PPM1A mRNA transcript variant 2 corresponding to NCBI Reference Sequence NM_177951.3 (SEQ ID NO: 1911)) or a portion thereof. In some embodiments described herein, the PPM1A gene product comprises a sequence that shares at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, 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 100% identity with a PPM1A mRNA isoform transcript (e.g., PPM1A mRNA transcript variant 2 corresponding to NCBI Reference Sequence NM_177951.3 (SEQ ID NO: 1911)) or a portion thereof.

[0405] In some embodiments described herein, the PPM1A gene product is a PPM1A mRNA isoform transcript (e.g., PPM1A mRNA transcript variant 3 corresponding to NCBI reference sequence NM_177952.3 (SEQ ID NO: 1912)) or a portion thereof. In some embodiments described herein, the PPM1A gene product comprises a sequence that shares at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, 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 100% identity with a PPM1A mRNA isoform transcript (e.g., PPM1A mRNA transcript variant 3 corresponding to NCBI reference sequence NM_177952.3 (SEQ ID NO: 1912)) or a portion thereof.

[0406] In some embodiments described herein, the PPM1A gene product is a Mus musculus PPM1A mRNA isoform transcript (e.g., a Mus musculus PPM1A mRNA alpha isoform transcript corresponding to NCBI reference sequence NM_008910.3 (sequence number 1913)) or a portion thereof. In some embodiments described herein, the PPM1A gene product comprises a sequence that shares at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, 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%, or 100% identity with a PPM1A mRNA isoform transcript (e.g., the Mus musculus PPM1A mRNA alpha isoform transcript corresponding to NCBI reference sequence NM_008910.3 (SEQ ID NO: 1913)) or a portion thereof.

[0407] In some embodiments of the present disclosure, the PPM1A gene product is a PPM1A mRNA transcript variant other than the PPM1A transcripts described above (e.g., PPM1A mRNA transcript variant 1 corresponding to NCBI Reference Sequence NM_021003.5 (SEQ ID NO: 1910), PPM1A mRNA transcript variant 2 corresponding to NCBI Reference Sequence NM_177951.3 (SEQ ID NO: 1911), PPM1A mRNA transcript variant 3 corresponding to NCBI Reference Sequence NM_177952.3 (SEQ ID NO: 1912), or the Mus musculus PPM1A mRNA alpha isoform transcript corresponding to NCBI Reference Sequence NM_008910.3 (SEQ ID NO: 1913)). In some embodiments, the PPM1A gene product is PPM1A mRNA transcript variant 1, which corresponds to NCBI Reference Sequence NM_021003.5 (SEQ ID NO: 1910), PPM1A mRNA transcript variant 2, which corresponds to NCBI Reference Sequence NM_177951.3 (SEQ ID NO: 1911), PPM1A mRNA transcript variant 3, which corresponds to NCBI Reference Sequence NM_177952.3 (SEQ ID NO: 1912), or Mus musculus PPM1A, which corresponds to NCBI Reference Sequence NM_008910.3 (SEQ ID NO: 1913). This includes sequences that share at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, 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% identity, or 100% identity with a sequence homologous to the sequence of an mRNA alpha isoform transcript.In some embodiments, the PPM1A gene product comprises a sequence sharing at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, 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 100% identity to a sequence homologous to nucleotides 457-1429 of PPM1A mRNA transcript variant 1 corresponding to NCBI reference sequence NM_021003.5 (i.e., nucleotides 457-1429 of SEQ ID NO: 1910), or a portion thereof.

[0408] ATXN2 In some embodiments of the present disclosure, the ATXN2 gene product is an ATNX2 mRNA or ATNX2 pre-mRNA comprising a sequence derived from an ATXN2 gene sequence (e.g., the ATXN2 gene sequence in NCBI Reference Sequence NG_011572.3 (SEQ ID NO: 149355), the ATXN2 gene sequence in NCBI Reference Sequence NC_000012.12 (Reference GRCh38.p13 Primary Assembly), or the ATXN2 coding sequence), or a portion thereof. In some embodiments of the present disclosure, the ATXN2 gene product comprises a sequence sharing at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, 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 100% identity with an ATXN2 mRNA or ATXN2 pre-mRNA sequence (e.g., the ATXN2 gene sequence in NCBI Reference Sequence NG_011572.3 (SEQ ID NO: 149355), the ATXN2 gene sequence in NCBI Reference Sequence NC_000012.12 (Reference GRCh38.p13 Primary Assembly), or the ATXN2 coding sequence) or a portion thereof.

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[0453] In various embodiments, the ATXN2 gene product is an ATXN2 mRNA isoform comprising the NCBI reference sequence NM_001310121.1 (SEQ ID NO: 149357). In various embodiments, the ATXN2 gene product comprises a sequence that shares at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, 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 100% identity with the ATXN2 mRNA isoform comprising the NCBI reference sequence NM_001310121.1 (SEQ ID NO: 149357).

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[0456] In various embodiments, the ATXN2 gene product is an ATXN2 mRNA isoform comprising the NCBI reference sequence NM_001310123.1 (SEQ ID NO: 149358). In various embodiments, the ATXN2 gene product comprises a sequence that shares at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, 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 100% identity with the ATXN2 mRNA isoform comprising the NCBI reference sequence NM_001310123.1 (SEQ ID NO: 149358).

[0457] In various embodiments, the ATXN2 gene product is an ATXN2 mRNA isoform comprising the NCBI reference sequence NM_001372574.1 (SEQ ID NO: 149359). In various embodiments, the ATXN2 gene product comprises a sequence that shares at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, 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 100% identity with the ATXN2 mRNA isoform comprising the NCBI reference sequence NM_001372574.1 (SEQ ID NO: 149359).

[0458] In various embodiments, the ATXN2 gene product is an ATXN2 mRNA isoform comprising the NCBI reference sequence NM_002973.4 (SEQ ID NO: 149360). In various embodiments, the ATXN2 gene product comprises a sequence that shares at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, 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 100% identity with the ATXN2 mRNA isoform comprising the NCBI reference sequence NM_002973.4 (SEQ ID NO: 149360).

[0459] In various embodiments, the ATXN2 gene product is an ATXN2 mRNA transcript variant comprising the NCBI reference sequence NR_132311.2 (SEQ ID NO: 149361). In various embodiments, the ATXN2 gene product comprises a sequence that shares at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, 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 100% identity with the ATXN2 mRNA transcript variant comprising the NCBI reference sequence NR_132311.2 (SEQ ID NO: 149361).

[0460] SOD1 In some embodiments of the present disclosure, the SOD1 gene product is SOD1 mRNA or SOD1 pre-mRNA comprising a sequence derived from the SOD1 gene sequence (e.g., the SOD1 gene sequence of NCBI Reference Sequence NG_008689.1 (SEQ ID NO: 167802), the SOD1 gene sequence of NCBI Reference Sequence NC_000021.9 (Reference GRCh38.p13 Primary Assembly) (SEQ ID NO: 167803), or the SOD1 coding sequence), or a portion thereof. In some embodiments of the present disclosure, the SOD1 gene product comprises a sequence sharing at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, 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 100% identity with an SOD1 mRNA or SOD1 pre-mRNA sequence (e.g., the SOD1 gene sequence in NCBI Reference Sequence NG_008689.1 (SEQ ID NO: 167802), the SOD1 gene sequence in NCBI Reference Sequence NC_000021.9 (Reference GRCh38.p13 Primary Assembly) (SEQ ID NO: 167803), or an SOD1 coding sequence) or a portion thereof.

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[0466] In various embodiments, the SOD1 gene product is an SOD1 mRNA sequence comprising the NCBI reference sequence NM_000454.5 (SEQ ID NO: 167804). In various embodiments, the SOD1 gene product comprises a sequence sharing at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, 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 100% identity with the SOD1 mRNA sequence comprising the NCBI reference sequence NM_000454.5 (SEQ ID NO: 167804).

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[0468] MAPT In some embodiments of the present disclosure, the MAPT gene product is a MAPT mRNA or MAPT pre-mRNA that comprises a sequence derived from a MAPT gene sequence (e.g., the MAPT gene sequence of NCBI Reference Sequence NG_007398.2 (SEQ ID NO: 301567), the MAPT gene sequence of NCBI Reference Sequence NC_000017.11 (Reference GRCh38.p13 Primary Assembly) (SEQ ID NO: 301568), or a MAPT coding sequence), or a portion thereof. In some embodiments of the present disclosure, the MAPT gene product comprises a sequence sharing at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, 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 100% identity with a MAPT mRNA or MAPT pre-mRNA sequence (e.g., the MAPT gene sequence in NCBI Reference Sequence NG_007398.2 (SEQ ID NO: 301567), the MAPT gene sequence in NCBI Reference Sequence NC_000017.11 (Reference GRCh38.p13 Primary Assembly) (SEQ ID NO: 301568), or a MAPT coding sequence) or a portion thereof.

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[0507] In various embodiments, the MAPT gene product is a MAPT mRNA sequence comprising the NCBI reference sequence NM_001123066.4 (SEQ ID NO: 301569). In various embodiments, the MAPT gene product comprises a sequence sharing at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, 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 100% identity with the MAPT mRNA sequence comprising the NCBI reference sequence NM_001123066.4 (SEQ ID NO: 301569).

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[0510] In various embodiments, the MAPT gene product is a MAPT mRNA sequence comprising the NCBI reference sequence NM_001123067.4 (SEQ ID NO: 301570). In various embodiments, the MAPT gene product comprises a sequence sharing at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, 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 100% identity with the MAPT mRNA sequence comprising the NCBI reference sequence NM_001123067.4 (SEQ ID NO: 301570).

[0511] In various embodiments, the MAPT gene product is a MAPT mRNA sequence comprising the NCBI reference sequence NM_001203251.2 (SEQ ID NO: 301571). In various embodiments, the MAPT gene product comprises a sequence sharing at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, 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 100% identity with the MAPT mRNA sequence comprising the NCBI reference sequence NM_001203251.2 (SEQ ID NO: 301571).

[0512] In various embodiments, the MAPT gene product is a MAPT mRNA sequence comprising the NCBI reference sequence NM_001203252.2 (SEQ ID NO: 301572). In various embodiments, the MAPT gene product comprises a sequence that shares at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, 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 100% identity with the MAPT mRNA sequence comprising the NCBI reference sequence NM_001203252.2 (SEQ ID NO: 301572).

[0513] In various embodiments, the MAPT gene product is a MAPT mRNA sequence comprising the NCBI reference sequence NM_001377265.1 (SEQ ID NO: 301573). In various embodiments, the MAPT gene product comprises a sequence that shares at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, 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 100% identity with the MAPT mRNA sequence comprising the NCBI reference sequence NM_001377265.1 (SEQ ID NO: 301573).

[0514] In various embodiments, the MAPT gene product is a MAPT mRNA sequence comprising the NCBI reference sequence NM_001377266.1 (SEQ ID NO: 301574). In various embodiments, the MAPT gene product comprises a sequence that shares at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, 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 100% identity with the MAPT mRNA sequence comprising the NCBI reference sequence NM_001377266.1 (SEQ ID NO: 301574).

[0515] In various embodiments, the MAPT gene product is a MAPT mRNA sequence comprising the NCBI reference sequence NM_001377267.1 (SEQ ID NO: 301575). In various embodiments, the MAPT gene product comprises a sequence that shares at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, 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 100% identity with the MAPT mRNA sequence comprising the NCBI reference sequence NM_001377267.1 (SEQ ID NO: 301575).

[0516] In various embodiments, the MAPT gene product is a MAPT mRNA sequence comprising the NCBI reference sequence NM_001377268.1 (SEQ ID NO: 301576). In various embodiments, the MAPT gene product comprises a sequence that shares at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, 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 100% identity with the MAPT mRNA sequence comprising the NCBI reference sequence NM_001377268.1 (SEQ ID NO: 301576).

[0517] In various embodiments, the MAPT gene product is a MAPT mRNA sequence comprising the NCBI reference sequence NM_005910.6 (SEQ ID NO: 301577). In various embodiments, the MAPT gene product comprises a sequence sharing at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, 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 100% identity with the MAPT mRNA sequence comprising the NCBI reference sequence NM_005910.6 (SEQ ID NO: 301577).

[0518] In various embodiments, the MAPT gene product is a MAPT mRNA sequence comprising the NCBI reference sequence NM_016834.5 (SEQ ID NO: 301578). In various embodiments, the MAPT gene product comprises a sequence that shares at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, 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 100% identity with the MAPT mRNA sequence comprising the NCBI reference sequence NM_016834.5 (SEQ ID NO: 301578).

[0519] In various embodiments, the MAPT gene product is a MAPT mRNA sequence comprising the NCBI reference sequence NM_016835.5 (SEQ ID NO: 301579). In various embodiments, the MAPT gene product comprises a sequence that shares at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, 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 100% identity with the MAPT mRNA sequence comprising the NCBI reference sequence NM_016835.5 (SEQ ID NO: 301579).

[0520] In various embodiments, the MAPT gene product is a MAPT mRNA sequence comprising the NCBI reference sequence NM_016841.5 (SEQ ID NO: 301580). In various embodiments, the MAPT gene product comprises a sequence sharing at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, 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 100% identity with the MAPT mRNA sequence comprising the NCBI reference sequence NM_016841.5 (SEQ ID NO: 301580).

[0521] In various embodiments, the MAPT gene product is a MAPT RNA sequence comprising the NCBI reference sequence NR_165166.1 (SEQ ID NO: 301581). In various embodiments, the MAPT gene product comprises a sequence sharing at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, 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 100% identity with the MAPT RNA sequence comprising the NCBI reference sequence NR_165166.1 (SEQ ID NO: 301581).

[0522] In various embodiments, the MAPT gene product is a MAPT mRNA transcript variant comprising the NCBI reference sequence XM_005257370.4 (SEQ ID NO: 301582). In various embodiments, the MAPT gene product comprises a sequence that shares at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, 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 100% identity with the MAPT mRNA transcript variant comprising the NCBI reference sequence XM_005257370.4 (SEQ ID NO: 301582).

[0523] In various embodiments, the MAPT gene product is a MAPT mRNA transcript variant comprising the NCBI reference sequence XM_005257366.3 (SEQ ID NO: 301583). In various embodiments, the MAPT gene product comprises a sequence that shares at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, 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 100% identity with the MAPT mRNA transcript variant comprising the NCBI reference sequence XM_005257366.3 (SEQ ID NO: 301583).

[0524] In various embodiments, the MAPT gene product is a MAPT mRNA transcript variant comprising the NCBI reference sequence XM_005257362.4 (SEQ ID NO: 301584). In various embodiments, the MAPT gene product comprises a sequence that shares at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, 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 100% identity with the MAPT mRNA transcript variant comprising the NCBI reference sequence XM_005257362.4 (SEQ ID NO: 301584).

[0525] In various embodiments, the MAPT gene product is a MAPT mRNA transcript variant comprising the NCBI reference sequence XM_005257365.4 (SEQ ID NO: 301585). In various embodiments, the MAPT gene product comprises a sequence that shares at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, 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 100% identity with the MAPT mRNA transcript variant comprising the NCBI reference sequence XM_005257365.4 (SEQ ID NO: 301585).

[0526] In various embodiments, the MAPT gene product is a MAPT mRNA transcript variant comprising the NCBI reference sequence XM_005257367.4 (SEQ ID NO: 301586). In various embodiments, the MAPT gene product comprises a sequence that shares at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, 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 100% identity with the MAPT mRNA transcript variant comprising the NCBI reference sequence XM_005257367.4 (SEQ ID NO: 301586).

[0527] In various embodiments, the MAPT gene product is a MAPT mRNA transcript variant comprising the NCBI reference sequence XM_005257368.4 (SEQ ID NO: 301587). In various embodiments, the MAPT gene product comprises a sequence that shares at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, 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 100% identity with the MAPT mRNA transcript variant comprising the NCBI reference sequence XM_005257368.4 (SEQ ID NO: 301587).

[0528] In various embodiments, the MAPT gene product is a MAPT mRNA transcript variant comprising the NCBI reference sequence XM_005257369.4 (SEQ ID NO: 301588). In various embodiments, the MAPT gene product comprises a sequence that shares at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, 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 100% identity with a MAPT mRNA transcript variant comprising the NCBI reference sequence XM_005257369.4 (SEQ ID NO: 301588).

[0529] In various embodiments, the MAPT gene product is a MAPT mRNA transcript variant comprising the NCBI reference sequence XM_005257371.4 (SEQ ID NO: 301589). In various embodiments, the MAPT gene product comprises a sequence that shares at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, 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 100% identity with a MAPT mRNA transcript variant comprising the NCBI reference sequence XM_005257371.4 (SEQ ID NO: 301589).

[0530] AONs targeting gene products PPM1A AON In various embodiments, a PPM1A AON disclosed herein, e.g., a PPM1A AON comprising the sequence of any one of SEQ ID NOs: 1-954, or the sequence of any one of SEQ ID NOs: 1-954 in which one or more nucleosides have been replaced with one or more spacers, e.g., SEQ ID NOs: 301692-301742 or any one of the sequences in Tables 4A-4C, targets a specific portion of a PPM1A gene product, e.g., a PPM1A mRNA or pre-mRNA transcript (e.g., any one of SEQ ID NOs: 1909-1913). In some embodiments, a PPM1A AON can be an oligonucleotide 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 100% complementary to a portion of a PPM1A gene product or a PPM1A gene sequence (e.g., any one of SEQ ID NOs: 1909-1913). In certain embodiments, a PPM1A AON can be an oligonucleotide sequence that is at least 85% complementary to a portion of a PPM1A gene product or a PPM1A gene sequence (e.g., any one of SEQ ID NOs: 1909-1913). In certain embodiments, a PPM1A AON can be an oligonucleotide sequence that is at least 90% complementary to a portion of a PPM1A gene product or a PPM1A gene sequence (e.g., any one of SEQ ID NOs: 1909-1913). In certain embodiments, a PPM1A AON can be an oligonucleotide sequence that is at least 95% complementary to a portion of a PPM1A gene product or a PPM1A gene sequence (e.g., any one of SEQ ID NOs: 1909-1913). In certain embodiments, a PPM1A AON can be an oligonucleotide sequence that is at least 98% complementary to a portion of a PPM1A gene product or a PPM1A gene sequence (e.g., any one of SEQ ID NOs: 1909-1913).In certain embodiments, a PPM1A AON can be an oligonucleotide sequence that is at least 99% complementary to a portion of a PPM1A gene product or a PPM1A gene sequence (e.g., any one of SEQ ID NOs: 1909-1913). In certain embodiments, a PPM1A AON can be an oligonucleotide sequence that is 100% complementary to a portion of a PPM1A gene product or a PPM1A gene sequence (e.g., any one of SEQ ID NOs: 1909-1913).

[0531] In some embodiments described herein, the PPM1A AON targets a specific portion of a PPM1A gene product, e.g., a PPM1A mRNA transcript or a PPM1A pre-mRNA transcript. For example, as described herein, the PPM1A 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 portion of a PPM1A gene product, e.g., a PPM1A mRNA transcript or a PPM1A pre-mRNA transcript (e.g., any one of SEQ ID NOs: 1909-1913).

[0532] In some embodiments, a PPM1A 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 nucleotide sequence in an untranslated region (UTR) of a PPM1A mRNA sequence, e.g., the 5' UTR or 3' UTR of a PPM1A mRNA sequence. In some embodiments, a PPM1A AON comprises a sequence that is 100% complementary to a nucleotide sequence in an untranslated region (UTR) of a PPM1A mRNA sequence, e.g., the 5' UTR or 3' UTR of a PPM1A mRNA sequence.

[0533] ATXN2 AON In various embodiments, the ATXN2 AON disclosed herein, e.g., an ATXN2 AON comprising any one of SEQ ID NOs: 1914-149354, or any one of SEQ ID NOs: 1914-149354 in which one or more nucleosides have been replaced with one or more spacers, e.g., any one of SEQ ID NOs: 301692-301742 or any one of the sequences in Tables 4A-4C, targets a specific portion of an ATXN2 gene product, e.g., an ATXN2 mRNA transcript (e.g., any one of SEQ ID NOs: 149355-149361). In some embodiments, the ATXN2 AON can be an oligonucleotide 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 100% complementary to a portion of the ATXN2 gene product or to an ATXN2 gene sequence (e.g., any one of SEQ ID NOs: 149355-149361). In certain embodiments, the ATXN2 AON can be an oligonucleotide sequence that is at least 85% complementary to a portion of the ATXN2 gene product or to an ATXN2 gene sequence (e.g., any one of SEQ ID NOs: 149355-149361). In certain embodiments, the ATXN2 AON can be an oligonucleotide sequence that is at least 90% complementary to a portion of the ATXN2 gene product or to the ATXN2 gene sequence (e.g., any one of SEQ ID NOs: 149355-149361). In certain embodiments, the ATXN2 AON can be an oligonucleotide sequence that is at least 95% complementary to a portion of the ATXN2 gene product or to the ATXN2 gene sequence (e.g., any one of SEQ ID NOs: 149355-149361). In certain embodiments, the ATXN2 AON can be an oligonucleotide sequence that is at least 98% complementary to a portion of the ATXN2 gene product or to the ATXN2 gene sequence (e.g., any one of SEQ ID NOs: 149355-149361).In certain embodiments, the ATXN2 AON can be an oligonucleotide sequence that is at least 99% complementary to a portion of the ATXN2 gene product or to an ATXN2 gene sequence (e.g., any one of SEQ ID NOs: 149355-149361). In certain embodiments, the ATXN2 AON can be an oligonucleotide sequence that is 100% complementary to a portion of the ATXN2 gene product or to an ATXN2 gene sequence (e.g., any one of SEQ ID NOs: 149355-149361).

[0534] In some embodiments described herein, the ATXN2 AON targets a specific portion of an ATXN2 gene product, such as an ATXN2 mRNA transcript or an ATXN2 pre-mRNA transcript. For example, as described herein, the ATXN2 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 portion of an ATXN2 gene product, such as an ATXN2 mRNA transcript or an ATXN2 pre-mRNA transcript (e.g., any one of SEQ ID NOs: 149355-149361).

[0535] SOD1 AON In various embodiments, an SOD1 AON disclosed herein, e.g., an SOD1 AON having the sequence of any one of SEQ ID NOs: 149362-158581, or an SOD1 AON having one or more spacers (e.g., one or more nucleosides are replaced with one or more spacers), e.g., an SOD1 AON having the sequence of any one of SEQ ID NOs: 149362-158581 replaced with the sequence of any one of SEQ ID NOs: 301610-301741), targets a specific portion of an SOD1 gene product, e.g., an SOD1 mRNA transcript (e.g., any one of SEQ ID NOs: 167802-167804). In some embodiments, the SOD1 AON can be an oligonucleotide 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 100% complementary to a portion of the SOD1 gene product or to an SOD1 gene sequence (e.g., any one of SEQ ID NOs: 167802-167804). In certain embodiments, the SOD1 AON can be an oligonucleotide sequence that is at least 85% complementary to a portion of the SOD1 gene product or to an SOD1 gene sequence (e.g., any one of SEQ ID NOs: 167802-167804). In certain embodiments, the SOD1 AON can be an oligonucleotide sequence that is at least 90% complementary to a portion of the SOD1 gene product or to the SOD1 gene sequence (e.g., any one of SEQ ID NOs: 167802-167804). In certain embodiments, the SOD1 AON can be an oligonucleotide sequence that is at least 95% complementary to a portion of the SOD1 gene product or to the SOD1 gene sequence (e.g., any one of SEQ ID NOs: 167802-167804). In certain embodiments, the SOD1 AON can be an oligonucleotide sequence that is at least 98% complementary to a portion of the SOD1 gene product or to the SOD1 gene sequence (e.g., any one of SEQ ID NOs: 167802-167804).In certain embodiments, the SOD1 AON can be an oligonucleotide sequence that is at least 99% complementary to a portion of the SOD1 gene product or to the SOD1 gene sequence (e.g., any one of SEQ ID NOs: 167802-167804). In certain embodiments, the SOD1 AON can be an oligonucleotide sequence that is 100% complementary to a portion of the SOD1 gene product or to the SOD1 gene sequence (e.g., any one of SEQ ID NOs: 167802-167804).

[0536] In some embodiments described herein, the SOD1 AON targets a specific portion of an SOD1 gene product, such as an SOD1 mRNA transcript or an SOD1 pre-mRNA transcript. For example, as described herein, the SOD1 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 portion of an SOD1 gene product, such as an SOD1 mRNA transcript or an SOD1 pre-mRNA transcript (e.g., any one of SEQ ID NOs: 167802-167804).

[0537] MAPT AON In various embodiments, a MAPT AON disclosed herein, e.g., a MAPT AON having the sequence of any one of SEQ ID NOs: 167805-301566, or a MAPT AON having one or more spacers (e.g., one or more nucleosides are replaced with one or more spacers), e.g., a MAPT AON having the sequence of any one of SEQ ID NOs: 301692-301742 or any one of the sequences in Tables 4A-4C), targets a specific portion of a MAPT gene product, e.g., a MAPT mRNA or pre-mRNA transcript (e.g., any one of SEQ ID NOs: 301567-301589). In some embodiments, a MAPT AON can be an oligonucleotide 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 100% complementary to a portion of a MAPT gene product or a MAPT gene sequence (e.g., any one of SEQ ID NOs: 301567-301589). In certain embodiments, a MAPT AON can be an oligonucleotide sequence that is at least 85% complementary to a portion of a MAPT gene product or a MAPT gene sequence (e.g., any one of SEQ ID NOs: 301567-301589). In certain embodiments, a MAPT AON can be an oligonucleotide sequence that is at least 90% complementary to a portion of a MAPT gene product or a MAPT gene sequence (e.g., any one of SEQ ID NOs: 301567-301589). In certain embodiments, the MAPT AON can be an oligonucleotide sequence that is at least 95% complementary to a portion of the MAPT gene product or to a MAPT gene sequence (eg, any one of SEQ ID NOs: 301567-301589).In certain embodiments, a MAPT AON can be an oligonucleotide sequence that is at least 98% complementary to a portion of the MAPT gene product or to a MAPT gene sequence (e.g., any one of SEQ ID NOs: 301567-301589). In certain embodiments, a MAPT AON can be an oligonucleotide sequence that is at least 99% complementary to a portion of the MAPT gene product or to a MAPT gene sequence (e.g., any one of SEQ ID NOs: 301567-301589). In certain embodiments, a MAPT AON can be an oligonucleotide sequence that is 100% complementary to a portion of the MAPT gene product or to a MAPT gene sequence (e.g., any one of SEQ ID NOs: 301567-301589).

[0538] In some embodiments described herein, the MAPT AON targets a specific portion of a MAPT gene product, such as a MAPT mRNA transcript or a MAPT pre-mRNA transcript. For example, as described herein, the MAPT 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 portion of a MAPT gene product, such as a MAPT mRNA transcript or a MAPT pre-mRNA transcript (e.g., any one of SEQ ID NOs: 301567-301589).

[0539] Nuclease-mediated inhibition In one aspect, the present disclosure provides a nuclease that reduces expression of a target gene product (e.g., a gene product of any one of PPM1A, ATXN2, SOD1, or MAPT). In some embodiments, the nuclease can be an RNAse (e.g., RNAse H), a zinc finger nuclease (ZFN), a meganuclease, a transcription activator-like effector nuclease (TALEN), or a clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein.

[0540] In certain embodiments, target gene products are inhibited, eliminated, or reduced in amount using zinc finger nucleases (ZFNs). Synthetic ZFNs are composed of, for example, a zinc finger binding domain fused with a FokI DNA cleavage domain. ZFNs can be designed / engineered for editing cellular genomes, including but not limited to, knocking out or knocking in gene expression in a wide range of organisms. Meganucleases, TALENs, or CRISPR-associated proteins can be used for genome manipulation in cells of patients suffering from or at risk of neurological diseases, including neurons, e.g., motor neurons, and other cells of the nervous system. The described reagents can be used to target promoters, protein-coding regions (exons), introns, 5' and 3' UTRs, and others.

[0541] CRISPR genome editing typically involves two distinct components: (1) guide RNA, and (2) endonuclease, specifically CRISPR-associated (Cas) nuclease (e.g., Cas9). The guide RNA combines endogenous bacterial crRNA and tracrRNA into a single chimeric guide RNA (gRNA) transcript. Without being bound by theory, it is believed that when gRNA and Cas are expressed in a cell, the genomic target sequence can be modified and permanently destroyed.

[0542] The gRNA / Cas complex can be recruited to a target sequence, e.g., a target gene (e.g., any one of PPM1A, ATXN2, SOD1, or MAPT), through base pairing between the gRNA sequence and the complement of the target DNA sequence in the target gene. A suitable genomic target sequence contains a protospacer adjacent motif (PAM) sequence immediately following the target sequence. Binding of the gRNA / Cas complex localizes Cas to the target sequence, allowing wild-type Cas to cleave both strands of DNA, resulting in a double-strand break. Double-strand breaks are repaired through one of two general repair pathways: (1) the nonhomologous end-joining DNA repair pathway or (2) the homology-directed repair pathway. The nonhomologous repair pathway results in insertions / deletions at the double-strand break, which can cause frameshifts and / or premature stop codons, effectively disrupting the open reading frame of the target gene. The homology-directed repair pathway requires the presence of a repair template to repair the double-strand break.

[0543] In certain embodiments, target gene expression is reduced using CRISPR genome editing. In some embodiments, a gRNA pair is used to target a target gene, thereby reducing and / or eliminating the expression of the target gene. In certain embodiments, one gRNA pair is used to reduce the expression of the target gene. In certain other embodiments, multiple gRNA pairs are used to reduce the expression of the target gene. The gRNA pair can be designed based on the gene sequence using known techniques. In certain embodiments, the gRNA sequence can include modifications in the terminal three nucleotides at both the 5' and 3' ends of the gRNA, such as 2'O-methyl analogs and 3' phosphorothioate internucleotide linkages.

[0544] Neurological disorders The methods described herein are directed to treating conditions including 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 or neuropathies, such as, but not limited to, chemotherapy-induced neuropathy, spinocerebellar ataxia (SCA), SCA type 2, spinal muscular atrophy (SMA), parkinsonism, Niemann-Pick disease type C (NPC), Charcot-Marie-Tooth disease (CMT), mucopolysaccharidosis type II (MPSIIA), mucolipidosis IV, GM1 gangliosidosis, sporadic inclusion body myositis (sIBM), Henoch-Schönlein purpura (HSP), Gaucher disease, as well as facial-onset sensorimotor neuropathy, Guam Parkinsonism-Dementia Complex, multisystem proteinopathy, Perry disease, and autism.

[0545] Motor neuron diseases are a group of disorders characterized by the loss of motor neuron function, which coordinates voluntary muscle movements with the brain. Motor neuron diseases can affect upper and / or lower motor neurons and can be sporadic or familial in origin. Examples of motor neuron diseases include amyotrophic lateral sclerosis (ALS or Lou Gehrig's disease), progressive bulbar palsy, pseudobulbar palsy, progressive muscular atrophy, primary lateral sclerosis, spinal muscular atrophy, post-polio syndrome, and ALS with frontotemporal dementia.

[0546] Symptoms of motor neuron disease include muscle breakdown or weakness, muscle pain, spasms, slurred speech, difficulty swallowing, loss of muscle control, joint pain, stiffness of the limbs, difficulty breathing, salivation, and complete loss of muscle control, including for basic functions such as breathing, swallowing, feeding, speaking, and limb movement. These symptoms may also be accompanied by depression, memory loss, difficulty planning, language deficits, altered behavior, and difficulty assessing spatial relationships, and / or personality changes.

[0547] Motor neuron disease can be evaluated and diagnosed by a skilled clinician, such as a neurologist, using various tools and tests.For example, the presence of motor neuron disease or the risk of developing it can be evaluated or diagnosed using blood and urine tests (such as tests that assay for the presence of creatine kinase), magnetic resonance imaging (MRI), electromyography (EMG), nerve conduction studies (NCS), spinal tap, lumbar puncture, and / or muscle biopsy.Motor neuron disease can be diagnosed using physical and / or neurological examinations to evaluate motor and sensory abilities, nerve function, hearing and speech, vision, coordination and balance, mental status, and mood or behavior changes.

[0548] A patient suffering from ALS, FTD, ALS with FTD, or another neurological or motor neuron disease may be a patient who has been diagnosed with the disease or who exhibits symptoms of the disease.A patient suffering from ALS, FTD, ALS with FTD, or another neurological or motor neuron disease may be a patient who has previously suffered from the disease and / or disease symptoms, who has recovered from the disease and / or disease symptoms or experienced a complete or partial improvement thereof, and then experiences a complete or partial recurrence of the disease or disease symptoms.A patient suffering from ALS, FTD, ALS with FTD, or another neurological or motor neuron disease or condition may be a patient who has a gene mutation associated with the symptoms of the disease or condition.For example, patients with ALS have been reported to have deficiencies in SOD1, C9orf72, ataxin 2 (ATXN2), charged multivesicular body protein 2B (CHMP2B), dynactin 1 (DCTN1), human epidermal growth factor receptor 4 (ERBB4), FIG4 phosphoinositide 5-phosphatase (FIG4), NIMA-related kinase 1 (NEK1), heterogeneous nuclear ribonucleoprotein HA1 (HNRNPA1), neurofilament heavy chain (NEFH), peripherin (PRPH), TAR DNA-binding protein 43 (TDP43 or TARDP), and fused in sarcoma. Sarcoma) (FUS), ubiquilin-2 (UBQLN2), kinesin family member 5A (KIF5A), valosin-containing protein (VCP), arsine (ALS2), senataxin (SETX), sigma non-opioid intracellular receptor 1 (SIGMAR1), survival of motor neuron 1 (SMN1), spastic paraplegia 11 (SPG11), autosomal recessive, transient receptor potential cation channel subfamily M member 7 (TRPM7), vesicle-associated membrane protein-associated protein B / C (VAPB), angiogenin (ANG), profile The patient may have a genetic mutation in any of: Phyllin-1 (PFN1), Matrin-3 (MATR3), Coiled-coil-helix-coiled-coil-helix domain containing 10 (CHCHD10), Tubulin, alpha 4A (TUBA4A), TBK1, C21orf2, Sequestosome-1 (SQSTM1, also known as ubiquitin-binding protein p62), and / or Optineurin (OPTN), particularly where the mutation is associated with ALS or a high risk of developing ALS.

[0549] Patients at risk for ALS, FTD, ALS with FTD, or another neurological or motor neuron disease can include patients with a family history of the disease or a genetic predisposition to the disease (e.g., patients with a gene mutation associated with an increased risk of the disease, for example), or patients exposed to environmental factors that increase the risk of the disease. For example, a patient may be at risk for ALS if they have a mutation in any of SOD1, C9orf72, ATXN2, CHMP2B, DCTN1, ERBB4, FIG4, HNRNPA1, NEFH, PRPH, NEK1, TDP43, FUS, UBQLN2, KIF5A, VCP, ALS2, SETX, SIGMAR1, SMN1, SPG11, TRPM7, VAPB, ANG, PFN1, MATR3, CHCHD10, TUBA4A, TBK1, SQSTM1, C21orf2, and / or OPTN, particularly where the mutation is associated with ALS or an elevated risk of developing ALS. At-risk patients may also include patients diagnosed with ALS, FTD, ALS with FTD, or a disease or condition that has high comorbidity with another neurological or motor neuron disease (e.g., patients suffering from dementia, which is significantly associated with a family history of ALS, FTD, and a higher likelihood of developing bulbar ALS (see Trojsi, F., et al. (2017) "Comorbidity of dementia with amyotrophic lateral sclerosis (ALS): insights from a large multicenter Italian cohort" J Neurol 264: 2224-31)).

[0550] Amyotrophic lateral sclerosis ALS is a progressive motor neuron disease that disrupts signals to all voluntary muscles. ALS results in atrophy of both upper and lower motor neurons. Symptoms of ALS include weakness and wasting of bulbar muscles, generalized and bilateral loss of muscle strength, spasticity, muscle spasms, muscle cramps, fasciculations, slurred speech, and difficulty breathing or loss of ability to breathe. Some individuals with ALS also suffer from cognitive decline. At the molecular level, ALS is characterized by protein and RNA aggregates in the cytoplasm of motor neurons, including aggregates of the RNA-binding protein TDP43.

[0551] ALS is most common in men over 40 years of age, but can also occur in women and children. The risk of ALS is also higher in individuals who smoke, are exposed to chemicals such as lead, or have served in the military. Most cases of ALS are sporadic, but only about 10% of cases are familial. Causes of ALS include sporadic or inherited genetic mutations, high levels of glutamate, and protein mishandling. Genetic mutations associated with ALS include mutations in the genes SOD1, C9orf72, TARDP, FUS, ANG, ATXN2, CHCHD10, CHMP2B, DCTN1, ERBB4, FIG4, HNRPA1, MATR3, NEFH, OPTN, PFN1, PRPH, SETX, SIGMAR1, SMN1, SPG11, SQSTM1, TBK1, TRPM7, TUBA4A, UBQLN2, VAPB, and VCP.

[0552] frontotemporal dementia Frontotemporal dementia (FTD) is a form of dementia affecting the frontal and temporal lobes of the brain. It has an average age of onset of 40 years, earlier than Alzheimer's disease. Symptoms of FTD include extreme changes in behavior and personality, speech and language problems, and motor symptoms such as tremor, stiffness, muscle spasms, weakness, and difficulty swallowing. Subtypes of FTD include behavioral variant frontotemporal dementia (bvFTD), characterized by personality and behavioral changes, and primary progressive aphasia (PPA), which affects language skills, speech, writing, and comprehension. FTD is associated with tau protein accumulation (Pick bodies) and altered TDP43 function. Approximately 30% of FTD cases are familial, with no known risk factors other than a family history of the disease. Genetic mutations associated with FTD include mutations in the genes C9orf72, progranulin (GRN), microtubule-associated protein tau (MAPT), UBQLN2, VPC, CHMP2B, TARDP, FUS, ITM2B, CHCHD10, SQSTM1, PSEN1, PSEN2, CTSF, CYP27A1, TBK1, and TBP.

[0553] Amyotrophic lateral sclerosis with frontotemporal dementia (ALS with FTD) is a clinical syndrome in which FTD and ALS occur in the same individual. Interestingly, mutations in C9orf72 are the most common cause of familial forms of ALS and FTD. In addition, mutations in TBK1, VCP, SQSTMI, UBQLN2, and CHMP2B have also been associated with ALS with FTD. Symptoms of ALS with FTD include personality changes, muscle weakness, muscle atrophy, fasciculations, spasticity, dysphagia, and degeneration of the spinal cord, motor neurons, and frontal and temporal lobes of the brain. At the molecular level, ALS with FTD is characterized by the accumulation of TDP-43 and / or FUS proteins. TBK1 mutations are associated with ALS, FTD, and ALS with FTD.

[0554] Functions of TBK1 and RIPK1 In one aspect, the method described herein includes exposing a cell to a PPM1A AON to alter the activity, function, or other characteristics of a gene or gene product, such as mRNA or protein. For example, the method described herein includes methods for increasing, decreasing, or inhibiting the activity, function, or other characteristics of a gene or gene product. For example, a method for increasing the phosphorylation of a residue of TANK-binding kinase 1 (serine / threonine protein kinase TBK1, also known as "TBK1") is described herein. For example, a method for increasing the phosphorylation of TBK1 serine residue 172 (ser172) in a cell is described herein, the method comprising exposing the cell to a PPM1A AON. In some embodiments, TBK1 ser172 phosphorylation is increased in cells of a patient suffering from ALS, FTD, or ALS with FTD. In some embodiments, a method for increasing TBK1 ser172 phosphorylation comprises exposing a cell to a PPM1A antisense oligonucleotide of any one of SEQ ID NOs: 1-954, or any one of SEQ ID NOs: 1-954 in which one or more nucleosides have been replaced with one or more spacers, e.g., SEQ ID NOs: 301692-301742, or any one of the sequences in Tables 4A-4C.

[0555] Also described herein are methods for increasing TBK1 function in cells, comprising exposing the cells to a PPM1A AON. For example, described herein are methods for increasing TBK1 function in cells, comprising exposing the cells to a PPM1A AON. In some embodiments, TBK1 function is increased in cells of a patient suffering from ALS, FTD, or ALS with FTD. In some embodiments, the method for increasing TBK1 function comprises exposing the cells to a PPM1A antisense oligonucleotide of any one of SEQ ID NOs: 1-954, or any one of SEQ ID NOs: 1-954 in which one or more nucleosides are replaced with one or more spacers, e.g., SEQ ID NOs: 301692-301742, or any one of the sequences in Tables 4A-4C.

[0556] Tank-binding kinase 1 (TBK1) is an IKK family kinase that induces type 1 interferon activity and plays a key role in phosphorylating autophagy adaptors. Mutations in TBK1 are thought to impair autophagy, contributing to the accumulation of protein aggregates and ALS pathology. At least 92 mutations in TBK1 have been identified in patients with ALS, FTD, or ALS with FTD (see Oakes et al., (2017) "TBK1: a new player in ALS linking autophagy and neuroinflammation," Molecular Brain 10:5, pg. 1-10). Furthermore, mutations in TBK1, along with mutations in C9orf72, OPTN, SQSTM1 / p62, UBQLN2, and TDP43, account for approximately 15% of ALS and FTD patients. Furthermore, TBK1 haploinsufficiency associated with loss-of-function mutations has been identified as a major driver of familial ALS (see Freischmidt et al., (2015) "Haploinsufficiency of TBK1 causes familial ALS and fronto-temporal dementia" Nature Neuroscience, 18(5):631-6).

[0557] Autophagy is the process by which proteins and damaged organelles are degraded and recycled. Abnormal protein aggregates are a hallmark of ALS pathology, and mutations in several genes involved in regulating autophagy have been associated with ALS (e.g., SQSTM1, SOD1, OPTN, VCP, UBQLN2, and TBK1). Therefore, disruption of autophagy appears to contribute to ALS pathology.

[0558] Phosphorylation of residue Ser172 in TBK1 leads to a conformational change in TBK1, allowing substrate binding by the protein's kinase domain. TBK1 phosphorylates several autophagy adaptors, and several TBK1 mutations identified in ALS patients inhibit TBK1's ability to phosphorylate these adaptors. Other TBK1 mutations result in decreased mRNA and protein levels. In addition, individuals with mutations in TBK1 also exhibit TDP43-positive aggregates in various brain regions. Thus, TBK1 mutations may result in decreased autophagy and the accumulation of protein aggregates in motor neurons.

[0559] PPM1A is a member of the PP2C family of Ser / Thr protein phosphatases. PP2C family members are negative regulators of cellular stress response pathways and are involved in regulating the cell cycle and the NF-κB pathway. PPM1A also dephosphorylates and inactivates TBK1. Specifically, PPM1A dephosphorylates Ser172 of TBK1. Activated TBK1 can phosphorylate RIPK1 in such a way that RIPK1 is deactivated. Therefore, PPM1A activity indirectly activates RIPK1, and therefore, reducing PPM1A expression indirectly inactivates RIPK1.

[0560] The present disclosure relates to increasing TBK1 activity, e.g., increasing TBK1 activity in an individual or cell of an individual suffering from TBK1 haploinsufficiency, to treat a variety of neurological diseases, such as 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 (MSA), and the like. The present invention is based, in part, on the discovery that the compounds of the present invention can be used as a mechanism for treating neuropathies, including ataxia, amnesic mild cognitive impairment, corticobasal degeneration (CBD), and / or neuropathy, such as chemotherapy-induced neuropathy, spinocerebellar ataxia (SCA), SCA type 2, spinal muscular atrophy (SMA), parkinsonism, Niemann-Pick disease type C (NPC), Charcot-Marie-Tooth disease (CMT), mucopolysaccharidosis type II (MPSIIA), mucolipidosis IV, GM1 gangliosidosis, sporadic inclusion body myositis (sIBM), Henoch-Schönlein purpura (HSP), Gaucher disease, and facial-onset sensorimotor neuropathy, Guam Parkinsonism-Dementia Complex, multisystem proteinopathy, Perry disease, and autism.

[0561] The present disclosure is also based in part on the discovery that increasing TBK1 activity, for example, increasing residual TBK1 activity in an individual and / or cell of an individual suffering from TBK1 haploinsufficiency, can be achieved, for example, by increasing the amount of phosphorylated TBK1, for example, by increasing the amount of phosphorylated Ser172 TBK1, in an individual and / or cell of an individual suffering from TBK1 haploinsufficiency.The present disclosure is also based in part on the discovery that increasing TBK1 activity, for example, increasing residual TBK1 activity in an individual and / or cell of an individual suffering from TBK1 haploinsufficiency, can be achieved, for example, by increasing the ratio of phosphorylated TBK1 to total TBK1, for example, by increasing the ratio of phosphorylated Ser172 TBK1 to unphosphorylated Ser172 TBK1, in an individual and / or cell of an individual suffering from TBK1 haploinsufficiency.

[0562] The present disclosure is further based in part on the discovery that increasing TBK1 activity (e.g., increasing residual TBK1 activity in an individual and / or cells of an individual suffering from TBK1 haploinsufficiency), increasing the amount of phosphorylated TBK1 (e.g., increasing the amount of phosphorylated Ser172 TBK1, for example, in an individual and / or cells of an individual suffering from TBK1 haploinsufficiency), and / or increasing the ratio of phosphorylated to unphosphorylated TBK1 (e.g., increasing the ratio of phosphorylated to unphosphorylated Ser172 TBK1, for example, in an individual and / or cells of an individual suffering from TBK1 haploinsufficiency) can be achieved, for example, by inhibiting PPM1A activity and / or decreasing PPM1A protein levels in an individual and / or cells of an individual suffering from TBK1 haploinsufficiency. Without being bound by theory, inhibiting PPM1A activity and / or reducing PPM1A protein levels may be achieved by administering to the patient or to the patient's cells a PPM1A AON, such as a PPM1A AON described herein.In certain embodiments, the disclosure provides a method of inhibiting PPM1A activity and / or decreasing the amount of PPM1A protein, comprising administering to a patient or a patient's cells (e.g., a patient suffering from a neurological disease, or a patient suffering from a neurological disease, e.g., 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, Huntin...

Claims

1. A gapmer oligonucleotide or a compound comprising the gapmer oligonucleotide, wherein the gapmer oligonucleotide comprises a nucleoside substitution group containing a non-sugar substitute that cannot be linked to a nucleotide base.

2. The compound or gapmer oligonucleotide according to claim 1, wherein the gapmer oligonucleotide comprises a second nucleoside substitution group which includes a non-sugar substitute that is not adjacent to the nucleoside substitution group and cannot be linked to a nucleotide base.

3. The compound or gapmer oligonucleotide according to claim 1, wherein the gapmer oligonucleotide comprises a sequence that is 85-98% complementary to an equal-length portion of PPM1A mRNA or premRNA transcript, ATXN2 mRNA or premRNA transcript, SOD1 mRNA or premRNA transcript, or MAPT mRNA or premRNA transcript.

4. The compound or gapmer oligonucleotide according to claim 1, wherein the gapmer oligonucleotide comprises one equal-length portion of any one of sequence numbers 1909-1913, 149355-149361, 167802-167804, or 301567-301589, a sequence having 90% identity therewith, or a sequence that is 85-98% complementary to 15-50 consecutive nucleic acid base portions thereof.

5. The compound or gapmer oligonucleotide according to claim 1, wherein the gapmer oligonucleotide includes a sequence that shares at least 85% identity, at least 90% identity, at least 95% identity, or 100% identity with any one equal-length portion of any one of the sequences in Tables 5A to 5B, 6, 7, or 8.

6. The compound or gapmer oligonucleotide according to claim 1, wherein the gapmer oligonucleotide comprises a segment having at least 11, 10, 9, 8, 7, 6, 5, 4, 3, or 2 linked nucleosides, and the gapmer oligonucleotide comprises a sequence that shares at least 85% identity or at least 90% identity with any one equal-length portion of the sequences in Tables 5A to 5B, 6, 7, or 8.

7. The compound or gapmer oligonucleotide according to claim 1, wherein the gapmer oligonucleotide is an oligonucleotide unit having a length of 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, and optionally at least one (i.e., one or more) nucleoside linkage of the gapmer oligonucleotide is a non-natural linkage.

8. The compound or gapmer oligonucleotide according to claim 1, wherein the nucleoside substitution group is positioned between the 5th and 11th positions of the gapmer oligonucleotide, or optionally between the 7th and 11th positions.

9. The compound or gapmer oligonucleotide according to claim 2, wherein the second nucleoside substitution group is located between the 15th and 19th positions of the gapmer oligonucleotide, and optionally the nucleoside substitution group and the second nucleoside substitution group are separated in the gapmer oligonucleotide by at least five nucleic acid bases, at least six nucleic acid bases, or at least seven nucleic acid bases, and optionally the nucleoside substitution group is located between the 5th and 11th positions of the gapmer oligonucleotide, the second nucleoside substitution group is located between the 15th and 19th positions of the gapmer oligonucleotide, and optionally the nucleoside substitution group is located at the 8th position of the gapmer oligonucleotide, and the second nucleoside substitution group is located at the 16th position of the gapmer oligonucleotide.

10. The compound or gapmer oligonucleotide according to claim 1, wherein the nucleoside substitution group is a nucleoside substitution group containing a non-sugar substitute, and the non-sugar substitute does not contain a ketone, aldehyde, ketal, hemiketal, acetal, hemiacetal, aminal, or hemiaminal moiety and cannot form a covalent bond with a nucleotide base.

11. The nucleoside substitution group is represented by formula (X), 【Chemistry 1】 During the ceremony, Ring A is an optionally substituted 4- to 8-membered monocyclic cycloalkyl group or a 4- to 8-membered monocyclic heterocyclyl group, wherein the heterocyclyl group contains one or two heteroatoms selected from O, S, and N, provided that A cannot form a covalent bond with a nucleic acid base. 【Chemistry 2】 This represents a connection point between nucleoside links, Optionally, the nucleoside substitution group is represented by formula (Xa), 【Transformation 3】 In the formula, ring A is an optionally substituted 4- to 8-membered monocyclic cycloalkyl group selected from cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl, or a 4- to 8-membered monocyclic heterocyclil group selected from oxetanyl, tetrahydrofuranil, tetrahydropyranil, 1,4-dioxanil, pyrrolidinyl, piperidinyl, piperazinyl, morpholinil, and azepanil, and optionally ring A is tetrahydrofuranil, or optionally ring A is tetrahydropyranil. Optionally, the nucleoside substitution group is represented by formula I, 【Chemistry 4】 During the ceremony, X is selected from -CH2- and -O-, n is 0, 1, 2, or 3. Optionally, the nucleoside substitution group is represented by formula I', 【Transformation 5】 During the ceremony, X is selected from -CH2- and -O-, n is 0, 1, 2, or 3. Optionally, the nucleoside substitution group is represented by formula (Ia), 【Transformation 6】 During the ceremony, n is 0, 1, 2, or 3. Optionally, the nucleoside substitution group is represented by formula (Ia'), 【Transformation 7】 During the ceremony, n is 0, 1, 2, or 3. Optionally, the nucleoside substitution group is represented by formula II, 【Transformation 8】 During the ceremony, X is selected from -CH2- and -O-, Optionally, the nucleoside substitution group is represented by formula II', 【Chemistry 9】 During the ceremony, X is selected from -CH2- and -O-, Optionally, the nucleoside substitution group is represented by formula (IIIa), 【Chemistry 10】 Optionally, the nucleoside substitution group is represented by formula (Iia'), 【Chemistry 11】 Optionally, the nucleoside substitution group is represented by formula III, 【Chemistry 12】 During the ceremony, X is selected from -CH2- and -O-, Optionally, the nucleoside substitution group is represented by formula III', 【Chemistry 13】 During the ceremony, X is selected from -CH2- and -O-, Optionally, the nucleoside substitution group is represented by formula (IIIa), 【Chemistry 14】 Alternatively, the nucleoside substitution group may be represented by formula (IIIa'), 【Chemistry 15】 The compound or gapmer oligonucleotide according to claim 1.

12. At least one nucleoside linkage in the nucleotide sequence is a phosphorothioate linkage, Optionally, the phosphorothioate nucleoside linkage is located in one of the Rp or Sp stereostructures. Optionally, all internucleoside links in the nucleotide sequence are phosphorothioate links. Optionally, the gapmer oligonucleotide comprises at least one modified nucleic acid base, wherein the at least one modified nucleic acid base is 5-methylcytosine, pseudouridine, or 5-methoxyuridine. Optionally, the gapmer oligonucleotide comprises at least one nucleoside having a modified sugar moiety. Optionally, the modified sugar moiety is one of the following: 2'-OMe modified sugar moiety, bicyclic sugar moiety, 2'-O-(2-methoxyethyl)(2'-MOE), 2'-O-(N-methylacetamide), 2'-deoxy-2'-fluoronucleoside, 2'-fluoro-β-D-arabinonucleoside, locked nucleic acid (LNA), restricted ethyl 2'-4'-crosslinked nucleic acid (cEt), S-cEt, hexitol nucleic acid (HNA), and tricyclic analog (e.g., tcDNA). Optionally, the gapmer oligonucleotide comprises two, three, four, five, six, seven, eight, nine, or ten nucleosides having a modified sugar moiety. Optionally, the modified sugar moiety is independently one of the following: 2'-OMe modified sugar moiety, bicyclic sugar moiety, 2'-O-(2-methoxyethyl)(2'-MOE), 2'-O-(N-methylacetamide), 2'-deoxy-2'-fluoronucleoside, 2'-fluoro-β-D-arabinonucleoside, locked nucleic acid (LNA), restricted ethyl 2'-4'-crosslinked nucleic acid (cEt), S-cEt, hexitol nucleic acid (HNA), and tricyclic analog (e.g., tcDNA). Optionally, the gapmer oligonucleotide comprises 10 2'-O-(2-methoxyethyl)(2'-MOE) nucleosides, Optionally, five of the 2'-O-(2-methoxyethyl)(2'-MOE) nucleosides are positioned at the 3' end of the gapmer oligonucleotide, and five of the 2'-O-(2-methoxyethyl)(2'-MOE) nucleosides are positioned at the 5' end of the gapmer oligonucleotide, Alternatively, optionally, the gapmer oligonucleotide contains at least one, two, three, four, five, six, seven, eight, nine, or ten deoxyribonucleosides. The gapmer oligonucleotide according to claim 1.

13. The gapmer oligonucleotide is A gap segment comprising one or more of linked deoxyribonucleosides, 2'-fluoroarabinonucleotides (FANA), and fluorocyclohexenylnucleotides (F-CeNA), A 5' wing region containing linked nucleosides, The 3' wing region contains linked nucleosides and Includes, The central region comprises a region of at least eight oligonucleotide units, each containing at least four consecutive nucleic acid bases, positioned between the 5' wing segment and the 3' wing segment, wherein the region has at least 80% identity with any one equal-length portion of sequence numbers 1-954, 1914-149354, 149362-158581, 167805-301566, sequence numbers 301692-301719, 301721-301724, 301729-301735, 301737-301738, and 301741, or any one of the sequences in Tables 5A-5B, 6, 7, or 8, and the 5' wing region and the 3' wing region each contain at least two linked nucleosides, and at least one nucleoside in each wing region contains a modified sugar. The gapmer oligonucleotide according to claim 1.

14. The gapmer oligonucleotide modifies sugars in the following patterns: eeeeee-d10-eeeeee, eeeeee-d8-eeeeee, eeeeee-d11-eeeeee, eee-d8-eee, eee-d10-eee, eeee-d10-eeee, eeeeee-d10-eeee, a eeee-d8-eeee It contains one of the following, where "e" is a 2'-MOE nucleoside and "d" is a deoxyribonucleoside, and at least one "e" or at least one "d" is replaced by a spacer. Optionally, the gapmer oligonucleotide may have nucleoside linkages in the following patterns: sssssoooooooooossss; oooooossssssssoooooo; ooooooooooooooooossss; soosssssssssssssss; sossssssssssssosos; sssssssssssssssssoos; sssssoooooooooooooooo; sssssssssssssssssss; sssssssssssssssssssss; ssssoooooooss; ooosssssssooo; sssssssssssss; sossssssssss; sossssssssss; ssssssssssss; ssssssssssooo; ooosssssssss; ssssssooooooooooss; ooosssssssssooo; sssssssssssssss; ssssssssssssooo; ooosssssssssss; sossssssssssss; sossssssssssss; ssssssssssssss; ssssoooooooooosss; ooooosssssssssoooo; ssssssssssssssss; ssssssssssssssssssoooo; soosssssssssssssss; soosssssssssssss; sssssssssssssssoos; ooooosssssssssss; ssssoooooooosss; ooooosssssssoooo; ssssssssssssssssoooo; ooooosssssssss; soosssssssssssss; soosssssssssss; sssssssssssssoos; soooossssssssssooos; soooosssssssssssooss; sossssssssssssssss; stysssssssssssssssssss; sssssssssssssss It contains one of the following, where "s" is a phosphorothioate linkage and "o" is a phosphodiester linkage. The gapmer oligonucleotide according to claim 13.

15. A pharmaceutical composition comprising a gapmer oligonucleotide according to Claim 1 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

16. A gapmer oligonucleotide according to claim 1 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 15, for use in the treatment of neurological disorders.

17. The neurological disorder is amyotrophic lateral sclerosis (ALS), frontotemporal dementia (FTD), ALS with FTD, Alzheimer's disease (AD), Parkinson's disease (PD), Parkinson's disease with dementia, Lewy body dementia, synucleinopathies, Huntington's disease, brachial plexus injury, peripheral nerve injury, progressive supranuclear palsy (PSP), traumatic brain injury, spinal cord injury, tuberous sclerosis complex, Pick's disease, tauopathy, primary age-related tauopathy, Down syndrome, epilepsy / paroxysmal disorder, depression, traumatic brain injury (TBI), chronic traumatic encephalopathy (CTE), HIV-related neurocognitive disorder (HAND), multiple system atrophy, amnesic mild cognitive impairment, corticobasal degeneration (CBD) and / or neuropathy, For example, a gapmer oligonucleotide or pharmaceutical composition for use according to claim 16, selected from the group consisting of chemotherapy-induced neuropathy, spinocerebellar ataxia (SCA), SCA type 2, spinal muscular atrophy (SMA), parkinsonism, Niemann-Pick disease type C (NPC), Charcot-Marie-Tooth disease (CMT), mucopolysaccharidosis type II (MPSIIA), mucolipidosis IV, GM1 gangliosidosis, sporadic inclusion body myositis (sIBM), Henoch-Schönlein purpura (HSP), Gaucher disease, as well as facial sensorimotor neuropathy, Guam-island Parkinson's dementia complex, multiple system proteinosis, Perry disease, and autism.

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