GAPMER antisense oligonucleotides with modified backbone chemistry

Abstract

Disclosed herein are antisense oligonucleotides, such as gapmer antisense oligonucleotides having a modified backbone chemistry (e.g., having one or more spacers), and the like. Such gapmer antisense oligonucleotides with modified backbone chemistry can be used to treat various diseases, such as nervous system diseases and the like.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of U.S. Provisional Application No. 63 / 285,705 filed on December 3, 2021, U.S. Provisional Application No. 63 / 285,888 filed on December 3, 2021, U.S. Provisional Application No. 63 / 285,692 filed on December 3, 2021, U.S. Provisional Application No. 63 / 285,696 filed on December 3, 2021, and U.S. Provisional Application No. 63 / 285,665 filed on December 3, 2021, each of which is hereby incorporated by reference in its entirety for all purposes.

[0003] Reference to Sequence Listing XML

[0004] This application contains a sequence listing submitted electronically in XML format. The sequence listing XML is incorporated herein by reference. The XML file was created on November 17, 2022, is named QRL-011WO_SL.xml, and is 418,206,230 bytes in size. Background Art

[0005] Antisense oligonucleotide is a compound based on nucleic acid, which can be used to suppress the expression of certain genes related to disease. Although antisense oligonucleotide can be designed to hybridize with target gene usually, conventional antisense oligonucleotide shows poor effect usually. Therefore, it is necessary to develop the modified antisense oligonucleotide that shows improved performance and effect, for preventing, improving and treating disease, and the example of these diseases comprises neurological disease. Summary of the Invention

[0006] Disclosed herein are compounds comprising gapmer oligonucleotides, and wherein the gapmer oligonucleotide comprises a spacer. Additionally disclosed herein are gapmer oligonucleotides, wherein the gapmer oligonucleotide comprises a spacer. In various embodiments, the gapmer oligonucleotide comprises a second spacer that is not adjacent to the spacer.

[0007] In various embodiments, the gapmer oligonucleotide comprises a sequence that is 85% to 98% complementary to an equal length portion of a transcript, the presence of which results in a neurological disease. In various embodiments, the gapmer oligonucleotide comprises a sequence that is 85% to 98% complementary to an equal length portion of a PPM1A mRNA or pre-mRNA transcript, an ATXN2 mRNA or pre-mRNA transcript, a SOD1 mRNA or pre-mRNA transcript, or a MAPT mRNA or pre-mRNA transcript. In various embodiments, the gapmer oligonucleotide comprises a sequence that is 85% to 98% complementary to an equal length portion of any one of SEQ ID NOs: 1909-1913, 149355-149361, 167802-167804, or 301567-301589, a sequence with 90% identity thereof, or a 15 to 50 contiguous nucleobase portion thereof.

[0008] In various embodiments, the gapmer oligonucleotide comprises a sequence that is at least 85% identical 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 a sequence in Tables 4A-4C. In various embodiments, the gapmer oligonucleotide comprises a sequence that is at least 90% identical 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 a sequence in Tables 4A-4C. In various embodiments, the gapmer oligonucleotide comprises a sequence that is at least 95% identical 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 a sequence in Tables 4A-4C. In various embodiments, the gapmer oligonucleotide comprises a sequence that is at least 100% identical 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 a sequence in Tables 4A-4C.

[0009] In various embodiments, the gapmer oligonucleotide comprises a stretch of up to 11 linked nucleosides, and wherein the gapmer oligonucleotide comprises a sequence at least 85% identical 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 a sequence in Tables 4A-4C. In various embodiments, the gapmer oligonucleotide comprises a stretch of up to 10, 9, 8, 7, 6, 5, 4, 3, or 2 linked nucleosides, and wherein the gapmer oligonucleotide comprises a sequence at least 85% identical to an equal length portion of any one of SEQ ID NOs: 301692-301742, or a sequence in Tables 4A-4C. In various embodiments, the gapmer oligonucleotide comprises a segment of up to 11 linked nucleosides, and wherein the gapmer oligonucleotide comprises a sequence at least 90% identical to an equal length portion of any one of SEQ ID NOs: 301692-301742, or a sequence in Tables 4A-4C. In various embodiments, the gapmer oligonucleotide comprises a segment of up to 10, 9, 8, 7, 6, 5, 4, 3, or 2 linked nucleosides, and wherein the gapmer oligonucleotide comprises a sequence at least 90% identical to an equal length portion of any one of SEQ ID NOs: 301692-301742, or a sequence in Tables 4A-4C.

[0010] In various embodiments, the length of 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 various embodiments, at least one (i.e., one or more) nucleoside linkage of the gapmer oligonucleotide is a non-natural linkage. In various embodiments, the length of the gapmer oligonucleotide is at least 19 oligonucleotide units.

[0011] In various embodiments, the spacer is a nucleoside replacement group comprising a non-sugar substituent that cannot be connected to a nucleotide base. In various embodiments, the spacer is located between positions 5 and 11 of the gapmer oligonucleotide. In various embodiments, the spacer is located between positions 7 and 11 of the gapmer oligonucleotide. In various embodiments, the gapmer oligonucleotide further comprises a second spacer, and 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 located between positions 15 and 19 of the gapmer oligonucleotide. In various embodiments, the spacer and the second spacer are separated by at least 5 core bases, at least 6 core bases, or at least 7 core bases in the gapmer oligonucleotide. In various embodiments, the spacer is located between positions 5 and 11 of the gapmer oligonucleotide, and wherein the second spacer is located between positions 15 and 19 of the gapmer oligonucleotide. In various embodiments, the spacer is located at position 8 of the gapmer oligonucleotide, and wherein the second spacer is located at position 16 of the gapmer oligonucleotide. In various embodiments, the spacer is located at position 5 of the gapmer oligonucleotide, and wherein the second spacer is located at position 17 of the gapmer oligonucleotide. In various embodiments, the spacer is located at position 7 of the gapmer oligonucleotide, and wherein the second spacer is located at position 15 of the gapmer oligonucleotide. In various embodiments, the spacer is located at position 11 of the gapmer oligonucleotide, and wherein the second spacer is located at position 19 of the gapmer oligonucleotide.

[0012] In various embodiments, each of the spacer or the second spacer is a nucleoside replacement group comprising a non-sugar substituent, wherein the non-sugar substituent does not comprise a ketone, aldehyde, ketal, hemiketal, acetal, hemiacetal, aminal, or hemiaminal moiety and is incapable of forming a covalent bond with a nucleotide base. In various embodiments, each of the spacer or the second spacer is independently represented by formula (Xa), wherein:

[0013]

[0014] Ring A is an optionally substituted 4-8 membered monocyclic cycloalkyl or 4-8 membered monocyclic heterocyclyl, wherein the heterocyclyl contains 1 or 2 heteroatoms selected from O, S, and N, provided that A cannot form a covalent bond with a nucleobase; and

[0015] The symbol indicates the point of attachment of the internucleoside linkage.

[0016] In various embodiments, each of the spacer or the second spacer is independently represented by formula (Xa), wherein:

[0017]

[0018] In various embodiments, Ring A is an optionally substituted 4-8 membered monocyclic cycloalkyl selected from cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl; or a 4-8 membered monocyclic heterocyclyl 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.

[0019] In various embodiments, each of the spacer or the second spacer is independently represented by formula (I), wherein:

[0020]

[0021] X is selected from -CH2- and -O-; and

[0022] n is 0, 1, 2, or 3.

[0023] In various embodiments, each of the spacer or the second spacer is independently represented by Formula I', wherein:

[0024]

[0025] X is selected from -CH2- and -O-; and

[0026] n is 0, 1, 2, or 3.

[0027] In various embodiments, each of the spacer or the second spacer is independently represented by formula (Ia), wherein:

[0028] and

[0029] n is 0, 1, 2, or 3.

[0030] In various embodiments, each of the spacer or the second spacer is independently represented by formula (Ia'), wherein:

[0031] and

[0032] n is 0, 1, 2, or 3.

[0033] In various embodiments, each of the spacer or the second spacer is independently of the formula II Indicates that:

[0034] and

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

[0036] In various embodiments, each of the spacer or the second spacer is independently represented by Formula II', wherein:

[0037] and

[0038] X is selected from -CH2- and -O-.

[0039] In various embodiments, each of the spacer or the second spacer is independently represented by formula (Iia), wherein:

[0040]

[0041] In various embodiments, each of the spacer or the second spacer is independently represented by formula (Iia'), wherein:

[0042]

[0043] In various embodiments, each of the spacer or the second spacer is independently represented by Formula III, wherein:

[0044] and

[0045] X is selected from -CH2- and -O-.

[0046] In various embodiments, each of the spacer and the second spacer is independently represented by Formula III', wherein:

[0047] and

[0048] X is selected from -CH2- and -O-.

[0049] In various embodiments, each of the spacer and the second spacer is independently represented by formula (IIIa), wherein:

[0050]

[0051] In various embodiments, each of the spacer and the second spacer is independently represented by formula (IIIa'), wherein:

[0052]

[0053] In various embodiments, the gapmer oligonucleotide comprising a spacer has a GC content of at least 10%. In various embodiments, the gapmer oligonucleotide comprising a spacer has a GC content of at least 20%. In various embodiments, the gapmer oligonucleotide comprising a spacer has a GC content of at least 25%. In various embodiments, the gapmer oligonucleotide comprising a spacer has a GC content of at least 30%. In various embodiments, the gapmer oligonucleotide comprising a spacer has a GC content of at least 40%. In various embodiments, the gapmer oligonucleotide comprising a spacer has a GC content of at least 50%. In various embodiments, at least one (i.e. one or more) nucleoside linkages of a gapmer oligonucleotide are independently selected from the group consisting of: phosphodiester linkages, phosphorothioate linkages, alkyl phosphate linkages, phosphorodithioate linkages, phosphotriester linkages, alkyl phosphate linkages, 3-methoxypropyl phosphate linkages, methyl phosphate linkages, aminoalkyl phosphotriester linkages, alkylene phosphonates linkages, phosphites linkages, phosphoramidate linkages, thiophosphoramidate linkages, thiophosphoradiamide linkages, phosphoradiamide linkages, aminoalkyl phosphoramidate linkages, thiophosphoramidate linkages, thioalkyl phosphate linkages, thioalkyl phosphotriester linkages, phosphorothioate linkages, selenophosphate linkages, and boronyl phosphate linkages. In various embodiments, at least one nucleoside linkage of a nucleotide sequence is a phosphorothioate linkage. In various embodiments, the linkage between phosphorothioate nucleosides is one of Rp configuration or Sp configuration.

[0054] In various embodiments, the gapmer oligonucleotide comprises 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 bonds of the nucleotide sequence are phosphorothioate bonds. In various embodiments, the gapmer oligonucleotide comprises at least one modified core base. In various embodiments, at least one modified core base is 5-methylcytosine, pseudouridine, or 5-methoxyuridine. In various embodiments, the gapmer oligonucleotide comprises at least one nucleoside with 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'-fluoro nucleoside, a 2'-fluoro-β-D-arabinonucleoside, a locked nucleic acid (LNA), a constrained ethyl 2'-4'-bridged nucleic acid (cEt), S-cEt, a hexitol nucleic acid (HNA), and a tricyclic analog (e.g., tcDNA).

[0055] In various embodiments, the gapmer oligonucleotide comprises 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleosides with modified sugar moieties. In various embodiments, the modified sugar moieties are independently 2'-OMe modified sugar moieties, bicyclic sugar moieties, 2'-O-(2-methoxyethyl) (2'-MOE), 2'-O-(N-methylacetamide), 2'-deoxy-2'-fluoro nucleosides, 2'-fluoro-β-D-arabinoside, locked nucleic acids (LNA), constrained ethyl 2'-4'-bridged nucleic acids (cEt), S-cEt, hexitol nucleic acids (HNA), and any of 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 located at the 3' end of the gapmer oligonucleotide, and wherein five of the 2'-O-(2-methoxyethyl) (2'-MOE) nucleosides are located at the 5' end of the gapmer oligonucleotide. In various embodiments, at least one nucleoside having a modified sugar moiety or a plurality of nucleosides 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.

[0056] In various embodiments, the gapmer oligonucleotide comprises: a gap segment comprising one or more linked deoxyribonucleosides, 2'-fluoroarabinoic acid (FANA), and fluorocyclohexenyl nucleic acid (F-CeNA); a 5' wing comprising linked nucleosides; and a 3' wing comprising linked nucleosides; wherein the central region comprises a region of at least 8 oligonucleotide units comprising at least 4 contiguous nucleobases, said region being at least 80% identical 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 any one of the sequences of Tables 4A-4C located between the 5' wing and the 3' wing; wherein each of the 5' wing and the 3' wing comprises at least two linked nucleosides; and wherein at least one nucleoside of each wing comprises a modified sugar.

[0057] In various embodiments, the at least two connected nucleosides of the 5' wing region are connected by a phosphorothioate internucleoside bond and / or wherein the at least two connected nucleosides of the 3' wing region are independently connected by a phosphorothioate internucleoside bond. In various embodiments, each internucleoside bond of the 5' wing region and / or each internucleoside bond of the 3' wing region are independently phosphorothioate internucleoside bonds. In various embodiments, the 5' wing region further comprises at least one phosphodiester internucleoside bond. In various embodiments, the 3' wing region further comprises at least one phosphodiester internucleoside bond. In various embodiments, the at least two connected nucleosides of the 5' wing region are connected by a phosphodiester internucleoside bond and / or wherein the at least two connected nucleosides of the 3' wing region are independently connected by a phosphodiester internucleoside bond. In various embodiments, at least one internucleoside bond of the central region is a phosphodiester bond. In various embodiments, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8 or at least 9 nucleoside bonds in the central region are phosphodiester bonds. In various embodiments, at least one nucleoside bond in the central region is a phosphorothioate nucleoside bond. In various embodiments, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8 or at least 9 nucleoside bonds in the central region are phosphorothioate nucleoside bonds. In various embodiments, all nucleoside bonds in the gapmer oligonucleotide are phosphorothioate nucleoside bonds. In various embodiments, any one or all of the bonds in the phosphorothioate nucleoside bonds are Rp configuration, Sp configuration or any combination of Rp and Sp configuration.

[0058] In various embodiments, gapmer oligonucleotide comprises the sugar moiety of at least one modification.In various embodiments, 5' wing region or 3' wing region comprise the sugar moiety of at least one modification.In various embodiments, central region comprises the sugar moiety of at least one modification.In various embodiments, the sugar moiety of at least one modification is the sugar moiety, bicyclic sugar moiety, 2'-O-(2-methoxyethyl) (2'-MOE) modified by 2'-OMe, 2'-O-(N-methylacetamide), 2'-deoxy-2'-fluoro nucleoside, 2'-fluoro-β-D-arabinoside, locked nucleic acid (LNA), constrained ethyl '-4'-bridged nucleic acid (cEt), S-cEt, tcDNA, hexitol nucleic acid (HNA) and tricyclic analog (such as tcDNA) one.In various embodiments, gapmer oligonucleotide comprises one or more 2'-MOE nucleosides.In various embodiments, 5' wing region or 3' wing region comprise one or more 2'-MOE nucleosides. In various embodiments, the 5' wing or 3' wing comprises 2, 3, 4, 5, or 6 2'-MOE nucleosides. In various embodiments, each nucleoside in the 5' wing or 3' wing is a 2'-MOE nucleoside. 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, each nucleoside in the central region is a 2'-MOE nucleoside. In various embodiments, one or more 2'-MOE nucleosides are linked by phosphorothioate internucleoside linkages.

[0059] In various embodiments, the gapmer oligonucleotide comprises any of the following patterns of sugar modifications: eeeee-d10-eeeee, eeeee-d8-eeeee, eeeeee-d11-eeeeee, eee-d8-eee, eee-d10-eee, eeee-d10-eeee, and eeee-d8-eeee, wherein e=2'-MOE nucleoside and d=deoxyribonucleoside, and wherein at least one "e" or at least one "d" is replaced with a spacer.In various embodiments, the gapmer oligonucleotide comprises internucleoside linkages of any of the following patterns: ssssssooooooooosssss; oooosssssssssooooo; ooooooooooooooosssss; soossssssssssssssss; sosososssssssssssosos; sssssssssssssssssoos; sssssoooooooooooooo; ssssssssssssssssssss; sssssssssssssssssssssss; sssooooooosss; ooosssssssooo; ssssssssssssss; Sosssssssssos; sosssssssssss; sssssssssssos; sssssssssssooo; ooossssssssss; sssooooooooosss; ooosssssssssooo; ssssssssssssssss; sssssssssssssooo; ooossssssssssss; sosssssssssssos; sosssssssssssss; ssssssssssssssos; ssssooooooooossss; oooosssssssssoooo; ssssssssssssssssss; ssssssssssssssoooo; soosssssssssssoos; soossssssssssssss; sssssssssssssssoos; ooosssssssssssss; ssssooooooossss; ooosssssssoooo; ssssssssssssoooo; ooosssssssssss; soosssssssssoos; soossssssssssss; sssssssssssssoos; soooossssssssssooos; soooossssssssssooss; sossssssssssssoss; sosssssssssssssosss; or ssssssssssssssss; where s = phosphorothioate linkage and o = phosphodiester linkage.

[0060] In various embodiments, the gapmer oligonucleotide comprises any of the following combinations of sugar modifications and internucleoside linkages, respectively:

[0061] a) eeeee-d10-eeeeee and ssssssooooooooossss;

[0062] b) eeeee-d10-eeeeee and oooooosssssssssooooo;

[0063] c) eeeee-d10-eeeeee and ssssssssssssssssss;

[0064] d) eee-d8-eee and sssooooooosss;

[0065] e) eee-d8-eee and ooosssssssooo

[0066] f) eee-d8-eee and ssssssssssssss;

[0067] g) eee-d10-eee and sssooooooooosss;

[0068] h) eee-d10-eee and ooosssssssssooo;

[0069] i) eee-d10-eee and sssssssssssssss;

[0070] j) eeee-d10-eeee and ssssooooooossss;

[0071] k) eeee-d10-eeee and oooosssssssssoooo;

[0072] l) eeee-d10-eeee and ssssssssssssssssss;

[0073] m) eeee-d8-eeee and ssssooooooossss;

[0074] n) eeee-d8-eeee and oooosssssssoooo;

[0075] o) eeee-d8-eeee and ssssssssssssssss;

[0076] p) eeeeee-d11-eeeeee and sssssssssssssssssssssss;

[0077] q) eeeee-d10-eeeeee and sosososssssssssssosos;

[0078] r) eeeee-d10-eeeeee and soooossssssssssooos;

[0079] s)eeeee-d10-eeeeee and soooossssssssssooss;

[0080] t)eeeee-d8-eeeeee and sosssssssssssoss;

[0081] u)eeeee-d10-eeeeee and sosssssssssssssssss;

[0082] V)eeeeee-d10-eeee and sssssssssssssssssss;

[0083] wherein e=2'-MOE nucleoside and d=deoxyribonucleoside, and wherein s=phosphorothioate linkage, and o=phosphodiester linkage, and wherein at least one "e" or at least one "d" of the gapmer oligonucleotide is replaced with a spacer.

[0084] In various embodiments, the gapmer oligonucleotide comprises at least one modified nucleobase. In various embodiments, the 5' wing or the 3' wing comprises at least one modified nucleobase. In various embodiments, the central region comprises at least one modified nucleobase. In various embodiments, at least one modified nucleobase is 5-methylcytosine, pseudouridine, or 5-methoxyuridine. In various embodiments, each cytosine in the 5' wing or the 3' wing is 5-methylcytosine. In various embodiments, each cytosine in the central region is 5-methylcytosine. In various embodiments, the gapmer oligonucleotide comprises the following sugar modification and internucleoside linkage combination: eeeee-d10-eeeee and ssssssssssssssssssss, wherein e=2'-MOE nucleoside and d=deoxyribonucleoside, wherein s=phosphorothioate linkage, wherein at least one "e" or at least one "d" of the gapmer oligonucleotide is replaced with a spacer, and wherein each cytosine of the 2'-MOE nucleoside is 5-methylcytosine.

[0085] In various embodiments, the gapmer oligonucleotide further comprises a conjugate moiety. In various embodiments, the conjugate moiety is a cholesterol conjugate located at the 3' end of the gapmer oligonucleotide.

[0086] Additionally disclosed herein are pharmaceutical compositions comprising a gapmer oligonucleotide disclosed herein, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0087] Further disclosed herein is a method for treating a neurological disease in a patient in need thereof, the method 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 selected from 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, synucleinopathy, Huntington's disease, brachial plexus injury, peripheral nerve injury, progressive supranuclear palsy (PSP), brain trauma, spinal cord injury, tuberous sclerosis, Pick's disease, tauopathy, primary age-related tauopathy, Down syndrome, epilepsy / seizure disorder, depression, traumatic brain injury (TBI), chronic traumatic encephalopathy (CTE), HIV-related neurocognitive disorder (HAND), multiple system atrophy, amnestic mild cognitive impairment, cortical basal ganglia Degeneration (CBD) and / or neuropathies such as chemotherapy-induced neuropathy, spinocerebellar ataxia (SCA), type 2 SCA, spinal muscular atrophy (SMA), Parkinson's disease, Niemann-Pick disease type C (NPC), Charcot-Marie-Tooth disease (CMT), mucopolysaccharidosis type II (MPSIIA), mucolipidosis type IV, GM1 gangliosidosis, sporadic inclusion body myositis (sIBM), Henoch-Schonlein purpura (HSP), limbic system age-related TDP-43 encephalopathy (LATE), cerebral age-related TDP-43 sclerosis (CARTS), Gaucher disease, facial onset sensory and motor neuron disease, Guam-type parkinsonism and dementia syndrome, multisystem proteinopathy, Perry disease, and synaptic diseases such as autism.

[0088] In various embodiments, the gapmer oligonucleotide is administered topically, parenterally, intrathecally, orally, to the lungs, intratrachea, intranasally, transdermally, orally, intrathalamus, intraventricularly, intraocularly, sublingually, rectum, vaginally, or intraduodenally. In various embodiments, the gapmer oligonucleotide is administered intrathecally. In various embodiments, the gapmer oligonucleotide is administered in a therapeutically effective amount. In various embodiments, the patient is a human being. In various embodiments, the pharmaceutical composition is suitable for topical, parenteral, intrathecally, orally, to the lungs, intratrachea, intranasally, transdermally, orally, intrathalamus, intraventricularly, intraocularly, sublingually, rectum, vaginally, or intraduodenally.

[0089] Further disclosed herein is the use of gapmer oligonucleotides in the preparation of a medicament for treating a neurological disease. In various embodiments, the neurological disease is selected from 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, synucleinopathy, Huntington's disease, brachial plexus injury, peripheral nerve injury, progressive supranuclear palsy (PSP), brain trauma, spinal cord injury, tuberous sclerosis, Pick's disease, tauopathy, primary age-related tauopathy, Down syndrome, epilepsy / seizure disorder, depression, traumatic brain injury (TBI), chronic traumatic encephalopathy (CTE), HIV-related neurocognitive disorder (HAND), multiple system atrophy, amnestic mild cognitive impairment, cortical basal ganglia Degeneration (CBD) and / or neurological diseases such as chemotherapy-induced neuropathy, spinocerebellar ataxia (SCA), type 2 SCA, spinal muscular atrophy (SMA), Parkinson's disease, Niemann-Pick disease type C (NPC), Charcot-Marie-Tooth disease (CMT), mucopolysaccharidosis type II (MPSIIA), mucolipidosis type IV, GM1 gangliosidosis, sporadic inclusion body myositis (sIBM), Henoch-Schonlein purpura (HSP), limbic system age-related TDP-43 encephalopathy (LATE), brain age-related TDP-43 sclerosis (CARTS), Gaucher disease, facial onset sensory and motor neuron disease, Guam type parkinsonism syndrome, multisystem protein disease, Perry disease, and synaptic diseases such as autism, etc. In various embodiments, the gapmer oligonucleotide is a gapmer oligonucleotide disclosed herein.

[0090] Further disclosed herein is a method for treating a neurological disease in a patient in need thereof, the method comprising administering to a 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 selected from 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, synucleinopathy, Huntington's disease, brachial plexus injury, peripheral nerve injury, progressive supranuclear palsy (PSP), brain trauma, spinal cord injury, tuberous sclerosis, Pick's disease, tauopathy, primary age-related tauopathy, Down syndrome, epilepsy / seizure disorder, depression, traumatic brain injury (TBI), chronic traumatic encephalopathy (CTE), HIV-related neurocognitive disorder (HAND), multiple system atrophy, amnestic mild cognitive impairment, cortical basal ganglia Degeneration (CBD) and / or neuropathies such as chemotherapy-induced neuropathy, spinocerebellar ataxia (SCA), type 2 SCA, spinal muscular atrophy (SMA), Parkinson's disease, Niemann-Pick disease type C (NPC), Charcot-Marie-Tooth disease (CMT), mucopolysaccharidosis type II (MPSIIA), mucolipidosis type IV, GM1 gangliosidosis, sporadic inclusion body myositis (sIBM), Henoch-Schonlein purpura (HSP), limbic system age-related TDP-43 encephalopathy (LATE), brain age-related TDP-43 sclerosis (CARTS), Gaucher disease, facial onset sensory and motor neuron disease, Guam-type Parkinson's disease syndrome, multisystem protein disease, Perry disease, and synaptic diseases such as autism, etc. 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, rectal, buccal, sublingual, vaginally, intratracheally, intranasally, intrathecally, intracisternal, transdermally, or intraduodenally. In various embodiments, the pharmaceutical composition is administered intrathecally. In various embodiments, the patient is a human.

[0091] Further disclosed herein are gapmer oligonucleotides or pharmaceutically acceptable salts thereof for use as a medicament. Further disclosed herein are gapmer oligonucleotides or pharmaceutically acceptable salts thereof for use in treating a neurological disease. In various embodiments, the neurological disease is selected from 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, synucleinopathy, Huntington's disease, brachial plexus injury, peripheral nerve injury, progressive supranuclear palsy (PSP), brain trauma, spinal cord injury, tuberous sclerosis, 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 (HIV-associated encephalopathy), and the like. ANND), multiple system atrophy, amnestic mild cognitive impairment, corticobasal degeneration (CBD), and / or neuropathies such as chemotherapy-induced neuropathy, spinocerebellar ataxia (SCA), SCA type 2, spinal muscular atrophy (SMA), Parkinson's disease, Niemann-Pick disease type C (NPC), Charcot-Marie-Tooth disease (CMT), mucopolysaccharidosis type II (MPSIIA), mucolipidosis type IV, GM1 gangliosidosis, sporadic inclusion body myositis (sIBM), Henoch-Schonlein purpura (HSP), Gaucher disease, facial-onset sensory and motor neuronopathy, Guam-type parkinsonism, multisystem proteinopathy, Perry disease, and autism.

[0092] Further disclosed herein are gapmer oligonucleotides comprising the sequence of any one of SEQ ID NOs: 301692-301742 or a sequence in Tables 4A-4C, or a pharmaceutically acceptable salt thereof; wherein the gapmer oligonucleotide further comprises: a gap segment comprising one or more of linked deoxyribonucleosides, 2'-fluoroarabinoic acid (FANA), and fluorocyclohexenyl nucleic acid (F-CeNA); a 5' wing comprising linked nucleosides; and a 3' wing comprising linked nucleosides; wherein the gapmer oligonucleotide comprises a sugar modification of eeeee-d10-eeeeee, wherein e=2'-MOE nucleoside and d=deoxyribonucleoside, wherein at least one "e" or at least one "d" is replaced with a spacer, and wherein each cytosine of the 2'MOE nucleoside is a 5-methylcytosine.

[0093] In various embodiments, at least one internucleoside linkage of a gapmer oligonucleotide is a phosphorothioate linkage. In various embodiments, the phosphorothioate internucleoside linkage is in one of an Rp configuration or an Sp configuration. In various embodiments, all internucleoside linkages of a gapmer oligonucleotide are phosphorothioate linkages. In various embodiments, a gapmer oligonucleotide comprises any of the following patterns of sugar modifications and internucleoside linkage combinations:

[0094] a) eeeee-d10-eeeeee and sssssooooooooossss;

[0095] b)eeeee-d10-eeeeee and ooooosssssssssooooo;

[0096] c)eeeee-d10-eeeeee and ssssssssssssssssss;

[0097] d)eee-d8-eee and ssssooooooosss;

[0098] e)eee-d8-eee and ooosssssssooo

[0099] f)eee-d8-eee and sssssssssssss;

[0100] g)eee-d10-eee and ssssooooooooosss;

[0101] h)eee-d10-eee and ooosssssssssooo;

[0102] i)eee-d10-eee and ssssssssssssss;

[0103] j)eeee-d10-eeee and ssssooooooossss;

[0104] k)eeee-d10-eeee andoooossssssssssoooo;

[0105] l)eeee-d10-eeee and sssssssssssssssss;

[0106] m)eeee-d8-eeee and ssssooooooossss;

[0107] n)eeee-d8-eeee andoooosssssssoooo;

[0108] o)eeee-d8-eeee and sssssssssssssss;

[0109] p)eeeeee-d11-eeeeee and ssssssssssssssssssssss;

[0110] q)eeeee-d10-eeeeee and sososssssssssssosos;

[0111] r)eeeee-d10-eeeeee and soooossssssssssooos;

[0112] s)eeeee-d10-eeeeee and soooossssssssssooss;

[0113] t)eeeee-d8-eeeeee and sosssssssssssoss;

[0114] u)eeeee-d10-eeeeee and sosssssssssssssssss,

[0115] wherein at least one "e" or at least one "d" of the gapmer oligonucleotide is replaced with a spacer.

[0116] Additionally disclosed herein are pharmaceutical compositions comprising the antisense gapmer oligonucleotides disclosed herein or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient. In various embodiments, the spacer is a nucleoside replacement group comprising a non-sugar substituent, wherein the non-sugar substituent does not comprise 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, the spacer is represented by formula (X), wherein:

[0117]

[0118] Ring A is an optionally substituted 4-8 membered monocyclic cycloalkyl or 4-8 membered monocyclic heterocyclyl, wherein the heterocyclyl contains 1 or 2 heteroatoms selected from O, S, and N, provided that A cannot form a covalent bond with a nucleobase; and

[0119] The symbol indicates the point of attachment of the internucleoside linkage.

[0120] In various embodiments, the spacer is represented by formula (Xa), wherein:

[0121]

[0122] In various embodiments, Ring A is an optionally substituted 4-8 membered monocyclic cycloalkyl selected from cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl; or a 4-8 membered monocyclic heterocyclyl 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.

[0123] In various embodiments, the spacer is represented by formula (I), wherein:

[0124]

[0125] X is selected from -CH2- and -O-; and

[0126] n is 0, 1, 2, or 3.

[0127] In various embodiments, the spacer is represented by formula (I'), wherein:

[0128]

[0129] In various embodiments, the spacer is represented by Formula (Ia), wherein:

[0130]

[0131] In various embodiments, the spacer is represented by Formula (Ia'), wherein:

[0132]

[0133] In various embodiments, the spacer is represented by formula (II), wherein:

[0134] and

[0135] X is selected from -CH2- and -O-.

[0136] In various embodiments, the spacer is represented by Formula (Ia'), wherein:

[0137] and

[0138] X is selected from -CH2- and -O-.

[0139] In various embodiments, the spacer is represented by Formula (Iia), wherein:

[0140]

[0141] In various embodiments, the spacer is represented by formula (Iia'), wherein:

[0142]

[0143] In various embodiments, the spacer is represented by formula (III), wherein:

[0144] and

[0145] X is selected from -CH2- and -O-.

[0146] In various embodiments, the spacer is represented by formula (III'), wherein:

[0147] and

[0148] X is selected from -CH2- and -O-.

[0149] In various embodiments, the spacer is represented by formula (IIIa), wherein:

[0150]

[0151] In various embodiments, the spacer is represented by formula (IIIa'), wherein:

[0152]

[0153] Further disclosed herein are methods of treating a neurological disease and / or a neurological disorder 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 a second therapeutic agent selected from the group consisting of riluzole (Rilutek), troriluzole, edaravone (Radicava), rivastigmine, donepezil, QRL-101, QRL-201, galantamine, a selective serotonin reuptake inhibitor, an antipsychotic, a cholinesterase inhibitor, memantine, a benzodiazepine anxiolytic, AMX0035 ZILUCOPLAN (RA101495), dual AON intrathecal administration (e.g., BIIB067, BIIB078), BIIB100, levodopa / carbidopa, dopaminergic drugs (e.g., ropinirole, pramipexole, rotigotine), medroxyprogesterone, KCNQ2 / KCNQ3 openers, pridopidine, PrimeC (a combination of ciprofloxacin and Celebrex), olanzapine (Zyprexa), quetiapine (Seroquel), SSRI, sodium valproate (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 of amyotrophic lateral sclerosis (ALS), frontotemporal dementia (FTD), or ALS with FTD.

[0154] Further disclosed herein are methods of treating a neurological disease and / or a neurological disorder 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 a second therapeutic agent selected from the group consisting of memantine, rivastigmine, galantamine, donepezil, QRL-101, QRL-201, (Rivastigmine), Aducanumab, BAN2401, BIIB091 (gosuranemab), BIIB076, BIIB080 (IONIS-MAPTRx), Elayta (CT1812), MK1942, allogeneic hMSCs, nilotinib, ABT-957, acitretin, ABT-354, GV1001, riluzole, CAD106, CNP520, AD-35, rilapladib, DHP1401, T-817MA, TC-5619, TPl-287, RVT-101, LY450139, JNJ - 54861911, dapagliflozin, GSK239512, PF-04360365, ASP0777, SB-742457 (5-HT6 receptor antagonist), PF-03654746 (H3 receptor antagonist), GSK933776 (Fc-inactivated anti-beta amyloid (Aβ) monoclonal antibody (mAb)), Posiphen ((+)-phenylserine tartrate), AMX0035 Coenzyme Q10, aducanumab (ADUHLEM), memantine (NAMENDA), Namzeric, Suvorexant (belsomra), lecanemab, or any combination thereof. In various embodiments, the neurological disease is Alzheimer's disease.

[0155] Further disclosed herein are methods 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 in combination with a second therapeutic agent selected from the group comprising levodopa, carbidopa-levodopa, 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, BIIB118, 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, Nourianz, Xadago, Cogentin, Ongentys, Exenatide, Ganoderma lucidum, caffeine, Thalizotan, embryonic dopamine cell implantation, or any combination thereof. In various embodiments, the neurological disease is Parkinson's disease.

[0156] Further disclosed herein are methods of treating a neurological disease and / or a neurological disorder 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 a second therapeutic agent selected from the group consisting of UCB0107, ABBV-8E12, F-18AV1451, BIIB092, C2N-8E12, tideglusib, deep transcranial magnetic stimulation, lipoic acid, tolfenamic acid, lithium, AZP2006, glial cell line-derived neurotrophic factor, NBMI, suvorxant, zolpidem, TPI 287, davunetide, pimavanserin, levodopa, carbidopa-levodopa, pramipexole, ropinirole, rotigotine, apomorphine, selegiline, rasagiline, entacapone, tolcapone, amantadine, trihexyphenidyl, BIIB054 (Cenipamab), B IIB094, BIIB118, 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, or any combination thereof. In various embodiments, the neurological disease is progressive supranuclear palsy (PSP).

[0157] Further disclosed herein are methods of treating a neurological disease and / or a 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 in combination with a second therapeutic agent selected from the group consisting of tetrabenazine, deutetrabenazine, physical therapy, risperidone, haloperidol, chlorpromazine, clonazepam, diazepam, benzodiazepines, selective serotonin reuptake inhibitors, quetiapine, carbatrol, valproic acid, lamotrigine, pridopidine, Delta-9-tetrahydrocannabinol, cannabidiol, stem cell therapy, ISIS-443139, nilotinib, resveratrol, neflamapimod, fenofibrate, creatine, RO7234292, SAGE-718, WVE-120102, WVE-120101, dimebon, minocycline, deep brain stimulation, ursodeoxycholic acid, coenzyme Q10, OMS643762, VX15 / 2503, PF-02545920, BN82451B, SEN0014196, olanzapine, tiapride, or any combination thereof. In various embodiments, the neurological disease is Huntington's disease.

[0158] Disclosed herein are additional methods for treating a neurological disease and / or a neuropathy in a patient in need thereof, comprising administering to a patient in need thereof a combination of a therapeutically effective amount of a gapmer oligonucleotide, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition and a second therapeutic agent selected from the group consisting of an anticoagulant, an antidepressant, a muscle relaxant, a stimulant, an anticonvulsant, an antianxiety drug, erythropoietin, hyperbaric therapy, rehabilitation therapy (e.g., physical, occupational, speech, psychological, or vocational counseling), or any combination thereof. In various embodiments, the neurological disease is a brain injury.

[0159] Additionally disclosed herein are methods for 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 in combination with a second therapeutic agent selected from the group consisting of AXER-204, glibenclamide, 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 exercise strategies, and a healthy diet), or any combination thereof. In various embodiments, the neurological disease is spinal cord injury.

[0160] Further disclosed herein are methods of treating a neurological disease and / or neuropathy in a patient in need thereof, the methods 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 in combination with a second therapeutic agent selected from the group consisting of TPI-287, lithium, occupational, physical and speech therapy, or any combination thereof may be selected as an additional therapy. In various embodiments, the neurological disease is corticobasal degeneration.

[0161] Disclosed herein are additional methods for treating a neurological disease and / or a neuropathy in a patient in need thereof, the methods 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 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, autoantibodies targeting neuronal antigens (TS-HDS and FGFR3), or any combination thereof. In various embodiments, the neuropathy is chemotherapy-induced neuropathy.

[0162] Disclosed herein are additional methods for 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 in combination with a second therapeutic agent selected from the group consisting of amantadine, armodafinil, baclofen, buspirone, carbamazepine, citalopram, clonazepam, desvenlafaxine, diazepam, duloxetine, escitalopram, flunarizine, fluoxetine, fluvoxamine, gabapentin, isoniazid, levetiracetam, levodopa, memantine, modafinil, ondansetron, paroxetine, pramipexole, primidone, riluzole, ropinirole, sertraline, tizanidine, topiramate, benzhexol, valproic acid, venlafaxine, BHV-4157, or a combination thereof. In various embodiments, the neurological disease is spinocerebellar ataxia.

[0163] Further disclosed herein are methods of treating a neurological disease and / or a 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 a second therapeutic agent selected from the group consisting of brivaracetam (briviact), cannabidiol (epidiolex), carbamazepine (Tegretol), xcopri, diazepam (valium), lorazepam (Ativan), clonazepam (klonopin), aptiom, ethosuximide (zarontin), felbatol, fenfluramine (fintepla), lacosamide (VIMPAT), lamotrigine (Lamictal), levetiracetam (Keppra), oxcarbazepine (oxtellar), fenfluramine (fintepla), lacosamide (VIMPAT), lamotrigine (Lamictal), levetiracetam (Keppra), oxcarbazepine (oxtellar), fenfluramine (fentepla), lacosamide (VIMPAT ... lamotrigine (fentepla), lacosamide (VIMPAT), lamotrigine (fentepla), lacosamide (VIMPAT), lamotrigine (fentepla), lacosamide (VIMPAT), lamotrigine (fentepla), lacosamide (VIMPAT), lacosamide (VIMPAT), lacosamide (VIMPAT), lacosamide (VIMPAT), lacosamide xr, Trileptal), perampanel (fycompa), phenobarbital, phenytoin (dilantin), pregabalin (lyrica), tiagabine (gabitril), topiramate (topamax), valproic acid (depakene, depakote), zonisamide (zonegran), or any combination thereof. In various embodiments, the neurological disease is epilepsy.

[0164] Further disclosed herein are methods for treating a neurological disease and / or a 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 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.

[0165] Additionally disclosed herein are methods for 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 in combination with a second therapeutic agent selected from the group consisting of an anti-epileptic 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.

[0166] Additionally disclosed herein are methods for 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 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).

[0167] Further disclosed herein are methods for treating a neurological disease and / or neuropathy in a patient in need thereof, the methods 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 in combination with a second therapeutic agent selected from the group consisting of: enzyme replacement therapy: 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).

[0168] Further disclosed herein are methods of treating a neurological disease and / or neuropathy in a patient in need thereof, the methods 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 in combination with a second therapeutic agent selected from the group consisting of physical, occupational, and speech therapy, contact lenses and artificial tears, genetic counseling, or any combination thereof. In various embodiments, the neurological disease is type IV mucolipidosis.

[0169] Further disclosed herein are methods of treating a neurological disease and / or a neuropathy in a patient in need thereof, the methods 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 in combination with a second therapeutic agent selected from the group consisting of an anticonvulsant, physical and occupational therapy, β-galactosidase, gene delivery of β-galactosidase, LYS-GM101 gene therapy, or any combination thereof. In various embodiments, the neurological disease is GM1 gangliosidosis.

[0170] Further disclosed herein are methods of treating a neurological disease and / or neuropathy in a patient in need thereof, the methods 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 in combination with a second therapeutic agent selected from the group consisting of physical and occupational therapy, use of a device such as a brace, a walker, a wheelchair, etc., an immunosuppressant, BYM338, or any combination thereof. In various embodiments, the neurological disease is sporadic inclusion body myositis (sIBM).

[0171] Additionally disclosed herein are methods for 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, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition in combination with a second therapeutic agent selected from the group consisting of a corticosteroid, colchicine, dapsone, azathioprine, or any combination thereof. In various embodiments, the neurological disease is Henoch-Schonlein purpura (HSP).

[0172] Additionally disclosed herein are methods for 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 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 DESCRIPTION OF THE DRAWINGS

[0173] Figure 1 Examples of antisense oligonucleotides (AONs) are shown, which are partially complementary to an mRNA transcript or pre-mRNA transcript. Dashed lines indicate the positions of AONs, which may or may not be complementary to corresponding positions in the transcript. DETAILED DESCRIPTION

[0174] The features and other details of the present disclosure will now be described in more detail. Before further describing the present invention, certain terms used in the specification, examples, and appended claims are collected here. These definitions should be read and understood by those skilled in the art in light of the remainder of this disclosure. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art.

[0175] definition

[0176] As used herein, the terms "treat," "treatment," "treatment," and the like generally mean obtaining a desired pharmacological and / or physiological effect. The effect may be therapeutic in the sense of partially or completely curing a disease and / or side effects attributable to the disease. As used herein, the term "treat" encompasses any treatment of a disease in mammals, particularly humans, and includes: (a) inhibiting the disease, i.e., preventing an increase in the severity or extent of the disease; (b) alleviating the disease, i.e., causing a partial or complete improvement in the disease; and (c) preventing the recurrence of the disease, i.e., preventing the disease from returning to an active state after a previous successful treatment of disease symptoms or treatment of the disease.

[0177] "Prevention" includes delaying the onset of clinical symptoms, complications, or biochemical markers of a state, disorder, disease, or condition in a subject who may be susceptible to or susceptible to the state, disorder, disease, or condition but who has not yet experienced or displayed clinical or subclinical symptoms of the state, disorder, disease, or condition. "Prevention" encompasses prophylactic treatment of a state, disorder, disease, or condition in a subject, including prophylactic treatment of clinical symptoms, complications, or biochemical markers of a state, disorder, disease, or condition in a subject.

[0178] The terms "pharmaceutically acceptable carrier" or "pharmaceutically acceptable excipient," as used interchangeably herein, refer to any and all solvents, dispersion media, coatings, isotonicity agents, absorption delaying agents, and the like, compatible with pharmaceutical administration. The use of such media and agents for pharmaceutically active substances is well known in the art. The compositions may also contain other active compounds that provide supplemental, additional, or enhanced therapeutic function.

[0179] As used herein, the term "pharmaceutical composition" refers to a composition comprising at least one biologically active compound disclosed herein (eg, antisense oligonucleotide (AON)) formulated together with one or more pharmaceutically acceptable excipients.

[0180] "Individual," "patient," or "subject" are used interchangeably and include any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cattle, sheep, horses, or non-human primates, and most preferably humans. The compounds of the present invention can be administered to mammals, such as humans, but can also be administered to other mammals, such as animals requiring veterinary treatment, for example, livestock (such as dogs, cats, etc.), farm animals (such as cattle, sheep, pigs, horses, etc.), and experimental animals (such as rats, mice, guinea pigs, non-human primates, etc.). In some embodiments, the mammal treated in the methods of the present invention is ideally a mammal in which modulation of target expression and / or activity is desired.

[0181] As used herein, "PPM1A" (also known as 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., protein encoded by the gene or mRNA transcript (including pre-mRNA)) identified by Entrez Gene ID No. 5494 and its allelic variants, as well as orthologs found in non-human species (e.g., non-human primate or mouse).

[0182] 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, Olivopontocerebellar Ataxia 2, autosomal dominant, Ataxin 2, and trinucleotide repeat-containing 13) refers to the gene or gene product (e.g., gene-encoded protein or mRNA transcript (including pre-mRNA)) identified by Entrez Gene ID No. 6311 and its allelic variants, as well as orthologs found in non-human species (e.g., non-human primates or mice).

[0183] 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, Cystolic, Cu / Zn superoxide dismutase, indophenol oxidase A, SOD, soluble, homodimer, HEL-S-44, STAHP, and SOD) refers to the gene or gene product (e.g., protein encoded by the gene or mRNA transcript (including pre-mRNA)) identified by Entrez Gene ID No. 6647 and its allelic variants, as well as orthologs found in non-human species (e.g., non-human primate or mouse).

[0184] As used herein, "MAPT" (also known as microtubule-associated protein Tau, MTBT1, PPP1R103, FTDP-17, MTBT2, MAPTL, PPND, MSTD, TAU, G protein β1 / γ2, subunit interacting factor, protein phosphatase 1, regulatory protein subunit 103, microtubule-associated protein Tau, neurofibrillary tangle paired helical filament-Tau, MGC138549, FLJ31424, PHF-Tau, Tau-40, DDPAC, Tau and microtubule-associated protein Tau, isoform 4) refers to the gene identified by Entrez Gene ID No. 4137 or gene products (e.g., protein encoded by the gene or mRNA transcript (including pre-mRNA)) and allelic variants thereof, as well as orthologs found in non-human species (e.g., non-human primates or mice).

[0185] As used herein, the term "therapeutically effective amount" refers to the amount of an AON in a subject that will elicit the biological or medical response in a tissue, system, animal, or human that is being sought 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 state, such as ALS, FTD, ALS with FTD, or another motor neuron disease or nervous system disease or condition. Alternatively, a therapeutically effective amount of an AON is the amount required to achieve the desired therapeutic and / or preventive effect, such as an amount that results in the prevention or reduction of symptoms associated with a disease disclosed herein.

[0186] As used herein, the term "antisense oligonucleotide" or "AON" encompasses antisense oligonucleotides that target any of the genes or gene products of PPM1A, ATXN2, SOD1, or MAPT. "Antisense oligonucleotide" or "AON" encompasses any parent oligonucleotide, oligonucleotide variant, oligonucleotide with one or more spacers, oligonucleotide variant with one or more spacers, gapmer antisense oligonucleotides (AON), and gapmer AON with one or more spacers. Examples of antisense oligonucleotides include oligonucleotides comprising the sequence of any of the antisense oligonucleotides of any 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 of the sequence of any of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, or 167805-301566, such as any of the sequences in SEQ ID NOs: 301692-301742 or in Tables 4A-4C, etc.

[0187] The terms "parent oligonucleotide," "parent antisense oligonucleotide," or "parent AON" refer to an antisense oligonucleotide that is complementary to a portion of a target gene product, such as any of PPM1A, ATXN2, SOD1, or MAPT mRNA or pre-mRNA transcripts. The parent oligonucleotide does not comprise a spacer. In specific embodiments, the parent oligonucleotide comprises 20 linked nucleosides. In such embodiments, the parent oligonucleotide is a 20mer. 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 below, oligonucleotides with spacers, oligonucleotide variants, and gapmer AONs are described relative to the corresponding parent oligonucleotides.

[0188] 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, the oligonucleotide variant is any one of a 15mer, 16mer, 17mer, 18mer, 19mer, 20mer, 21mer, 22mer, or 23mer. For example, an AON variant can be a shorter or longer 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. Examples of oligonucleotide variants include oligonucleotides comprising a sequence of any one of SEQ ID NOs: 301590-301594. In various embodiments, the oligonucleotide variant comprises one or more spacers.

[0189] The term "oligonucleotide with one or more spacers" or "oligonucleotide comprising a spacer" refers to an oligonucleotide having at least one spacer. In various embodiments, the oligonucleotide with one or more spacers can comprise 1 spacer, 2 spacers, 3 spacers, 4 spacers, 5 spacers, 6 spacers, 7 spacers, 8 spacers, 9 spacers, or 10 spacers. In various embodiments, the oligonucleotide comprising one or more spacers comprises at least one segment with up to 10 connected nucleosides. For example, as described in the 5' to 3' direction, the oligonucleotide comprising a spacer can comprise a segment with 10 connected nucleosides, followed by a spacer, followed by a second segment of 9 connected nucleosides. In various embodiments, the oligonucleotide comprising one or more spacers comprises at least one segment with up to 7 connected nucleosides. For example, as described on the 5' to 3' direction, the oligonucleotide comprising spacer can comprise the section with 7 nucleosides connected, followed by a spacer, the second section with 9 nucleosides connected, followed by the second spacer and the third section with 7 nucleosides connected.Here, the first section of 7 nucleosides connected and the third section of 7 nucleosides connected each represent the section with 7 nucleosides connected.In various embodiments, the oligonucleotide comprising one or more spacers comprises at least one section with at most 6 nucleosides connected.For example, as described on the 5' to 3' direction, the oligonucleotide comprising spacer can comprise the section with 6 nucleosides connected, followed by a spacer, the second section with 6 nucleosides connected, followed by the second spacer and the third section with 6 nucleosides connected.Here, the first section of 6 nucleosides connected, the second section of 6 nucleosides connected and the third section of 6 nucleosides connected each represent the section with 6 nucleosides connected.Here, each fragment of the oligonucleotide with one or more spacers has at most 6 nucleosides connected. In various embodiments, a gapmer oligonucleotide comprises a segment of up to 10, 9, 8, 7, 6, 5, 4, 3, or 2 linked nucleosides.

[0190] As another example, the oligonucleotide comprising a spacer can comprise a section with 10 nucleosides connected, followed by a spacer, a second section with 10 nucleosides connected, followed by a second spacer and a third section with 3 nucleosides connected. Here, the third section of the nucleosides of 3 connections represents a section with 6 or 7 nucleosides connected at the most. In various embodiments, the oligonucleotide with one or more spacers comprises a plurality of sections with 6 or 7 nucleosides connected at the most. In various embodiments, each section of the oligonucleotide with one or more spacers has 6 or 7 nucleosides connected at the most. For example, an oligonucleotide can be a 20mer and comprise two spacers, and these two spacers are divided into three separate sections of 20mer, and each section has 6 nucleosides connected. Therefore, each section of an oligonucleotide has a maximum of 6 nucleosides connected. For example, an oligonucleotide can be a 23mer and comprise two spacers, and these two spacers are divided into three separate sections of 23mer, and each section has 7 nucleosides connected. Thus, each segment of the oligonucleotide has a maximum of 7 linked nucleosides. In general, oligonucleotides comprising one or more spacers are described with reference to the corresponding parent oligonucleotide or a corresponding oligonucleotide variant. Exemplary oligonucleotides comprising one or more spacers include any of the sequences in SEQ ID NOs: 301692-301742 or in Tables 4A-4C.

[0191] In various embodiments, one or more spacers can be located at one or more positions of an oligonucleotide. A spacer can be located between the first position and the second position of an oligonucleotide. As used herein, the spacer located between the first position and the second position encompasses a spacer located at the first position, located at the second position, or located at any position of the oligonucleotide that is sandwiched between the first position and the second position.

[0192] The term "gapmer antisense oligonucleotide", "gapmer AON" or "gapmer AON variant" refers to an AON having at least three different structural regions in the "5→3" direction, including a 5' wing, a central region, and a 3'-wing. The central region comprises a stretch of nucleosides capable of recruiting and activating RNAseH. For example, the central region comprises linked DNA nucleosides, 2'-fluoroarabinoic acid (FANA), and fluorocyclohexenyl nucleic acid (F-CeNA). Example gapmer AONs or gapmer oligonucleotide variants comprise any one of SEQ ID NOs: 301595-301607. In various embodiments, the gapmer AON comprises a sequence that is at least 85%, at least 90%, at least 95%, at least 98%, or 100% identical to an equal length portion of any one of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, or 167805-301566. In various embodiments, the gapmer AON can comprise one or more spacers. Exemplary gapmer AONs comprising one or more spacers comprise sequences that are at least 85%, at least 90%, at least 95%, at least 98%, or 100% identical to an equal length portion of any one of SEQ ID NOs: 301692-301742 or a sequence in Tables 4A-4C.

[0193] The term "pharmaceutically acceptable salt" as used herein refers to salts of acidic or basic groups that may be present in the antisense oligonucleotides used in the compositions of the present invention. The naturally basic antisense oligonucleotides included in the compositions of the present invention are capable of forming a variety of salts with various inorganic and organic acids. Acids that can be used to prepare pharmaceutically acceptable acid addition salts of such basic compounds are those that form non-toxic acid addition salts, for example, salts containing pharmacologically acceptable anions, including but not limited to malate, oxalate, chloride, bromide, iodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, isonicotinate, acetate, lactate, salicylate, citrate, tartrate, oleate, tannate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisate, fumarate, gluconate, glucuronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, and pamoate (e.g., 1,1'-methylene-bis-(2-hydroxy-3-naphthoate)). The antisense oligonucleotides containing amino moieties contained in the compositions of the present invention can form pharmaceutically acceptable salts with various amino acids in addition to the acids listed above. The naturally acidic compounds included in the compositions of the present invention are capable of forming base salts with various pharmacologically acceptable cations. Examples of such salts include alkali metal salts or alkaline earth metal salts, particularly calcium salts, magnesium salts, sodium salts, lithium salts, zinc salts, potassium salts, and iron salts. Pharmaceutically acceptable salts of the present disclosure include, for example, pharmaceutically acceptable salts of AONs comprising the nucleotide sequence of any of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, 167805-301566, SEQ ID NOs: 301692-301742, or the sequences in Tables 4A-4C.

[0194] The antisense oligonucleotides of the present disclosure can contain one or more chiral centers, groups, bonds and / or double bonds, and therefore exist as stereoisomers, such as geometric isomers, enantiomers or diastereomers.Term used herein " stereoisomer " is made up of all geometric isomers, enantiomers or diastereomers.These compounds can be represented by symbol " R " or " S " (or " Rp " or " Sp "), depending on the configuration of the substituent around stereoatoms (such as stereocarbon, phosphorus or sulphur atom).In some embodiments, one or more bonds of compound can have Rp or Sp configuration (such as, one or more thiophosphate bonds have Rp or Sp configuration).The configuration of each thiophosphate bond can be independent of another thiophosphate bond (such as, a thiophosphate bond has Rp configuration, and second thiophosphate bond has Sp configuration).The present invention covers various stereoisomers and mixtures thereof of these compounds.Stereoisomers comprise enantiomers and diastereomers. A mixture of enantiomers or diastereomers may be designated as "(±)" in the nomenclature, but the skilled artisan will recognize that a structure may implicitly represent a chiral center. Individual stereoisomers of the antisense oligonucleotides of the invention may be prepared synthetically from commercially available starting materials containing asymmetric or stereogenic centers, or by preparing a racemic mixture followed by resolution methods well known to those skilled in the art. Examples of such resolution methods are as follows: (1) attaching the enantiomeric mixture to a chiral auxiliary, separating the resulting diastereomeric mixture by recrystallization or chromatography, and liberating the optically pure product from the auxiliary, (2) forming a salt using an optically active resolving agent, or (3) directly separating the optical enantiomer mixture on a chiral chromatographic column. Stereoisomeric mixtures may also be resolved into their component stereoisomers by well-known methods, such as chiral gas chromatography, chiral supercritical fluid chromatography, chiral simulated moving bed chromatography, chiral high performance liquid chromatography, 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.

[0195] The individual stereoisomeric antisense oligonucleotides of the present invention can be synthesized from commercially available starting materials containing asymmetric or stereogenic centers, or by preparing a racemic mixture followed by resolution methods well known to those skilled in the art. Examples of these resolution methods are as follows: (1) attaching the enantiomeric mixture to a chiral auxiliary, separating the resulting diastereomeric mixture by recrystallization or chromatography, and releasing the optically pure product from the auxiliary, (2) forming a salt using an optically active resolving agent, or (3) directly separating the optical enantiomer mixture on a chiral chromatographic column. Stereoisomeric mixtures can also be resolved into their component stereoisomers by well-known methods, such as chiral supercritical fluid chromatography, chiral simulated moving bed chromatography, chiral gas chromatography, chiral 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 stereoisomerically pure intermediates, reagents, and catalysts by well-known asymmetric synthesis methods.

[0196] 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 it is intended that the present invention embrace both solvated and unsolvated forms.

[0197] The present invention also encompasses isotopically labeled compounds of the invention (e.g., isotopically labeled antisense oligonucleotides) that are identical to those described herein except that one or more atoms are replaced by an atom having an atomic mass or mass number different from the most abundant atomic mass or mass number in nature. Examples of isotopes that can be incorporated into the compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, and chlorine, for example, 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 Cl.

[0198] Certain isotopically labeled disclosed compounds (e.g., 3 H and 14 C-labeled) for use in compound and / or substrate tissue distribution assays. 3 H), carbon 14 (i.e. 14 C) or 35S phosphorothioate isotopes are particularly preferred for their ease of preparation and detectability. Further, substitution with heavier isotopes such as deuterium (i.e., 2H) may afford certain therapeutic advantages resulting from greater metabolic stability (e.g., increased in vivo half-life or reduced dosage requirements) and therefore may be preferred in some circumstances.

[0199] As used herein, "2'-O-(2-methoxyethyl)" (also known as 2'-MOE and 2'-O(CH2)2OCH3 and MOE) refers to an O-methoxyethyl modification of the 2' position of the furanose ring. In this disclosure, 2'-O-(2-methoxyethyl) is used interchangeably as "2'-O-methoxyethyl." The sugar moiety in a nucleoside modified with 2'-MOE is a modified sugar.

[0200] As used herein, "2'-MOE nucleoside" (also known as 2'-O-(2-methoxyethyl) nucleoside) refers to a nucleoside comprising a 2'-MOE modified sugar moiety.

[0201] As used herein, "2'-substituted nucleoside" refers to a nucleoside comprising a substitution other than H or OH at the 2' position of the furanose ring. In certain embodiments, the 2'-substituted nucleoside comprises a nucleoside having a bicyclic sugar modification.

[0202] As used herein, "5-methylcytosine" (5-MeC) refers to cytosine modified with a methyl group attached to position 5. 5-Methylcytosine (5-MeC) is a modified nucleobase.

[0203] As used herein, "bicyclic sugar" refers to a furanose ring modified by bridging two atoms. Bicyclic sugars are modified sugars.

[0204] As used herein, "bicyclic nucleoside" (also known as BNA) refers to a nucleoside having a sugar moiety comprising a bridge connecting the two carbon atoms of the sugar ring, thereby forming a bicyclic ring system. In certain embodiments, the bridge connects the 4'-carbon and the 2'-carbon of the sugar ring.

[0205] As used herein, "cEt" or "contrained ethyl" refers to a bicyclic nucleoside having a sugar moiety comprising a bridge connecting the 4'-carbon and the 2'-carbon, wherein the bridge has the formula: 4'-CH(CH3)-O-2'.

[0206] As used herein, "constrained ethyl nucleoside" (also referred to as cEt nucleoside) refers to a nucleoside comprising a bicyclic sugar moiety comprising a 4'-CH(CH3)-O-2' bridge. In some embodiments, cEt can be modified. In some embodiments, cEt can be S-cEt (in S-constrained ethyl 2'-4'-bridged nucleic acids). In some other embodiments, cEt can be R-cEt.

[0207] As used herein, "internucleoside linkage" refers to an atom or group linking 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 a "modified internucleoside linkage".

[0208] As used herein, "continuous" in the context of an oligonucleotide refers to nucleosides, nucleobases, sugar moieties, or internucleoside linkages that are immediately adjacent to one another. For example, "continuous nucleobases" refers to nucleobases that are immediately adjacent to one another in a sequence. As a negative example, two nucleosides separated by a spacer are not continuous.

[0209] As used herein, "modified nucleobase" means any nucleobase other than adenine, cytosine, guanine, thymine, or uracil. Examples of modified nucleobases include 5-methylcytosine, pseudouridine, or 5-methoxyuridine. "Unmodified nucleobase" refers to the purine bases adenine (A) and guanine (G), and the pyrimidine bases thymine (T), cytosine (C), and uracil (U).

[0210] As used herein, " spacer " refers to a nucleoside replacement group (for example, a non-nucleoside group that replaces the nucleoside present in the parent oligonucleotide). Spacer is characterized in that it lacks a nucleotide base and replaces the nucleoside sugar moiety with a non-sugar substitute. The non-sugar substitute of spacer lacks an aldehyde, ketone, acetal, ketal, hemiacetal or hemiketal group. Therefore, the non-sugar substitute of spacer can be connected to the 3' and 5' positions of the nucleoside adjacent to the spacer by an internucleoside linker as described herein, but can not form a covalent bond with the nucleotide base (that is, core base can not be connected to another group, such as the terminal group in internucleoside bond, conjugated group or oligonucleotide). Generally speaking, the antisense oligonucleotide with a spacer is described relative to the parent antisense oligonucleotide, and wherein the spacer replaces the nucleoside of the parent antisense oligonucleotide. In all embodiments of the present disclosure, the spacer cannot hybridize to a nucleoside comprising a nucleobase at a corresponding position of a transcript (e.g., a pre-mRNA or mRNA transcript) within the numerical sequential length of the AON (i.e., if the spacer is located after nucleoside 4 of the AON (i.e., the 5th position 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).

[0211] As used herein, "modified nucleosides" refers to nucleosides that independently have a modified sugar moiety and / or a modified nucleobase. A universal base is a modified nucleobase that can pair with any of the five unmodified nucleobases. Modified nucleosides include abasic nucleosides that lack a nucleobase. However, modified nucleosides do not include spacers or other groups that cannot connect nucleobases.

[0212] As used herein, " nucleosides connected " are nucleosides connected in continuous sequence (that is, there is no extra nucleosides between those nucleosides connected). In various embodiments, oligonucleotide can have the nucleoside segments of different connections connected by spacers. Here, spacers (that is, nucleosides replace) are not considered to be nucleosides, so oligonucleotide is divided into the segment of two nucleosides connected. Oligonucleotide can have the first segment of the nucleosides connected by Y (for example, the Y nucleosides connected in continuous sequence), followed by a spacer, then the second segment of the nucleosides connected by Z. Herein, the nucleosides connected by Y and Z are described in 5' to 3' direction or 3' to 5' direction. In various embodiments, the first segment is made up of 7 or less nucleosides connected (for example, Y=7 or less), and the second segment comprises 8 or more nucleosides connected (for example, Z=8 or more).

[0213] As used herein, "locked nucleic acid" or "LNA" or "LNA nucleoside" refers to a nucleic acid monomer having a bridge (e.g., a methylene, ethylene, aminooxy, or oxyimino bridge) connecting the two carbon atoms between the 4' and 2' positions of the nucleoside sugar unit to form a bicyclic sugar. Examples of such bicyclic sugars include, but are not limited to, (A) α-L-methyleneoxy (4'-CH2-O-2') LNA, (B) β-D-methyleneoxy (4'-CH2-O-2') LNA, (C) ethoxy (4'-(CH2)2-O-2') LNA, (D) aminooxy 4'-CH2-ON(R)-2') LNA, and (E) oxyamino (4'-CH2-N(R)-O-2') LNA; wherein R is H, C1-C 12 an alkyl group, or a protecting group (see U.S. Patent No. 7,427,672 issued September 23, 2008).

[0214] As used herein, LNA compounds include, but are not limited to, compounds having at least one bridge between the 4' and 2' positions of the sugar, wherein each bridge independently comprises 1 or 2 to 4 linking groups independently selected from: -[C(R1)(R2)] n -, -C(R1)=C(R2)-, -C(R1)=N-, -C(=NR1)-, -C(=O)-, -C(=S)-, -O-, -Si(R1)2-, -S(=O) x -, and -N(R1)-; wherein: x is 0, 1, or 2; n is 1, 2, 3, or 4; each R1 and R2 is independently H, a protecting group, a hydroxyl group, a C1-C 12 Alkyl, substituted C 1- C 12 Alkyl, C2-C 12 Alkenyl, substituted C2-C 12 Alkenyl, C2-C 12Alkynyl, substituted C2-C 12 Alkynyl, C5-C 20 Aryl, substituted C5-C 20 aryl, heterocyclic, substituted heterocyclic, heteroaryl, substituted heteroaryl, C5-C7 alicyclic, substituted C5-C7 alicyclic, halogen, OJ1, NJ1J2, SJ1, N3, COOJ1, acyl (C(=O)-H), substituted acyl, CN, sulfonyl (S(=O)2-J1), or sulfinyl (S(=O)-J1); and each J1 and J2 are independently H, C1-C 12 Alkyl, substituted C1-C 12 Alkyl, C2-C 12 Alkenyl, substituted C2-C 12 Alkenyl, C2-C 12 Alkynyl, substituted C2-C 12 Alkynyl, C5-C 20 Aryl, substituted C5-C 20 Aryl, acyl (C (= O) -H), substituted acyl, heterocyclic, substituted heterocyclic, C1-C 12 Aminoalkyl, substituted C1-C 12 aminoalkyl, or a protecting group.

[0215] Examples of 4'-2' bridges encompassed within the definition of LNA include, but are not limited to, one of the following formulas: -[C(R1)(R2)] n -、-[C(R1)(R2)] n -O-, -C(R1R2)-N(R1)-O-, or -C(R1R2)-ON(R1)-. In addition, other bridge groups encompassed by the definition of LNA are 4'-CH2-2', 4'-(CH2)2-2', 4'-(CH2)3-2', 4'-CH2-O-2', 4'-(CH2)2-O-2', 4'-CH2-ON(R1)-2', and 4'-CH2-N(R1)-O-2'-bridges, wherein each R1 and R2 are independently H, a protecting group, or a C1-C 12 alkyl.

[0216] Also included in the definition of LNA according to the present invention is that wherein the 2 '-hydroxyl of ribosyl sugar ring is connected to 4 ' carbon atom of sugar ring, thus forms bridge to form the LNA of dicyclic sugar moiety.Bridge can be the methylene (-CH2-) group connecting 2 ' oxygen atom and 4 ' carbon atom, wherein used term methyleneoxy (4 '-CH2-O-2 ') LNA.In addition, in the case of the dicyclic sugar moiety with ethylene bridge group in this position, use term ethyleneoxy (4 '-CH2CH2-O-2 ') LNA.As used herein, the isomer of AL-methyleneoxy (4 '-CH2-O-2 '), methyleneoxy (4 '-CH2-O-2 ') LNA is also encompassed within the definition of LNA.

[0217] As used herein, "hybridization" refers to the pairing or annealing of complementary oligonucleotides and / or nucleic acids. Although not limited to a particular mechanism, the most common hybridization mechanism involves hydrogen bonding, which can be Watson-Crick, Hoosteen, or reversed Hoosteen hydrogen bonding between complementary nucleobases.

[0218] As used herein, "increasing the amount of activity" refers to greater transcript expression, more accurate splicing resulting in full-length mature mRNA and / or protein expression, and / or greater activity relative to transcript expression or activity in an untreated or control sample.

[0219] As used herein, "mismatch" or "non-complementary nucleobase" refers to the situation when a group (e.g., nucleobase) of a first nucleic acid is unable to pair with a corresponding group (e.g., nucleobase) of a second or target nucleic acid.

[0220] As used herein, a "modified internucleoside linkage" refers to a substitution or any alteration from a naturally occurring internucleoside linkage (e.g., a phosphodiester internucleoside linkage). A "phosphorothioate linkage" is a modified internucleoside linkage in which one of the non-bridging oxygen atoms of the phosphodiester internucleoside linkage is replaced by a sulfur atom.

[0221] As used herein, a "modified oligonucleotide" refers to an oligonucleotide comprising at least one (ie, one or more) modified internucleoside linkage, modified sugar, and / or modified nucleobase.

[0222] As used herein, "modified sugar" or "modified sugar moiety" refers to a modified furanosyl sugar moiety or a sugar moiety having a modified moiety other than a furanosyl moiety that can link a nucleobase to another group, such as an internucleoside linkage, a conjugate group, or a terminal group in an oligonucleotide.

[0223] As used herein, "monomer" refers to a single unit of an oligomer. Monomers include, but are not limited to, nucleosides and nucleotides, whether naturally occurring or modified.

[0224] As used herein, "motif" refers to the pattern of unmodified and modified nucleosides in an antisense compound.

[0225] As used herein, "natural sugar moiety" refers to a sugar moiety found in DNA (2'-H) or RNA (2'-OH).

[0226] As used herein, "naturally occurring internucleoside linkage" refers to a 3' to 5' phosphodiester linkage.

[0227] As used herein, "nucleobase" refers to a heterocyclic moiety capable of base pairing with another nucleic acid.

[0228] As used herein, "nucleobase complementarity" refers to a nucleobase that can base pair with another nucleobase. For example, in DNA, adenine (A) is complementary to thymine (T). For example, in RNA, adenine (A) is complementary to uracil (U). In certain embodiments, complementary nucleobases refer to nucleobases of antisense compounds that can base pair with nucleobases of their target nucleic acids. For example, if a nucleobase at a certain position of the antisense compound can hydrogen bond with a nucleobase at a certain position of the target nucleic acid, the hydrogen bonding position between the oligonucleotide and the target nucleic acid is considered to be complementary at that nucleobase pair.

[0229] As used herein, "non-complementary nucleobases" refers to a pair of nucleobases that do not hydrogen bond, or otherwise support hybridization, with each other.

[0230] As used herein, "nucleic acid" refers to a molecule composed of monomeric nucleotides. Nucleic acids include, but are not limited to, ribonucleic acid (RNA), deoxyribonucleic acid (DNA), single-stranded nucleic acid, double-stranded nucleic acid, non-coding RNA, small interfering ribonucleic acid (siRNA), short hairpin RNA (shRNA), circular RNA, circular DNA, and microRNA (miRNA).

[0231] As used herein, "nucleobase sequence" refers to the order of nucleobases independent of any sugar, linkage and / or nucleobase modifications.

[0232] As used herein, "nucleoside" refers to a nucleobase linked to a sugar. The term "nucleoside" also encompasses "modified nucleosides" independently having a modified sugar moiety and / or a modified nucleobase.

[0233] As used herein, " oligonucleotide unit " refers to the nucleoside (for example, the nucleoside comprising sugar and / or core base) or nucleoside replacement group (for example, spacer) of oligonucleotide.Oligonucleotide unit covers nucleoside, modified nucleoside and spacer.For example, oligonucleotide can have 20 oligonucleotide units, wherein one of oligonucleotide unit is spacer, and other 19 oligonucleotide units are nucleoside or modified nucleoside.As another example, oligonucleotide can have 20 oligonucleotide units, wherein two in oligonucleotide unit are spacer, and other 18 oligonucleotide units are nucleoside or modified nucleoside.

[0234] As used herein, " nucleoside mimics " comprises those structures for replacing sugar or sugar and base, and is not necessarily the bond at one or more positions of oligomeric compound, for example, has morpholino, cyclohexenyl, cyclohexyl, tetrahydropyranyl, dicyclo or tricyclic sugar mimics, (for example, the nucleoside mimics of non-furanose unit). Nucleotide mimics comprises those structures for replacing nucleosides at one or more positions of oligomeric compound and bond, for example peptide nucleic acid or morpholino (morpholino connected by phosphorodiamidate or other non-phosphodiester bonds). Sugar substitute overlaps with slightly broad term " nucleoside mimics ", but is intended to only represent the replacement of sugar unit (furanose ring). Tetrahydropyranyl ring provided herein is the example of sugar substitute, and wherein furanose group is replaced by tetrahydropyranyl ring system." stand-ins " refers to the group substituted by sugar, core base and / or nucleoside bond. Typically, stand-ins are used to replace sugar or sugar-nucleoside bond combination, and retain core base to hybridize with selected target.

[0235] As used herein, "nucleotide" refers to a nucleoside having a phosphate group covalently linked to the sugar portion of the nucleoside.

[0236] As used herein, "oligomeric compound" or "oligomer" refers to a polymer of linked monomeric subunits capable of hybridizing to at least a region of a nucleic acid molecule.

[0237] As used herein, "oligonucleotide" refers to a polymer of one or more linked nucleoside segments, wherein each segment may be modified or unmodified independently of the others.

[0238] The present disclosure provides methods for treating, ameliorating, or preventing a disease in a patient, such as, but not limited to, 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, synucleinopathies, Huntington's disease, brachial plexus injury, peripheral nerve injury, progressive supranuclear palsy (PSP), traumatic brain injury, spinal cord injury, tuberous sclerosis, Pick's disease, tauopathies, primary age-related tauopathies, Down syndrome, epilepsy / seizure disorders, depression, traumatic brain injury (TBI), chronic traumatic encephalopathy (CTE), HIV-associated neurocognitive disorder (HAND), multiple system atrophy, hereditary Amnesiac mild cognitive impairment, corticobasal degeneration (CBD) and / or neuropathy such as chemotherapy-induced neuropathy, spinocerebellar ataxia (SCA), type 2 SCA, spinal muscular atrophy (SMA), Parkinson's disease, Niemann-Pick disease type C (NPC), Charcot-Marie-Tooth disease (CMT), mucopolysaccharidosis type II (MESIIA), mucolipidosis type IV, GM1 gangliosidosis, sporadic inclusion body myositis (sIBM), Henoch-Schonlein purpura (HSP), Gaucher disease, facial onset sensory and motor neuron disease, Guam-type parkinsonism, multisystem proteinopathy, Perry disease, and autism, the method comprising administering to the patient a composition comprising a therapeutically effective amount of an antisense oligonucleotide and a pharmaceutically acceptable excipient.

[0239] Also provided herein are methods for treating, improving 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, improving or preventing a disease comprises administering to a patient a pharmaceutically acceptable composition comprising one or more antisense oligonucleotides, such as a pharmaceutically acceptable formulation. Antisense oligonucleotides disclosed herein can target transcripts (e.g., mRNA or pre-mRNA transcripts) of any one of PPM1A, ATXN2, SOD1, or MAPT. Antisense oligonucleotides can regulate the expression of transcripts to treat diseases disclosed herein.

[0240] The disclosure also provides pharmaceutical compositions, it comprises the antisense oligonucleotide as disclosed herein prepared together with one or more pharmaceutically or cosmetically acceptable excipients.These preparations comprise those suitable for oral, sublingual, intratracheal, intranasal, vaginal, rectal, local, percutaneous, pulmonary, intrathecal, intracerebral cistern, intrathecal, intrathalamic, intraventricular, intraocular, oral and parenteral (such as subcutaneous, intramuscular, intradermal, intraduodenal or intravenous) use, or for topical use, such as as the part of the composition suitable for topical application to skin and / or mucous membrane, such as gel, paste, wax, cream, spray, liquid, foam, lotion, ointment, external solution, transdermal patch, powder, steam or tincture form.Although the most suitable form of application in any given case will depend on the degree of the treated patient's condition and the property of the specific antisense oligonucleotide used of severity and.

[0241] The present invention also provides pharmaceutical compositions comprising antisense oligonucleotides, or pharmaceutically acceptable salts thereof. The present disclosure also provides methods comprising using an AON comprising an antisense oligonucleotide as disclosed herein (e.g., an AON comprising the sequence of any one of the antisense oligonucleotides 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 the sequence of any one of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, or 167805-301566, such as 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 antisense oligonucleotides as described above and one or more pharmaceutically acceptable excipients. Formulations include those suitable for oral, sublingual, intratracheal, intranasal, rectal, vaginal, topical, transdermal, pulmonary, intrathecal, intracisternal, intrathecal, intrathalamic, intracerebroventricular, intraocular, oral, and parenteral (e.g., subcutaneous, intramuscular, intradermal, intraduodenal, or intravenous) administration or topical use. The most appropriate form of administration in any given case will depend on the clinical symptoms, complications, or biochemical indicators of the condition, disorder, disease, or condition of the subject that one is trying to prevent; the condition, disorder, disease, or condition of the subject that one is trying to prevent; and / or the nature of the specific compound and / or composition used.

[0242] Antisense therapeutics

[0243] Antisense therapeutics are a class of nucleic acid-based compounds that can be used to inhibit gene expression. Antisense therapeutics can be based on single-stranded or double-stranded deoxyribonucleic acid (DNA), based on ribonucleic acid (RNA) or DNA / RNA chemical analog compounds. Generally speaking, antisense therapeutics are designed to include nucleotide sequences that are complementary or almost complementary to the mRNA or pre-mRNA sequence of a given gene to promote the binding between the antisense therapeutic and the pre-mRNA or mRNA. Examples of given genes disclosed herein include, but are not limited to, PPM1A, ATXN2, SOD1, or MAPT.

[0244] Without being bound by theory, it is believed that in most cases, antisense therapeutics act by binding to mRNA or pre-mRNA, thereby inhibiting protein translation, altering pre-mRNA splicing into mature mRNA, and / or causing mRNA destruction. In most cases, the antisense therapeutic nucleotide sequence is complementary to a portion of the sense sequence of the target gene, mRNA, or pre-mRNA. The antisense therapeutics described herein are oligonucleotide-based compounds comprising an oligonucleotide sequence that is complementary to a gene sense sequence, a pre-mRNA sense sequence, and / or an mRNA sense sequence, or a portion thereof. The antisense therapeutics described herein can also be compounds based on nucleotide chemical analogs that are capable of binding to a gene sense sequence, a pre-mRNA sense sequence, and / or an mRNA sense sequence, or a portion thereof. Antisense therapeutics include antisense oligonucleotides, shRNAs, siRNAs, PNAs, LNAs, and morpholino oligomers.

[0245] Antisense oligonucleotide (AON) is a short sequence based on oligonucleotide, comprising an oligonucleotide sequence complementary to a target RNA sequence. The length of AON is generally between 8 to 50 nucleotides, for example, 18 nucleotides in length, 20 nucleotides in length or 23 nucleotides in length. AON can comprise chemically modified nucleosides (for example, 2'-O-methylated nucleosides or 2'-O-(2-methoxyethyl) nucleosides) and modified internucleoside bonds (for example, phosphorothioate bonds). AON as herein described comprises an oligonucleotide sequence complementary to an RNA sequence, for example, mRNA or pre-mRNA transcripts. AON as herein described can comprise chemically modified nucleosides and modified internucleoside bonds (for example, phosphorothioate bonds).

[0246] Peptide nucleic acids (PNAs) are short, artificially synthesized polymers with a structure similar to DNA or RNA. PNAs contain a backbone composed of repeating N-(2-aminoethyl)-glycine units linked by peptide bonds. The PNAs described herein can be used as antisense therapeutics, binding to mRNA or pre-mRNA sequences with high specificity and inhibiting target gene expression.

[0247] Locked nucleic acids (LNA) are oligonucleotide sequences containing one or more modified RNA nucleotides in which the ribose moiety is modified with an additional bridge connecting the 2' oxygen and the 4' carbon. LNA is believed to have a higher Tm than similar oligonucleotide sequences. The LNA described herein can be used as antisense therapeutics, which bind to RNA sequences with high specificity and inhibit target gene expression.

[0248] Morpholino oligomers are oligonucleotide compounds comprising a DNA base attached to a methylenemorpholine ring backbone connected by a phosphorodiamidate group. The morpholino oligomers of the present invention can be designed to bind to a specific RNA sequence of interest (e.g., an mRNA or pre-mRNA sequence of interest), thereby preventing gene expression. The morpholino oligomers described herein can be used as antisense therapeutics 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, changing pre-mRNA splicing and gene expression.

[0249] Small hairpin RNA (shRNA) is typically an RNA molecule with a hairpin-like structure that can be used to silence gene expression. shRNA is typically expressed by a plasmid encoding the shRNA sequence, and can be expressed by a viral vector to allow slow virus, adenovirus or adeno-associated virus to express. Without being bound by theory, it is believed that shRNA inhibits gene expression by utilizing RNA interference (RNAi) steps. In short, shRNA transcripts are processed by Drosha and Dicer, and are then loaded onto the RNA-induced silencing complex (RISC), allowing targeting specific mRNA, and degrading mRNA or inhibiting protein translation. shRNA as described herein can inhibit target gene expression.

[0250] Small interfering RNA (siRNA) is a double-stranded RNA molecule of about 20-25 base pairs in length that uses RNAi mechanisms (such as Drosha and RISC) to bind and target mRNA for degradation. siRNA does not rely on plasmids or vectors for expression and can usually be delivered directly to target cells, such as by transfection. siRNA is a double-stranded RNA sequence that contains an RNA sequence complementary to the mRNA sequence and prevents protein translation.

[0251] In some embodiments, the length of the antisense oligonucleotide is 12 to 15 oligonucleotide units. In some embodiments, the length of the antisense oligonucleotide is 15 to 20 oligonucleotide units. In some embodiments, the length of the antisense oligonucleotide is 20 to 40 oligonucleotide units. In some embodiments, the length of the antisense oligonucleotide is 20 to 22 oligonucleotide units. In some embodiments, the length of the antisense oligonucleotide is 22 to 40 oligonucleotide units. In some embodiments, the length of the antisense oligonucleotide is 20 to 30, 25 to 35 or 30 to 40 oligonucleotide units. In specific embodiments, the length of the antisense oligonucleotide is 18 oligonucleotide units. In specific embodiments, the length of the antisense oligonucleotide is 20 oligonucleotide units. In specific embodiments, the length of the antisense oligonucleotide is 23 oligonucleotide units.

[0252] antisense oligonucleotides

[0253] The antisense oligonucleotides (AONs) described herein are short synthetic oligonucleotide sequences that are complementary to a portion of a gene product, such as any 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).

[0254] In various embodiments, AON comprises 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, such as an mRNA or pre-mRNA sequence. In some embodiments, AON can comprise a non-double-stranded oligonucleotide. In some embodiments, AON can comprise a double strand of two oligonucleotides, wherein the first oligonucleotide comprises a nucleotide sequence that is completely or almost completely complementary to the target mRNA sequence, and the second oligonucleotide comprises a nucleotide sequence that is complementary to the nucleic acid sequence of the first oligonucleotide. AON binding specificity can be assessed by measuring parameters (such as dissociation constant, melting temperature (Tm)), or other standards (such as changes in protein or RNA expression levels) or other assays that measure target activity or expression.

[0255] AON, such as disclosed herein, can be an oligonucleotide sequence of 5 to 100 oligonucleotide units in length, for example, 10 to 40 oligonucleotide units in length, for example, 14 to 40 oligonucleotide units in length, for example, 10 to 30 oligonucleotide units in length, for example, 14 to 30 oligonucleotide units in length, for example, 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 a specific embodiment, the AON is 18 oligonucleotide units in length. In a specific embodiment, the AON is 20 oligonucleotide units in length. In a specific embodiment, the AON is 23 oligonucleotide units in length. In a specific embodiment, the AON comprises 20 nucleosides connected. As used herein, " parent oligonucleotide " refers to an AON comprising 20 nucleosides connected.

[0256] The AONs described herein also include antisense oligonucleotides comprising the oligonucleotide sequences listed in Tables 1A-1D. Specifically, Table 1A shows example PPM1A AONs. In various embodiments, the PPM1A AON comprises the sequence of any one of SEQ ID NOs: 1-954. Table 1B shows example ATXN2 AONs. In various embodiments, the ATXN2 AON comprises the sequence of any one of SEQ ID NOs: 1914-149354. Table 1C shows example SOD1 AONs. In various embodiments, the SOD1 AON comprises the sequence of any one of SEQ ID NOs: 149362-158581. Table 1D shows example MAPT AONs. In various embodiments, the MAPT AON comprises the sequence of any one of SEQ ID NOs: 167805-301566.

[0257] In various embodiments, the AONs shown in Tables 1A-1D comprise at least one nucleoside linkage selected from the group consisting of phosphorothioate linkages, alkylphosphate linkages, alkylphosphate linkages, 3-methoxypropylphosphate linkages, phosphorodithioate linkages, phosphotriester linkages, methylphosphate linkages, aminoalkylphosphotriester linkages, alkylenephosphonate linkages, phosphite linkages, phosphoramidate linkages, thiophosphoramidate linkages, phosphorodiamidate (e.g., comprising phosphorodiamidate morpholino (PMO), 3' aminoribose, or 5' aminoribose) linkages, aminoalkylphosphodiamidate linkages, thiophosphoramidate linkages, thioalkylphosphate linkages, thioalkylphosphotriester linkages, phosphorothioate linkages, phosphoroselenoate linkages, and phosphoroboronate linkages.

[0258] In specific embodiments, disclosed herein are gapmer oligonucleotides comprising a sequence that is at least 85% identical 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 Tables 4A-4C. In specific embodiments, disclosed herein are gapmer oligonucleotides comprising a sequence that is at least 90% identical 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 Tables 4A-4C. In specific embodiments, disclosed herein are gapmer oligonucleotides comprising a sequence that is at least 95% identical 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 Tables 4A-4C. In specific embodiments, disclosed herein are gapmer oligonucleotides comprising a sequence that is at least 98% identical 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 Tables 4A-4C. In specific embodiments, disclosed herein are gapmer oligonucleotides comprising a sequence 100% identical 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 Tables 4A-4C.

[0259] Oligonucleotide variants

[0260] In various embodiments, AON comprises different variants (e.g., AON of different lengths), hereinafter referred to as AON variants. AON variants can be oligonucleotide sequences of 5 to 100 core bases in length, for example, 10 to 40 core bases in length, for example, 14 to 40 core bases in length, 10 to 30 core bases in length, for example, 14 to 30 core bases in length, for example, 16 to 28 core bases in length, for example, 19 to 23 core bases in length, for example, 18 to 21 core bases in length, or for example, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 core bases 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.

[0261] In various embodiments, the AON variant represents a modified form of the 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, the AON variant comprises a nucleobase sequence that represents a shortened form of the nucleobase sequence of a nucleoAON selected from any one of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, or 167805-301566. As an example, if the parent oligonucleotide comprises a 20mer (e.g., 20 nucleotide bases in length), the variant (e.g., AON variant) can comprise a shorter version (e.g., 15mer, 16mer, 17mer, 18mer, or 19mer) of the 20mer parent oligonucleotide or a longer version (e.g., 21mer, 22mer, 23mer, 24mer, 25mer, 26mer, 27mer, 28mer, 29mer, or 30mer) of the 20mer parent oligonucleotide. 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 are removed or added from one or both of the 5' and 3' ends of the nucleobase sequence of the parent oligonucleotide.

[0262] In one embodiment, the corresponding AON variant can comprise an 18mer wherein one nucleotide base is removed from each of the 3' and 5' ends of the 20mer comprised in the parent oligonucleotide. For example, an 18mer AON variant can comprise a sequence wherein one nucleotide base is 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.

[0263] In one embodiment, the corresponding AON variant may comprise an 18mer wherein two nucleotide bases are removed from the 3' end of the 20mer contained in the parent oligonucleotide. For example, an 18met AON variant may comprise a sequence wherein two nucleotide bases are 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 18mer wherein two nucleotide bases are removed from the 5' end of the 20mer contained in the parent oligonucleotide. For example, an 18mer AON variant may comprise a sequence wherein two nucleotide bases are removed from the 5' end of any one of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, or 167805-301566.

[0264] In one embodiment, the corresponding AON variant can comprise a 19mer wherein one nucleotide base is removed from the 3' or 5' end of the 20mer contained in the parent oligonucleotide. For example, a 19mer AON variant can comprise a sequence wherein one nucleotide base is removed from the 3' end of any one of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, or 167805-301566. For example, a 19mer AON variant can comprise a sequence wherein one nucleotide base is removed from the 5' end of any one of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, or 167805-301566.

[0265] In one embodiment, the corresponding AON variant can comprise a 23mer wherein three nucleotide bases are added from the 3' or 5' end of the 20mer comprised in the parent oligonucleotide. For example, a 19mer AON variant can comprise a sequence wherein three nucleotide bases are added from the 3' or 5' end of any one of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, or 167805-301566.

[0266] Exemplary sequences of AON variants are shown below in Tables 2A-2B.

[0267] Table 2A. Examples of MAPT AON variant sequences.

[0268]

[0269] Table 2B. Examples of PPM1A AON variant sequences

[0270]

[0271]

[0272] In Tables 2A and 2B, "*" indicates that at least one nucleoside linkage of the nucleobase sequence is selected from the group consisting of phosphorothioate linkages, alkylphosphate linkages, phosphorodithioate linkages, phosphotriester linkages, alkylphosphate linkages, 3-methoxypropylphosphate linkages, methylphosphonate linkages, aminoalkylphosphotriester linkages, alkylenephosphonate linkages, phosphite linkages, phosphoramidate linkages, thiophosphoramidate linkages, phosphorodiamidate linkages, phosphorodiamidate (e.g., comprising phosphorodiamidate morpholino (PMO), 3' aminoribose, or 5' aminoribose) linkages, aminoalkylphosphoramidate linkages, thiophosphoramidate linkages, thioalkylphosphate linkages, thioalkylphosphotriester linkages, phosphorothioate linkages, phosphoroselenoate linkages, and phosphoroboronate linkages.

[0273] Gapmer AON

[0274] In specific embodiments, the AONs disclosed herein have a gapmer design or structure, also referred to herein as a "gapmer" or "gapmerAON." In a gapmer structure, an AON comprises at least three distinct structural regions, including a 5' wing, a central region, and a 3' wing, in a "5→3" orientation.

[0275] In various embodiments, the central region comprises 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 oligonucleotide units. In various embodiments, at least one oligonucleotide unit of 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 connected nucleosides. In various embodiments, the central region comprises 1, 2, 3, 4, 3, 6, 7, 8, 9 or 10 oligonucleotide units. In various embodiments, at least one oligonucleotide unit of 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 connected nucleosides. The 5' and 3' wing regions (also referred to as flanking regions) comprise at least one oligonucleotide unit adjacent to the central region, and in some embodiments, the central region comprises a continuous nucleoside. The 5' and 3' wings may be symmetrical or asymmetrical with respect to the number of oligonucleotide units or linked nucleosides they comprise.

[0276] 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 RNA nucleosides 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 RNA nucleosides and DNA nucleosides.

[0277] 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 connected nucleosides. In some embodiments, the central region comprises a section of nucleosides that can recruit and activate RNAseH. In some embodiments, the central region comprises one or more of connected DNA nucleosides, 2'-fluoroarabinoic 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 continuous segment of 5-16 DNA nucleosides. In some embodiments, the central region comprises a continuous segment of 6-15, 7-14, 8-13 or 9-11 DNA nucleosides. In various embodiments, the central region comprises a mixture of DNA nucleosides 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.

[0278] In some embodiments, all 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 (2'-fluoroarabinoic acid (FANA) and fluorocyclohexenyl nucleic acid (F-CeNA)) that can mediate RNase H cleavage. In some embodiments, at least 50% of the nucleosides in the central region are DNA nucleosides, for example, 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% DNA nucleosides.

[0279] In a specific embodiment, the AON comprises a 5' wing region of 5 connected nucleosides, a central region of 10 connected nucleosides, and a 3' wing region of 5 connected nucleosides, also referred to as a 5-10-5 gapmer. In a specific embodiment, the AON comprises a 5' wing region of 3 connected nucleosides, a central region of 8 connected nucleosides, and a 3' wing region of 3 connected nucleosides, also referred to as a 3-8-3 gapmer. In a specific embodiment, the AON comprises a 5' wing region of 3 connected nucleosides, a central region of 10 connected nucleosides, and a 3' wing region of 3 connected nucleosides, also referred to as a 3-10-3 gapmer. In a specific embodiment, the AON comprises a 5' wing region of 4 connected nucleosides, a central region of 10 connected nucleosides, and a 3' wing region of 4 connected nucleosides, also referred to as a 4-10-4 gapmer. In a specific embodiment, the AON comprises a 5' wing region of 4 connected nucleosides, a central region of 8 connected nucleosides, and a 3' wing region of 4 connected nucleosides, also referred to as a 4-8-4 gapmer. In various embodiments, at least one nucleoside in any of the 3' wing, the 5' wing, or the center is substituted with a spacer. Embodiments of gapmer AONs having one or more spacers are described in further detail herein.

[0280] In specific embodiments, the gapmer AON disclosed herein comprises the sequence of any one of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, or 167805-301566, wherein the gapmer AON comprises a 5' wing comprising one or more RNA nucleosides, a central region comprising one or more DNA nucleosides, and a 3' wing comprising one or more RNA nucleosides. In specific embodiments, the gapmer AON disclosed herein comprises the sequence of any one of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, or 167805-301566, which is a 5-10-5 gapmer (e.g., a 5' wing of 5 linked nucleosides, a central region of 10 linked nucleosides, and a 3' wing of 5 linked nucleosides). In specific embodiments, the gapmer AON disclosed herein comprises the sequence of any one of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, or 167805-301566, which is a sequence of any one of 6-11-61-954, 1914-149354, 149362-158581, or 167805-301566, which is a 6-11-6 gapmer (e.g., a 5' wing of 6 linked nucleosides, a central region of 11 linked nucleosides, and a 3' wing of 6 linked nucleosides). In specific embodiments, the gapmer AON disclosed herein comprises the sequence of any one of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, or 167805-301566, which is a 5-8-51-954, 1914-149354, 149362-158581, or 167805-301566, which is a 5-8-5 gapmer (e.g., a 5' wing of 5 linked nucleosides, a central region of 8 linked nucleosides, and a 3' wing of 5 linked nucleosides).

[0281] 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).

[0282] Table 3A: Examples of PPM1A Gapmer AONs and Gapmer variant AONs. In the gapmer structure, "e" refers to a ribonucleoside, such as a 2'-MOE modified ribonucleoside, and "d10" or "d11" refers to 10 or 11 DNA deoxyribonucleosides, respectively.

[0283]

[0284]

[0285] Table 3B: Examples of ATXN2 Gapmer AONs. In the gapmer structure, "e" refers to a ribonucleoside, such as a 2'-MOE modified ribonucleoside, and "d10" indicates 10 deoxyribonucleosides.

[0286]

[0287] Table 3C: Examples of SOD1 Gapmer AONs In the gapmer structure, "e" refers to a ribonucleoside, such as a 2'-MOE modified ribonucleoside, and "d10" indicates 10 deoxyribonucleosides.

[0288]

[0289] Table 3D: Examples of MAPT Gapmer AONs and Gapmer variant AONs In the gapmer structure, "e" refers to a ribonucleoside, such as a 2'-MOE modified ribonucleoside, and "d8" or "d10" refers to 8 or 10 deoxyribonucleosides, respectively.

[0290]

[0291] In various embodiments, the gapmer AON, e.g., a gapmer AON comprising the sequence of 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), comprises a mixture of ribonucleosides and deoxyribonucleosides (including modified ribonucleosides and / or modified deoxyribonucleosides) in the 5' wing and / or the 3' wing.

[0292] In specific embodiments, a gapmer AON, e.g., a gapmer AON comprising the sequence of 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), comprises a modified ribonucleoside in the 5' wing and / or the 3' wing. In specific embodiments, each nucleoside in the 5' wing and the 3' wing is a modified nucleoside, e.g., a 2'-O-(2-methoxyethyl) (2'-MOE) nucleoside. For example, each guanosine in the 5' wing or the 3' wing can be a modified 2'-O-(2-methoxyethyl) guanosine. For example, each adenosine in the 5' wing or the 3' wing can be a modified 2'-O-(2-methoxyethyl) adenosine. For example, each cytosine in the 5' wing and the 3' wing can be 2'-O-(2-methoxyethyl)-5-methylcytosine. For example, each thymidine in the 5' wing and the 3' wing can be 2'-O-(2-methoxyethyl)thymidine.

[0293] In various embodiments, exemplary gapmer AONs have one or more modified internucleoside linkages, such as phosphorothioate linkages, alkylphosphate linkages, alkylphosphate linkages, 3-methoxypropylphosphate linkages, phosphorodithioate linkages, phosphotriester linkages, methylphosphonate linkages, aminoalkylphosphotriester linkages, alkylenephosphonate linkages, phosphite linkages, phosphoramidate linkages, thiophosphoramidate linkages, phosphorodiamidate (e.g., comprising phosphorodiamidate morpholino (PMO), 3' aminoribose, or 5' aminoribose) linkages, aminoalkylphosphodiamidate linkages, thiophosphoramidate linkages, thioalkylphosphate linkages, thioalkylphosphotriester linkages, phosphorothioate linkages, phosphoroselenoate linkages, and phosphoroboronate linkages.

[0294] Antisense oligonucleotides with one or more spacers

[0295] Embodiments disclosed herein include antisense oligonucleotides (AONs) comprising one or more spacers. In a specific embodiment, the AON comprises one spacer. In a specific embodiment, the AON comprises two spacers. In a specific embodiment, the AON comprises three spacers. Generally speaking, a spacer refers to a nucleoside replacement group lacking a nucleobase, and wherein the nucleoside sugar moiety is replaced with a non-sugar substitute. The non-sugar substitute cannot be connected to the nucleobase, but can be connected to the 3' and 5' positions of the nucleoside adjacent to the spacer through an internucleoside linking group.

[0296] In various embodiments, the AON with one or more spacers comprises a parent AON (e.g., an antisense oligonucleotide complementary to a portion of a target gene product, such as any of PPM1A, ATXN2, SOD1, or MAPT mRNA transcripts), wherein one or more nucleosides of the parent oligonucleotide (e.g., any of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, or 167805-301566) are replaced with a spacer.

[0297] In various embodiments, the AON having one or more spacers comprises an oligonucleotide variant (e.g., an antisense oligonucleotide that represents a modified version (e.g., shorter or longer) of a corresponding parent oligonucleotide), wherein one or more nucleosides of the oligonucleotide variant (e.g., a shorter or longer 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) are replaced with a spacer.

[0298] In various embodiments, the AON with one or more spacers comprises a gapmer AON comprising at least three distinct structural regions (e.g., a 5' wing, a central region, and a 3' wing, in a "5→3" orientation), wherein one or more nucleosides of the gapmer AON (e.g., a gapmer AON comprising a sequence of any one of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, or 167805-301566, or any gapmer AON sequence shown in Tables 3A-3D (e.g., SEQ ID NOs: 301595-301607)) are replaced with a spacer. In specific embodiments, for gapmer AONs with one or more spacers, the nucleosides in the 5' wing are replaced with a spacer. In specific embodiments, for gapmer AONs with one or more spacers, the nucleosides in the 3' wing are replaced with a spacer. In specific embodiments, for gapmer AONs with one or more spacers, the nucleosides in the central region are replaced with spacers. In specific embodiments, for gapmer AONs with one or more spacers, the nucleosides in the 5' wing region are replaced with spacers and the nucleosides in the central region are replaced with spacers. In specific embodiments, for gapmer AONs with one or more spacers, the nucleosides in the 5' wing region are replaced with spacers and the nucleosides in the 3' wing region are replaced with spacers. In specific embodiments, for gapmer AONs with one or more spacers, the nucleosides in the central region are replaced with spacers and the nucleosides in the 3' wing region are replaced with spacers.

[0299] In specific embodiments, the gapmer oligonucleotide comprises any of the following patterns of sugar modifications: eeeee-d10-eeeee, eeeee-d8-eeeee, eeeeee-d11-eeeeee, eee-d8-eee, eee-d10-eee, eeee-d10-eeee, and eeee-d8-eeee, wherein e=2'-MOE nucleoside and d=deoxyribonucleoside. In such embodiments, the spacer can replace at least one "e" in the gapmer oligonucleotide (e.g., 2'-MOE nucleoside) or can replace at least one "d" in the gapmer oligonucleotide (e.g., deoxyribonucleoside).

[0300] As used herein, "AON with one or more spacers" refers to any parent AON with one or more spacers (e.g., any of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, or 167805-301566]), an AON variant with one or more spacers (e.g., a shorter or longer version of the corresponding parent oligonucleotide comprising a nucleobase sequence selected from any one of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, or 167805-301566), or a gapmer AON with one or more spacers (e.g., a gapmer comprising the sequence of any one of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, or 167805-301566). AON or any gapmer AON sequence shown in Tables 3A-3D (e.g., SEQ ID NOs: 301595-301607). In certain embodiments, the 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 a nucleoside (e.g., a nucleoside comprising a sugar and / or a nucleobase) or a nucleoside replacement group (e.g., a spacer) of an oligonucleotide.

[0301] In a specific embodiment, the length of the AON with one or more spacers is 25 oligonucleotide units. In a specific embodiment, the length of the AON with one or more spacers is 23 oligonucleotide units. In a specific embodiment, the length of the AON with one or more spacers is 21 oligonucleotide units. In a specific embodiment, the length of the AON with one or more spacers is 20 oligonucleotide units. In a specific embodiment, the length of the AON with one or more spacers is 19 oligonucleotide units. In a specific embodiment, the length of the AON with one or more spacers is 18 oligonucleotide units. In various embodiments, the length of the AON with one or more spacers is 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 various embodiments, the length of the AON with one or more spacers is at least 15 oligonucleotide units. In various embodiments, the length of the AON with one or more spacers is at least 16 oligonucleotide units. In various embodiments, the length of the AON with one or more spacers is at least 17 oligonucleotide units. In various embodiments, the length of the AON with one or more spacers is at least 18 oligonucleotide units. In various embodiments, the length of the AON with one or more spacers is at least 19 oligonucleotide units. In various embodiments, the length of the AON with one or more spacers is at least 20 oligonucleotide units. In various embodiments, the length of the AON with one or more spacers is at least 21 oligonucleotide units. In various embodiments, the length of the AON with one or more spacers is at least 22 oligonucleotide units. In various embodiments, the length of the AON with one or more spacers is at least 23 oligonucleotide units. In various embodiments, the length of the AON with one or more spacers is at least 24 oligonucleotide units. In various embodiments, the length of the AON with one or more spacers is at least 25 oligonucleotide units.

[0302] In various embodiments, the AON with one or more spacers comprises a sequence that is at least 80% identical to an equal length portion of any one of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, 167805-301566, SEQ ID NOs: 167805-301566. In various embodiments, the AON with one or more spacers comprises a sequence that is at least 85% identical to an equal length portion of any one of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, 167805-301566, SEQ ID NOs: 167805-301566. In various embodiments, the AON with one or more spacers comprises a sequence that is at least 90% identical to an equal length portion of any one of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, 167805-301566, SEQ ID NOs: 167805-301566. In various embodiments, the AON with one or more spacers comprises a sequence that is at least 95% identical to an equal length portion of any one of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, 167805-301566, SEQ ID NOs: 167805-301566. In various embodiments, the AON comprises a sequence that is 98% identical to an equal length portion of any one of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, or 167805-301566.

[0303] In some embodiments, the spacer has formula (X):

[0304]

[0305] wherein Ring A is as defined herein.

[0306] In some embodiments, the spacer has formula (Xa):

[0307]

[0308] wherein Ring A is as defined herein and the -CH2-O- group is located on the Ring A atom adjacent to the -O- group.

[0309] As generally defined herein, Ring A of Formula (X) and (Xa) is an optionally substituted 4-8 membered monocyclic cycloalkyl (e.g., Ring A is cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl) or a 4-8 membered monocyclic heterocyclyl, wherein the heterocyclyl contains 1 or 2 heteroatoms selected from O, S, and N (e.g., Ring A is oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, 1,4-dioxanyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, azepanyl). In some embodiments, Ring A is tetrahydrofuranyl. In some embodiments, Ring A is tetrahydropyranyl. In some embodiments, Ring A is pyrrolidinyl. In some embodiments, Ring A is cyclopentyl. In some embodiments, the monocyclic cycloalkyl or monocyclic heterocyclyl is not further substituted. In some embodiments, the cycloalkyl or heterocyclyl is further substituted with 0, 1, 2, or 3 substituents selected from halogen (e.g., -F, -Cl), -OMe, -OEt-O(CH2)OMe, -O(CH2)2OMe, and CN. In some embodiments, the spacer is represented by formula (I), wherein:

[0310]

[0311] X is selected from -CH2- and -O-; and

[0312] n is 0, 1, 2, or 3.

[0313] In some embodiments, the spacer is represented by formula (I'), wherein:

[0314]

[0315] X is selected from -CH2- and -O-; and

[0316] n is 0, 1, 2, or 3.

[0317] In some embodiments, the spacer is represented by Formula (Ia), wherein:

[0318]

[0319] n is 0, 1, 2, or 3.

[0320] In some embodiments, the spacer is represented by formula (Ia'), wherein:

[0321]

[0322] n is 0, 1, 2, or 3.

[0323] As generally defined herein, X is selected from -CH2- and -O-. In some embodiments, X is -CH2-. In other embodiments, X is -O-.

[0324] 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.

[0325] In some embodiments, the spacer is represented by formula (II), wherein:

[0326]

[0327] X is selected from -CH2- and -O-.

[0328] In some embodiments, the spacer is represented by formula (II'), wherein:

[0329]

[0330] X is selected from -CH2- and -O.

[0331] In some embodiments, the spacer is represented by formula (Iia), wherein:

[0332]

[0333] In some embodiments, the spacer is represented by formula (Iia'), wherein:

[0334]

[0335] In some embodiments, the spacer is represented by formula (III), wherein:

[0336]

[0337] X is selected from -CH2- and -O-.

[0338] In some embodiments, the spacer is represented by formula (III'), wherein:

[0339]

[0340] X is selected from -CH2- and -O.

[0341] In some embodiments, the spacer is represented by formula (IIIa), wherein:

[0342]

[0343] In some embodiments, the spacer is represented by formula (IIIa'), wherein:

[0344]

[0345] In some embodiments, the open positions (i.e., positions not specifically described as bearing only hydrogen atoms, -CH2- groups containing X) of Formula (I), (I'), (Ia), (Ia'), (II), (II'), (Iia), (Iia'), (III), (III'), (IIIa), and (IIIa') are further substituted with 0-3 substituents selected from halogen (e.g., -F, -Cl), -OMe, -OEt, -O(CH2)OMe, -O(CH2)2OMe, and CN. In some embodiments, Formula (I), (I'), (Ia), (Ia'), (II), (II'), (Iia), (Iia'), (III), (III'), (IIIa), and (IIIa') are not further substituted.

[0346] As further described below, the AON with one or more spacers is described with reference to corresponding parent oligonucleotide, corresponding oligonucleotide variant or corresponding gapmer oligonucleotide.In various embodiments, the difference between the oligonucleotide with one or more spacers and parent oligonucleotide, oligonucleotide variant or gapmer oligonucleotide is that each in the one or more spacers replaces the nucleoside in parent oligonucleotide, oligonucleotide variant or gapmer oligonucleotide.As used below, " position " of oligonucleotide refers to the specific position counting from the 5 ' end of oligonucleotide.In various embodiments, spacer replaces nucleoside at any one in position 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 parent oligonucleotide, oligonucleotide variant or gapmer oligonucleotide.In specific embodiments, spacer replaces the nucleoside at position 7 of parent oligonucleotide, oligonucleotide variant or gapmer oligonucleotide. In a specific embodiment, the spacer replaces the nucleoside at position 8 of the parent oligonucleotide, oligonucleotide variant or gapmer oligonucleotide. In a specific embodiment, the spacer replaces the nucleoside at position 11 of the parent oligonucleotide, oligonucleotide variant or gapmer oligonucleotide. In a specific embodiment, the spacer replaces the nucleoside at position 14 of the parent oligonucleotide, oligonucleotide variant or gapmer oligonucleotide. In a specific embodiment, the spacer replaces the nucleoside at position 16 of the parent oligonucleotide, oligonucleotide variant or gapmer oligonucleotide. In a specific embodiment, the spacer replaces the nucleoside at position 19 of the parent oligonucleotide, oligonucleotide variant or gapmer oligonucleotide. In a specific embodiment, the spacer replaces the nucleoside at position 22 of the oligonucleotide variant or gapmer oligonucleotide.

[0347] In various embodiments, the oligonucleotide comprises a spacer that replaces a nucleoside in a parent oligonucleotide, oligonucleotide variant or gapmer oligonucleotide (e.g., a spacer replaces a nucleoside in a parent oligonucleotide, oligonucleotide variant or gapmer oligonucleotide). Exemplary oligonucleotides with a spacer are shown in Table 4A below. In specific embodiments, the spacer replaces the nucleoside between position 9 and 15 of the parent oligonucleotide, oligonucleotide variant or gapmer oligonucleotide. In specific embodiments, the spacer replaces the nucleoside between position 9 and 12 of the parent oligonucleotide, oligonucleotide variant or gapmer oligonucleotide. In specific embodiments, the spacer replaces the nucleoside at position 9 of the parent oligonucleotide, oligonucleotide variant or gapmer oligonucleotide. In specific embodiments, the spacer replaces the nucleoside at position 10 of the parent oligonucleotide, oligonucleotide variant or gapmer oligonucleotide. In specific embodiments, the spacer replaces the nucleoside at position 11 of the parent oligonucleotide, oligonucleotide variant or gapmer oligonucleotide. In specific embodiments, the spacer replaces the nucleoside at position 12 of the parent oligonucleotide, oligonucleotide variant or gapmer oligonucleotide. In specific embodiments, the spacer replaces the nucleoside of the parent oligonucleotide, oligonucleotide variant or gapmer oligonucleotide between positions 12 and 16. In specific embodiments, the spacer replaces the nucleoside at position 15 of the parent oligonucleotide, oligonucleotide variant or gapmer oligonucleotide.

[0348] In various embodiments, an oligonucleotide comprising a spacer has two fragments, wherein at least one of the two fragments has at most 11 connected nucleosides. For example, the length of the oligonucleotide can be 23 oligonucleotide units, and the spacer can be located at position 12. Thus, the oligonucleotide has two fragments separated by a spacer, wherein the length of these two fragments is 11 nucleosides. In various embodiments, an oligonucleotide comprising a spacer has two fragments, wherein at least one of the two fragments has at most 10 connected nucleosides. For example, the length of the oligonucleotide can be 21 oligonucleotide units, and the spacer can be located at position 11. Thus, the oligonucleotide has two fragments separated by a spacer, wherein the length of these two fragments is 10 nucleosides. As another example, the length of the oligonucleotide can be 25 oligonucleotide units, and the spacer can be located at position 15. Thus, the oligonucleotide has two fragments separated by a spacer, wherein one of the two fragments is 14 nucleobases in length, and the second of the two fragments 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. Thus, the oligonucleotide has two segments separated by a spacer, wherein one of the two segments is 10 nucleobases in length and the second of the two segments 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. Thus, the oligonucleotide has two segments separated by a spacer, wherein one of the two segments is 10 nucleobases in length and the second of the two segments is 7 nucleobases in length. As another example, the oligonucleotide can be 18 oligonucleotide units in length, and the spacer can be located at position 9. Thus, the oligonucleotide has two segments separated by a spacer, wherein one of the two segments is 8 nucleobases in length and the second of the two segments is 9 nucleobases in length.

[0349] In various embodiments, oligonucleotide comprises 2 spacers, and it replaces the nucleosides in parent oligonucleotide, oligonucleotide variant or gapmer oligonucleotide separately (for example, 2 spacers replace 2 independent nucleosides of parent oligonucleotide, oligonucleotide variant or gapmer oligonucleotide).Exemplary oligonucleotide with 2 spacers is as shown in following table 4B and table 4C.In various embodiments, the first spacer and the second spacer are separated by at least 5 core bases, at least 6 core bases, at least 7 core bases, at least 8 core bases, at least 9 core bases or at least 10 core bases in oligonucleotide.In specific embodiments, the first spacer and the second spacer are separated by at least 5 core bases, at least 6 core bases or at least 7 core bases.In specific embodiments, the first spacer and the second spacer are not adjacent to each other in oligonucleotide.

[0350] In specific embodiments, the first spacer replaces the nucleosides between positions 7 and 11 of the parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide. In various embodiments, the first spacer replaces the nucleosides between positions 8 and 11, positions 9 and 11, positions 10 and 11, positions 7 and 10, positions 7 and 9, positions 7 and 8, positions 8 and 10, positions 8 and 9, or positions 9 and 10 of the parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide. In specific embodiments, the second spacer replaces the nucleosides between positions 14 and 22 of the parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide. In various embodiments, the second spacer replaces the nucleoside between positions 15 and 22, positions 16 and 22, positions 17 and 22, positions 18 and 22, positions 19 and 22, positions 20 and 22, positions 21 and 22, positions 15 and 21, positions 16 and 21, positions 17 and 21, positions 18 and 21, positions 19 and 21, positions 20 and 21, positions 15 and 20, positions 16 and 20, positions 17 and 20, positions 18 and 20, positions 19 and 20, positions 15 and 19, positions 16 and 19, positions 17 and 19, positions 18 and 19, positions 15 and 18, positions 16 and 18, positions 17 and 18, positions 15 and 17, positions 16 and 17, or positions 15 and 16 of the parent oligonucleotide, oligonucleotide variant or gapmer oligonucleotide.

[0351] In a preferred embodiment, the first spacer replaces the nucleoside at position 7 of the parent oligonucleotide, oligonucleotide variant or gapmer oligonucleotide and the second spacer replaces the nucleoside at position 14 of the parent oligonucleotide, oligonucleotide variant or gapmer oligonucleotide. In a preferred embodiment, the first spacer replaces the nucleoside at position 7 of the parent oligonucleotide, oligonucleotide variant or gapmer oligonucleotide and the second spacer replaces the nucleoside at position 15 of the parent oligonucleotide, oligonucleotide variant or gapmer oligonucleotide. In a preferred embodiment, the first spacer replaces the nucleoside at position 7 of the parent oligonucleotide, oligonucleotide variant or gapmer oligonucleotide and the second spacer replaces the nucleoside at position 19 of the parent oligonucleotide, oligonucleotide variant or gapmer oligonucleotide. In a preferred embodiment, the first spacer replaces the nucleoside at position 8 of the parent oligonucleotide, oligonucleotide variant or gapmer oligonucleotide and the second spacer replaces the nucleoside at position 16 of the parent oligonucleotide, oligonucleotide variant or gapmer oligonucleotide. In a preferred embodiment, the first spacer replaces the nucleoside at position 11 of the parent oligonucleotide, oligonucleotide variant or gapmer oligonucleotide and the second spacer replaces the nucleoside at position 19 of the parent oligonucleotide, oligonucleotide variant or gapmer oligonucleotide. In a preferred embodiment, the first spacer replaces the nucleoside at position 11 of the parent oligonucleotide, oligonucleotide variant or gapmer oligonucleotide and the second spacer replaces the nucleoside at position 22 of the parent oligonucleotide, oligonucleotide variant or gapmer oligonucleotide. In a preferred embodiment, the first spacer replaces the nucleoside at position 9 of the parent oligonucleotide, oligonucleotide variant or gapmer oligonucleotide and the second spacer replaces the nucleoside at position 19 of the parent oligonucleotide, oligonucleotide variant or gapmer oligonucleotide. In a preferred embodiment, the first spacer replaces the nucleoside at position 5 of the parent oligonucleotide, oligonucleotide variant or gapmer oligonucleotide and the second spacer replaces the nucleoside at position 17 of the parent oligonucleotide, oligonucleotide variant or gapmer oligonucleotide.

[0352] In various embodiments, the oligonucleotide comprises 3 spacers, each of which replaces a nucleoside in a parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide (e.g., 3 spacers replace 3 separate nucleosides of a parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide). In specific embodiments, the first spacer replaces the nucleoside between position 7 and 11 of the parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide. In specific embodiments, the second spacer replaces the nucleoside between position 14 and 22 of the parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide. In specific embodiments, the third spacer replaces the nucleoside between position 21 and 24 of the parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide. In specific embodiments, the first spacer replaces the nucleoside between position 2 and 5 of the parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide. In specific embodiments, the second spacer replaces the nucleoside between position 8 and 12 of the parent oligonucleotide, oligonucleotide variant, or gapmer oligonucleotide. In specific embodiments, the third spacer replaces nucleosides between positions 18 and 22 of the parent oligonucleotide, oligonucleotide variant or gapmer oligonucleotide.

[0353] In various embodiments, the three spacers in the oligonucleotide are positioned so that each of the four segments of the oligonucleotide is at most 7 linked nucleosides in length. For example, an oligonucleotide can have a first segment of 7 linked nucleosides linked to a first spacer, followed by a second segment of 7 linked nucleosides linked at one end to the first spacer and at the other end to the second spacer, followed by a third segment of 6 linked nucleosides linked at one end to the second spacer and at the other end to the third spacer, followed by a fourth segment of 6 linked nucleosides linked to the third spacer.

[0354] In various embodiments, one or more spacers are positioned in oligonucleotide to replace one or more adenosine or thymidine (relative to guanine or cytidine). For example, one or more spacers can replace 1, 2, 3, 4, 5, 6, 7, 8 or 9 adenosine or thymidine in oligonucleotide. In various embodiments, one or more spacers are positioned in oligonucleotide to replace one or more guanine or cytidine (relative to adenosine or thymidine). For example, one or more spacers can replace 1, 2, 3, 4, 5, 6, 7, 8 or 9 guanine or cytidine in oligonucleotide. In various embodiments, spacers are positioned in oligonucleotide to replace same number of adenosine / thymidine and guanine / cytidine. For example, the first spacer in oligonucleotide can replace adenosine / thymidine, and the second spacer in oligonucleotide can replace guanine / cytidine.

[0355] In various embodiments, one or more spacers are positioned in the oligonucleotide to control the sequence content in the oligonucleotide.For example, one or more spacers are positioned so that at least one spacer is positioned near the guanine group.In various embodiments, the oligonucleotide with spacer can comprise a spacer adjacent to the guanine group, two spacers adjacent to the guanine group, three spacers adjacent to the guanine group, four spacers adjacent to the guanine group or five spacers adjacent to the guanine group.In one embodiment, if counting from the 5 ' end of the oligonucleotide, the spacer is immediately before the guanine group in the sequence.Therefore, in various embodiments, the oligonucleotide with spacer can comprise a spacer immediately before the guanine group, two spacers immediately before the guanine group respectively, three spacers immediately before the guanine group respectively, four spacers immediately before the guanine group respectively or five spacers immediately before the guanine group respectively.In one embodiment, if counting from the 5 ' end of the oligonucleotide, the guanine group is followed by a spacer. Thus, in various embodiments, an oligonucleotide with a spacer can include one spacer immediately following a guanine group, two spacers each immediately following a guanine group, three spacers each immediately following a guanine group, four spacers each immediately following a guanine group, or five spacers each immediately following a guanine group. In various embodiments, the spacers in the oligonucleotide can be positioned so as to maximize the number of spacers adjacent to the guanine group.

[0356] In various embodiments, one or more spacers are positioned in an oligonucleotide to replace one or more adenosine or thymidine nucleosides such that the one or more spacers are positioned adjacent to a guanine group. For example, two spacers can replace an adenosine or thymidine nucleoside in an oligonucleotide, with each of the two spacers being positioned adjacent to a guanine group.

[0357] In various embodiments, the oligonucleotide with one or more spacers has specific GC content.As used herein, GC content (or guanine-cytosine content) is the percentage of nitrogenous base guanine (G) or cytosine (C) in the oligonucleotide.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 specific embodiments, the oligonucleotide with one or more spacers has at least 30% GC content.In specific 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, rather than selecting guanine or cytosine to be replaced by a spacer, thymine or adenine in the oligonucleotide can be selected to be replaced by a spacer.

[0358] In various embodiments, the oligonucleotides with spacers are designed such that 1) each fragment of the oligonucleotide has up to 7 connected nucleosides, and 2) at least two, three, or four spacers are positioned adjacent to a guanine group. In some embodiments, the oligonucleotides with spacers are designed such that 1) each fragment of the oligonucleotide has up to 7 connected nucleosides, and 2) each of the two spacers is positioned before a guanine group.

[0359] In various embodiments, the oligonucleotide comprises one or more spacers that do 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 the corresponding parent oligonucleotide, oligonucleotide variants, or gapmer oligonucleotides. Exemplary mRNA or pre-mRNA transcripts comprise a gene product of any one of PPM1A, ATXN2, SOD1, or MAPT. In various embodiments, the oligonucleotide comprises one or more spacers that increase the effectiveness of the oligonucleotide with spacers in reducing the frequency of target mRNA or pre-mRNA transcripts compared to the effect of the corresponding parent oligonucleotide, oligonucleotide variants, or gapmer oligonucleotides.

[0360] Table 4A, 4B and 4C have recorded the example AON with one or more spacers and the relationship between them and corresponding parent oligonucleotide, corresponding oligonucleotide variants and / or corresponding gapmer oligonucleotide.Each oligonucleotide is all assigned a sequence name.As used hereinafter, the nomenclature of sequence name is expressed as " X_spA " (for the AON with a spacer), " X_spA_spB " (for the AON with two spacers) or " X_spA_spB_spC " (for the AON with three spacers).Here, " X ​​" refers to the length of AON, " A " refers to the position of the first spacer in the AON, " B " refers to the position of the second spacer in the AON, and if present, " C " refers to the position of the 3rd spacer in the AON.

[0361] Example AONs with one spacer are recorded in Table 4A below.

[0362] Table 4A: Identification of AONs with one spacer. Here, each AON has 2 segments.

[0363]

[0364]

[0365]

[0366]

[0367] *The at least one nucleoside linkage of the nucleobase sequence is selected from the group consisting of phosphorothioate linkage, alkylphosphate linkage, phosphorodithioate linkage, phosphotriester linkage, alkylphosphate linkage, 3-methoxypropylphosphate linkage, methylphosphonate linkage, aminoalkylphosphotriester linkage, alkylenephosphonate linkage, phosphite linkage, phosphoramidate linkage, thiophosphoramidate linkage, phosphorodiamidate (e.g., comprising phosphorodiamidate morpholino (PMO), 3' aminoribose, or 5' aminoribose) linkage, aminoalkylphosphoramidate linkage, thiophosphoramidate linkage, thioalkylphosphate linkage, thioalkylphosphotriester linkage, phosphorothioate linkage, phosphoroselenoate linkage, and phosphoroboronate linkage.

[0368] In various embodiments, the oligonucleotides disclosed herein comprise two spacers.In various embodiments, the inclusion of two spacers divides the oligonucleotide into three separate segments, wherein at least one segment is at most 7 linked nucleosides in length.

[0369] In specific embodiments, the first spacer is located between positions 5 and 11 of the oligonucleotide. In various embodiments, the first spacer is located between positions 7 and 11 of the oligonucleotide. In various embodiments, the second spacer is located between positions 15 and 19 of the oligonucleotide. In various embodiments, the first spacer and the second spacer are separated in the oligonucleotide by at least 5 nucleobases, at least 6 nucleobases, or at least 7 nucleobases.

[0370] In various embodiments, the first spacer is located between positions 5 and 11 of the oligonucleotide, and the second spacer is located between positions 15 and 19 of the oligonucleotide. In various embodiments, the first spacer is located at position 8 of the oligonucleotide, and wherein the second spacer is located at position 16 of the oligonucleotide. In various embodiments, the first spacer is located at position 5 of the oligonucleotide, and wherein the second spacer is located at position 17 of the oligonucleotide. In various embodiments, the first spacer is located at position 7 of the oligonucleotide, and wherein the second spacer is located at position 14 of the oligonucleotide. In various embodiments, the first spacer is located at position 7 of the oligonucleotide, and wherein the second spacer is located at position 15 of the oligonucleotide. In various embodiments, the first spacer is located at position 11 of the oligonucleotide, and wherein the second spacer is located at position 19 of the oligonucleotide.

[0371] Example AONs with two spacers are reported in Table 4B below.

[0372] Table 4B: Identification of AONs with two spacers. Here, each AON has 3 segments.

[0373]

[0374]

[0375]

[0376]

[0377]

[0378]

[0379]

[0380]

[0381]

[0382] *The at least one nucleoside linkage of the nucleobase sequence is selected from the group consisting of phosphorothioate linkage, alkylphosphate linkage, phosphorodithioate linkage, phosphotriester linkage, alkylphosphate linkage, 3-methoxypropylphosphate linkage, methylphosphonate linkage, aminoalkylphosphotriester linkage, alkylenephosphonate linkage, phosphite linkage, phosphoramidate linkage, thiophosphoramidate linkage, phosphorodiamidate (e.g., comprising phosphorodiamidate morpholino (PMO), 3' aminoribose, or 5' aminoribose) linkage, aminoalkylphosphoramidate linkage, thiophosphoramidate linkage, thioalkylphosphate linkage, thioalkylphosphotriester linkage, phosphorothioate linkage, phosphoroselenoate linkage, and phosphoroboronate linkage.

[0383] In various embodiments, compared with the AON described in Table 4A and 4B above, the length of the AON with one or more spacers reduces or the length increases.For example, such AON can be the oligonucleotide variant with one or more spacers.In various embodiments, the oligonucleotide variant with one or more spacers is 23mer, 21mer, 19mer or 18mer.In various embodiments, oligonucleotide variant comprises two spacers so that oligonucleotide variant comprises three sections separated by two spacers.In various embodiments, at least one of the three sections has 7 nucleosides connected at the most.In various embodiments, each of the three sections has 7 nucleosides connected at the most.Exemplary oligonucleotide variant with one or more spacers is as shown in Table 4C below.

[0384] Table 4C: AON variants with two spacers. Here, each AON variant has three segments.

[0385]

[0386]

[0387]

[0388]

[0389] *The at least one nucleoside linkage of the nucleobase sequence is selected from the group consisting of phosphorothioate linkage, alkylphosphate linkage, phosphorodithioate linkage, phosphotriester linkage, alkylphosphate linkage, 3-methoxypropylphosphate linkage, methylphosphonate linkage, aminoalkylphosphotriester linkage, alkylenephosphonate linkage, phosphite linkage, phosphoramidate linkage, thiophosphoramidate linkage, phosphorodiamidate (e.g., comprising phosphorodiamidate morpholino (PMO), 3' aminoribose, or 5' aminoribose) linkage, aminoalkylphosphoramidate linkage, thiophosphoramidate linkage, thioalkylphosphate linkage, thioalkylphosphotriester linkage, phosphorothioate linkage, phosphoroselenoate linkage, and phosphoroboronate linkage.

[0390] In various embodiments, the antisense oligonucleotides disclosed herein (e.g., PPM1A AON, ATXN2 AON, SOD1AON, or MAPT AON) comprise one or more locked nucleic acids (LNAs). In specific embodiments, the antisense oligonucleotide comprises one LNA. In specific embodiments, the antisense oligonucleotide comprises two LNAs. In specific embodiments, the antisense oligonucleotide comprises three LNAs. Generally speaking, LNAs refer to nucleic acid monomers having a bridge (e.g., a methylene, ethylene, aminooxy, or oxyimino bridge) connecting two carbon atoms between the 4' and 2' positions of the nucleoside sugar unit, thereby forming a bicyclic sugar.

[0391] In various embodiments, the antisense oligonucleotides disclosed herein (e.g., PPM1A AON, ATXN2 AON, SOD1 AON, or MAPT AON) comprise one or more spacers and one or more locked nucleic acids (LNAs). In some embodiments, the antisense oligonucleotides disclosed herein (e.g., PPM1A AON, ATXN2 AON, SOD1 AON, or MAPT AON) comprise two spacers and two LNAs. In some embodiments, the antisense oligonucleotides disclosed herein comprise two spacers and three LNAs.

[0392] In various embodiments, the spacer and LNA are positioned adjacent to each other in the antisense oligonucleotide. For example, the spacer may be located at position M of the antisense oligonucleotide. Alternatively, the LNA may be located at position M+1 or position M-1 of the antisense oligonucleotide. In various embodiments, in the antisense oligonucleotide, the first spacer is located adjacent to the first LNA, and the second spacer is located adjacent to the second LNA. For example, the first spacer may be located at position M of the antisense oligonucleotide, and the second spacer may be located at position N of the antisense oligonucleotide. The first LNA may be located at position M+1 or position M-1 of the antisense oligonucleotide, and the second LNA may be located at position N+1 or position N-1 of the antisense oligonucleotide. In other embodiments, one or more spacers and one or more LNAs are not adjacent to each other in the 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.

[0393] Chemical modification of AON

[0394] As described herein, AONs, such as AONs having a sequence of 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 of a sequence of any one of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, or 167805-301566, such as any of SEQ ID NOs: 301692-301742 or the sequences in Tables 4A-4C, can comprise one or more chemical modifications to one or more nucleosides and / or to one or more internucleoside linkages. A nucleoside is a base-sugar combination. The nucleobase (also referred to as a base) portion of a nucleoside is typically a heterocyclic base portion. A nucleotide is a nucleoside that further comprises a phosphate group covalently linked to the sugar portion of the nucleoside. For those nucleosides containing pentofuranosyl sugars, the phosphate group can be connected to the 2', 3' or 5' hydroxyl moiety of the sugar. Oligonucleotides are formed by covalently linking adjacent nucleosides to each other to form a linear polymeric oligonucleotide. Within the oligonucleotide structure, the phosphate group is generally referred to as the internucleoside linkage that forms the oligonucleotide.

[0395] Modifications to AON encompass substitutions or changes to internucleoside linkages and / or nucleosides (e.g., sugar moieties or core bases of nucleosides). Due to desired properties, such as enhanced cellular uptake, enhanced affinity for nucleic acid targets, increased stability in the presence of nucleases, or increased inhibitory activity, modified AONs can be superior to native forms. Chemically modified nucleosides, core bases, and internucleoside linkages are described in Agrawal and Gait, History and Development of NucleotideAnalogues in Nucleic Acids Drugs, in Drug Discovery Series No.68, Advances inNucleic Acid Therapeutics, 1-21 (Agrawal and Gait eds., 2019), the contents of which are incorporated herein by reference.

[0396] Modified internucleoside linkages

[0397] In various embodiments, the AON, e.g., an AON comprising the 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 the sequence 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 of the sequences in Tables 4A-4C, comprises one or more modified internucleoside linkages. The internucleoside linkage naturally occurring in RNA and DNA is a 3' to 5' phosphodiester linkage. AONs having one or more modified, i.e., non-naturally occurring, internucleoside linkages can be selected relative to antisense compounds having naturally occurring internucleoside linkages because of desirable properties, such as enhanced cellular uptake, enhanced affinity for nucleic acid targets, increased stability in the presence of nucleases.

[0398] The internucleoside linkage of modification comprises the internucleoside linkage that retains phosphorus atom and the internucleoside linkage that does not have phosphorus atom.Representational phosphorus-containing internucleoside linkage includes but is not limited to phosphodiester, phosphotriester, methylphosphonate, phosphoramidate and thiophosphate.The method for preparing phosphorus-containing and non-phosphorus-containing linkage is well-known. Examples of modified internucleoside linkages include phosphorothioate linkages, alkylphosphate linkages, alkylphosphate linkages, 3-methoxypropylphosphate linkages, phosphorodithioate linkages, phosphotriester linkages, methylphosphonate linkages, aminoalkylphosphotriester linkages, alkylenephosphonate linkages, phosphite linkages, phosphoramidate linkages, thiophosphoramidate linkages, phosphorodiamidate (e.g., comprising phosphorodiamidate morpholino (PMO), 3' aminoribose, or 5' aminoribose) linkages, aminoalkylphosphoramidate linkages, thiophosphoramidate linkages, thioalkylphosphate linkages, thioalkylphosphotriester linkages, phosphorothioate linkages, phosphoroselenoate linkages, and phosphoroboronate linkages.

[0399] In various embodiments, the AON comprises one or more modified internucleoside linkages connecting the oligonucleotide units. In 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 connecting the oligonucleotide units.

[0400] In various embodiments, each modified internucleoside linkage of an AON can be designed independently of other modified internucleoside linkages of an AON. In other words, the modified internucleoside linkages of an AON do not all need to be modified internucleoside linkages of the same type. In various embodiments, the modified internucleoside linkages are dispersed throughout the antisense compound.

[0401] 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 specific embodiments, the AON comprises 13, 14, 15, 16, 17, 18, 19, 20, 21, or 22 phosphorothioate linkages. In specific embodiments, the AON comprises 17 phosphorothioate linkages. In specific embodiments, the AON comprises 19 phosphorothioate linkages. In specific embodiments, the AON comprises 22 phosphorothioate linkages. In specific embodiments, all internucleoside linkages of the AON are phosphorothioate linkages.

[0402] In various embodiments, AON comprises a mixture of modified internucleoside linkages and naturally occurring phosphodiester linkages. For example, AON comprises at least one phosphodiester linkage and at least one phosphorothioate linkage. In various embodiments, AON comprises 6 to 10, 6 to 9, 6 to 8, 7 to 10, 7 to 9 or 6, 7 or 8 phosphorothioate linkages. In some embodiments, AON comprises 6, 7, 8, 9 or 10 phosphorothioate linkages. In some embodiments, AON comprises 6 to 10, 6 to 9, 6 to 8, 7 to 10, 7 to 9 or 6, 7 or 8 phosphodiester linkages. In some embodiments, AON comprises 6, 7, 8, 9 or 10 phosphodiester linkages.

[0403] In a specific embodiment, the AON comprises 10 phosphorothioate linkages and 9 phosphodiester linkages. In a specific embodiment, the AON comprises 6 phosphorothioate linkages and 7 phosphodiester linkages. In a specific embodiment, the AON comprises 6 phosphorothioate linkages and 9 phosphodiester linkages. In a specific embodiment, the AON comprises 8 phosphorothioate linkages and 9 phosphodiester linkages. In a specific embodiment, the AON comprises 8 phosphorothioate linkages and 7 phosphodiester linkages. In a specific embodiment, the AON comprises 12 phosphorothioate linkages and 7 phosphodiester linkages. In a specific embodiment, the AON comprises 15 phosphorothioate linkages and 4 phosphodiester linkages. In a specific embodiment, the AON comprises 15 phosphorothioate linkages and 2 phosphodiester linkages. In a specific embodiment, the AON comprises 17 phosphorothioate linkages and 2 phosphodiester linkages.

[0404] In some embodiments, AON includes internucleoside linkage designed according to the gapmer design of AON. In some embodiments, 5' wing region includes at least one modified internucleoside linkage (e.g., internucleoside linkage modification from naturally occurring 3' to 5' phosphodiester linkage). In some embodiments, 5' wing region includes 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. In some embodiments, 3' wing region includes at least one modified internucleoside linkage. In some embodiments, 3' wing region includes 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. In some embodiments, central region includes 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.

[0405] In a specific embodiment, all internucleoside linkages in the 5' wing are modified internucleoside linkages, such as phosphorothioate linkages. In a specific embodiment, all internucleoside linkages in the 3' wing are modified internucleoside linkages, such as phosphorothioate linkages. In a specific embodiment, all internucleoside linkages in the central region are modified internucleoside linkages, such as phosphorothioate linkages. In a specific embodiment, all internucleoside linkages in each of the 5' wing, the 3' wing, and the central region are modified internucleoside linkages, such as phosphorothioate linkages.

[0406] In various embodiments, the two nucleoside bonds in the 5' wing are phosphorothioate bonds. In various embodiments, the two nucleoside bonds in the 5' wing are phosphodiester bonds. In various embodiments, the two nucleoside bonds in the 5' wing are phosphorothioate bonds, and the other two nucleoside bonds in the 5' wing are phosphodiester bonds. In various embodiments, the two nucleoside bonds in the 3' wing are phosphorothioate bonds. In various embodiments, the two nucleoside bonds in the 3' wing are phosphodiester bonds. In various embodiments, the two nucleoside bonds in the 3' wing are phosphorothioate bonds, and the other two nucleoside bonds in the 3' wing are phosphodiester bonds.

[0407] In various embodiments, the four internucleoside linkages in the 5' wing are phosphorothioate linkages. In various embodiments, the four internucleoside linkages in the 5' wing are phosphorothioate linkages, and the two internucleoside linkages in the 3' wing are phosphorothioate linkages. In various embodiments, one internucleoside linkage in the 5' wing is a phosphorothioate linkage. In various embodiments, one internucleoside linkage in the 3' wing is a phosphorothioate linkage. In various embodiments, one internucleoside linkage in the 5' wing is a phosphorothioate linkage, and one internucleoside linkage in the 3' wing is a phosphorothioate linkage.

[0408] In some embodiments, one or more modified internucleoside linkages in the 5' wing, 3' wing, or central region are phosphorothioate internucleoside linkages. In some embodiments, the phosphorothioate linkage is a stereochemically pure phosphorothioate linkage. In some embodiments, the phosphorothioate linkage is an Sp phosphorothioate linkage. In other embodiments, the phosphorothioate linkage is an Rp phosphorothioate linkage.

[0409] In some embodiments, one or more modified internucleoside linkages in the 5' wing, 3' wing, or central region can be any alkyl phosphate linkage, alkyl phosphate linkage, 3-methoxypropyl phosphate linkage, phosphorodithioate linkage, phosphotriester linkage, methylphosphonate linkage, aminoalkylphosphotriester linkage, alkylenephosphonate linkage, phosphite linkage, phosphoramidate linkage, thiophosphoramidate linkage, phosphorodiamidate (e.g., comprising phosphorodiamidate morpholino (PMO), 3' aminoribose, or 5' aminoribose) linkage, aminoalkylphosphoramidate linkage, thiophosphoramidate linkage, thioalkylphosphate linkage, thioalkylphosphotriester linkage, phosphorothioate linkage, phosphoroselenoate linkage, and borophosphate linkage. In various embodiments, each modified internucleoside linkage in the 5' wing, 3' wing, or central region can be designed independently of the other modified internucleoside linkages. In other words, the modified internucleoside linkages of the 5' wing, the 3' wing, and the central region need not all be the same type of modified internucleoside linkage.In various embodiments, the modified internucleoside linkages are interspersed throughout the antisense compound.

[0410] In various embodiments, one or more internucleoside linkages in the 5' wing, 3' wing, 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).

[0411] In various embodiments, the internucleoside linkages of one region (e.g., the 5' wing, the 3' wing, or the central region) may be different from the internucleoside linkages of another region. In specific embodiments, the 5' wing comprises at least one modified internucleoside linkage, the 3' wing 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 an oligonucleotide comprises a phosphodiester linkage, and each of the 5' and 3' wing regions comprises one or more phosphorothioate linkages. In specific embodiments, all internucleoside linkages in the 5' wing are modified internucleoside linkages, all internucleoside linkages in the 3' wing are modified internucleoside linkages, and all internucleoside linkages in the central region are unmodified internucleoside linkages (e.g., phosphodiester linkages).

[0412] In a specific embodiment, the gapmer AON is a 5-10-5 gapmer and the internucleoside linkage of the gapmer AON is represented by: Ssssssssssssssssssss (wherein "s" refers to a phosphorothioate linkage). In a specific embodiment, the gapmer AON is a 5-10-5 gapmer and the gapmer The internucleoside linkages of AON are represented by any of the following: ssssssssssssssssss, sossssssssssssssosss, sssssooooooooossssss, sosossssssssssssosos, sooooosssssssssssooos, sooooossssssssssoos, sooooossssssssssooss, ooooossssssssssssoooos, oooooooooooooooosssss, soosssssssssssssssoos, soosssssssssssssssss, sssssssssssssssssoos, and ssssssoooooooooooooo (wherein "s" refers to a phosphorothioate linkage and "o" refers to a phosphodiester linkage). In specific embodiments, the gapmer AON is a 3-8-3 gapmer and the internucleoside linkage of the gapmer AON is represented by: sssssssssssss (wherein "s" refers to a phosphorothioate linkage).

[0413] In specific embodiments, the gapmer AON is a 3-8-3 gapmer and the internucleoside linkage of the gapmer AON is represented by any of the following: sssooooooosss, ooossssssssooo, sssssssssssooo, sosssssssssos, sossssssssssss, sssssssssssos and ooosssssssssss (wherein "s" refers to a phosphorothioate bond and "o" refers to a phosphodiester bond). In specific embodiments, the gapmer AON is a 3-10-3 gapmer and the internucleoside linkage of the gapmer AON is represented by: ssssssssssssss (wherein "s" refers to a phosphorothioate bond).

[0414] In specific embodiments, the gapmer AON is a 3-10-3 gapmer and the internucleoside linkage of the gapmer AON is represented by any of the following: sssooooooooosss, ooossssssssssooo, sssssssssssssooo, sosssssssssssos, sossssssssssssss, ssssssssssssos and ooosssssssssssss (wherein "s" refers to a phosphorothioate bond and "o" refers to a phosphodiester bond). In specific embodiments, the gapmer AON is a 4-10-4 gapmer and the internucleoside linkage of the gapmer AON is represented by: ssssssssssssssss (wherein "s" refers to a phosphorothioate bond).

[0415] In specific embodiments, the gapmer AON is a 4-10-4 gapmer and the internucleoside linkage of the gapmer AON is represented by any of the following: ssssooooooooossss, oooosssssssssssoooo, sssssssssssssssoooo, soosssssssssssssoos, soosssssssssssssss, ssssssssssssssoos and oooossssssssssssss (wherein "s" refers to a phosphorothioate bond and "o" refers to a phosphodiester bond). In specific embodiments, the gapmer AON is a 4-8-4 gapmer and the internucleoside linkage of the gapmer AON is represented by: ssssssssssssss (wherein "s" refers to a phosphorothioate bond).

[0416] In specific embodiments, the gapmer AON is a 4-8-4 gapmer and the internucleoside linkages of the gapmer AON are represented by any of the following: ssssooooooosssss, oooossssssssoooo, ssssssssssssoooo, soosssssssssssoos, soossssssssssss, ssssssssssssoos and oooosssssssssss (wherein "s" refers to a phosphorothioate bond and "o" refers to a phosphodiester bond).

[0417] In specific embodiments, the gapmer AON is a 5-8-5 gapmer, and the internucleoside linkage of the gapmer AON is represented by any of the following: sssssssssssssssss or sosssssssssssssoss (wherein "s" refers to a phosphorothioate linkage and "o" refers to a phosphodiester linkage).

[0418] Modified sugar moiety

[0419] AONs, such as those having the sequence of any one of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, or 167805-301566, or those having one or more spacers replacing one or more nucleosides of any one of the sequences of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, or 167805-301566, such as SEQ ID NOs: 301692-301742 or any of the sequences in Tables 4A-4C, can contain one or more nucleosides in which the sugar group has been modified. Such sugar-modified nucleosides can confer enhanced nuclease stability, increased binding affinity, or some other beneficial biological property on the antisense compounds.

[0420] In various embodiments, the nucleoside with a modified sugar moiety comprises a ribose sugar in which the 2'-OH group can be replaced by any one selected from the group consisting of OR, R, R'OR, SH, SR, NH2, NR2, N3, CN, F, Cl, Br and I (wherein R is alkyl or aryl and R' is alkylene), 2'-O-methyl (2'-OMe) nucleosides, 2'-O-(2-methoxyethyl) (2'-MOE) nucleosides, peptide nucleic acids (PNAs), bicyclic nucleic acids (BNAs), 2'-deoxy-2'-fluoro nucleosides, 2'-fluoro-β-D-arabinoside, locked nucleic acids (LNAs), constrained ethyl 2'-4'-bridged nucleic acids (cEt), S-cEt, morpholino oligomers, tcDNAs, 2'-O,4'-C-ethylene-linked nucleic acids (ENA), hexitol nucleic acids (HNAs), and tricyclic analogs (e.g., tcDNAs).

[0421] In certain embodiments, the nucleoside comprises a chemically modified ribofuranose ring portion. Examples of chemically modified ribofuranose rings include, but are not limited to, the addition of substituents (including 5' and 2' substituents, bridging non-twin ring atoms to form bicyclic nucleic acids (BNAs), the use of S, N (R), or C (R1) (R2) (R, R1, and R2 are each independently H, C1-C 12alkyl, or protecting group), and combinations thereof to replace the ribosyl ring oxygen atom. Examples of chemically modified sugars include 2'-F-5'-methyl substituted nucleosides (other disclosed 5', 2'-disubstituted nucleosides are described in PCT International Application WO 2008 / 101157 published on August 21, 2008), or replacement of the ribosyl ring oxygen atom with S or CF2 and further substitution at the 2'-position (see U.S. Patent Application US2005-0130923 published on June 16, 2005), or 5'-substitution of BNA (see PCT International Application WO 2007 / 134181 published on November 22, 2007, in which LNA is, for example, 5'-methyl or 5'-vinyl substituted).

[0422] Examples of nucleosides with modified sugar moieties include, but are not limited to, nucleosides containing 5'-vinyl, 5'-methyl (R or S), 4'-S, 2'-F, 2'-OCH3, 2'-OCH2CH3, 2'-O CH2CH2F, and 2'-O(CH2)2OCH3 substituents. The substituent at the 2' position can also be selected from allyl, amino, azido, thio, O-allyl, OC1-C 10 Alkyl, OCF3, OCH2F, O(CH2)2SCH3, 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 R1, R m , and R n are independently H or substituted or unsubstituted C1-C 10 alkyl.

[0423] Other 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(CH3)-O-2'), S-constrained ethyl (S-cEt) 2'-4'-bridged nucleic acids, 4'-CH2-O-CH2-2', 4'-CH2-N(R)-2', 4'-CH(CH2OCH3)-O-2' ("constrained MOE" or "cMOE"), hexitol nucleic acids (HNA), and tricyclic analogs (e.g., tcDNA).

[0424] In some embodiments, the AON comprises a 2'-O-methyl nucleoside (2'OMe) (e.g., an AON comprising one or more 2'OMe-modified sugars), a 2'-O-(2-methoxyethyl) (2'-MOE) (e.g., an AON comprising one or more 2'-MOE-modified sugars (e.g., 2'-MOE)), a peptide nucleic acid (PNA) (e.g., an AON comprising one or more N-(2-aminoethyl)-glycine units linked by amide bonds or carbonyl methylene linkages as repeating units in place of a sugar-phosphate backbone), a locked nucleic acid (LNA) (e.g., an AON comprising one or more locked ribose sugars and can be a mixture of 2'-deoxynucleotides or 2'OMe nucleotides), a constrained ethyl 2'-4'-nucleotide. '-bridged nucleic acid (c-ET) (e.g., an AON comprising one or more cET sugars), cMOE (e.g., an AON comprising one or more cMOE sugars), a morpholino oligomer (e.g., an AON comprising a backbone comprising one or more PMOs), a deoxy-2'-fluoro nucleoside (e.g., an AON comprising one or more 2'-fluoro-β-D-arabinoside), a 2'-O,4'-C-ethylene-linked nucleic acid (ENA) (e.g., an AON comprising one or more ENA-modified sugars), a hexitol nucleic acid (HNA) (e.g., an AON comprising one or more HNA-modified sugars), or a tricyclic analog (tcDNA) (e.g., an AON comprising one or more tcDNA-modified sugars).

[0425] As used herein, "bicyclic nucleoside" refers to a modified nucleoside comprising a bicyclic sugar moiety. Examples of bicyclic nucleosides include, but are not limited to, nucleosides comprising a bridge between 4' and 2' ribosyl ring atoms. In certain embodiments, provided herein are antisense compounds comprising one or more bicyclic nucleosides comprising a 4' to 2' bridge. Examples of such 4' to 2' bridged bicyclic nucleosides include, but are not limited to, one of the following formulae: 4'-(CH2)-O-2'(LNA); 4'-(CH2)-S-2'; 4'-(CH2)2-O-2'(ENA); 4'-CH(CH3)-O-2' and 4'-CH(CH2OCH3)-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 International Application Publication No. WO / 2009 / 006478, published on January 8, 2009); 4'-CH2-N(OCH3)-2' (and its analogs (see International Application No. WO / 2008 / 150729, published on December 11, 2008); 4'-CH2-ON(CH3)-2' (see U.S. Patent Application Publication No. US2004-0171570, published on September 2, 2004); 4'-CH2-N(R)-O-2', wherein R is H, C1-C 12Alkyl, or protecting group (see U.S. Patent No. 7,427,672, issued September 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 International Application Publication No. WO 2008 / 154401 published December 8, 2008)).

[0426] Further reports related to bicyclic nucleosides can also be found in the published literature (see, for example: 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 Serial Nos. 60 / 989,574; 61 / 026,995; 61 / 026,998; 61 / 056,564; 61 / 086,231; 61 / 097,787; and 61 / 099,844; PCT International Application Publication No. WO 1994 / 014226; WO 2004 / 106356; WO 2005 / 021570; WO 2007 / 134181; WO 2008 / 150729; WO 2008 / 154401; and WO 2009 / 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 March 25, 1999 as WO 99 / 14226).

[0427] In certain embodiments, the bicyclic sugar moiety of the 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, wherein such bridge independently comprises 1, or 2 to 4 independently selected from -[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 )-linking group;

[0428] in:

[0429] x is 0, 1, or 2;

[0430] n is 1, 2, 3, or 4;

[0431] Each R a and R b are independently H, a protecting group, a hydroxyl group, a 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, substituted heterocyclic, heteroaryl, substituted heteroaryl, C5-C7 alicyclic, substituted C5-C7 alicyclic, halogen, OJ1, NJ1J2, SJ1, N3, COOJ1, acyl (C(=O)-H), substituted acyl, CN, sulfonyl (S(=O)2-J1), or sulfinyl (S(=O)-J1); and each J1 and J2 are independently H, C1-C 12 Alkyl, substituted C1-C 12 Alkyl, C2-C 12 Alkenyl, substituted C2-C 12 Alkenyl, C2-C 12 Alkynyl, substituted C2-C 12 Alkynyl, C5-C 20 Aryl, substituted C5-C 20 Aryl, acyl (C (= O) -H), substituted acyl, heterocyclic, substituted heterocyclic, C1-C 12 Aminoalkyl, substituted C1-C 12 aminoalkyl, or a protecting group.

[0432] In certain embodiments, the bridge of the bicyclic sugar moiety is -[C(R a )(Rb )] n -、-[-[C(R a )(R b )] n -O-、-C(R a R b )-N(R)-O-, or -C(R a R b In certain embodiments, the bridge is 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'-, wherein each R is independently H, a protecting group or a C1-C 12 Alkyl, each R a and R b are independently H, a protecting group, a hydroxyl group, a 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, substituted heterocyclic, heteroaryl, substituted heteroaryl, C5-C7 alicyclic, substituted C5-C7 alicyclic, halogen, OJ1, NJ1J2, SJ1, N3, COOJ1, acyl (C(=O)--H), substituted acyl, CN, sulfonyl (S(=O)2-J1), or sulfonyl (S(=O)-J1).

[0433] In certain embodiments, bicyclic nucleosides are further defined by isomeric configurations. For example, nucleosides comprising a 4'-2' methyleneoxy bridge can be in an α-L configuration or a β-D configuration. Previously, α-L-methyleneoxy (4'-CH2-O-2') BNAs have been incorporated into antisense oligonucleotides that exhibit antisense activity (Frieden et al., Nucleic Acids Research, 2003, 21, 6365-6372).

[0434] In certain embodiments, bicyclic nucleosides include, but are not limited to, α-L-methyleneoxy (4′-CH2-O-2′) BNA, β-D-methyleneoxy (4′-CH2-O-2′) BNA, ethoxy (4′-(CH2)2-O-2) BNA, aminooxy (4′-CH2-ON(R)-2′) BNA, oxyamino (4′-CH2-N(R)-O-2′) BNA, methyl(methyleneoxy) (4′-CH(CH3)-O-2′) BNA, methylenethio (4′-CH2-S-2′) BNA, methyleneamino (4′-CH2-N(R)-2′) BNA, methylcarbocycle (4′-CH2-CH(CH3)-2′) BNA, and propylenecarbocycle (4′-(CH2)3-2′) BNA.

[0435] 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 oxyimino bridge) connecting the two carbon atoms between the 4' and 2' positions of the nucleoside sugar unit to form a bicyclic sugar. Examples of such bicyclic sugars include, but are not limited to, (A) α-L-methyleneoxy (4'-CH2-O-2') LNA, (B) β-D-methyleneoxy (4'-CH2-O-2') LNA, (C) ethoxy (4'-(CH2)2-O-2') LNA, (D) aminooxy 4'-CH2-ON(R)-2') LNA, and (E) oxyamino (4'-CH2-N(R)-O-2') LNA; wherein R is H, C1-C 12 an alkyl group, or a protecting group (see U.S. Patent No. 7,427,672 issued September 23, 2008).

[0436] As used herein, LNA nucleosides include, but are not limited to, nucleosides having at least one bridge between the 4' and 2' positions of the sugar, wherein each bridge independently comprises 1 or 2 to 4 linking groups independently selected from: -[C(R1)(R2)] n -, -C(R1)=C(R2)-, -C(R1)=N-, -C(=NR1)-, -C(=O)-, -C(=S)-, -O-, -Si(R1)2-, -S(=O) x - and -N(R1)-; wherein: x is 0, 1, or 2; n is 1, 2, 3, or 4; each R1 and R2 is independently H, a protecting group, a hydroxyl group, a C1-C 12 Alkyl, substituted C 1- C 12 Alkyl, C2-C 12 Alkenyl, substituted C2-C 12 Alkenyl, C2-C 12 Alkynyl, substituted C2-C 12 Alkynyl, C5-C20 Aryl, substituted C5-C 20 Aryl, heterocyclic, substituted heterocyclic, heteroaryl, substituted heteroaryl, C5-C7 alicyclic, substituted C5-C7 alicyclic, halogen, OJ1, NJ1J2, SJ1, N3, COOJ1, acyl (C(=O)-H), substituted acyl, CN, sulfonyl (S(=O)2-J1), or sulfinyl (S(=O)-J1); and each J1 and J2 are independently H, C1-C 12 Alkyl, substituted C1-C 12 Alkyl, C2-C 12 Alkenyl, substituted C2-C 12 Alkenyl, C2-C 12 Alkynyl, substituted C2-C 12 Alkynyl, C5-C 20 Aryl, substituted C5-C 20 Aryl, acyl (C (= O) -H), substituted acyl, heterocyclic, substituted heterocyclic, C1-C 12 Aminoalkyl, substituted C1-C 12 aminoalkyl, or a protecting group.

[0437] Examples of 4'-2' bridges encompassed within the definition of LNA include, but are not limited to, one of the following formulae: -[C(R1)(R2)] n -、-[C(R1)(R2)] n -O-, -C(R1R2)-N(R1)-O-, or -C(R1R2)-ON(R1)-. In addition, other bridge groups encompassed by the definition of LNA are 4'-CH2-2', 4'-(CH2)2-2', 4'-(CH2)3-2', 4'-CH2-O-2', 4'-(CH2)2-O-2', 4'-CH2-ON(R1)-2', and 4'-CH2-N(R1)-O-2'-bridges, wherein each R1 and R2 are independently H, a protecting group, or C1-C 12 alkyl.

[0438] In the definition of LNA according to the present invention, also include wherein 2 '-hydroxyl of ribosyl sugar ring is connected to 4 ' carbon atom of sugar ring so as to form bridge to form the LNA of bicyclic sugar moiety.Bridge can be the methylene (-CH2-) group connecting 2 ' oxygen atom and 4 ' carbon atom, wherein used term methyleneoxy (4 '-CH2-O-2 ') LNA.In addition, in the case of the bicyclic sugar moiety with ethylene abutment in this position, use term ethyleneoxy (4 '-CH2CH2-O-2 ') LNA.As used herein, the definition of LNA also contains α-L-methyleneoxy (4 '-CH2-O-2 '), the isomer of methyleneoxy (4 '-CH2-O-2 ') LNA.

[0439] In various embodiments, AON disclosed herein comprises one or more 2'-O-(2-methoxyethyl) (2'-MOE) nucleosides. In various embodiments, AON disclosed herein comprises 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 2'-O-(2-methoxyethyl) (2'-MOE) nucleosides. In various embodiments, AON disclosed herein comprises more than 25 2'-O-(2-methoxyethyl) (2'-MOE) nucleosides.

[0440] In some embodiments, the AON comprises a modified sugar moiety 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 comprises at least one modified sugar moiety. In various embodiments, the 3' wing comprises at least one modified sugar moiety. In various embodiments, the 5' wing 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 sugar moieties. In various embodiments, the 3' wing 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 sugar moieties. In some embodiments, each of the 5' wing and / or 3' wing comprises 1 to 7 modified sugar moieties, for example, 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 a specific embodiment, the 5' wing comprises 3 modified sugar moieties and the 3' wing comprises 3 modified sugar moieties. In a specific embodiment, the 5' wing comprises 4 modified sugar moieties and the 3' wing comprises 4 modified sugar moieties. In a specific embodiment, the 5' wing comprises 5 modified sugar moieties and the 3' wing comprises 5 modified sugar moieties.

[0441] In various embodiments, the nucleoside having modified sugar moieties at the 5' and 3' wings is any of a ribose sugar, wherein the 2'-OH group of the ribose sugar can be selected from the group consisting of OR, R, R'OR, SH, SR, OR, R, R'OR, SH, SR, NH2, NR2, N3, CN, F, Cl, Br, and I (wherein R is alkyl or aryl and R' is alkylene), 2'-O-methyl (2'-OMe) nucleoside, 2'-O-(2-methoxyethyl) (2'-OMe) nucleoside, '-MOE) nucleosides, peptide nucleic acids (PNAs), bicyclic nucleic acids (BNAs), 2'-deoxy-2'-fluoro nucleosides, 2'-fluoro-β-D-arabinoside, locked nucleic acids (LNAs), constrained ethyl 2'-4'-bridged nucleic acids (cEt), S-cEt, morpholino oligomers, tcDNAs, 2'-O,4'-C-ethylene-linked nucleic acids (ENA), hexitol nucleic acids (HNAs), and tricyclic analogs (e.g., tcDNAs) are substituted with any one of the group consisting of.

[0442] In some embodiments, the 5' wing and / or 3' wing comprises at least one 2'-MOE nucleoside. In some embodiments, both the 5' and 3' wing comprise at least one 2'-MOE nucleoside. In some embodiments, each of the 5' and 3' wing comprises 2, 3, 4, 5, 6, 7, 8, 9, or 10 2'-MOE nucleosides. In specific embodiments, each of the 5' and 3' wing comprises 4 2'-MOE nucleosides. In specific embodiments, each of the 5' and 3' wing comprises 5 2'-MOE nucleosides. In some embodiments, all nucleosides in each of the 5' and 3' wing are 2'-MOE nucleosides.

[0443] In other embodiments, the wings can comprise 2'-MOE nucleosides and other nucleosides (mixed wings), such as DNA nucleosides and / or non-MOE modified nucleosides, such as bicyclic nucleosides (BNA) (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.

[0444] In some embodiments, the 5'-wing or 3'-wing comprises at least one BNA (e.g., at least one LNA nucleoside or cET nucleoside). In some embodiments, each of the 5'-wing and 3'-wing comprises a BNA. In some embodiments, all nucleosides in the 5'- and 3'-wings are BNAs. In further embodiments, the BNAs in the 5'- and / or 3'-wings are independently selected from the group consisting of oxy-LNA, thio-LNA, amino-LNA, cET, and / or ENA, whether in β-D or α-L configuration, or a combination thereof.

[0445] 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 comprise 2'-O-methyl nucleosides. In some embodiments, all nucleosides in the wing are 2'-O-methyl nucleosides.

[0446] Modified nucleobases

[0447] AONs, such as those having the sequence of 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 of any one of the sequences of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, or 167805-301566, such as SEQ ID NOs: 301692-301742 or any of the sequences in Tables 4A-4C, comprise 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.

[0448] 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.

[0449] In various embodiments, the AON comprises modified and unmodified nucleobases. For example, the AON can comprise cytosine and 5-methylcytosine. In some embodiments, the AON can comprise 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 cytosines and further comprise 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 5-methylcytosines.

[0450] In various embodiments, each specific type of nucleobase in the AON is replaced by a corresponding modified nucleobase. For example, each guanine in the AON is replaced by 5-methylguanine. As another example, each cytosine in the AON is replaced by 5-methylcytosine.

[0451] In some embodiments, AON comprises the core base of the modification designed according to the gapmer design of gapmer AON.In various embodiments, the nucleoside of the connection of 5' wing region, the nucleoside of the connection of 3' wing region or the nucleoside of the connection of central region comprise the core base of one or more modifications.In some embodiments, 5' wing region and / or 3' wing region comprise 1 to 10 core bases of modification, for example, 2 to 8 core bases of modification, 3 to 6 core bases of modification or 4 to 5 core bases of modification.In some embodiments, 5' wing region and / or 3' wing region comprise 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 core bases of modification.In some embodiments, central region comprises 1 to 10 core bases of modification, for example, 2 to 8 core bases of modification, 3 to 6 core bases of modification or 4 to 5 core bases of modification. 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, and 5-methylpurines such as 5-methylguanine.

[0452] In various embodiments, at least one cytosine in the 5' wing and / or 3' wing of the AON is replaced by a modified nucleobase, such as 5-methylcytosine. In various embodiments, at least one cytosine in the 5' wing is replaced by a modified nucleobase, such as 5-methylcytosine. In various embodiments, at least one cytosine in the 3' wing is replaced by a modified nucleobase, such as 5-methylcytosine. In various embodiments, at least one cytosine in the central region is replaced by a modified nucleobase, such as 5-methylcytosine. In various embodiments, all cytosines in the 5' wing are replaced by a modified nucleobase, such as 5-methylcytosine. In various embodiments, all cytosines in the 3' wing are replaced by a modified nucleobase, such as 5-methylcytosine. In various embodiments, all cytosines in the central region are replaced by a modified nucleobase, such as 5-methylcytosine.

[0453] In a specific embodiment, all cytosines in the 5' wing, all cytosines in the 3' wing, and all cytosines in the central region are replaced with modified nucleobases, such as 5-methylcytosine. In a specific embodiment, all cytosines in the 5' wing, all cytosines in the 3' wing are replaced with modified nucleobases, such as 5-methylcytosine; however, all cytosines in the central region are unmodified nucleobases.

[0454] Modified oligonucleotides

[0455] Additional embodiments of modified oligonucleotides are described herein, which can comprise any of the modified internucleoside linkages and / or modified nucleosides (eg, modified sugar moieties and / or modified nucleobases) described above.

[0456] In some embodiments, the AON or a pharmaceutically acceptable salt thereof comprises the 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 the sequence of any one of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, or 167805-301566, such as SEQ ID NOs: NO: 301692-301742 or any of the sequences in Tables 4A-4C, etc., wherein at least one nucleoside in the sequence is substituted with 2'-O-(2-methoxyethyl) nucleoside, 2'-O-methyl nucleoside, 2'-O-(N-methylacetamide) nucleoside, 2'-deoxy-2'-fluoro nucleoside, 2'-fluoro-β-D-arabinoside, bicyclic nucleic acid, bridged nucleic acid, locked nucleic acid (LNA), constrained ethyl (cET) nucleic acid, tricyclic DNA (tcDNA), 2'-O, 4'-C-ethylene-linked nucleic acid (ENA) or peptide nucleic acid (PNA). In specific embodiments, at least one internucleoside bond of the AON is a phosphorothioate bond. In some embodiments, all internucleoside bonds of the AON are phosphorothioate bonds. Also described herein are pharmaceutical compositions comprising any of the aforementioned antisense oligonucleotides or pharmaceutically acceptable salts thereof and a pharmaceutically acceptable excipient.

[0457] AON as described herein can include chemically modified nucleosides, including modified ribonucleosides and / or modified deoxyribonucleosides.Chemically modified nucleosides include 2'-substituted nucleosides, wherein the 2' position of the sugar ring includes a part (e.g., -F or O-alkyl) that is different from-H or -OH. 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.

[0458] In some embodiments, the AON may comprise chemically modified nucleosides, such as 2'O-methyl ribonucleosides, such as 2'O-methyl cytidine, 2'O-methyl guanosine, 2'O-methyl uridine, and / or 2'O-methyl adenosine. The AON described herein may also comprise one or more chemically modified bases, including 5-methyl pyrimidines, such as 5-methyl cytosine, and / or 5-methyl purines, such as 5-methyl guanine. The AON described herein may also comprise any of the following chemically modified nucleosides: 5-methyl-2'-O-methyl cytidine, 5-methyl-2'-O-methyl thymidine, 5-methyl cytidine, 5-methyl uridine, and / or 5-methyl 2'-deoxycytidine.

[0459] It is contemplated that in some embodiments, the disclosed AONs may optionally have at least one modified nucleobase, such as 5-methylcytosine and / or at least one methylphosphate nucleotide, located, for example, at only one of the 5' or 3' termini or both the 5' and 3' termini or along the oligonucleotide sequence.

[0460] In certain embodiments, the present disclosure provides AONs having a combination of various modified nucleosides, such as a combination of peptide nucleic acid (PNA) and locked nucleic acid (LNA). 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.

[0461] AON as described herein can include chemical modifications that promote the stabilization of the 5'-phosphate at the end of the oligonucleotide and phosphatase-resistant analogs of 5'-phosphate. Chemical modifications that promote the stabilization of the 5'-phosphate at the end of the oligonucleotide or chemical modifications that serve as phosphatase-resistant analogs of 5'-phosphate include but are not limited to 5'-methyl phosphate, 5'-methylene phosphate, 5'-methylene phosphate analogs, 5'-E-vinyl phosphate (5'-E-VP), 5'-thiophosphate, and 5'-C-methyl analogs. Chemical modifications that promote the stabilization of the 5'-phosphate at the end of the AON and phosphatase-resistant analogs of 5'-phosphate are described in the following literature: 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.

[0462] In some embodiments described herein, the AON or a 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 an AON having 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, such as SEQ ID NOs: NO: 301692-301742 or any of the sequences in Tables 4A-4C, and the like, and at least one nucleoside linkage of the nucleotide sequence is a phosphorothioate linkage, an alkylphosphate linkage, an alkylphosphate linkage, a 3-methoxypropylphosphate linkage, a phosphorodithioate linkage, a phosphotriester linkage, a methylphosphonate linkage, an aminoalkylphosphotriester linkage, an alkylenephosphonate linkage, a phosphite linkage, a phosphoramidate linkage, a thiophosphoramidate linkage, a phosphorodiamidate (e.g., comprising phosphorodiamidate morpholino (PMO), 3' aminoribose or 5' aminoribose) linkage, an aminoalkylphosphodiamidate linkage, a thiophosphoramidate linkage, a thioalkylphosphate linkage, a thioalkylphosphotriester linkage, a phosphorothioate linkage, a phosphoroselenoate linkage, and a borophosphate linkage. In specific 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, the internucleoside linkages of any of the following AON sequences are phosphorothioate linkages: sequences of 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 of any one of the sequences of SEQ ID NOs: 1-954, 1914-149354, 149362-158581, or 167805-301566, e.g., any one of SEQ ID NOs: 301692-301742 or the sequences in Tables 4A-4C.

[0463] The contemplated AONs may optionally comprise at least one modified sugar. For example, the sugar moiety of at least one nucleotide constituting the oligonucleotide is ribose, wherein the 2'-OH group may be selected from OR, R, R'OR, SH, SR, OR, R, R'OR, SH, SR, NH2, NR2, N3, CN, F, Cl, Br, and I (wherein R is alkyl or aryl and R' is alkylene).

[0464] In a specific embodiment, the AON has a structure of eeeee-d10-eeeee (wherein "e" represents a 2'-MOE modified nucleoside, and wherein "d10" represents a continuous sequence of 10 DNA nucleobases). In this embodiment, the 5' wing region comprises 5 2'-MOE modified nucleosides, the gap region comprises 10 continuous DNA nucleobases, and the 3' wing region comprises 5 2'-MOE modified nucleosides. In various embodiments, the internucleoside linkage of the 5-10-5 gapmer AON can have a sequence of sssssooooooooosssss (wherein "s" refers to a phosphorothioate bond and "o" refers to a phosphodiester bond), wherein all phosphorothioate bonds are located in the 5' wing region or the 3' wing region, and all phosphodiester bonds are located in the central region of the AON. In various embodiments, the AON comprises unmodified cytosine. In various embodiments, the AON comprises a modified cytosine (e.g., 5-methylcytosine). In various embodiments, all cytosines in the 5' wing and the 3' wing are modified cytosines (eg, 5-methylcytosine). In various embodiments, all cytosines in the central region are modified cytosines (eg, 5-methylcytosine).

[0465] In a specific embodiment, the AON has a structure of eeeee-d10-eeeee (wherein "e" represents a 2'-MOE modified nucleoside, and wherein "d10" represents a continuous sequence of 10 DNA nucleobases). In this embodiment, the 5' wing region comprises 5 2'-MOE modified nucleosides, the gap region comprises 10 continuous DNA nucleobases, and the 3' wing region comprises 5 2'-MOE modified nucleosides. The internucleoside linkage of the AON can have a sequence of ooooosssssssssooooo (wherein "s" refers to a phosphorothioate bond and "o" refers to a phosphodiester bond). In various embodiments, the AON comprises unmodified cytosine. In various embodiments, the AON comprises a modified cytosine (e.g., 5-methylcytosine). In various embodiments, all cytosines in the 5' wing region and the 3' wing region are modified cytosines (e.g., 5-methylcytosine). In various embodiments, all cytosines in the central region are modified cytosines (e.g., 5-methylcytosine).

[0466] In a specific embodiment, the AON has a structure of eeeee-d10-eeeee (wherein "e" represents a 2'-MOE modified nucleoside, and wherein "d10" represents a continuous sequence of 10 DNA nucleobases). In this embodiment, the 5' wing region comprises 5 2'-MOE modified nucleosides, the gap region comprises 10 continuous DNA nucleobases, and the 3' wing region comprises 5 2'-MOE modified nucleosides. The internucleoside linkages of the AON can have a ssssssssssssssssss sequence (wherein "s" refers to a phosphorothioate bond), wherein all internucleoside linkages of the AON are phosphorothioate bonds. In various embodiments, the AON comprises unmodified cytosine. In various embodiments, the AON comprises modified cytosine (e.g., 5-methylcytosine). In various embodiments, all cytosines in the 5' wing region and the 3' wing region are modified cytosine (e.g., 5-methylcytosine). In various embodiments, all of the cytosines in the central region are modified cytosines (eg, 5-methylcytosine).

[0467] In a specific embodiment, AON has the structure of eee-d8-eee (wherein "e" represents a 2'-MOE modified nucleoside, and wherein "d8" represents a continuous sequence of 8 DNA nucleobases). In this embodiment, the 5' wing region comprises 3 2'-MOE modified nucleosides, the gap region comprises 8 oligonucleotide units comprising at least 4 continuous nucleobases, and the 3' wing region comprises 3 2'-MOE modified nucleosides. The internucleoside linkage of AON can have a sequence of sssooooooosss (wherein "s" refers to a thiophosphate bond, and "o" refers to a phosphodiester bond), wherein all thiophosphate bonds are in the 5' wing region or the 3' wing region and all phosphodiester bonds are located in the central region of the AON. In various embodiments, AON comprises unmodified cytosine. In various embodiments, AON comprises modified cytosine (e.g., 5-methylcytosine). In various embodiments, all cytosines in the 5' wing region and the 3' wing region are modified cytosine (e.g., 5-methylcytosine). In various embodiments, all of the cytosines in the central region are modified cytosines (eg, 5-methylcytosine).

[0468] In a specific embodiment, AON has the structure of eee-d8-eee (wherein "e" represents a 2'-MOE modified nucleoside, and wherein "d8" represents a continuous sequence of 8 DNA nucleobases). In this embodiment, the 5' wing region comprises 3 2'-MOE modified nucleosides, the gap region comprises 8 oligonucleotide units comprising 4 continuous DNA nucleobases, and the 3' wing region comprises 3 2'-MOE modified nucleosides. The internucleoside linkage of AON can have a sequence of ooosssssssooo (wherein "s" refers to a thiophosphate bond, and "o" refers to a phosphodiester bond), wherein all phosphodiester bonds are in the 5' wing region or the 3' wing region and all thiophosphate bonds are located in the central region of the AON. In various embodiments, AON comprises unmodified cytosine. In various embodiments, AON comprises modified cytosine (e.g., 5-methylcytosine). In various embodiments, all cytosines in the 5' wing region and the 3' wing region are modified cytosine (e.g., 5-methylcytosine). In various embodiments, all of the cytosines in the central region are modified cytosines (eg, 5-methylcytosine).

[0469] In a specific embodiment, the AON has a structure of eee-d8-eee (wherein "e" represents a 2'-MOE modified nucleoside, and wherein "d8" represents a sequence of 8 consecutive DNA nucleobases). In this embodiment, the 5' wing region comprises 3 2'-MOE modified nucleosides, the gap region comprises 8 oligonucleotide units (the oligonucleotide comprises 4 consecutive DNA nucleobases), and the 3' wing region comprises 3 2'-MOE modified nucleosides. The internucleoside linkages of the AON can have a sequence of sssssssssssss (wherein "s" refers to a thiophosphate bond), wherein all internucleoside linkages of the AON are thiophosphate bonds. In various embodiments, the AON comprises unmodified cytosine. In various embodiments, the AON comprises modified cytosine (e.g., 5-methylcytosine). In various embodiments, all cytosines in the 5' wing region and the 3' wing region are modified cytosine (e.g., 5-methylcytosine). In various embodiments, all of the cytosines in the central region are modified cytosines (eg, 5-methylcytosine).

[0470] In a specific embodiment, AON has the structure of eee-d10-eee (wherein "e" represents a 2'-MOE modified nucleoside, and wherein "d10" represents a continuous sequence of 10 DNA nucleobases). In this embodiment, the 5' wing region comprises 3 2'-MOE modified nucleosides, the gap region comprises 10 continuous DNA nucleobases, and the 3' wing region comprises 3 2'-MOE modified nucleosides. The internucleoside linkage of AON can have a sequence of sssooooooooosss (wherein "s" refers to a thiophosphate bond, and "o" refers to a phosphodiester bond), wherein all thiophosphate bonds are in the 5' wing region or the 3' wing region and all phosphodiester bonds are located in the central region of the AON. In various embodiments, AON comprises unmodified cytosine. In various embodiments, AON comprises modified cytosine (e.g., 5-methylcytosine). In various embodiments, all cytosines in the 5' wing region and the 3' wing region are modified cytosine (e.g., 5-methylcytosine). In various embodiments, all of the cytosines in the central region are modified cytosines (eg, 5-methylcytosine).

[0471] In a specific embodiment, the AON has a structure of eee-d10-eee (wherein "e" represents a 2'-MOE modified nucleoside, and wherein "d10" represents a sequence of 10 consecutive DNA nucleobases). In this embodiment, the 5' wing region comprises 3 2'-MOE modified nucleosides, the gap region comprises 10 consecutive DNA nucleobases, and the 3' wing region comprises 3 2'-MOE modified nucleosides. The internucleoside linkage of the AON can have a sequence of ooosssssssssooo (wherein "s" refers to a phosphorothioate bond and "o" refers to a phosphodiester bond). In various embodiments, the AON comprises unmodified cytosine. In various embodiments, the AON comprises a modified cytosine (e.g., 5-methylcytosine). In various embodiments, all cytosines in the 5' wing region and the 3' wing region are modified cytosines (e.g., 5-methylcytosine).

[0472] In a specific embodiment, the AON has a structure of eee-d10-eee (wherein "e" represents a 2'-MOE modified nucleoside, and wherein "d10" represents a continuous sequence of 10 DNA nucleobases). In this embodiment, the 5' wing region comprises 3 2'-MOE modified nucleosides, the gap region comprises 10 continuous DNA nucleobases, and the 3' wing region comprises 3 2'-MOE modified nucleosides. The internucleoside linkages of the AON can have a sequence of sssssssssssssss (wherein "s" refers to a thiophosphate bond), wherein all internucleoside linkages of the AON are thiophosphate bonds. In various embodiments, the AON comprises unmodified cytosine. In various embodiments, the AON comprises modified cytosine (e.g., 5-methylcytosine). In various embodiments, all cytosines in the 5' wing region and the 3' wing region are modified cytosine (e.g., 5-methylcytosine). In various embodiments, all of the cytosines in the central region are modified cytosines (eg, 5-methylcytosine).

[0473] In a specific embodiment, AON has the structure of eeee-d10-eeee (wherein "e" represents a 2'-MOE modified nucleoside, and wherein "d10" represents a continuous sequence of 10 DNA nucleobases). In this embodiment, the 5' wing region comprises 4 2'-MOE modified nucleosides, the gap region comprises 10 continuous DNA nucleobases, and the 3' wing region comprises 4 2'-MOE modified nucleosides. The internucleoside linkage of AON can have a sequence of ssssooooooooossss (wherein "s" refers to a thiophosphate bond, and "o" refers to a phosphodiester bond), wherein all thiophosphate bonds are in the 5' wing region or the 3' wing region and all phosphodiester bonds are located in the central region of the AON. In various embodiments, AON comprises unmodified cytosine. In various embodiments, AON comprises modified cytosine (e.g., 5-methylcytosine). In various embodiments, all cytosines in the 5' wing region and the 3' wing region are modified cytosine (e.g., 5-methylcytosine). In various embodiments, all of the cytosines in the central region are modified cytosines (eg, 5-methylcytosine).

[0474] In a specific embodiment, the AON has a structure of eeee-d10-eeee (wherein "e" represents a 2'-MOE modified nucleoside, and wherein "d10" represents a continuous sequence of 10 DNA nucleobases). In this embodiment, the 5' wing region comprises 4 2'-MOE modified nucleosides, the gap region comprises 10 continuous DNA nucleobases, and the 3' wing region comprises 4 2'-MOE modified nucleosides. The internucleoside linkage of the AON can have a sequence of oooosssssssssoooo (wherein "s" refers to a phosphorothioate bond and "o" refers to a phosphodiester bond). In various embodiments, the AON comprises unmodified cytosine. In various embodiments, the AON comprises a modified cytosine (e.g., 5-methylcytosine). In various embodiments, all cytosines in the 5' wing region and the 3' wing region are modified cytosines (e.g., 5-methylcytosine). In various embodiments, all cytosines in the central region are modified cytosines (e.g., 5-methylcytosine).

[0475] In a specific embodiment, the AON has a structure of eeee-d10-eeee (wherein "e" represents a 2'-MOE modified nucleoside, and wherein "d10" represents a sequence of 10 consecutive DNA nucleobases). In this embodiment, the 5' wing region comprises 4 2'-MOE modified nucleosides, the gap region comprises 10 consecutive DNA nucleobases, and the 3' wing region comprises 4 2'-MOE modified nucleosides. The internucleoside linkages of the AON can have a sequence of sssssssssssssssss (wherein "s" refers to a thiophosphate bond), wherein all internucleoside linkages of the AON are thiophosphate bonds. In various embodiments, the AON comprises unmodified cytosine. In various embodiments, the AON comprises modified cytosine (e.g., 5-methylcytosine). In various embodiments, all cytosines in the 5' wing region and the 3' wing region are modified cytosine (e.g., 5-methylcytosine). In various embodiments, all of the cytosines in the central region are modified cytosines (eg, 5-methylcytosine).

[0476] In a specific embodiment, AON has the structure of eeee-d8-eeee (wherein "e" represents a 2'-MOE modified nucleoside, and wherein "d8" represents a continuous sequence of 8 DNA nucleobases). In this embodiment, the 5' wing region comprises 4 2'-MOE modified nucleosides, the gap region comprises 8 oligonucleotide units comprising 4 continuous DNA nucleobases, and the 3' wing region comprises 4 2'-MOE modified nucleosides. The internucleoside linkage of AON can have a sequence of ssssooooooossss (wherein "s" refers to a thiophosphate bond and "o" refers to a phosphodiester bond), wherein all thiophosphate bonds are in the 5' wing region or the 3' wing region and all phosphodiester bonds are located in the central region of the AON. In various embodiments, AON comprises unmodified cytosine. In various embodiments, AON comprises modified cytosine (e.g., 5-methylcytosine). In various embodiments, all cytosines in the 5' wing and the 3' wing are modified cytosines (eg, 5-methylcytosine). In various embodiments, all cytosines in the central region are modified cytosines (eg, 5-methylcytosine).

[0477] In a specific embodiment, AON has the structure of eeee-d8-eeee (wherein "e" represents a 2'-MOE modified nucleoside, and wherein "d8" represents a continuous sequence of 8 DNA nucleobases). In this embodiment, the 5' wing region comprises 4 2'-MOE modified nucleosides, the gap region comprises 8 oligonucleotide units comprising 4 consecutive DNA nucleobases, and the 3' wing region comprises 4 2'-MOE modified nucleosides. The internucleoside linkage of AON can have a sequence of oooosssssssoooo (wherein "s" refers to a thiophosphate bond and "o" refers to a phosphodiester bond). In various embodiments, AON comprises unmodified cytosine. In various embodiments, AON comprises modified cytosine (e.g., 5-methylcytosine). In various embodiments, all cytosines in the 5' wing region and the 3' wing region are modified cytosines (e.g., 5-methylcytosine). In various embodiments, all cytosines in the central region are modified cytosines (e.g., 5-methylcytosine).

[0478] In a specific embodiment, the AON has a structure of eeee-d8-eeee (wherein "e" represents a 2'-MOE modified nucleoside, and wherein "d8" represents a sequence of 8 consecutive DNA nucleobases). In this embodiment, the 5' wing region comprises 4 2'-MOE modified nucleosides, the gap region comprises 8 oligonucleotide units comprising 4 consecutive DNA nucleobases, and the 3' wing region comprises 4 2'-MOE modified nucleosides. The internucleoside linkages of the AON can have a sequence of sssssssssssssss (wherein "s" refers to a thiophosphate bond), wherein all internucleoside linkages of the AON are thiophosphate bonds. In various embodiments, the AON comprises unmodified cytosine. In various embodiments, the AON comprises modified cytosine (e.g., 5-methylcytosine). In various embodiments, all cytosines in the 5' wing region and the 3' wing region are modified cytosine (e.g., 5-methylcytosine).

[0479] In a specific embodiment, the AON has the structure of eeeeee-d11-eeeeee (wherein "e" represents a 2'-MOE modified nucleoside, and wherein "d11" represents a continuous sequence of 11 DNA nucleobases). In this embodiment, the 5' wing region comprises 6 2'-MOE modified nucleosides, the gap region comprises 11 consecutive DNA nucleobases, and the 3' wing region comprises 6 2'-MOE modified nucleosides. In various embodiments, the internucleoside linkage of the 6-11-6gapmer AON can have the sequence sssssssssssssssssssss (wherein "s" refers to a phosphorothioate bond). In various embodiments, the AON comprises unmodified cytosine. In various embodiments, the AON comprises a modified cytosine (e.g., 5-methylcytosine). In various embodiments, all cytosines in the 5' wing region and the 3' wing region are modified cytosines (e.g., 5-methylcytosine). In various embodiments, all of the cytosines in the central region are modified cytosines (eg, 5-methylcytosine).

[0480] In a specific embodiment, the AON has a structure of eeeee-d10-eeeee (wherein "e" represents a 2'-MOE modified nucleoside, and wherein "d10" represents a sequence of 10 consecutive DNA nucleobases). In this embodiment, the 5' wing region comprises 5 2'-MOE modified nucleosides, the gap region comprises 10 consecutive DNA nucleobases, and the 3' wing region comprises 5 2'-MOE modified nucleosides. In various embodiments, the internucleoside linkage of the 5-10-5gapmer AON can have a sequence of sososssssssssssosos (wherein "s" refers to a phosphorothioate bond and "o" refers to a phosphodiester bond). In various embodiments, the AON comprises unmodified cytosine. In various embodiments, the AON comprises a modified cytosine (e.g., 5-methylcytosine). In various embodiments, all cytosines in the 5' wing region and the 3' wing region are modified cytosines (e.g., 5-methylcytosine). In various embodiments, all of the cytosines in the central region are modified cytosines (eg, 5-methylcytosine).

[0481] In a specific embodiment, the AON has a structure of eeeee-d10-eeeee (wherein "e" represents a 2'-MOE modified nucleoside, and wherein "d10" represents a sequence of 10 consecutive DNA nucleobases). In this embodiment, the 5' wing region comprises 5 2'-MOE modified nucleosides, the gap region comprises 10 consecutive DNA nucleobases, and the 3' wing region comprises 5 2'-MOE modified nucleosides. In various embodiments, the internucleoside linkage of the 5-10-5gapmerAON can have a sequence of sooooossssssssssooos (wherein "s" refers to a phosphorothioate bond and "o" refers to a phosphodiester bond). In various embodiments, the AON comprises unmodified cytosine. In various embodiments, the AON comprises a modified cytosine (e.g., 5-methylcytosine). In various embodiments, all cytosines in the 5' wing region and the 3' wing region are modified cytosines (e.g., 5-methylcytosine). In various embodiments, all of the cytosines in the central region are modified cytosines (eg, 5-methylcytosine).

[0482] In a specific embodiment, the AON has a structure of eeeee-d10-eeeee (wherein "e" represents a 2'-MOE modified nucleoside, and wherein "d10" represents a sequence of 10 consecutive DNA nucleobases). In this embodiment, the 5' wing region comprises 5 2'-MOE modified nucleosides, the gap region comprises 10 consecutive DNA nucleobases, and the 3' wing region comprises 5 2'-MOE modified nucleosides. In various embodiments, the internucleoside linkage of the 5-10-5gapmerAON can have a sequence of sooooossssssssssooss (wherein "s" refers to a phosphorothioate bond and "o" refers to a phosphodiester bond). In various embodiments, the AON comprises unmodified cytosine. In various embodiments, the AON comprises a modified cytosine (e.g., 5-methylcytosine). In various embodiments, all cytosines in the 5' wing region and the 3' wing region are modified cytosines (e.g., 5-methylcytosine). In various embodiments, all of the cytosines in the central region are modified cytosines (eg, 5-methylcytosine).

[0483] In a specific embodiment, the AON has a structure of eeeee-d8-eeeee (wherein "e" represents a 2'-MOE modified nucleoside, and wherein "d8" represents a sequence of 8 consecutive DNA nucleobases). In this embodiment, the 5' wing region comprises 5 2'-MOE modified nucleosides, the gap region comprises 8 oligonucleotide units comprising 4 consecutive DNA nucleobases, and the 3' wing region comprises 5 2'-MOE modified nucleosides. In various embodiments, the internucleoside linkage of the 5-8-5gapmer AON can have a sequence of sossssssssssssss (wherein "s" refers to a phosphorothioate bond and "o" refers to a phosphodiester bond). In various embodiments, the AON comprises unmodified cytosine. In various embodiments, the AON comprises a modified cytosine (e.g., 5-methylcytosine). In various embodiments, all cytosines in the 5' wing region and the 3' wing region are modified cytosines (e.g., 5-methylcytosine). In various embodiments, all of the cytosines in the central region are modified cytosines (eg, 5-methylcytosine).

[0484] In a specific embodiment, the AON has the structure of eeeee-d10-eeeee (wherein "e" represents a 2'-MOE modified nucleoside, and wherein "d10" represents a sequence of 10 consecutive DNA nucleobases). In this embodiment, the 5' wing comprises 5 2'-MOE modified nucleosides, the gap comprises 10 consecutive DNA nucleobases, and the 3' wing comprises 5 2'-MOE modified nucleosides. In various embodiments, the internucleoside linkages of the 5-10-5 gapmer AON can have a sequence of sosssssssssssssssss (wherein "s ” refers to a phosphorothioate bond, and “o” refers to a phosphodiester bond). In various embodiments, the AON comprises unmodified cytosines. In various embodiments, the AON comprises modified cytosines (e.g., 5-methylcytosine). In various embodiments, all cytosines in the 5' wing and the 3' wing are modified cytosines (e.g., 5-methylcytosine). In various embodiments, all cytosines in the central region are modified cytosines (e.g., 5-methylcytosine).

[0485] In a specific embodiment, the AON has a structure of eeeeee-d10-eeee (wherein "e" represents a 2'-MOE modified nucleoside, and wherein "d10" represents a sequence of 10 consecutive DNA nucleobases). In this embodiment, the 5' wing region comprises 6 2'-MOE modified nucleosides, the gap region comprises 10 consecutive DNA nucleobases, and the 3' wing region comprises 4 2'-MOE modified nucleosides. The internucleoside linkages of the AON can have a sequence of sssssssssssssss (wherein "s" refers to a thiophosphate bond), wherein all internucleoside linkages of the AON are thiophosphate bonds. In various embodiments, the AON comprises unmodified cytosine. In various embodiments, the AON comprises modified cytosine (e.g., 5-methylcytosine). In various embodiments, all cytosines in the 5' wing region and the 3' wing region are modified cytosine (e.g., 5-methylcytosine). In various embodiments, all of the cytosines in the central region are modified cytosines (eg, 5-methylcytosine).

[0486] target gene product

[0487] In general, 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 specific embodiments, 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 specific 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 an ATXN2 gene product. In specific 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 an SOD1 gene product. In specific embodiments, the AON disclosed herein comprises 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 MAPT gene product.

[0488] In embodiments of the invention described herein, AONs can target gene products of genes of one or more species. For example, AONs can 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 genes or macaque monkey genes). In specific embodiments, AONs target human gene products.

[0489] The gene product can be, for example, an RNA gene product such as an mRNA gene product, or a protein product of a 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 a gene sequence or RNA sequence such as an mRNA or pre-mRNA sequence or a portion thereof. In some embodiments, the AON comprises a nucleobase sequence that is partially complementary to a sequence shared between genes or RNA (e.g., mRNA) of multiple species. For example, 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 a gene sequence or RNA sequence such as an mRNA or pre-mRNA sequence or a portion thereof. N Antisense therapeutics are antisense therapeutics, such as AONs, that are complementary to sequences common to human, mouse, and / or primate genes or mRNAs.

[0490] PPM1A

[0491] In some embodiments of the present disclosure, the PPM1A gene product is a PPM1A mRNA or a PPM1A pre-mRNA comprising a sequence from nucleotide 41932 to nucleotide 42787 and from nucleotide 44874 to nucleotide 44990 of a PPM1A gene sequence (e.g., the PPM1A gene sequence of NCBI reference sequence NG_029698.1 (SEQ ID NO: 1909) or a PPM1A coding sequence), or a portion thereof. In some embodiments of the present disclosure, the PPM1A gene product comprises a sequence that is 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% identical to a PPM1A mRNA or PPM1A pre-mRNA comprising the sequence from nucleotide 41932 to nucleotide 42787 and from nucleotide 44874 to nucleotide 44990 of a PPM1A gene sequence, such as the PPM1A gene sequence of NCBI Reference Sequence NG_029698.1 (SEQ ID NO: 1909) or a PPM1A coding sequence, or a portion thereof.

[0492]

[0493]

[0494]

[0495]

[0496]

[0497]

[0498]

[0499]

[0500]

[0501]

[0502]

[0503]

[0504]

[0505]

[0506]

[0507]

[0508]

[0509]

[0510]

[0511]

[0512]

[0513]

[0514]

[0515]

[0516]

[0517]

[0518]

[0519]

[0520]

[0521]

[0522]

[0523]

[0524]

[0525]

[0526]

[0527] In some embodiments of the present disclosure, the PPM1A gene product is a PPM1A mRNA or PPM1A pre-mRNA comprising a sequence of one or more of nucleotides 8470-8926, 41933-42787, 44874-45990, 49055-49164, 50647-50704, and 51703-58336 from a PPM1A gene sequence (e.g., the PPM1A gene sequence of 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 a PPM1A pre-mRNA comprising a sequence from the coding region of the PPM1A gene sequence, such as a coding region comprising nucleotides 8470-8926, 41933-42787, 44874-45990, 49055-49164, 50647-50704, and 51703-58336 of the PPM1A gene sequence (e.g., 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 1, corresponding to NCBI reference sequence NM_021003.5 (SEQ ID NO: 1910).

[0528] In some embodiments of the present disclosure, the PPM1A gene product is a PPM1A mRNA or PPM1A pre-mRNA comprising a sequence of one or more of nucleotides 8470-8926, 9629-9730, 41933-42787, and 44874-47804 from a PPM1A gene sequence (e.g., the PPM1A gene sequence of 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 comprising a sequence from the coding region of a PPM1A gene sequence, such as 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., 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 2, corresponding to NCBI reference sequence NM_177951.3 (SEQ ID NO: 1911).

[0529] In some embodiments of the present disclosure, the PPM1A gene product is a PPM1A mRNA or PPM1A pre-mRNA comprising a sequence of one or more of nucleotides 4999-5295, 41933-42787, 44874-44990, 49055-49164, 50647-50704, 51703-58336 from a PPM1A gene sequence (e.g., the PPM1A gene sequence of 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 a PPM1A pre-mRNA comprising a sequence from one or more coding regions of a PPM1A gene sequence, such as a coding region comprising nucleotides 4999-5295, 41933-42787, 44874-44990, 49055-49164, 50647-50704, 51703-58336 of the PPM1A gene sequence (e.g., 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, corresponding to NCBI reference sequence NM_177952.3 (SEQ ID NO: 1912).

[0530] In some embodiments of the present disclosure, the PPM1A gene product comprises a sequence comprising nucleotides 457-1429 of PPM1A mRNA transcript variant 1 (i.e., for example, nucleotides 457-1429 of PPM1A mRNA transcript variant 1, corresponding to 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 that is 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% identical to nucleotides 457-1429 of PPM1A mRNA transcript variant 1 (i.e., for example, nucleotides 457-1429 of PPM1A mRNA transcript variant 1, corresponding to NCBI reference sequence NM_021003.5 (SEQ ID NO: 1910)) or a portion thereof.

[0531] In some embodiments described herein, the PPM1A gene product is a PPM1A mRNA isoform transcript (eg, 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 is 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% identical to 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.

[0532] PPM1A mRNA transcript variant 1, corresponding to NCBI reference sequence NM_021003.5 (SEQ ID NO: 1910)

[0533]

[0534]

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[0536]

[0537]

[0538]

[0539] In some embodiments described herein, the PPM1A gene product is a PPM1A mRNA isoform transcript (eg, 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 is 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% identical to 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.

[0540] In some embodiments described herein, the PPM1A gene product is a PPM1A mRNA isoform transcript (eg, 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 is 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% identical to 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.

[0541] In some embodiments described herein, the PPM1A gene product is a mouse (Mus musculus) PPM1A mRNA isoform transcript (e.g., mouse PPM1A mRNA alpha isoform transcript corresponding to NCBI reference sequence NM_008910.3 (SEQ ID NO: 1913)) or a portion thereof. In some embodiments described herein, the PPM1A gene product comprises a sequence that is 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% identical to a PPM1A mRNA isoform transcript, such as mouse PPM1A mRNA alpha isoform transcript corresponding to NCBI reference sequence NM_008910.3 (SEQ ID NO: 1913), or a portion thereof.

[0542] In some embodiments of the present disclosure, the PPM1A gene product is a PPM1A mRNA transcript variant other than the above-mentioned PPM1A transcript (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 mouse PPM1A mRNA alpha isoform transcript, corresponding to NCBI reference sequence NM_008910.3 (SEQ ID NO: 1913)). In some embodiments, the PPM1A gene product comprises a sequence that is 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% identical to a sequence homologous to 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: 1913), NO: 1912), or mouse PPM1A mRNA alpha isoform transcript corresponding to NCBI Reference Sequence NM_008910.3 (SEQ ID NO: 1913). In some embodiments, the PPM1A gene product comprises a sequence having 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 nucleotides 457-1429 of SEQ ID NO: 1910, or a portion thereof.

[0543] ATXN2

[0544] In some embodiments of the present disclosure, the ATXN2 gene product is an ATNX2 mRNA or ATNX2 pre-mRN comprising a sequence from an ATXN2 gene sequence (e.g., the ATXN2 gene sequence of NCBI Reference Sequence NG_011572.3 (SEQ ID NO: 149355), the ATXN2 gene sequence of NCBI Reference Sequence NC_000012.12 (Reference GRCh38.p13 Primary Assembly), or an ATXN2 coding sequence) or a portion thereof. In some embodiments of the present disclosure, the ATXN2 gene product comprises a sequence that is 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% identical to an ATXN2 mRNA or ATXN2 pre-mRNA sequence, such as the ATXN2 gene sequence of NCBI Reference Sequence NG_011572.3 (SEQ ID NO: 149355), the ATXN2 gene sequence of NCBI Reference Sequence NC_000012.12 (Reference GRCh38.p13 Primary Assembly), or an ATXN2 coding sequence, or a portion thereof.

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[0637] NCBI Reference NG_011572.3 (SEQ ID NO: 149355)

[0638] 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 some embodiments, the ATXN2 gene product comprises a sequence having 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 the ATXN2 mRNA isoform comprising the NCBI reference sequence NM_001310121.1 (SEQ ID NO: 149357).

[0639]

[0640]

[0641]

[0642] NCBI Reference NM_001310121.1 (SEQ ID NO: 149357)

[0643] 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 some embodiments, the ATXN2 gene product comprises a sequence having 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 the ATXN2 mRNA isoform comprising the NCBI Reference Sequence NM_001310123.1 (SEQ ID NO: 149358).

[0644] 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 having 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 the ATXN2 mRNA isoform comprising the NCBI reference sequence NM_001372574.1 (SEQ ID NO: 149359).

[0645] 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 having 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 the ATXN2 mRNA isoform comprising the NCBI reference sequence NM_002973.4 (SEQ ID NO: 149360).

[0646] 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 having 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 the ATXN2 mRNA transcript variant comprising the NCBI Reference Sequence NR_132311.2 (SEQ ID NO: 149361).

[0647] SOD1

[0648] In some embodiments of the present disclosure, the SOD1 gene product is a SOD1 mRNA or SOD1 pre-mRN comprising a sequence from a 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 a SOD1 coding sequence) or a portion thereof. In some embodiments of the present disclosure, the SOD1 gene product comprises a sequence that is 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% identical to a SOD1 mRNA or SOD1 pre-mRNA sequence, such as 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.pl3 Primary Assembly) (SEQ ID NO: 167803), or a SOD1 coding sequence, or a portion thereof.

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[0658]

[0659] NCBI reference sequence NG_008689.1 (SEQ ID NO: 167802)

[0660] In various embodiments, the SOD1 gene product is a 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 that is 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% identical to the SOD1 mRNA sequence comprising the NCBI reference sequence NM_000454.5 (SEQ ID NO: 167804).

[0661]

[0662] NM_000454.5(SEQ ID NO:167804)

[0663] MAPT

[0664] In some embodiments of the present disclosure, the MAPT gene product is a MAPT mRNA or MAPT pre-mRN comprising a sequence 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 that is 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% identical to a MAPT mRNA or MAPT pre-mRNA sequence, such as 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 the MAPT coding sequence, or a portion thereof.

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[0744] NG_007398.2(SEQ ID NO:301567)

[0745] 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 that is 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% identical to the MAPT mRNA sequence comprising the NCBI reference sequence NM_001123066.4 (SEQ ID NO: 301569).

[0746]

[0747]

[0748]

[0749]

[0750] NCBI Reference Sequence NM_001123066.4 (SEQ ID NO: 301569).

[0751] 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 that is 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% identical to the MAPT mRNA sequence comprising the NCBI reference sequence NM_001123067.4 (SEQ ID NO: 301570).

[0752] 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 that is 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% identical to the MAPT mRNA sequence comprising the NCBI reference sequence NM_001203251.2 (SEQ ID NO: 301571).

[0753] 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 is 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% identical to the MAPT mRNA sequence comprising the NCBI reference sequence NM_001203252.2 (SEQ ID NO: 301572).

[0754] 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 is 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% identical to the MAPT mRNA sequence comprising the NCBI reference sequence NM_001377265.1 (SEQ ID NO: 301573).

[0755] 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 is 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% identical to the MAPT mRNA sequence comprising the NCBI reference sequence NM_001377266.1 (SEQ ID NO: 301574).

[0756] 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 is 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% identical to the MAPT mRNA sequence comprising the NCBI reference sequence NM_001377267.1 (SEQ ID NO: 301575).

[0757] 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 is 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% identical to the MAPT mRNA sequence comprising the NCBI reference sequence NM_001377268.1 (SEQ ID NO: 301576).

[0758] 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 that is 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% identical to the MAPT mRNA sequence comprising the NCBI reference sequence NM_005910.6 (SEQ ID NO: 301577).

[0759] 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 is 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% identical to the MAPT mRNA sequence comprising the NCBI reference sequence NM_016834.5 (SEQ ID NO: 301578).

[0760] 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 is 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% identical to the MAPT mRNA sequence comprising the NCBI reference sequence NM_016835.5 (SEQ ID NO: 301579).

[0761] 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 that is 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% identical to the MAPT mRNA sequence comprising the NCBI reference sequence NM_016841.5 (SEQ ID NO: 301580).

[0762] 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 that is 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% identical to the MAPT RNA sequence comprising the NCBI reference sequence NR_165166.1 (SEQ ID NO: 301581).

[0763] 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 is 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% identical to the MAPT mRNA transcript variant comprising the NCBI reference sequence XM_005257370.4 (SEQ ID NO: 301582).

[0764] 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 is 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% identical to the MAPT mRNA transcript variant comprising the NCBI reference sequence XM_005257366.3 (SEQ ID NO: 301583).

[0765] 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).

[0766] In various embodiments, the MAPT gene product comprises a sequence that is 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% identical to a MAPT mRNA transcript variant comprising the NCBI reference sequence XM_005257362.4 (SEQ ID NO: 301584).

[0767] 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 is 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% identical to the MAPT mRNA transcript variant comprising the NCBI reference sequence XM_005257365.4 (SEQ ID NO: 301585).

[0768] 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 is 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% identical to the MAPT mRNA transcript variant comprising the NCBI reference sequence XM_005257367.4 (SEQ ID NO: 301586).

[0769] 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 is 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% identical to the MAPT mRNA transcript variant comprising the NCBI reference sequence XM_005257368.4 (SEQ ID NO: 301587).

[0770] 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 is 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% identical to the MAPT mRNA transcript variant comprising the NCBI reference sequence XM_005257369.4 (SEQ ID NO: 301588).

[0771] 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 is 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% identical to the MAPT mRNA transcript variant comprising the NCBI reference sequence XM_005257371.4 (SEQ ID NO: 301589).

[0772] AONs targeting gene products

[0773] PPM1A AON

[0774] In various embodiments, the PPM1A AONs 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 are replaced by one or more spacers, such as the sequence of any one of SEQ ID NOs: 301692-301742, or the sequences in Tables 4A-4C, target 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, the 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 specific embodiments, the 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 specific embodiments, the 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 specific embodiments, the PPM1A AON may 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 specific embodiments, the PPM1AAON may 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 specific embodiments, the PPM1AAON may 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 specific embodiments, the PPM1A AON may 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).

[0775] In some embodiments described herein, the PPM1A AON targets a specific portion of a PPM1A gene product, such as a PPM1A mRNA transcript or a PPM1A pre-mRNA transcript. For example, in some embodiments, 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).

[0776] For example, in some embodiments, 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 sequence of an untranslated region (UTR) of a PPM1A mRNA sequence (e.g., a 5'UTR or 3'UTR of a PPM1A mRNA sequence). In some embodiments, the PPM1A AON comprises a sequence that is 100% complementary to a nucleotide sequence of an untranslated region (UTR) of a PPM1A mRNA sequence (e.g., a 5'UTR or 3'UTR of a PPM1A mRNA sequence).

[0777] ATXN2 AON

[0778] In various embodiments, the ATXN2 AONs disclosed herein, e.g., an ATXN2 AON having the sequence of any one of SEQ ID NOs: 1914-149354, or an ATXN2 AON having the sequence of any one of SEQ ID NOs: 1914-149354 in which one or more nucleosides are replaced by one or more spacers, e.g., the sequence of any one of SEQ ID NOs: 301692-301742, or the sequences in Tables 4A-4C), target 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 an ATXN2 gene product or a portion of an ATXN2 gene sequence (e.g., any one of SEQ ID NOs: 149355-149361). In specific embodiments, the ATXN2 AON can be an oligonucleotide sequence that is at least 85% complementary to a portion of an ATXN2 gene product or an ATXN2 gene sequence (e.g., any one of SEQ ID NOs: 149355-149361). In a specific embodiment, the ATXN2 AON may be an oligonucleotide sequence that is at least 90% complementary to a portion of the ATXN2 gene product or the ATXN2 gene sequence (e.g., any one of SEQ ID NOs: 149355-149361). In a specific embodiment, the ATXN2 AON may be an oligonucleotide sequence that is at least 95% complementary to a portion of the ATXN2 gene product or the ATXN2 gene sequence (e.g., any one of SEQ ID NOs: 149355-149361). In a specific embodiment, the ATXN2 AON may be an oligonucleotide sequence that is at least 98% complementary to a portion of the ATXN2 gene product or the ATXN2 gene sequence (e.g., any one of SEQ ID NOs: 149355-149361). In a specific embodiment, the ATXN2 AON may be an oligonucleotide sequence that is at least 99% complementary to a portion of the ATXN2 gene product or the ATXN2 gene sequence (e.g., any one of SEQ ID NOs: 149355-149361). In specific embodiments, the ATXN2AON can be an oligonucleotide sequence that is 100% complementary to a portion of the ATXN2 gene product or the PATXN2 gene sequence (eg, any one of SEQ ID NOs: 149355-149361).

[0779] 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 (e.g., an ATXN2 mRNA transcript or an ATXN2 pre-mRNA transcript) (e.g., any one of SEQ ID NOs: 149355-149361).

[0780] SOD1 AON

[0781] In various embodiments, a SOD1 AON disclosed herein, e.g., a SOD1 AON having a sequence of any one of 149362-158581, or a SOD1 AON having one or more spacers (e.g., a SOD1 AON having a sequence of any one of SEQ ID NOs: 149362-158581 in which one or more nucleosides are replaced by one or more spacers), e.g., a sequence of any one of SEQ ID NOs: 301610-301741, targets a specific portion of a SOD1 gene product, e.g., a SOD1 mRNA transcript (e.g., any one of SEQ ID NOs: 167802-167804). In some embodiments, the SOD1 AON may 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 SOD1 gene product or a SOD1 gene sequence (e.g., any one of SEQ ID NOs: 167802-167804). In specific embodiments, the SOD1 AON may be an oligonucleotide sequence that is at least 85% complementary to a portion of a SOD1 gene product or a SOD1 gene sequence (e.g., any one of SEQ ID NOs: 167802-167804). In specific embodiments, the SOD1 AON may be an oligonucleotide sequence that is at least 90% complementary to a portion of a SOD1 gene product or a SOD1 gene sequence (e.g., any one of SEQ ID NOs: 167802-167804). In a specific embodiment, the SOD1 AON may be an oligonucleotide sequence that is at least 985% complementary to a portion of the SOD1 gene product or the SOD1 gene sequence (e.g., any one of SEQ ID NOs: 167802-167804). In a specific embodiment, the SOD1 AON may be an oligonucleotide sequence that is at least 98% complementary to a portion of the SOD1 gene product or the SOD1 gene sequence (e.g., any one of SEQ ID NOs: 167802-167804). In a specific embodiment, the SOD1 AON may be an oligonucleotide sequence that is at least 99% complementary to a portion of the SOD1 gene product or the SOD1 gene sequence (e.g., any one of SEQ ID NOs: 167802-167804). In a specific embodiment, the SOD1 AON may be an oligonucleotide sequence that is 100% complementary to a portion of the SOD1 gene product or the SOD1 gene sequence (e.g., any one of SEQ ID NOs: 167802-167804).

[0782] In some embodiments described herein, the SOD1 AON targets a specific portion of a SOD1 gene product, such as a SOD1 mRNA transcript or a 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 a SOD1 gene product (e.g., a SOD1 mRNA transcript or a SOD1 pre-mRNA transcript) (e.g., any one of SEQ ID NOs: 167802-167804).

[0783] MAPT AON

[0784] In various embodiments, the MAPT AON disclosed herein, such as a MAPT AON having a sequence of any one of SEQ ID NOs: 167805-301566, or a MAPT AON having one or more spacers (e.g., a MAPT AON having a sequence of any one of SEQ ID NOs: 167805-301566 in which one or more nucleosides are replaced by one or more spacers), such as a sequence of any one of SEQ ID NOs: 301692-301742 or a sequence in Tables 4A-4C, targets a specific portion of a MAPT gene product, such as a MAPT mRNA or pre-mRNA transcript (e.g., any one of SEQ ID NOs: 301567-301589). In some embodiments, the 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 specific embodiments, the 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 specific embodiments, the 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 a specific embodiment, the MAPT AON can be an oligonucleotide sequence that is at least 95% complementary to a portion of the MAPT gene product or the MAPT gene sequence (e.g., any one of SEQ ID NOs: 301567-301589). In a specific embodiment, the MAPT AON can be an oligonucleotide sequence that is at least 98% complementary to a portion of the MAPT gene product or the MAPT gene sequence (e.g., any one of SEQ ID NOs: 301567-301589). In a specific embodiment, the MAPT AON can be an oligonucleotide sequence that is at least 99% complementary to a portion of the MAPT gene product or the MAPT gene sequence (e.g., any one of SEQ ID NOs: 301567-301589). In a specific embodiment, the MAPT AON can be an oligonucleotide sequence that is 100% complementary to a portion of the MAPT gene product or the MAPT gene sequence (e.g., any one of SEQ ID NOs: 301567-301589).

[0785] In some embodiments described herein, 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, a 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 (e.g., a MAPT mRNA transcript or a MAPT pre-mRNA transcript) (e.g., any one of SEQ ID NOs: 301567-301589).

[0786] Nuclease-mediated inhibition

[0787] On the one hand, the disclosure provides a nuclease that reduces the 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 RNA enzyme (e.g., RNAseH), a zinc finger nuclease (ZFN), a meganuclease, a transcription activator-like effector nuclease (TALEN), or a clustered regular spacer short palindromic repeat (CRISPR)-associated protein.

[0788] In certain embodiments, zinc finger nucleases (ZFNs) are used to inhibit, eliminate, or reduce the amount of target gene products. Synthetic ZFNs are composed of zinc finger binding domains fused to, for example, FokI DNA cleavage domains. ZFNs can be designed / engineered for editing the genome of cells, including but not limited to knocking out or knocking in gene expression in a variety of organisms. Large-scale nucleases, TALENs, or CRISPR-associated proteins can be used for genome engineering in cells of patients with neurological diseases or at risk of neurological diseases, the cells comprising neurons, such as motor neurons and other cells of the nervous system. The reagents can be used for targeting promoters, protein coding regions (exons), introns, 5' and 3' UTRs, etc.

[0789] CRISPR genome editing typically involves two distinct components: (1) a guide RNA and (2) an endonuclease, particularly a CRISPR-associated (Cas) nuclease (e.g., Cas9). The guide RNA is a combination of endogenous bacterial crRNA and tracrRNA into a single chimeric guide RNA (gRNA) transcript. Without being bound by theory, it is believed that when the gRNA and Cas are expressed in cells, the genomic target sequence can be modified or permanently destroyed.

[0790] The gRNA / Cas complex can be recruited to a target sequence, such as a target gene (e.g., any one of PPM1A, ATXN2, SOD1, or MAPT) by base pairing the gRNA sequence with the target DNA sequence component in the target gene. An appropriate genomic target sequence comprises a protospacer adjacent motif (PAM) sequence immediately following the target sequence. The binding of the gRNA / Cas complex positions Cas to the target sequence, allowing wild-type Cas to cut both strands of DNA, resulting in double-strand breaks. Double-strand breaks are repaired through one of two conventional repair pathways: (1) non-homologous end joining DNA repair pathway or (2) homology directed repair pathway. Non-homologous repair pathways can result in insertions / deletions at the double-strand break, which can result in frameshifts and / or premature stop codons, effectively destroying the open reading frame of the target gene. Homology directed repair pathways require the presence of a repair template for repairing double-strand breaks.

[0791] In certain embodiments, CRISPR genome editing is used to reduce target gene expression. In some embodiments, gRNA pairs are used to target target genes to reduce and / or eliminate the expression of target genes. In certain embodiments, one gRNA pair is used to reduce the expression of target genes. In certain other embodiments, multiple gRNA pairs are used to reduce the expression of target genes. gRNA pairs can be designed using known techniques and based on gene sequences. In certain embodiments, the gRNA sequence can include modifications, such as 2'O-methyl analogs and 3' phosphorothioate internucleotide linkages in the terminal three nucleotides of both the 5' and 3' ends of the gRNA.

[0792] Neurological diseases

[0793] The methods described herein can be used to treat neurological diseases including, but not limited to, 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, synucleinopathies, Huntington's disease, brachial plexus injury, peripheral nerve injury, progressive supranuclear palsy (PSP), brain trauma, spinal cord injury, tuberous sclerosis, Pick's disease, tauopathies, primary age-related tauopathies, Down syndrome, epilepsy / seizure disorder, depression, traumatic brain injury (TBI), chronic traumatic encephalopathy (CTE), HIV-related neurocognitive disorders, and the like. and / or neuropathies such as hand and cognitive disorder (HAND), multiple system atrophy, amnestic mild cognitive impairment, corticobasal degeneration (CBD), and / or neuropathy such as chemotherapy-induced neuropathy, spinocerebellar ataxia (SCA), SCA type 2, spinal muscular atrophy (SMA), Parkinson's disease, Niemann-Pick disease type C (NPC), Charcot-Marie-Tooth disease (CMT), mucopolysaccharidosis type II (MPSIIA), mucolipidosis type IV, GM1 gangliosidosis, sporadic inclusion body myositis (sIBM), Henoch-Schonlein purpura (HSP), Gaucher disease, facial-onset sensory and motor neuronopathy, Guam-type parkinsonism and dementia syndrome, multisystem proteinopathy, Perry disease, and autism.

[0794] Motor neuron disease is a group of diseases characterized by the loss of motor neuron function that coordinates the autonomous movement of brain muscles. Motor neuron disease may affect upper motor neurons and / or lower motor neurons and may have a sporadic or familial origin. Motor neuron disease comprises amyotrophic lateral sclerosis (ALS or Lou Gehrig's disease), progressive bulbar palsy, pseudobulbar palsy, progressive muscular dystrophy, primary lateral sclerosis, spinal muscular atrophy, post-polio syndrome and ALS with frontotemporal dementia.

[0795] Symptoms of motor neurone disease include muscle weakness or degeneration, muscle pain, spasticity, slurred speech, difficulty swallowing, loss of muscle control, joint pain, stiffness in the limbs, difficulty breathing, drooling, and complete loss of muscle control, including loss of basic functions such as breathing, swallowing, eating, speaking, and movement. These symptoms are sometimes accompanied by depression, memory loss, difficulty planning, speech impairment, behavioral changes, difficulty assessing spatial relationships, and / or personality changes.

[0796] Motor neuron disease can be assessed and diagnosed using various tools and tests by a skilled clinician (such as a neurologist).For example, blood and urine tests (such as, measuring the test of the existence of creatinine kinase), magnetic resonance imaging (MRI), electromyography (EMG), nerve conduction studies (NCS), spinal puncture, lumbar puncture and / or muscle biopsy can be used to assess or diagnose the existence or development risk of motor neuron disease.Motor neuron disease can be diagnosed by physical examination and / or neurological examination to assess the change of movement and sensory skills, neurological function, hearing and speech, vision, coordination and balance, mental state and emotion or behavior.

[0797] A patient with ALS, FTD, ALS with FTD, or another neurological or motor neuron disease can be a patient diagnosed with the disease or exhibiting symptoms of the disease. A patient with ALS, FTD, ALS with FTD, or another neurological or motor neuron disease can be a patient who previously had the disease and who, after recovering or experiencing a complete or partial improvement in the disease and / or disease symptoms, experiences a complete or partial recurrence of the disease or disease symptoms. A patient with ALS, FTD, ALS with FTD, or another neurological or motor neuron disease or condition can be a patient who carries a genetic mutation associated with manifestation of the disease or condition. For example, patients with ALS may carry SOD1, C9orf72, Ataxin 2 (ATXN2), charged multivesicular body protein 2B (CHMP2B), dynamin activator protein 1 (DCTN1), human epidermal growth factor receptor 4 (ERBB4), FIG4 phosphoinositide 5-phosphatase (FIG4), NIMA-related kinase 1 (NEK1), heterogeneous nuclear ribonucleoprotein A1 (HNRNPA1), neurofilament heavy (NEFH), peripherin (PRPH), TAR DNA binding protein 43 (TDP43 or TARDP), sarcoma fusion protein (FUS), ubiquitin-like protein 2 (UBQLN2), kinesin family member 5A (KIF5A), valosin-containing protein (VCP), Alsin (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), angiopoietin (ANG), Profilin-1 (PFN1), matrix protein 3 (MATR3), coiled-coil domain-containing 10 (CHCHD10), tubulin, Alpha Patients with ALS (Tuba4a), TBK1, C21orf2, sequestrant 1 (SQSTM1, also known as ubiquitin-binding protein p62), and / or optineurin (OPTN), particularly when the mutation is associated with ALS or a high risk of developing ALS.

[0798] Patients at risk for ALS, FTD, ALS with FTD, or another nervous system or motor neuron disease can include patients with a family history of the disease or a genetic predisposition to the disease (e.g., patients carrying a genetic mutation associated with a high risk of the disease), or patients exposed to environmental factors that increase the risk of the disease. For example, if a patient carries 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 when the mutation is associated with ALS or a high risk of developing ALS, the patient may be at risk for ALS. Patients at risk may also include those diagnosed with a disease or condition that has a high comorbidity with ALS, FTD, ALS with FTD, or another nervous system or motor neuron disease (e.g., patients with dementia, which is significantly associated with a higher odds of a family history of ALS, FTD, and bulbar ALS (see Trojsi, F. et al., (2017) "Comorbidity of dementia with amyotrophiclateral sclerosis (ALS): insights from a large multicenter Italian cohort" J Neurol 264:2224-31).

[0799] Amyotrophic lateral sclerosis

[0800] ALS is a progressive motor neuron disease that disrupts signaling to all voluntary muscles. ALS causes atrophy of both upper and lower motor neurons. Symptoms of ALS include bulbar muscle weakness and atrophy, generalized and bilateral loss of strength, spasticity, muscle spasms, muscle cramps, fasciculations, slurred speech, and difficulty breathing or loss of the ability to breathe. Some people with ALS also experience 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.

[0801] ALS most commonly develops in men over 40 years of age, but it can also occur in women and children. People who smoke, are exposed to chemicals such as lead, or have served in the military are also at increased risk for ALS. Most cases of ALS are sporadic, while only about 10% are familial. Causes of ALS include sporadic or inherited gene mutations, high levels of glutamate, and improper protein processing. Genetic mutations associated with ALS include mutations in 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.

[0802] Frontotemporal dementia

[0803] Frontotemporal dementia (FTD) is a form of dementia that affects the frontal and temporal lobes of the brain. It has an average age of onset earlier than Alzheimer's disease - 40 years old. Symptoms of FTD include extreme changes in behavior and personality, speech and language problems, and movement-related symptoms such as tremors, rigidity, muscle spasms, weakness, and difficulty swallowing. Subtypes of FTD include behavioral variant frontotemporal dementia (bvFTD), which is characterized by changes in personality and behavior, and primary progressive aphasia (PPA), which affects language skills, speech, writing, and comprehension. FTD is associated with the accumulation of tau proteins (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 C9orf72, progranulin (GRN), microtubule-associated protein tau (MAPT), UBQLN2, VPC, CHMP2B, TARDP, FUS, ITM2B, CHCHD10, SQSTM1, PSEN1, PSEN2, CTSF, CYP27A1, TBK1, and TBP.

[0804] Amyotrophic lateral sclerosis with frontotemporal dementia (ALS with FTD) is a clinical syndrome in which FTD and ALS occur in the same individuals. Interestingly, mutations in C9orf72 are the most common cause of familial ALS and FTD. Furthermore, mutations in TBK1, VCP, SQSTMI, UBQLN2, and CHMP2B have also been associated with ALS with FTD. Symptoms of ALS with FTD include profound personality changes, as well as muscle weakness, atrophy, fasciculations, spasticity, dysarthria, 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 have been associated with ALS, FTD, and ALS with FTD.

[0805] TBK1 and RIPK1 functions

[0806] In one aspect, the methods described herein include exposing cells to PPM1A AON to modify the activity, function, or other characteristics of a gene or gene product, such as an mRNA or protein. For example, the methods described herein include methods of increasing or decreasing or inhibiting the activity, function, or other characteristics of a gene or gene product. For example, described herein are methods of increasing the phosphorylation of residues of TANK-binding kinase 1 (also known as serine / threonine protein kinase TBK1; "TBK1"). For example, described herein are methods of increasing the phosphorylation of TBK1 serine residue 172 (ser172) in a cell, wherein the method comprises exposing the cell to PPM1A AON. In some embodiments, TBK1 ser172 phosphorylation is increased in cells of a patient with ALS, FTD, or ALS with FTD. In some embodiments, the method of increasing TBK1 ser172 phosphorylation comprises exposing a cell to a PPM1A antisense oligonucleotide of any one of SEQ ID NOs: 1-954, or a sequence of any one of SEQ ID NOs: 1-954 in which one or more nucleosides are replaced by one or more spacers, e.g., SEQ ID NOs: 301692-301742, or any one of the sequences in Tables 4A-4C.

[0807] Also described herein are methods of increasing TBK1 function in a cell, wherein the method comprises exposing the cell to a PPM1A AON. For example, described herein are methods of increasing TBK1 function in a cell, wherein the method comprises exposing the cell 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 of increasing TBK1 function comprises exposing the 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 are replaced by one or more spacers, such as SEQ ID NOs: 301692-301742, or any one of the sequences in Tables 4A-4C.

[0808] Tank binding kinase 1 (TBK1) is a member of the IKK kinase family that induces type 1 interferon activity and plays an important role in the phosphorylation of autophagy adaptors. TBK1 mutations are thought to lead to impaired autophagy and contribute 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). In addition, together with C9orf72, OPTN, SQSTM1 / p62, UBQLN2, and TDP43 mutations, TBK1 mutations 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-temporaldementia” Nature Neuroscience, 18(5): 631-6).

[0809] 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 multiple genes involved in regulating autophagy are associated with ALS (e.g., SQSTM1, SOD1, OPTN, VCP, UBQLN2, and TBK1). Therefore, disruption of autophagy appears to contribute to ALS pathology.

[0810] Phosphorylation of TBK1 residue Ser172 results in a conformational change in TBK1, which allows substrate binding via the protein kinase domain. TBK1 phosphorylates numerous 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. Furthermore, individuals carrying TBK1 mutations also exhibit TDP43-positive aggregates in various brain regions. Therefore, TBK1 mutations may lead to decreased autophagy and accumulation of protein aggregates in motor neurons.

[0811] 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 NF-κB pathways. PPM1A also dephosphorylates and inactivates TBK1. Specifically, PPM1A dephosphorylates Ser172 of TBK1. Activated TBK1 can phosphorylate RIPK1, thereby inactivating RIPK1. Therefore, PPM1A activity indirectly activates RIPK1, and consequently, reduced PPM1A expression indirectly inactivates RIPK1.

[0812] The present disclosure is based, in part, on the discovery that increasing TBK1 activity, e.g., increasing TBK1 activity in an individual or a cell of an individual having TBK1 haploinsufficiency, can be used as a mechanism for treating 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, synucleinopathies, Huntington's disease, brachial plexus injury, peripheral nerve injury, progressive supranuclear palsy (PSP), traumatic brain injury, spinal cord injury, tuberous sclerosis, Pick's disease, tauopathies, primary age-related tauopathies, Down syndrome, epilepsy / seizure disorders, depression, traumatic brain injury (TBI), and other neurological disorders. BI), chronic traumatic encephalopathy (CTE), HIV-associated neurocognitive disorder (HAND), multiple system atrophy, amnestic mild cognitive impairment, corticobasal degeneration (CBD), and / or neuropathies such as chemotherapy-induced neuropathy, spinocerebellar ataxia (SCA), SCA type 2, spinal muscular atrophy (SMA), Parkinson's disease, Niemann-Pick disease type C (NPC), Charcot-Marie-Tooth disease (CMT), mucopolysaccharidosis type II (MPSIIA), mucolipidosis type IV, GM1 gangliosidosis, sporadic inclusion body myositis (sIBM), Henoch-Schonlein purpura (HSP), Gaucher disease, facial-onset sensory and motor neuronopathy, Guam-type parkinsonism, multisystem proteinopathy, Perry disease, and autism.

[0813] The present disclosure is...

Claims

1. A compound comprising a gapmer oligonucleotide, and wherein the gapmer oligonucleotide comprises a spacer.

2. A gapmer oligonucleotide, wherein the gapmer oligonucleotide comprises a spacer.

3. The compound or gapmer oligonucleotide according to claim 1 or 2, wherein the gapmer oligonucleotide comprises a second spacer that is not adjacent to the spacer.

4. The compound or gapmer oligonucleotide according to any one of claims 1 to 3, wherein the gapmer oligonucleotide comprises a sequence that is 85% to 98% complementary to an equal length portion of a transcript, the presence of which results in a neurological disease.

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

6. The compound or gapmer oligonucleotide of any one of claims 1-5, wherein the gapmer oligonucleotide comprises a sequence that is 85% to 98% complementary to any one of SEQ ID NOs: 1909-1913, 149355-149361, 167802-167804, or 301567-301589, a sequence having 90% identity thereof, or an equal length portion of a 15 to 50 consecutive nucleobase portion thereof.

7. The compound or gapmer oligonucleotide of any one of claims 1-6, wherein the gapmer oligonucleotide comprises a sequence that is at least 85% identical 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 a sequence in Tables 4A-4C.

8. The compound or gapmer oligonucleotide according to any one of claims 1 to 7, wherein the gapmer oligonucleotide comprises a sequence that is at least 90% identical 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 a sequence in Tables 4A-4C.

9. The compound or gapmer oligonucleotide according to any one of claims 1-7, wherein the gapmer oligonucleotide comprises a sequence that is at least 95% identical 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 a sequence in Tables 4A-4C.

10. The compound or gapmer oligonucleotide according to any one of claims 1-8, wherein the gapmer oligonucleotide comprises a sequence that is at least 100% identical 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 a sequence in Tables 4A-4C.

11. A compound or gapmer oligonucleotide according to any one of claims 1-9, wherein the gapmer oligonucleotide comprises a segment having up to 11 linked nucleosides, and wherein the gapmer oligonucleotide comprises a sequence having at least 85% 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 a sequence in Tables 4A-4C.

12. A compound or gapmer oligonucleotide according to any one of claims 1-10, wherein the gapmer oligonucleotide comprises a segment having up to 10, 9, 8, 7, 6, 5, 4, 3, or 2 linked nucleosides, and wherein the gapmer oligonucleotide comprises a sequence that is at least 85% identical to an equal length portion of any one of SEQ ID NOs: 301692-301742 or the sequences in Tables 4A-4C.

13. A compound or gapmer oligonucleotide according to any one of claims 1-10, wherein the gapmer oligonucleotide comprises a segment having up to 11 linked nucleosides, and wherein the gapmer oligonucleotide comprises a sequence having at least 90% identity to an equal length portion of any one of SEQ ID NOs: 301692-301742 or the sequences in Tables 4A-4C.

14. The compound or gapmer oligonucleotide of any one of claims 1-10, wherein the gapmer oligonucleotide comprises a segment having up to 10, 9, 8, 7, 6, 5, 4, 3 or 2 linked nucleosides, and wherein the gapmer oligonucleotide comprises a sequence having at least 90% identity to an equal length portion of any one of SEQ ID NOs: 301610-301741.

15. The compound or gapmer oligonucleotide of any one of claims 1-14, wherein 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.

16. The compound or gapmer oligonucleotide according to any one of claims 1-15, wherein at least one (ie, one or more) nucleoside linkage of the gapmer oligonucleotide is a non-natural linkage.

17. The compound or gapmer oligonucleotide according to any one of claims 1 to 16, wherein the gapmer oligonucleotide is at least 19 oligonucleotide units in length.

18. The oligonucleotide compound according to any one of claims 1 to 17, wherein the spacer is a nucleoside replacement group comprising a non-sugar substituent that cannot be linked to a nucleotide base.

19. The compound or gapmer oligonucleotide of claim 18, wherein the spacer is located between positions 5 and 11 of the gapmer oligonucleotide.

20. The compound or gapmer oligonucleotide of claim 18, wherein the spacer is located between positions 7 and 11 of the gapmer oligonucleotide.

21. The compound or gapmer oligonucleotide of claim 18, wherein the gapmer oligonucleotide further comprises a second spacer, and wherein the spacer and the second spacer are not adjacent to each other.

22. The compound or gapmer oligonucleotide of claim 21, wherein the gapmer oligonucleotide further comprises a second spacer, wherein the second spacer is located between positions 15 and 19 of the gapmer oligonucleotide.

23. The compound or gapmer oligonucleotide of claim 22, wherein the spacer and the second spacer are separated by at least 5 nucleobases, at least 6 nucleobases, or at least 7 nucleobases in the gapmer oligonucleotide.

24. The compound or gapmer oligonucleotide of claim 22 or 23, wherein the spacer is located between positions 5 and 11 of the gapmer oligonucleotide, and wherein the second spacer is located between positions 15 and 19 of the gapmer oligonucleotide.

25. The compound or gapmer oligonucleotide of any one of claims 22-24, wherein the spacer is located at position 8 of the gapmer oligonucleotide, and wherein the second spacer is located at position 16 of the gapmer oligonucleotide.

26. The compound or gapmer oligonucleotide of any one of claims 22-24, wherein the spacer is located at position 5 of the gapmer oligonucleotide, and wherein the second spacer is located at position 17 of the gapmer oligonucleotide.

27. The compound or gapmer oligonucleotide of any one of claims 22-24, wherein the spacer is located at position 7 of the gapmer oligonucleotide, and wherein the second spacer is located at position 15 of the gapmer oligonucleotide.

28. The compound or gapmer oligonucleotide of any one of claims 22-24, wherein the spacer is located at position 11 of the gapmer oligonucleotide, and wherein the second spacer is located at position 19 of the gapmer oligonucleotide.

29. The compound or gapmer oligonucleotide of any one of claims 18-28, wherein each of the spacer or second spacer is a nucleoside replacement group comprising a non-sugar substituent, wherein the non-sugar substituent does not contain a ketone, aldehyde, ketal, hemiketal, acetal, hemiacetal, aminal, or hemiaminal moiety and is incapable of forming a covalent bond with a nucleotide base.

30. The compound or gapmer oligonucleotide according to any one of claims 18 to 29, wherein each of the spacer or the second spacer is independently represented by formula (X), wherein: Ring A is an optionally substituted 4-8 membered monocyclic cycloalkyl or 4-8 membered monocyclic heterocyclyl, wherein the heterocyclyl contains 1 or 2 heteroatoms selected from O, S, and N, provided that A cannot form a covalent bond with a nucleobase; and The symbol indicates the point of attachment of the internucleoside linkage.

31. The compound or gapmer oligonucleotide of claim 30, wherein each of the spacer or the second spacer is independently represented by formula (Xa), wherein:

32. The compound or nucleotide according to claim 30 or 31, wherein Ring A is an optionally substituted 4-8 membered monocyclic cycloalkyl selected from cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl; or a 4-8 membered monocyclic heterocyclyl selected from oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, 1,4-dioxanyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, and azepanyl.

33. The compound or nucleotide of claim 32, wherein Ring A is tetrahydrofuranyl.

34. The compound or nucleotide of claim 32, wherein Ring A is tetrahydropyranyl.

35. The compound or gapmer oligonucleotide of any one of claims 18-29, wherein each of the spacer and the second spacer is independently represented by Formula I, wherein: X is selected from -CH2- and -O-; and n is 0, 1, 2, or 3.

36. The compound or gapmer oligonucleotide of any one of claims 18-29, wherein each of the spacer and the second spacer is independently represented by Formula I', wherein: X is selected from -CH2- and -O-; and n is 0, 1, 2, or 3.

37. The compound or gapmer oligonucleotide according to any one of claims 18 to 29, wherein each of the spacer and the second spacer is independently represented by formula (Ia), wherein: Know n is 0, 1, 2, or 3.

38. The compound or gapmer oligonucleotide according to any one of claims 18 to 29, wherein each of the spacer and the second spacer is independently represented by formula (Ia'), wherein: Know n is 0, 1, 2, or 3.

39. The compound or gapmer oligonucleotide of any one of claims 18-29, wherein each of the spacer and the second spacer is independently represented by Formula II, wherein: Know X is selected from -CH2- and -O-.

40. The compound or gapmer oligonucleotide of any one of claims 18-29, wherein each of the spacer and the second spacer is independently represented by Formula II', wherein: Know X is selected from -CH2- and -O-.

41. The compound or gapmer oligonucleotide of any one of claims 18-29, wherein each of the spacer and the second spacer is independently represented by formula (Iia), wherein:

42. The compound or gapmer oligonucleotide according to any one of claims 18 to 29, wherein each of the spacer and the second spacer is independently represented by formula (Iia'), wherein:

43. The compound or gapmer oligonucleotide of any one of claims 18-29, wherein each of the spacer and the second spacer is independently represented by Formula III, wherein: and X is selected from -CH2- and -O-.

44. The compound or gapmer oligonucleotide of any one of claims 18-29, wherein each of the spacer and the second spacer is independently represented by Formula III', wherein: Know X is selected from -CH2- and -O-.

45. The compound or gapmer oligonucleotide according to any one of claims 18 to 29, wherein each of the spacer and the second spacer is independently represented by formula (IIIa), wherein:

46. ​​The compound or gapmer oligonucleotide according to any one of claims 18 to 29, wherein each of the spacer and the second spacer is independently represented by formula (IIIa'), wherein:

47. The compound or gapmer oligonucleotide according to any one of the preceding claims, wherein the gapmer oligonucleotide comprising the spacer has a GC content of at least 10%.

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

49. The compound or gapmer oligonucleotide according to any one of the preceding claims, wherein the gapmer oligonucleotide comprising the spacer has a GC content of at least 25%.

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

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

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

53. A compound or gapmer oligonucleotide according to any of the preceding claims, wherein at least one (i.e., one or more) nucleoside linkages of the gapmer oligonucleotide are independently selected from the group consisting of phosphodiester linkages, phosphorothioate linkages, alkyl phosphate linkages, phosphorodithioate linkages, phosphotriester linkages, alkyl phosphonate linkages, 3-methoxypropylphosphonate linkages, methylphosphonate linkages, aminoalkylphosphotriester linkages, alkylenephosphonate linkages, phosphite linkages, phosphoramidate linkages, thiophosphoramidate linkages, phosphorodiamidate linkages, phosphorodiamidate linkages, aminoalkylphosphoramidate linkages, thiophosphoramidate linkages, alkylthiophosphonate linkages, alkylthiophosphotriester linkages, phosphorothioate linkages, selenophosphorothioate linkages, and borylphosphate linkages.

54. The gapmer oligonucleotide of any of the preceding claims, wherein at least one internucleoside linkage of the nucleotide sequence is a phosphorothioate linkage.

55. The gapmer oligonucleotide of claim 54, wherein the phosphorothioate internucleoside linkage is in one of an Rp configuration or an Sp configuration.

56. The gapmer oligonucleotide of any one of claims 1-55, wherein the gapmer oligonucleotide comprises one or more chiral centers and / or double bonds.

57. The gapmer oligonucleotide of claim 55, wherein the gapmer oligonucleotide exists as stereoisomers selected from geometric isomers, enantiomers, and diastereomers.

58. The gapmer oligonucleotide of any of the preceding claims, wherein all internucleoside linkages of the nucleotide sequence are phosphorothioate linkages.

59. The gapmer oligonucleotide of any of the preceding claims, wherein the gapmer oligonucleotide comprises at least one modified nucleobase.

60. The gapmer oligonucleotide of claim 59, wherein the at least one modified nucleobase is 5-methylcytosine, pseudouridine, or 5-methoxyuridine.

61. The gapmer oligonucleotide of any of the preceding claims, wherein the gapmer oligonucleotide comprises at least one nucleoside having a modified sugar moiety.

62. The gapmer oligonucleotide of claim 61, wherein 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), 2'-deoxy-2'-fluoro nucleoside, 2'-fluoro-β-D-arabinoside, locked nucleic acid (LNA), constrained ethyl 2'-4'-bridged nucleic acid (cEt), S-cEt, a hexitol nucleic acid (HNA), and a tricyclic analog (e.g., tcDNA).

63. The gapmer oligonucleotide of any of the preceding claims, wherein the gapmer oligonucleotide comprises 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleosides having modified sugar moieties.

64. The gapmer oligonucleotide of claim 63, wherein the modified sugar moiety is independently any one of a 2'-OMe modified sugar moiety, a bicyclic sugar moiety, 2'-O-(2-methoxyethyl) (2'-MOE), 2'-O-(N-methylacetamide), 2'-deoxy-2'-fluoro nucleoside, 2'-fluoro-β-D-arabinoside, locked nucleic acid (LNA), constrained ethyl 2'-4'-bridged nucleic acid (cEt), S-cEt, a hexitol nucleic acid (HNA), and a tricyclic analog (e.g., tcDNA).

65. The gapmer oligonucleotide of any one of the preceding claims, wherein the gapmer oligonucleotide comprises ten 2'-0-(2-methoxyethyl) (2'-MOE) nucleosides.

66. The gapmer oligonucleotide of claim 65, wherein five of the 2'-O-(2-methoxyethyl) (2'-MOE) nucleosides are located at the 3' end of the gapmer oligonucleotide, and wherein five of the 2'-O-(2-methoxyethyl) (2'-MOE) nucleosides are located at the 5' end of the gapmer oligonucleotide.

67. The gapmer oligonucleotide of claim 65, wherein the modified sugar moiety is a locked nucleic acid (LNA), and wherein two or more nucleosides of the 3' wing of the gapmer oligonucleotide are LNA, and wherein two or more nucleosides of the 5' wing of the gapmer oligonucleotide are LNA.

68. The gapmer oligonucleotide of any one of claims 61-66, wherein at least one nucleoside having the modified sugar moiety or a plurality of nucleosides having the modified sugar moiety is a ribonucleoside.

69. The gapmer oligonucleotide of any of the preceding claims, wherein the gapmer oligonucleotide comprises at least one deoxyribonucleoside.

70. The gapmer oligonucleotide of any of the preceding claims, wherein the gapmer oligonucleotide comprises 2, 3, 4, 5, 6, 7, 8, 9 or 10 deoxyribonucleosides.

71. The gapmer oligonucleotide of any one of claims 1-53, wherein the gapmer oligonucleotide comprises: a gap segment comprising one or more of linked deoxyribonucleosides, 2'-fluoroarabinoic acid (FANA), and fluorocyclohexenyl nucleic acid (F-CeNA); a 5' wing comprising linked nucleosides; and a 3' wing comprising linked nucleosides; in, The central region comprises a region of at least 8 oligonucleotide units, wherein the oligonucleotide units comprise at least 4 consecutive nucleobases, and the region has at least 80% identity to an isometric portion of any one of the sequences between the 5' wing segment and the 3' wing segment in SEQ ID NO: 1-954, 1914-149354, 149362-158581, 167805-301566, SEQ ID NO: 301692-301742, or Tables 4A-4C; wherein each of the 5' wing region and the 3' wing region comprises at least two connected nucleosides; and wherein at least one nucleoside in each wing region comprises a modified sugar.

72. The gapmer oligonucleotide of claim 71, wherein the at least two linked nucleosides of the 5' wing are linked by a phosphorothioate internucleoside linkage and / or wherein the at least two linked nucleosides of the 3' wing are independently linked by a phosphorothioate internucleoside linkage.

73. The gapmer oligonucleotide of claim 71 or 72, wherein each internucleoside linkage of the 5' wing and / or each internucleoside linkage of the 3' wing is independently a phosphorothioate internucleoside linkage.

74. The gapmer oligonucleotide of claim 71 or 72, wherein the 5' wing further comprises at least one phosphodiester internucleoside linkage.

75. The gapmer oligonucleotide of claim 71 or 72, wherein the 3' wing further comprises at least one phosphodiester internucleoside linkage.

76. The gapmer oligonucleotide of claim 71, wherein the at least two linked nucleosides of the 5' wing are linked by a phosphodiester internucleoside linkage and / or wherein the at least two linked nucleosides of the 3' wing are independently linked by a phosphodiester internucleoside linkage.

77. The gapmer oligonucleotide of any one of claims 71-76, wherein at least one of the internucleoside linkages of the central region is a phosphodiester linkage.

78. The gapmer oligonucleotide of claim 77, wherein at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8 or at least 9 of the internucleoside linkages of the central region are phosphodiester linkages.

79. The gapmer oligonucleotide of any one of claims 71-76, wherein at least one internucleoside linkage of the central region is a phosphorothioate internucleoside linkage.

80. The gapmer oligonucleotide of claim 79, wherein at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8 or at least 9 internucleoside linkages of the central region are phosphorothioate internucleoside linkages.

81. The gapmer oligonucleotide of any one of claims 71-72 or 79-80, wherein all of the internucleoside linkages of the gapmer oligonucleotide are phosphorothioate internucleoside linkages.

82. The gapmer oligonucleotide of any one of claims 71-81, wherein any or all of the phosphorothioate internucleoside linkages are in the Rp configuration, the Sp configuration, or any combination of the Rp and Sp configurations.

83. The gapmer oligonucleotide of any one of claims 71-82, wherein the gapmer oligonucleotide comprises at least one modified sugar moiety.

84. The gapmer oligonucleotide of claim 83, wherein the 5' wing or the 3' wing comprises the at least one modified sugar moiety.

85. The gapmer oligonucleotide of claim 83, wherein the central region comprises the at least one modified sugar moiety.

86. The gapmer oligonucleotide of any one of claims 83-85, wherein the at least one 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), 2'-deoxy-2'-fluoro nucleoside, 2'-fluoro-β-D-arabinoside, a locked nucleic acid (LNA), a constrained ethyl 2'-4'-bridged nucleic acid (cEt), S-cEt, tcDNA, a hexitol nucleic acid (HNA), and a tricyclic analog (e.g., tcDNA).

87. The gapmer oligonucleotide of any one of claims 83-86, wherein the gapmer oligonucleotide comprises one or more 2'-MOE nucleosides.

88. The gapmer oligonucleotide of claim 87, wherein the 5' wing or the 3' wing comprises one or more 2'-MOE nucleosides.

89. The gapmer oligonucleotide of claim 87 or 88, wherein the 5' wing or the 3' wing comprises 2, 3, 4, 5, or 6 2'-MOE nucleosides.

90. The gapmer oligonucleotide of claim 89, wherein each nucleoside of the 5' wing or the 3' wing is a 2'-MOE nucleoside.

91. The gapmer oligonucleotide of claim 87, wherein the central region comprises one or more 2'-MOE nucleosides.

92. The gapmer oligonucleotide of claim 91, wherein the central region comprises 2, 3, 4, 5, 6, 7, 8, 9, or 10 2'-MOE nucleosides.

93. The gapmer oligonucleotide of claim 92, wherein each nucleoside of the central region is a 2'-MOE nucleoside.

94. The gapmer oligonucleotide of any one of claims 87-93, wherein the one or more 2'-MOE nucleosides are linked by phosphorothioate internucleoside linkages.

95. The gapmer oligonucleotide of claim 71, wherein the gapmer oligonucleotide comprises any of the following patterns of sugar modifications: eeeee-d10-eeeee, eeeee-d8-eeeee, eeeeee-d11-eeeeee, eee-d8-eee, eee-d10-eee, eeeeee-d10-eeee, and eeee-d8-eeee, wherein e=2'-MOE nucleoside, and d=deoxyribonucleoside, and wherein at least one "e" or at least one "d" is replaced with a spacer.

96. The gapmer oligonucleotide of claim 95, wherein the gapmer oligonucleotide comprises any of the following patterns of internucleoside linkages: sssssooooooooosssss; ooooossssssssssooooo; oooooooooooooosssss; soosssssssssssssssss; soosossssssssssssosos; sssssssssssssssoos; sssssoooooooooooooo; sssssssssssssssssss ssssssssssssssssssssssssssssooooosssssssssssooo os;sssssssssssooo;ooosssssssssss;sssooooooooosss;ooossssssssssooo;ssssssssssssssss;ssssssssssssooo;ooossssssssss sss;sosssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssss; o ooossssssssoooo;sssssssssssssoooo;oooosssssssssss;soossssssssssoos;soosssssssssssss;sssssssssssssoos;soooosssssss ssssooos; soooosssssssssssooss; sosssssssssssssoss; sossssssssssssssssss; or sssssssssssssss; where s = phosphorothioate linkage, and o = phosphodiester linkage.

97. The gapmer oligonucleotide of claim 95 or 96, wherein the gapmer oligonucleotide comprises any of the following patterns of sugar modifications and internucleoside linkage combinations, respectively: a) eeeee-d10-eeeeee and sssssooooooooossss; b)eeeee-d10-eeeeee and ooooosssssssssooooo; c)eeeee-d10-eeeeee and ssssssssssssssssss; d)eee-d8-eee and ssssooooooosss; e)eee-d8-eee and ooosssssssooo; f)eee-d8-eee and sssssssssssss; g)eee-d10-eee and ssssooooooooosss; h)eee-d10-eee and ooosssssssssooo; i)eee-d10-eee and ssssssssssssss; j)eeee-d10-eeee and ssssooooooossss; k)eeee-d10-eeee andoooossssssssssoooo; l)eeee-d10-eeee and sssssssssssssssss; m)eeee-d8-eeee and ssssooooooossss; n)eeee-d8-eeee andoooosssssssoooo; o)eeee-d8-eeee and sssssssssssssss; p)eeeeee-d11-eeeeee and ssssssssssssssssssssss; q)eeeee-d10-eeeeee and sososssssssssssosos; r)eeeee-d10-eeeeee and soooossssssssssooos; s)eeeee-d10-eeeeee and soooossssssssssooss; t) eeeee-d8-eeeeee and sosssssssssssoss; u)eeeee-d10-eeeeee and sosssssssssssssssss; v)eeeeee-d10-eeee and sssssssssssssssssss, in, e=2'-MOE nucleoside, and d=deoxyribonucleoside, and wherein s=phosphorothioate linkage, and o=phosphodiester linkage, and wherein at least one "e" or at least one "d" of the gapmer oligonucleotide is replaced with a spacer.

98. The gapmer oligonucleotide of any one of claims 71-97, wherein the gapmer oligonucleotide comprises at least one modified nucleobase.

99. The gapmer oligonucleotide of claim 98, wherein the 5' wing or the 3' wing comprises the at least one modified nucleobase.

100. The gapmer oligonucleotide of claim 98, wherein the central region comprises the at least one modified nucleobase.

101. The gapmer oligonucleotide of any one of claims 98-100, wherein the at least one modified nucleobase is 5-methylcytosine, pseudouridine, or 5-methoxyuridine.

102. The gapmer oligonucleotide of any one of claims 98-101, wherein each cytosine in the 5' wing or the 3' wing is a 5-methylcytosine.

103. The gapmer oligonucleotide of any one of claims 98-102, wherein each cytosine in the central region is a 5-methylcytosine.

104. The gapmer oligonucleotide of claim 71, wherein the gapmer oligonucleotide comprises the following sugar modifications and internucleoside linkage combinations: eeeee-d10-eeeee and ssssssssssssssssssss, wherein e=2'-MOE nucleoside and d=deoxyribonucleoside, wherein s=phosphorothioate linkage, wherein at least one "e" or at least one "d" of the gapmer oligonucleotide is replaced with a spacer, and wherein each cytosine of the 2'-MOE nucleoside is a 5-methylcytosine.

105. The gapmer oligonucleotide of any of the preceding claims, wherein the gapmer oligonucleotide further comprises a conjugate moiety.

106. The gapmer oligonucleotide of claim 105, wherein the conjugate moiety is a cholesterol conjugate located at the 3' end of the gapmer oligonucleotide.

107. A pharmaceutical composition comprising the gapmer oligonucleotide or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 106, and a pharmaceutically acceptable excipient.

108. A method of treating a nervous system disease in a patient in need thereof, the method comprising administering to the patient the gapmer oligonucleotide of any one of claims 1-106, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 107.

109. The method of claim 108, wherein the neurological disease is selected from the group consisting of amyotrophic lateral sclerosis (ALS), frontotemporal dementia (FTD), ALS with FTD, Alzheimer's disease (AD), Parkinson's disease (PD), Parkinson's disease with dementia, dementia with Lewy bodies, synucleinopathy, Huntington's disease, brachial plexus injury, peripheral nerve injury, progressive supranuclear palsy (PSP), brain trauma, spinal cord injury, tuberous sclerosis, 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 and / or neuropathies such as chemotherapy-induced neuropathy, spinocerebellar ataxia (SCA), SCA type 2, spinal muscular atrophy (SMA), Parkinson's disease, Niemann-Pick disease type C (NPC), Charcot-Marie-Tooth disease (CMT), mucopolysaccharidosis type II (MPSIIA), mucolipidosis type IV, GM1 gangliosidosis, sporadic inclusion body myositis (sIBM), Henoch-Schonlein purpura (HSP), limbic age-related TDP-43 encephalopathy (LATE), brain age-related TDP-43 sclerosis (CARTS), Gaucher disease, facial onset sensory and motor neuronopathy, Guam Parkinson's syndrome, multisystem proteinopathy, Perry disease, and synaptic diseases such as autism.

110. The method of claims 108-109, wherein the gapmer oligonucleotide is administered topically, parenterally, intrathecally, orally, pulmonary, intratracheally, intranasally, transdermally, buccally, intrathalamically, intracerebroventricularly, intraocularly, sublingually, rectally, vaginally, or intraduodenally.

111. The method of claim 110, wherein the gapmer oligonucleotide is administered intrathecally.

112. The method of any one of claims 108-111, wherein a therapeutically effective amount of the gapmer oligonucleotide is administered.

113. The method of any one of claims 108-112, wherein the patient is a human.

114. The pharmaceutical composition of claim 107, wherein the pharmaceutical composition is suitable for topical, parenteral, intrathecal, oral, pulmonary, intratracheal, intranasal, transdermal, buccal, intrathalamic, intracerebroventricular, intraocular, sublingual, rectal, vaginal, or intraduodenal administration.

115. Use of gapmer oligonucleotides in the preparation of drugs for treating nervous system diseases.

116. The use according to claim 115, wherein the neurological disease is selected from 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, 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 Disorders, corticobasal degeneration (CBD), and / or neuropathies such as chemotherapy-induced neuropathy, spinocerebellar ataxia (SCA), SCA type 2, spinal muscular atrophy (SMA), Parkinson's disease, Niemann-Pick disease type C (NPC), Charcot-Marie-Tooth disease (CMT), mucopolysaccharidosis type II (MPSIIA), mucolipidosis type IV, GM1 gangliosidosis, sporadic inclusion body myositis (sIBM), Henoch-Schonlein purpura (HSP), limbic age-related TDP-43 encephalopathy (LATE), brain age-related TDP-43 sclerosis (CARTS), Gaucher disease, facial onset sensory and motor neuron disease, Guam Parkinson's disease syndrome, multisystem proteinopathy, Perry disease, and synaptic diseases such as autism.

117. The use according to claim 115 or 116, wherein the gapmer oligonucleotide is the gapmer oligonucleotide according to any one of claims 1-106.

118. A method for treating a neurological disease in a patient in need thereof, the method 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.

119. The method of claim 118, wherein the neurological disease is selected from the group consisting of amyotrophic lateral sclerosis (ALS), frontotemporal dementia (FTD), ALS with FTD, Alzheimer's disease (AD), Parkinson's disease (PD), Parkinson's disease with dementia, dementia with Lewy bodies, synucleinopathy, Huntington's disease, brachial plexus injury, peripheral nerve injury, progressive supranuclear palsy (PSP), brain trauma, spinal cord injury, tuberous sclerosis, 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 and / or neuropathies such as chemotherapy-induced neuropathy, spinocerebellar ataxia (SCA), SCA type 2, spinal muscular atrophy (SMA), Parkinson's disease, Niemann-Pick disease type C (NPC), Charcot-Marie-Tooth disease (CMT), mucopolysaccharidosis type II (MPSIIA), mucolipidosis type IV, GM1 gangliosidosis, sporadic inclusion body myositis (sIBM), Henoch-Schonlein purpura (HSP), limbic age-related TDP-43 encephalopathy (LATE), brain age-related TDP-43 sclerosis (CARTS), Gaucher disease, facial onset sensory and motor neuronopathy, Guam Parkinson's syndrome, multisystem proteinopathy, Perry disease, and synaptic diseases such as autism.

120. The method of claim 118 or 119, wherein the gapmer oligonucleotide is the gapmer oligonucleotide of any one of claims 1-106, a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 107.

121. The method of any one of claims 118-120, wherein the pharmaceutical composition is administered topically, parenterally, orally, pulmonary, rectal, buccal, sublingually, vaginally, intratracheally, intranasally, intrathecally, intracisternal, transdermally, or intraduodenally.

122. The method of any one of claims 118-121, wherein the pharmaceutical composition is administered intrathecally.

123. The method of any one of claims 118-122, wherein the patient is a human.

124. The gapmer oligonucleotide of any one of claims 1-106, or a pharmaceutically acceptable salt thereof, for use as a medicament.

125. The gapmer oligonucleotide or a pharmaceutically acceptable salt thereof of any one of claims 1-106 for use in treating a nervous system disease.

126. The gapmer oligonucleotide for use according to claim 124 or 125, wherein the neurological disease is selected from the group consisting of amyotrophic lateral sclerosis (ALS), frontotemporal dementia (FTD), ALS with FTD, Alzheimer's disease (AD), Parkinson's disease (PD), Parkinson's disease with dementia, dementia with Lewy bodies, synucleinopathy, Huntington's disease, brachial plexus injury, peripheral nerve injury, progressive supranuclear palsy (PSP), brain trauma, spinal cord injury, tuberous sclerosis, Pick's disease, tauopathy, primary age-related tauopathy, Down syndrome, epilepsy / seizure disorder, depression, traumatic brain injury (TBI), chronic traumatic encephalopathy (CTE) , HIV-associated neurocognitive disorder (HAND), multiple system atrophy, amnestic mild cognitive impairment, corticobasal degeneration (CBD), and / or neuropathies such as chemotherapy-induced neuropathy, spinocerebellar ataxia (SCA), SCA type 2, spinal muscular atrophy (SMA), Parkinson's disease, Niemann-Pick disease type C (NPC), Charcot-Marie-Tooth disease (CMT), mucopolysaccharidosis type II (MPSIIA), mucolipidosis type IV, GM1 gangliosidosis, sporadic inclusion body myositis (sIBM), Henoch-Schönlein purpura (HSP), Gaucher disease, facial-onset sensory and motor neuronopathy, Guam-type parkinsonism and dementia syndrome, multisystem proteinopathy, Perry disease, and autism.

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

128. The gapmer oligonucleotide of claim 127, wherein at least one internucleoside linkage of the gapmer oligonucleotide is a phosphorothioate linkage.

129. The gapmer oligonucleotide of claim 128, wherein the phosphorothioate internucleoside linkages are in one of an Rp configuration or an Sp configuration.

130. The gapmer oligonucleotide of claim 127 or 128, wherein all internucleoside linkages of the gapmer oligonucleotide are phosphorothioate linkages.

131. The gapmer oligonucleotide of claim 127 or 128, wherein the gapmer oligonucleotide comprises any of the following patterns of sugar modifications and internucleoside linkage combinations, respectively: a) eeeee-d10-eeeeee and sssssooooooooossss; b)eeeee-d10-eeeeee and ooooosssssssssooooo; c)eeeee-d10-eeeeee and ssssssssssssssssss; d)eee-d8-eee and ssssooooooosss; e)eee-d8-eee and ooosssssssooo f)eee-d8-eee and sssssssssssss; g)eee-d10-eee and ssssooooooooosss; h)eee-d10-eee and ooosssssssssooo; i)eee-d10-eee and ssssssssssssss; j)eeee-d10-eeee and ssssooooooossss; k)eeee-d10-eeee andoooossssssssssoooo; l)eeee-d10-eeee and sssssssssssssssss; m)eeee-d8-eeee and ssssooooooossss; n)eeee-d8-eeee andoooosssssssoooo; o)eeee-d8-eeee and sssssssssssssss; p)eeeeee-d11-eeeeee and ssssssssssssssssssssss; q)eeeee-d10-eeeeee and sososssssssssssosos; r)eeeee-d10-eeeeee and soooossssssssssooos; s)eeeee-d10-eeeeee and soooossssssssssooss; t)eeeee-d8-eeeeee and sosssssssssssoss; u)eeeee-d10-eeeeee and sosssssssssssssssss; v)eeeeee-d10-eeee and sssssssssssssssssss, Wherein at least one "e" or at least one "d" of the gapmer oligonucleotide is replaced with a spacer.

132. A pharmaceutical composition comprising the antisense gapmer oligonucleotide or a pharmaceutically acceptable salt thereof according to any one of claims 126 to 131, and a pharmaceutically acceptable excipient.

133. The gapmer oligonucleotide of any one of claims 127-131, wherein the spacer is a nucleoside replacement group comprising a non-sugar substituent, wherein the non-sugar substituent group does not contain a ketone, aldehyde, ketal, hemiketal, acetal, hemiacetal, aminal, or hemiaminal moiety and is incapable of forming a covalent bond with a nucleotide base.

134. The gapmer oligonucleotide of claim 133, wherein the spacer is represented by formula (X), wherein: Ring A is an optionally substituted 4-8 membered monocyclic cycloalkyl or 4-8 membered monocyclic heterocyclyl, wherein the heterocyclyl contains 1 or 2 heteroatoms selected from O, S and N, provided that A cannot form a covalent bond with a nucleobase; and The symbol indicates the point of attachment of the internucleoside linkage.

135. The gapmer oligonucleotide of claim 133, wherein the spacer is represented by formula (Xa), wherein:

136. The gapmer oligonucleotide of claim 134 or 135, wherein ring A is an optionally substituted 4-8 membered monocyclic cycloalkyl selected from cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl; or a 4-8 membered monocyclic heterocyclyl selected from oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, 1,4-dioxanyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, and azepanyl.

137. The method of claim 136, wherein Ring A is tetrahydrofuranyl.

138. The method of claim 136, wherein Ring A is tetrahydropyranyl.

139. The gapmer oligonucleotide of claim 133, wherein the spacer is represented by formula (I), wherein: X is selected from -CH2- and -O-; and n is 0, 1, 2, or 3.

140. The gapmer oligonucleotide of claim 133, wherein the spacer is represented by formula (I'), wherein:

141. The gapmer oligonucleotide of claim 133, wherein the spacer is represented by formula (Ia), wherein:

142. The gapmer oligonucleotide of claim 133, wherein the spacer is represented by formula (Ia'), wherein:

143. The gapmer oligonucleotide of claim 133, wherein the spacer is represented by Formula II, wherein: and X is selected from -CH2- and -O-.

144. The gapmer oligonucleotide of claim 133, wherein the spacer is represented by formula II', wherein: and X is selected from -CH2- and -O-.

145. The gapmer oligonucleotide of claim 133, wherein the spacer is represented by formula (IIa), wherein:

146. The gapmer oligonucleotide of claim 133, wherein the spacer is represented by formula (Iia'), wherein:

147. The gapmer oligonucleotide of claim 133, wherein the spacer is represented by Formula III, wherein: and X is selected from -CH2- and -O-.

148. The gapmer oligonucleotide of claim 133, wherein the spacer is represented by formula III', wherein: and X is selected from -CH2- and -O-.

149. The gapmer oligonucleotide of claim 133, wherein the spacer is represented by formula (IIIa), wherein:

150. The gapmer oligonucleotide of claim 133, wherein the spacer is represented by formula (IIIa'), wherein:

151. A method for treating a neurological disease and / or a neurological disorder in a patient in need thereof, the method comprising administering to a patient in need thereof a therapeutically effective amount of a gapmer oligonucleotide or a pharmaceutically acceptable salt thereof according to any one of claims 1-106 or claims 127-150, or a pharmaceutical composition according to claim 107 or 132, in combination with a second therapeutic agent selected from the group consisting of riluzole, troriluzole, edaravone (Radicava), rivastigmine, donepezil, QRRL-101, QRL-201, galantamine, selective serotonin reuptake inhibitors, antipsychotics, cholinesterase inhibitors, memantine, benzodiazepine anxiolytics, AMX0035 ZILUCOPLAN (RA101495), dual AON intrathecal administration (e.g., BIIB067, BIIB078), BIIB100, levodopa / carbidopa, dopaminergic drugs (e.g., ropinirole, pramipexole, rotigotine), medroxyprogesterone, KCNQ2 / KCNQ3 openers, pridopidine, PrimeC (a combination of ciprofloxacin and Celebrex), olanzapine (Zyprexa), quetiapine (Seroquel), SSRIs, sodium valproate (Depakote), carbamazepine (Tegretol), medroxyprogesterone, lithium, anticonvulsants and psychostimulants, respiratory care, physical therapy, occupational therapy, speech therapy, nutritional support, or any combination thereof.

152. The method of claim 151, wherein the neurological disease is any of amyotrophic lateral sclerosis (ALS), frontotemporal dementia (FTD), or ALS with FTD.

153. A method for treating a nervous system disease and / or a neuropathy in a patient in need thereof, the method comprising administering to a patient in need thereof a therapeutically effective amount of a gapmer oligonucleotide according to any one of claims 1-106 or claims 127-130, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 107 or 132, in combination with a second therapeutic agent selected from the group consisting of memantine, rivastigmine, galantamine, donepezil, QRL-101, QRL-201, (Rivastigmine), , Aducanumab, BAN2401, BIIB091 (gosuranemab), BIIB076, BIIB080 (IONIS-MAPTRx), Elayta (CT1812), MK1942, allogeneic hMSC, nilotinib, ABT-957, acitretin, ABT-354, GV1001, riluzole, CAD106, CNP520, AD-35, rilapladib, DHP1401 , T-817MA, 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-inactivated anti-β-amyloid protein (Aβ) monoclonal antibody (mAb)), Posiphen ((+)-tartrate phenylserine), AMX0035 Coenzyme Q10, aducanumab (ADUHLEM), memantine (NAMENDA), Namzeric, Suvorexant (belsomra), lecanemab, or any combination thereof.

154. The method of claim 153, wherein the neurological disease is Alzheimer's disease.

155. A method of treating a nervous system disease and / or a neuropathy in a patient in need thereof, the method comprising administering to a patient in need thereof a therapeutically effective amount of a gapmer oligonucleotide of any one of claims 1-106 or claims 127-130, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of claim 107 or 132, in combination with a second therapeutic agent selected from the group consisting of levodopa, carbidopa-levodopa, 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 118, ABBV-0805, zonisamide, deep brain stimulation, brain-derived neurotrophic factor, stem cell transplantation, niacin, brainstem stimulation, nicotine, nabilone, PF-06649751, DNL201, LRRK2 inhibitor, CK1 inhibitor, isradipine, CLR4001, IRX4204, Yohimbine, coenzyme Q10, OXB-102, duloxetine, pioglitazone, preladenant, nourianz, xadago, cogentin, ongentys, exenatide, ganoderma, caffeine, thalizotan, embryonic dopamine cell implantation, or any combination thereof.

156. The method of claim 155, wherein the neurological disease is Parkinson's disease.

157. A method for treating a nervous system disease and / or a neuropathy in a patient in need thereof, the method comprising administering to a patient in need thereof a therapeutically effective amount of a gapmer oligonucleotide or a pharmaceutically acceptable salt thereof as described in any one of claims 1-106 or claims 127-130, or a pharmaceutical composition as described in claims 107 or 132, in combination with a second therapeutic agent selected from the group consisting of UCB0107, ABBV-8E12, F-18AV1451, BIIB092, C2N-8E12, tideglusib, deep transcranial magnetic stimulation, lipoic acid, tolfenamic acid, lithium, AZP2006, glial cell line-derived neurotrophic factor, NBMI, suvorxant, zolpidem, TPI 287, davunetide, pimavanserin, levodopa, carbidopa-levodopa, pramipexole, ropinirole, rotigotine, apomorphine, selegiline, rasagiline, entacapone, tolcapone, amantadine, trihexyphenidyl, BIIB054 (cinipazumab), BIIB094, BIIB 118, ABBV-0805, zonisamide, deep brain stimulation, brain-derived neurotrophic factor, stem cell transplantation, niacin, brainstem stimulation, nicotine, nabilone, PF-06649751, DNL201, LRRK2 inhibitor, CK1 inhibitor, isradipine, CLR4001, IRX4204, yohimbine, coenzyme Q10, OXB-102, duloxetine, pioglitazone, preladenant, or any combination thereof.

158. The method of claim 157, wherein the neurological disease is progressive supranuclear palsy (PSP).

159. A method for treating a nervous system disease and / or a neuropathy in a patient in need thereof, the method comprising administering to a patient in need thereof a therapeutically effective amount of a gapmer oligonucleotide or a pharmaceutically acceptable salt thereof as described in any one of claims 1-106 or claims 127-130, or a pharmaceutical composition as described in claim 107 or 132, in combination with a second therapeutic agent selected from the group consisting of tetrabenazine, deutetrabenazine, physical therapy, risperidone, haloperidol, chlorpromazine, clonazepam, diazepam, benzodiazepines, selective serotonin reuptake inhibitors, quetiapine, carbatrol, valproic acid, lamotrigine (lamotrigine), pridopidine, delta-9-tetrahydrocannabinol, cannabidiol, stem cell therapy, ISIS-443139, nilotinib, resveratrol, neflamapimod, fenofibrate, creatine, RO7234292, SAGE-718, WVE-120102, WVE-120101, dimebon, minocycline, deep brain stimulation, ursodeoxycholic acid, coenzyme Q10, OMS643762, VX15 / 2503, PF-02545920, BN82451B, SEN0014196, olanzapine, tiapride, or any combination thereof.

160. The method of claim 159, wherein the neurological disease is Huntington's disease.

161. A method for treating a neurological disease and / or a neuropathy in a patient in need thereof, the method comprising administering to a patient in need thereof a therapeutically effective amount of a gapmer oligonucleotide or a pharmaceutically acceptable salt thereof as described in any one of claims 1-106 or claims 127-130, or a pharmaceutical composition as described in claim 107 or 132 in combination with a second therapeutic agent selected from the group consisting of an anticoagulant, an antidepressant, a muscle relaxant, a stimulant, an anticonvulsant, an anxiolytic, erythropoietin, hyperbaric therapy, rehabilitation therapy (e.g., physical, occupational, speech, psychological, or vocational counseling), or any combination thereof.

162. The method of claim 161, wherein the neurological disease is brain trauma.

163. A method for treating a nervous system disease and / or a neuropathy in a patient in need thereof, the method comprising administering to a patient in need thereof a therapeutically effective amount of a gapmer oligonucleotide or a pharmaceutically acceptable salt thereof as described in any one of claims 1-106 or claims 127-130, or a pharmaceutical composition as described in claim 107 or 132, in combination with a second therapeutic agent selected from the group comprising: AXER-204, glibenclamide, 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 exercise strategies, and healthy diet), or any combination thereof.

164. The method of claim 163, wherein the neurological disease is spinal cord injury.

165. A method for treating a neurological disease and / or a neuropathy in a patient in need thereof, the method comprising administering to a patient in need thereof a therapeutically effective amount of a gapmer oligonucleotide or a pharmaceutically acceptable salt thereof as described in any one of claims 1-106 or claims 127-130, or a pharmaceutical composition as described in claims 107 or 132 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 that can be selected as an additional therapy.

166. The method of claim 165, wherein the neurological disease is corticobasal degeneration.

167. A method for treating a neurological disease and / or a neuropathy in a patient in need thereof, the method comprising administering to a patient in need thereof a therapeutically effective amount of a gapmer oligonucleotide or a pharmaceutically acceptable salt thereof as described in any one of claims 1-106 or claims 127-130, or a pharmaceutical composition as described in claims 107 or 132, in combination with a second therapeutic agent selected from the group comprising gabapentin, pregabalin, lamotrigine, carbamazepine, duloxetine, gabapentinoids, tricyclic antidepressants, serotonin-norepinephrine reuptake inhibitors, opioids, neurotoxins, dextromethorphan, nicotinamide riboside, autoantibodies targeting neuronal antigens (TS-HDS and FGFR3), or any combination thereof.

168. The method of claim 167, wherein the neuropathy is chemotherapy-induced neuropathy.

169. A method of treating a nervous system disease and / or a neuropathy in a patient in need thereof, the method comprising administering to a patient in need thereof a therapeutically effective amount of a gapmer oligonucleotide or a pharmaceutically acceptable salt thereof according to any one of claims 1-106 or claims 127-130, or a pharmaceutical composition according to claim 107 or 132 in combination with a second therapeutic agent selected from the group consisting of amantadine, armodafinil, baclofen , buspirone, carbamazepine, citalopram, clonazepam, desvenlafaxine, diazepam, duloxetine, escitalopram, flunarizine, fluoxetine, 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.

170. The method of claim 169, wherein the neurological disease is spinocerebellar ataxia.

171. A method of treating a nervous system disease and / or a neuropathy in a patient in need thereof, the method comprising administering to a patient in need thereof a therapeutically effective amount of a gapmer oligonucleotide or a pharmaceutically acceptable salt thereof of any one of claims 1-106 or claims 127-130, or a pharmaceutical composition of claim 107 or 132 in combination with a second therapeutic agent selected from the group consisting of briviact, epidiolex, carbamazepine (Tegretol), xcopri, valium, ativan, klonopin, aptiom, zarontin, felbatol, fintepla, VIMPAT, Lamictal, Keppra, oxcarbazepine, xr, Trileptal), perampanel (fycompa), phenobarbital, phenytoin (dilantin), pregabalin (lyrica), tiagabine (gabitril), topiramate (topamax), valproic acid (depakene, depakote), zonisamide (zonegran), or any combination thereof.

172. The method of claim 171, wherein the neurological disease is epilepsy.

173. A method for treating a neurological disease and / or a neuropathy in a patient in need thereof, the method comprising administering to a patient in need thereof a therapeutically effective amount of a gapmer oligonucleotide or a pharmaceutically acceptable salt thereof as described in any one of claims 1-106 or claims 127-130, or a pharmaceutical composition as described in claim 107 or 132 in combination with a second therapeutic agent selected from the group consisting of nusinersen (SPINRAZA), onasemnogeneabeparvovec-xioi (ZOLGENSMA), risdiplam (EVRYSDI), or any combination thereof.

174. The method of claim 173, wherein the neurological disease is spinal muscular atrophy.

175. A method for treating a neurological disease and / or a neuropathy in a patient in need thereof, the method comprising administering to a patient in need thereof a therapeutically effective amount of a gapmer oligonucleotide or a pharmaceutically acceptable salt thereof as described in any one of claims 1-106 or claims 127-130, or a pharmaceutical composition as described in claim 107 or 132 in combination with a second therapeutic agent selected from the group consisting of anti-epileptic drugs, speech therapy, physical therapy, occupational therapy, Adrabetadex, Arimoclomol, N-acetyl-L-leucine, or any combination thereof.

176. The method of claim 175, wherein the neurological disease is Niemann-Pick disease type C.

177. A method for treating a neurological disease and / or a neuropathy in a patient in need thereof, the method comprising administering to the patient in need thereof a therapeutically effective amount of a gapmer oligonucleotide or a pharmaceutically acceptable salt thereof as described in any one of claims 1-106 or claims 127-130, or a pharmaceutical composition as described in claim 107 or 132 in combination with a second therapeutic agent selected from the group consisting of physical and occupational therapy, orthopedic surgery, orthopedic devices, PXT3003, or any combination thereof.

178. The method of claim 177, wherein the neurological disease is Charcot-Marie-Tooth Disease (CMT).

179. A method for treating a neurological disease and / or a neuropathy in a patient in need thereof, the method comprising administering to a patient in need thereof a therapeutically effective amount of a gapmer oligonucleotide or a pharmaceutically acceptable salt thereof as described in any one of claims 1-106 or claims 127-130, or a pharmaceutical composition as described in claim 107 or 132 in combination with a second therapeutic agent selected from the group consisting of: enzyme replacement therapy: Elaprase, surgical intervention (tonsillectomy and / or adenoidectomy), RGX-121 gene therapy, adalimumab, MT2013-31, or any combination thereof.

180. The method of claim 179, wherein the neurological disease is mucopolysaccharidosis type II (MPSIIA).

181. A method for treating a neurological disease and / or a neuropathy in a patient in need thereof, the method comprising administering to a patient in need thereof a therapeutically effective amount of a gapmer oligonucleotide or a pharmaceutically acceptable salt thereof as described in any one of claims 1-106 or claims 127-130, or a pharmaceutical composition as described in claim 107 or 132 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.

182. The method of claim 181, wherein the neurological disease is mucolipidosis type IV.

183. A method for treating a neurological disease and / or a neuropathy in a patient in need thereof, the method comprising administering to a patient in need thereof a therapeutically effective amount of a gapmer oligonucleotide or a pharmaceutically acceptable salt thereof as described in any one of claims 1-106 or claims 127-130, or a pharmaceutical composition as described in claim 107 or 132 in combination with a second therapeutic agent selected from the group consisting of anticonvulsants, physical and occupational therapy, β-galactosidase, gene delivery of β-galactosidase, LYS-GM101 gene therapy, or any combination thereof.

184. The method of claim 183, wherein the neurological disease is GM1 gangliosidosis.

185. A method for treating a neurological disease and / or a neuropathy in a patient in need thereof, the method comprising administering to a patient in need thereof a therapeutically effective amount of a gapmer oligonucleotide or a pharmaceutically acceptable salt thereof as described in any one of claims 1-106 or claims 127-130, or a pharmaceutical composition as described in claim 107 or 132 in combination with a second therapeutic agent selected from the group consisting of physical and occupational therapy, use of a device such as a brace, a walker, a wheelchair, an immunosuppressant, BYM338, or any combination thereof.

186. The method of claim 185, wherein the neurological disease is sporadic inclusion body myositis (sIBM).

187. A method for treating a neurological disease and / or a neuropathy in a patient in need thereof, the method comprising administering to a patient in need thereof a therapeutically effective amount of a gapmer oligonucleotide or a pharmaceutically acceptable salt thereof as described in any one of claims 1-106 or claims 127-130, or a pharmaceutical composition as described in claim 107 or 132 in combination with a second therapeutic agent selected from the group consisting of: corticosteroids, colchicine, dapsone, azathioprine, or any combination thereof.

188. The method of claim 187, wherein the neurological disease is Henoch-Schonlein purpura (HSP).

189. A method for treating a neurological disease and / or a neuropathy in a patient in need thereof, the method comprising administering to a patient in need thereof a therapeutically effective amount of a gapmer oligonucleotide or a pharmaceutically acceptable salt thereof as described in any one of claims 1-106 or claims 127-130, or a pharmaceutical composition as described in claim 107 or 132 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.

190. The method of claim 189, wherein the neurological disease is Gaucher disease.

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