WO2024201423A2 - Arnsi ciblant la protéine tau associée aux microtubules et leurs utilisations - Google Patents

Arnsi ciblant la protéine tau associée aux microtubules et leurs utilisations Download PDF

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WO2024201423A2
WO2024201423A2 PCT/IB2024/053120 IB2024053120W WO2024201423A2 WO 2024201423 A2 WO2024201423 A2 WO 2024201423A2 IB 2024053120 W IB2024053120 W IB 2024053120W WO 2024201423 A2 WO2024201423 A2 WO 2024201423A2
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seq
oligonucleotide
nucleotides
strand
antisense strand
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WO2024201423A3 (fr
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Jin-Hyeob RYU
Young Jin Park
Suhyeong Park
Yu Na Lim
Insun Kim
Mincheol Kwon
Jihye Kim
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Biorchestra Co Ltd
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Biorchestra Co Ltd
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7088—Compounds having three or more nucleosides or nucleotides
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/10—Type of nucleic acid
    • C12N2310/14—Type of nucleic acid interfering nucleic acids [NA]
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/30—Chemical structure
    • C12N2310/31—Chemical structure of the backbone
    • C12N2310/315—Phosphorothioates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/30—Chemical structure
    • C12N2310/32—Chemical structure of the sugar
    • C12N2310/321—2'-O-R Modification
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/30—Chemical structure
    • C12N2310/32—Chemical structure of the sugar
    • C12N2310/322—2'-R Modification

Definitions

  • the present disclosure relates to oligonucleotides, e.g., siRNAs, that target a Microtubule-associated protein tau (MAPT) transcript in a cell, leading to modified expression of Microtubule-associated protein tau (MAPT) protein, thereby treating a disease or disorder related to MAPT gene/protein expression.
  • oligonucleotides e.g., siRNAs
  • MRNAs Microtubule-associated protein tau
  • MAPT Microtubule-associated protein tau
  • MAPT is a small soluable protein that is expressed in many cell types. It is predominantly localized within the presynaptic terminals of neurons. While the precise function has yet to be fully elucidated, MAPT has been suggested to play an important role in the regulation of synaptic transmission.
  • tauopathies are neurodegenerative diseases characterized by abnormal accumulation of MAPT protein aggregates within the brain.
  • Tauopathies such as behavioral variant frontotemporal dementia, are highly prevalent neurodegenerative conditions, especially in the elderly. Dementia currently affects more than 55 million people worldwide and as of 2017, more than 5 million Americans had been diagnosed with dementia. In 2017, the average annual cost of care for a patient with dementia was $89,000. As the population ages the number of people living with the disease will increase placing an even greater burden on the healthcare system and the families of the patients.
  • the treatment options for this disease are scant, and there are no small molecule treatments for this disease on market.
  • the current potential best treatment for the disease, Lecanemab (currently pending before the FDA), is linked to increased brain bleeding and swelling, with potentially deadly contraindications that limit its usefulness. More cost effective and robust treatment options are necessary to properly handle this worsening health crisis.
  • AMT Microtubule-associated protein tau
  • 5’-AACGUCCAGUCCAAGUGUGGC-3’ (SEQ ID NO: 1) (sense strand) or 5’-GCCACACUUGGACUGGACGTT-3’ (SEQ ID NO: 2) (antisense strand); ii. 5’-AGCAACGUCCAGUCCAAG-3’ (SEQ ID NO: 3) (sense strand) or 5’-CUUGGACUGGACGUUGCU-3’ (SEQ ID NO: 4) (antisense strand); iii.
  • 5’-AGCGGCUACAGCAGCCCCG -3’ (SEQ ID NO: 9) (sense strand) or 5’-CGGGGCUGCUGUAGCCGCU-3’ (SEQ ID NO: 10) (antisense strand); vi. 5’-AAUUAAUAAGAAGCUGGAUCU-3’ (SEQ ID NO: 11) (sense strand) or 5’-AGAUCCAGCUUCUUAUUAATT-3’ (SEQ ID NO: 12) (antisense strand); vii.
  • 5’-GAGCCCAAGAAGGUGGCAG-3’ (SEQ ID NO: 13) (sense strand) or 5’-CUGCCACCUUCUUGGGCUC-3’ (SEQ ID NO: 14) (antisense strand); viii. 5’-ACCCGGGAGCCCAAGAAG-3’ (SEQ ID NO: 15) (sense strand) or 5’-CUUCUUGGGCUCCCGGGU-3’ (SEQ ID NO: 16) (antisense strand); ix. 5’-AGAAGCUGGAUCUUAGCAA-3’ (SEQ ID NO: 17) (sense strand) or 5’-UUGCUAAGAUCCAGCUUCU-3’ (SEQ ID NO: 18) (antisense strand); x.
  • 5’-AGGGCAGCCUGUGGGAGAA-3’ (SEQ ID NO: 35) (sense strand) or 5’-UUCUCCCACAGGCUGCCCU-3’ (SEQ ID NO: 36) (antisense strand); or xix.
  • 5’-GGUGCAGAUAAUUAAUAAG-3’ (SEQ ID NO: 64) (sense strand) or 5’-CUUAUUAAUUAUCUGCACC-3’ (SEQ ID NO: 65) (antisense strand).
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are capable of hybridizing to a nucleic acid sequence within an Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: i. 5’-AAGCUGGAUCUUAGCAACGUC-3’ (SEQ ID NO: 37) (sense strand) or 5’-GACGUUGCUAAGAUCCAGCTT-3’ (SEQ ID NO: 38) (antisense strand); ii.
  • MTT Microtubule-associated protein tau
  • 5’-AUCAAACACGUCCCGGGAG-3’ (SEQ ID NO: 49) (sense strand) or 5’-CUCCCGGGACGUGUUUGAU-3’ (SEQ ID NO: 50) (antisense strand); viii. 5’-AACACGUCCCGGGAGGCGGCA-3’ (SEQ ID NO: 51) (sense strand) or 5’-UGCCGCCUCCCGGGACGUGTT-3’ (SEQ ID NO: 52) (antisense strand); ix.
  • the present disclosure provides an oligonucleotide wherein the contiguous nucleotide sequence is complementary to the nucleic acid sequence.
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- AACGUCCAGUCCAAGUGUGGC-3’ (SEQ ID NO: 1) (sense strand) or 5’-GCCACACUUGGACUGGACGTT-3’ (SEQ ID NO: 2) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- AGCAACGUCCAGUCCAAG-3’ (SEQ ID NO: 3) (sense strand) or 5’- CUUGGACUGGACGUUGCU-3’ (SEQ ID NO: 4).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- AAGGUGCAGAUAAUUAAUAAG-3’ (SEQ ID NO: 5) (sense strand) or 5’-CUUAUUAAUUAUCUGCACCTT -3’ (SEQ ID NO: 6) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- UGAGAACCUGAAGCACCAG -3’ (SEQ ID NO: 7) (sense strand) or 5’-CUGGUGCUUCAGGUUCUCA-3’ (SEQ ID NO: 8) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- AGCGGCUACAGCAGCCCCG -3’ (SEQ ID NO: 9) (sense strand) or 5’-CGGGGCUGCUGUAGCCGCU-3’ (SEQ ID NO: 10) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- AAUUAAUAAGAAGCUGGAUCU-3’ (SEQ ID NO: 11) (sense strand) or 5’-AGAUCCAGCUUCUUAUUAATT-3’ (SEQ ID NO: 12) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- GAGCCCAAGAAGGUGGCAG-3’ (SEQ ID NO: 13) (sense strand) or 5’-CUGCCACCUUCUUGGGCUC-3’ (SEQ ID NO: 14) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- ACCCGGGAGCCCAAGAAG-3’ (SEQ ID NO: 15) (sense strand) or 5’-CUUCUUGGGCUCCCGGGU-3’ (SEQ ID NO: 16) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- AGAAGCUGGAUCUUAGCAA-3’ (SEQ ID NO: 17) (sense strand) or 5’-UUGCUAAGAUCCAGCUUCU-3’ (SEQ ID NO: 18) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- AAGGAUAAUAUCAAACACGUC-3’ (SEQ ID NO: 19) (sense strand) or 5’-GACGUGUUUGAUAUUAUCCTT-3’ (SEQ ID NO: 20) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- AUAAUAUCAAACACGUCCC-3’ (SEQ ID NO: 21) (sense strand) or 5’-GGGACGUGUUUGAUAUUAU-3’ (SEQ ID NO: 22) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- AGGAGGUGGCCAGGUGGAA-3’ (SEQ ID NO: 23) (sense strand) or 5’-UUCCACCUGGCCACCUCCU-3’ (SEQ ID NO: 24) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-AAUAUCACCCACGUCCCUGG-3’ (SEQ ID NO: 25) (sense strand) or 5’-CCAGGGACGUGGGUGAUATT-3’ (SEQ ID NO: 26) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- UGGCCAGGUGGAAGUAAAA-3’ (SEQ ID NO: 27) (sense strand) or 5’-UUUUACUUCCACCUGGCCA-3’ (SEQ ID NO: 28) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- CCAAAGCCAAGACAGACCA-3’ (SEQ ID NO: 29) (sense strand) or 5’-UGGUCUGUCUUGGCUUUGG-3’ (SEQ ID NO: 30) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- AAGCAGGGUUUGUGAUCAG-3’ (SEQ ID NO: 31) (sense strand) or 5’-CUGAUCACAAACCCUGCTT-3’ (SEQ ID NO: 32) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- CUCGGGACUUCAAAAUCAG-3’ (SEQ ID NO: 33) (sense strand) or 5’-CUGAUUUUGAAGUCCCGAG-3’ (SEQ ID NO: 34) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- AGGGCAGCCUGUGGGAGAA-3’ (SEQ ID NO: 35) (sense strand) or 5’-UUCUCCCACAGGCUGCCCU-3’ (SEQ ID NO: 36) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- AAGCUGGAUCUUAGCAACGUC-3’ (SEQ ID NO: 37) (sense strand) or 5’-GACGUUGCUAAGAUCCAGCTT-3’ (SEQ ID NO: 38) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-AGUGUGCAAAUAGUCUACAA-3’ (SEQ ID NO: 39) (sense strand) or 5’-UUGUAGACUAUUUGCACACU-3’ (SEQ ID NO: 40) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- AAGGACAGAGUCCAGUCGAAG-3’ (SEQ ID NO: 41) (sense strand) or 5’-CUUCGACUGGACUCUGUCCTT-3’ (SEQ ID NO: 42) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- AAGAUUGAAACCCACAAGC-3’ (SEQ ID NO: 43) (sense strand) or 5’-GCUUGUGGGUUUCAAUCTT-3’ (SEQ ID NO: 44) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- GAAACCCACAAGCUGACCU-3’ (SEQ ID NO: 45) (sense strand) or 5’-AGGUCAGCUUGUGGGUUUC-3’ (SEQ ID NO: 46) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- AAUAUCAAACACGUCCCGGGA-3’ (SEQ ID NO: 47) (sense strand) or 5’-UCCCGGGACGUGUUUGAUATT-3’ (SEQ ID NO: 48) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- AUCAAACACGUCCCGGGAG-3’ (SEQ ID NO: 49) (sense strand) or 5’-CUCCCGGGACGUGUUUGAU-3’ (SEQ ID NO: 50) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- AACACGUCCCGGGAGGCGGCA-3’ (SEQ ID NO: 51) (sense strand) or 5’-UGCCGCCUCCCGGGACGUGTT-3’ (SEQ ID NO: 52) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- ACGUCCCGGGAGGCGGCAG-3’ (SEQ ID NO: 53) (sense strand) or 5’-CUGCCGCCUCCCGGGACGU-3’ (SEQ ID NO: 54) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- UCCCGGGAGGCGGCAGUGU-3’ (SEQ ID NO: 55) (sense strand) or 5’-ACACUGCCGCCUCCCGGGA-3’ (SEQ ID NO: 56) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-GGUGCAGAUAAUUAAUAAG-3’ (SEQ ID NO: 64) (sense strand) or 5’-CUUAUUAAUUAUCUGCACC-3’ (SEQ ID NO: 65) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-GGUGCAGAUAAUUAAUAAG-3’ (SEQ ID NO: 64) (sense strand) or 5’-CUUAUUAAUUAUCUGCACC -3’ (SEQ ID NO: 65) (antisense strand); wherein the 1 st , 3 rd , 4 th , 7 th , 8 th , 9 th , 10 th , 11 th , 13 th , and 18 th nucleotides from 5’ to 3’of the sense strand is RNA and the 2 nd , 5 th , 6 th , 12 th , 14 th , 15 th , 16
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-GGUGCAGAUAAUUAAUAAG-3’ (SEQ ID NO: 64) (sense strand) or 5’-CUUAUUAAUUAUCUGCACC -3’ (SEQ ID NO: 65) (antisense strand); wherein the 1 st , 2 nd , 3 rd , 4 th , 5 th , 6 th , 8 th , 9 th , 10 th , 11 th , 12 th , 13 th , 14 th , 15 th , 16 th , 18 th , and 19 th nucleotides from 5’ to 3’of the antisense strand
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-GGUGCAGAUAAUUAAUAAG-3’ (SEQ ID NO: 64) (sense strand) or 5’-CUUAUUAAUUAUCUGCACC -3’ (SEQ ID NO: 65) (antisense strand); wherein at least one of the backbone of the sense and/or antisense strands is linked to a phosphodiester bond, a phosphothioate bond, or a combination thereof.
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-GGUGCAGAUAAUUAAUAAG-3’ (SEQ ID NO: 64) (sense strand) or 5’-CUUAUUAAUUAUCUGCACC -3’ (SEQ ID NO: 65) (antisense strand); wherein each of the backbone of the sense and antisense strands is linked by a phosphodiester bond.
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-GGUGCAGAUAAUUAAUAAG-3’ (SEQ ID NO: 64) (sense strand) or 5’-CUUAUUAAUUAUCUGCACC -3’ (SEQ ID NO: 65) (antisense strand); wherein the 1 st , 2 nd , 3 rd , 4 th , 5 th , 6 th , 8 th , 12 th , 13 th , 14 th , 15 th , 16 th , 17 th , 18 th and 19 th nucleotides from 5’ to 3’of the sense strand comprise 2’-OMe at the sugar position and the 7
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-GGUGCAGAUAAUUAAUAAG-3’ (SEQ ID NO: 64) (sense strand) or 5’-CUUAUUAAUUAUCUGCACC -3’ (SEQ ID NO: 65) (antisense strand); wherein the 1 st , 3 rd , 4 th , 5 th , 7 th , 8 th , 10 th , 11 th , 12 th , 13 th , 15 th , 17 th , 18 th , and 19 th nucleotides from 5’ to 3’of the antisense strand comprise 2’-OMe at the sugar position and the 2 nd
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-GGUGCAGAUAAUUAAUAAG-3’ (SEQ ID NO: 64) (sense strand) or 5’-CUUAUUAAUUAUCUGCACC -3’ (SEQ ID NO: 65) (antisense strand); wherein the nucleotides in the sense strand and/or the nucleotides in the antisense strand are linked by a phosphodiester bond, a phosphothioate bond, or any combination thereof.
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-GGUGCAGAUAAUUAAUAAG-3’ (SEQ ID NO: 64) (sense strand) or 5’-CUUAUUAAUUAUCUGCACC -3’ (SEQ ID NO: 65) (antisense strand); wherein the 1 st , 2 nd and 3 rd nucleotides from 5’ to 3’of the sense strand, the 1 st , 2 nd and 3 rd nucleotides from 5’ to 3’of the antisense strand, and the 17 th , 18 th and 19 th nucleotides from 5’ to 3’of the antisense strand are linked by phosphoroth
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-AGCGGCUACAGCAGCCCCG-3’ (SEQ ID NO: 9) (sense strand) or 5’-CGGGGCUGCUGUAGCCGCU-3’ (SEQ ID NO: 10) (antisense strand); wherein the 1 st , 3 rd , 4 th , 7 th , 8 th , 9 th , 10 th , 11 th , 13 th , and 18 th nucleotides from 5’ to 3’ of the sense strand is RNA and the 2 nd , 5 th , 6 th , 12 th , 14 th , 15 th , 16 th
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-AGCGGCUACAGCAGCCCCG-3’ (SEQ ID NO: 9) (sense strand) or 5’-CGGGGCUGCUGUAGCCGCU-3’ (SEQ ID NO: 10) (antisense strand); wherein the 1 st , 2 nd , 3 rd , 4 th , 5 th , 6 th , 8 th , 9 th , 10 th , 11 th , 12 th , 13 th , 14 th , 15 th , 16 th , 18 th , and 19 th nucleotides from 5’ to 3’of the antisense strand is
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-AGCGGCUACAGCAGCCCCG-3’ (SEQ ID NO: 9) (sense strand) or 5’-CGGGGCUGCUGUAGCCGCU-3’ (SEQ ID NO: 10) (antisense strand); wherein at least one of the backbone of the sense and/or antisense strands is linked to a phosphodiester bond, a phosphothioate bond, or a combination thereof.
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-AGCGGCUACAGCAGCCCCG-3’ (SEQ ID NO: 9) (sense strand) or 5’-CGGGGCUGCUGUAGCCGCU-3’ (SEQ ID NO: 10) (antisense strand); wherein each of the backbone of the sense and antisense strands is linked by a phosphodiester bond.
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-AGCGGCUACAGCAGCCCCG-3’ (SEQ ID NO: 9) (sense strand) or 5’-CGGGGCUGCUGUAGCCGCU-3’ (SEQ ID NO: 10) (antisense strand); wherein the 1 st , 2 nd , 3 rd , 4 th , 5 th , 6 th , 8 th , 12 th , 13 th , 14 th , 15 th , 16 th , 17 th , 18 th and 19 th nucleotides from 5’ to 3’of the sense strand comprise 2’-OMe at the sugar position and the 7 th
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-AGCGGCUACAGCAGCCCCG-3’ (SEQ ID NO: 9) (sense strand) or 5’-CGGGGCUGCUGUAGCCGCU-3’ (SEQ ID NO: 10) (antisense strand); wherein the 1 st , 3 rd , 4 th , 5 th , 7 th , 8 th , 10 th , 11 th , 12 th , 13 th , 15 th , 17 th , 18 th , and 19 th nucleotides from 5’ to 3’of the antisense strand comprise 2’-OMe at the sugar position and the 2 nd
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-AGCGGCUACAGCAGCCCCG-3’ (SEQ ID NO: 9) (sense strand) or 5’-CGGGGCUGCUGUAGCCGCU-3’ (SEQ ID NO: 10) (antisense strand); wherein at least one of the backbone of the sense and/or antisense strands is linked to a phosphodiester bond, a phosphothioate bond, or a combination thereof.
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-AGCGGCUACAGCAGCCCCG-3’ (SEQ ID NO: 9) (sense strand) or 5’-CGGGGCUGCUGUAGCCGCU-3’ (SEQ ID NO: 10) (antisense strand); wherein the 1 st , 2 nd and 3 rd nucleotides from 5’ to 3’of the sense strand, the 1 st , 2 nd and 3 rd nucleotides from 5’ to 3’of the antisense nucleotides, and the 17 th , 18 th and 19 th nucleotides from 5’ to 3’of the antisense strand are linked by phospho
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-AGGAGGUGGCCAGGUGGAA-3’ (SEQ ID NO: 23) (sense strand) or 5’-UUCCACCUGGCCACCUCCU-3’ (SEQ ID NO: 24) (antisense strand); wherein the 1 st , 3 rd , 4 th , 7 th , 8 th , 9 th , 10 th , 11 th , 13 th , and 18 th nucleotides from 5’ to 3’of the sense strand is RNA and the 2 nd , 5 th , 6 th , 12 th , 14 th , 15 th , 16 th
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-AGGAGGUGGCCAGGUGGAA-3’ (SEQ ID NO: 23) (sense strand) or 5’-UUCCACCUGGCCACCUCCU-3’ (SEQ ID NO: 24) (antisense strand); wherein at least one of the backbone of the sense and/or antisense strands is linked to a phosphodiester bond, a phosphothioate bond, or a combination thereof.
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-AGGAGGUGGCCAGGUGGAA-3’ (SEQ ID NO: 23) (sense strand) or 5’-UUCCACCUGGCCACCUCCU-3’ (SEQ ID NO: 24) (antisense strand); wherein each of the backbone of the sense and antisense strands is linked by a phosphodiester bond.
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-AGGAGGUGGCCAGGUGGAA-3’ (SEQ ID NO: 23) (sense strand) or 5’-UUCCACCUGGCCACCUCCU-3’ (SEQ ID NO: 24) (antisense strand); wherein the 1 st , 2 nd , 3 rd , 4 th , 5 th , 6 th , 8 th , 12 th , 13 th , 14 th , 15 th , 16 th , 17 th , 18 th and 19 th nucleotides from 5’ to 3’of the sense strand comprise 2’-OMe at the sugar position and the 7 th
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-AGGAGGUGGCCAGGUGGAA-3’ (SEQ ID NO: 23) (sense strand) or 5’-UUCCACCUGGCCACCUCCU-3’ (SEQ ID NO: 24) (antisense strand); wherein the 1 st , 3 rd , 4 th , 5 th , 7 th , 8 th , 10 th , 11 th , 12 th , 13 th , 15 th , 17 th , 18 th , and 19 th nucleotides from 5’ to 3’of the antisense strand comprise 2’-OMe at the sugar position and the 2 nd
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-AGGAGGUGGCCAGGUGGAA-3’ (SEQ ID NO: 23) (sense strand) or 5’-UUCCACCUGGCCACCUCCU-3’ (SEQ ID NO: 24) (antisense strand); wherein at least one of the backbone of the sense and/or antisense strands is linked to a phosphodiester bond, a phosphothioate bond, or a combination thereof.
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-AGGAGGUGGCCAGGUGGAA-3’ (SEQ ID NO: 23) (sense strand) or 5’-UUCCACCUGGCCACCUCCU-3’ (SEQ ID NO: 24) (antisense strand); wherein the 1 st , 2 nd and 3 rd nucleotides from 5’ to 3’of the sense strand, the 1 st , 2 nd and 3 rd nucleotides of the antisense strand, and the 17 th , 18 th and 19 th nucleotides from 5’ to 3’of the antisense strand are linked by phosphorothioate bonds, and every
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-UGGCCAGGUGGAAGUAAAA-3’ (SEQ ID NO: 27) (sense strand) or 5’-UUUUACUUCCACCUGGCCA-3’ (SEQ ID NO: 28) (antisense strand); wherein the 1 st , 3 rd , 4 th , 7 th , 8 th , 9 th , 10 th , 11 th , 13 th , and 18 th nucleotides from 5’ to 3’of the sense strand is RNA and the 2 nd , 5 th , 6 th , 12 th , 14 th , 15 th , 16
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-UGGCCAGGUGGAAGUAAAA-3’ (SEQ ID NO: 27) (sense strand) or 5’-UUUUACUUCCACCUGGCCA-3’ (SEQ ID NO: 28) (antisense strand); wherein the 1 st , 2 nd , 3 rd , 4 th , 5 th , 6 th , 8 th , 9 th , 10 th , 11 th , 12 th , 13 th , 14 th , 15 th , 16 th , 18 th , and 19 th nucleotides from 5’ to 3’ of the antisense strand
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-UGGCCAGGUGGAAGUAAAA-3’ (SEQ ID NO: 27) (sense strand) or 5’-UUUUACUUCCACCUGGCCA-3’ (SEQ ID NO: 28) (antisense strand); wherein at least one of the backbone of the sense and/or antisense strands is linked to a phosphodiester bond, a phosphothioate bond, or a combination thereof.
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-UGGCCAGGUGGAAGUAAAA-3’ (SEQ ID NO: 27) (sense strand) or 5’-UUUUACUUCCACCUGGCCA-3’ (SEQ ID NO: 28) (antisense strand); wherein each of the backbone of the sense and antisense strands is linked by a phosphodiester bond.
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-UGGCCAGGUGGAAGUAAAA-3’ (SEQ ID NO: 27) (sense strand) or 5’-UUUUACUUCCACCUGGCCA-3’ (SEQ ID NO: 28) (antisense strand); wherein the 1 st , 2 nd , 3 rd , 4 th , 5 th , 6 th , 8 th , 12 th , 13 th , 14 th , 15 th , 16 th , 17 th , 18 th , and 19 th nucleotides from 5’ to 3’of the sense strand comprises 2’ OMe at the sugar position and the contiguous nucleot
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-UGGCCAGGUGGAAGUAAAA-3’ (SEQ ID NO: 27) (sense strand) or 5’-UUUUACUUCCACCUGGCCA-3’ (SEQ ID NO: 28) (antisense strand); wherein the 1 st , 3 rd , 4 th , 5 th , 7 th , 8 th , 10 th , 11 th , 12 th , 13 th , 15 th , 17 th , 18 th , and 19 th nucleotides from 5’ to 3’of the antisense strand comprise 2’-OMe at the sugar position and the 2 nd
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-UGGCCAGGUGGAAGUAAAA-3’ (SEQ ID NO: 27) (sense strand) or 5’-UUUUACUUCCACCUGGCCA-3’ (SEQ ID NO: 28) (antisense strand); wherein the 1 st , 2 nd and 3 rd nucleotides from 5’ to 3’ of the sense strand, the 1 st , 2 nd and 3 rd nucleotides from 5’ to 3’of the antisense strand, and the 17 th , 18 th and 19 th nucleotides from 5’ to 3’of the antisense strand are linked by phosphoroth
  • the oligonucleotide in the present disclosure is double stranded and comprises: i. 5’-AACGUCCAGUCCAAGUGUGGC-3’ (SEQ ID NO: 1) (sense strand) and/or 5’-GCCACACUUGGACUGGACGTT-3’ (SEQ ID NO: 2) (antisense strand); ii. 5’-AGCAACGUCCAGUCCAAG-3’ (SEQ ID NO: 3) (sense strand) and/or 5’-CUUGGACUGGACGUUGCU-3’ (SEQ ID NO: 4) (antisense strand); iii.
  • 5’-AGCGGCUACAGCAGCCCCG -3’ (SEQ ID NO: 9) (sense strand) and/or 5’-CGGGGCUGCUGUAGCCGCU-3’ (SEQ ID NO: 10) (antisense strand); vi. 5’-AAUUAAUAAGAAGCUGGAUCU-3’ (SEQ ID NO: 11) (sense strand) and/or 5’-AGAUCCAGCUUCUUAUUAATT-3’ (SEQ ID NO: 12) (antisense strand); vii.
  • 5’-AAUAUCACCCACGUCCCUGG-3’ (SEQ ID NO: 25) (sense strand) and/or 5’-CCAGGGACGUGGGUGAUATT-3’ (SEQ ID NO: 26) (antisense strand); xiv. 5’-UGGCCAGGUGGAAGUAAAA-3’ (SEQ ID NO: 27) (sense strand) and/or 5’-UUUUACUUCCACCUGGCCA-3’ (SEQ ID NO: 28) (antisense strand); xv.
  • 5’-CCAAAGCCAAGACAGACCA-3’ (SEQ ID NO: 29) (sense strand) and/or 5’-UGGUCUGUCUUGGCUUUGG-3’ (SEQ ID NO: 30) (antisense strand); xvi. 5’-AAGCAGGGUUUGUGAUCAG-3’ (SEQ ID NO: 31) (sense strand) and/or 5’-CUGAUCACAAACCCUGCTT-3’ (SEQ ID NO: 32) (antisense strand); xvii.
  • 5’-CUCGGGACUUCAAAAUCAG-3’ (SEQ ID NO: 33) (sense strand) and/or 5’-CUGAUUUUGAAGUCCCGAG-3’ (SEQ ID NO: 34) (antisense strand); xviii. 5’-AGGGCAGCCUGUGGGAGAA-3’ (SEQ ID NO: 35) (sense strand) and/or 5’-UUCUCCCACAGGCUGCCCU-3’ (SEQ ID NO: 36) (antisense strand); or xix.
  • the oligonucleotide in the present disclosure is double stranded and comprises: i. 5’-AAGCUGGAUCUUAGCAACGUC-3’ (SEQ ID NO: 37) (sense strand) and/or 5’-GACGUUGCUAAGAUCCAGCTT-3’ (SEQ ID NO: 38) (antisense strand); ii.
  • 5’-AAGAUUGAAACCCACAAGC-3’ (SEQ ID NO: 43) (sense strand) and/or 5’-GCUUGUGGGUUUCAAUCTT-3’ (SEQ ID NO: 44) (antisense strand); v. 5’-GAAACCCACAAGCUGACCU-3’ (SEQ ID NO: 45) (sense strand) and/or 5’-AGGUCAGCUUGUGGGUUUC-3’ (SEQ ID NO: 46) (antisense strand); vi.
  • 5’-AACACGUCCCGGGAGGCGGCA-3’ (SEQ ID NO: 51) (sense strand) and/or 5’-UGCCGCCUCCCGGGACGUGTT-3’ (SEQ ID NO: 52) (antisense strand); ix. 5’-ACGUCCCGGGAGGCGGCAG-3’ (SEQ ID NO: 53) (sense strand) and/or 5’-CUGCCGCCUCCCGGGACGU-3’ (SEQ ID NO: 54) (antisense strand); or x.
  • the oligonucleotide in the present disclosure comprises: 5’- AACGUCCAGUCCAAGUGUGGC-3’ (SEQ ID NO: 1) (sense strand) and 5’- GCCACACUUGGACUGGACGTT-3’ (SEQ ID NO: 2) (antisense strand).
  • the oligonucleotide in the present disclosure comprises: 5’- UGAGAACCUGAAGCACCAG-3’ (SEQ ID NO: 7) (sense strand) and 5’- CUGGUGCUUCAGGUUCUCA-3’ (SEQ ID NO: 8) (antisense strand).
  • the oligonucleotide in the present disclosure comprises: 5’- AGCGGCUACAGCAGCCCCG-3’ (SEQ ID NO: 9) (sense strand) and 5’- CGGGGCUGCUGUAGCCGCU-3’ (SEQ ID NO: 10) (antisense strand).
  • the oligonucleotide in the present disclosure comprises: 5’- AAUUAAUAAGAAGCUGGAUCU-3’ (SEQ ID NO: 11) (sense strand) and 5’- AGAUCCAGCUUCUUAUUAATT-3’ (SEQ ID NO: 12) (antisense strand).
  • the oligonucleotide in the present disclose comprises: 5’- GAGCCCAAGAAGGUGGCAG-3’ (SEQ ID NO: 13) (sense strand) and 5’- CUGCCACCUUCUUGGGCUC-3’ (SEQ ID NO: 14) (antisense strand).
  • the oligonucleotide in the present disclosure comprises: 5’- ACCCGGGAGCCCAAGAAG-3’ (SEQ ID NO: 15) (sense strand) and 5’-CUUCUUGGGCUCCCGGGU- 3’ (SEQ ID NO: 16) (antisense strand).
  • the oligonucleotide in the present disclosure comprises: 5’- AGAAGCUGGAUCUUAGCAA-3’ (SEQ ID NO: 17) (sense strand) and 5’- UUGCUAAGAUCCAGCUUCU-3’ (SEQ ID NO: 18) (antisense strand).
  • the oligonucleotide in the present disclosure comprises: 5’- AAGGAUAAUAUCAAACACGUC-3’ (SEQ ID NO: 19) (sense strand) and 5’- GACGUGUUUGAUAUUAUCCTT-3’ (SEQ ID NO: 20) (antisense strand).
  • the oligonucleotide in the present disclosure comprises: 5’- AUAAUAUCAAACACGUCCC-3’ (SEQ ID NO: 21) (sense strand) and 5’- GGGACGUGUUUGAUAUUAU-3’ (SEQ ID NO: 22) (antisense strand).
  • the oligonucleotide in the present disclosure comprises: 5’- AGGAGGUGGCCAGGUGGAA-3’ (SEQ ID NO: 23) (sense strand) and 5’- UUCCACCUGGCCACCUCCU-3’ (SEQ ID NO: 24) (antisense strand).
  • the oligonucleotide in the present disclosure comprises: 5’- AAUAUCACCCACGUCCCUGG-3’ (SEQ ID NO: 25) (sense strand) and 5’- CCAGGGACGUGGGUGAUATT-3’ (SEQ ID NO: 26) (antisense strand).
  • the oligonucleotide in the present disclosure comprises: 5’- UGGCCAGGUGGAAGUAAAA-3’ (SEQ ID NO: 27) (sense strand) and 5’- UUUUACUUCCACCUGGCCA-3’ (SEQ ID NO: 28) (antisense strand). [0086] In some aspects, the oligonucleotide in the present disclosure comprises: 5’- CCAAAGCCAAGACAGACCA-3’ (SEQ ID NO: 29) (sense strand) and 5’- UGGUCUGUCUUGGCUUUGG-3’ (SEQ ID NO: 30) (antisense strand).
  • the oligonucleotide in the present disclosure comprises: 5’- AAGCAGGGUUUGUGAUCAG-3’ (SEQ ID NO: 31) (sense strand) and 5’- CUGAUCACAAACCCUGCTT-3’ (SEQ ID NO: 32) (antisense strand). [0088] In some aspects, the oligonucleotide in the present disclosure comprises: 5’- CUCGGGACUUCAAAAUCAG-3’ (SEQ ID NO: 33) (sense strand) and 5’- CUGAUUUUGAAGUCCCGAG-3’ (SEQ ID NO: 34) (antisense strand).
  • the oligonucleotide in the present disclosure comprises: 5’- AGGGCAGCCUGUGGGAGAA-3’ (SEQ ID NO: 35) (sense strand) and 5’- UUCUCCCACAGGCUGCCCU-3’ (SEQ ID NO: 36) (antisense strand).
  • the oligonucleotide in the present disclosure comprises: 5’- AAGCUGGAUCUUAGCAACGUC-3’ (SEQ ID NO: 37) (sense strand) and 5’- GACGUUGCUAAGAUCCAGCTT-3’ (SEQ ID NO: 38) (antisense strand).
  • the oligonucleotide in the present disclosure comprises: 5’- AGUGUGCAAAUAGUCUACAA-3’ (SEQ ID NO: 39) (sense strand) and 5’- UUGUAGACUAUUUGCACACU-3’ (SEQ ID NO: 40) (antisense strand).
  • the oligonucleotide in the present disclosure comprises: 5’- AAGGACAGAGUCCAGUCGAAG-3’ (SEQ ID NO: 41) (sense strand) and 5’- CUUCGACUGGACUCUGUCCTT-3’ (SEQ ID NO: 42) (antisense strand).
  • the oligonucleotide in the present disclosure comprises: 5’- AAGAUUGAAACCCACAAGC-3’ (SEQ ID NO: 43) (sense strand) and 5’- GCUUGUGGGUUUCAAUCTT-3’ (SEQ ID NO: 44) (antisense strand).
  • the oligonucleotide in the present disclosure comprises: 5’- GAAACCCACAAGCUGACCU-3’ (SEQ ID NO: 45) (sense strand) and 5’- AGGUCAGCUUGUGGGUUUC-3’ (SEQ ID NO: 46) (antisense strand).
  • the oligonucleotide in the present disclosure comprises: 5’- AAUAUCAAACACGUCCCGGGA-3’ (SEQ ID NO: 47) (sense strand) and 5’- UCCCGGGACGUGUUUGAUATT-3’ (SEQ ID NO: 48) (antisense strand).
  • the oligonucleotide in the present disclosure comprises: 5’- AUCAAACACGUCCCGGGAG-3’ (SEQ ID NO: 49) (sense strand) and 5’- CUCCCGGGACGUGUUUGAU-3’ (SEQ ID NO: 50) (antisense strand).
  • the oligonucleotide in the present disclosure comprises: 5’- AACACGUCCCGGGAGGCGGCA-3’ (SEQ ID NO: 51) (sense strand) and 5’- UGCCGCCUCCCGGGACGUGTT-3’ (SEQ ID NO: 52) (antisense strand).
  • the oligonucleotide in the present disclosure comprises: 5’- ACGUCCCGGGAGGCGGCAG-3’ (SEQ ID NO: 53) (sense strand) and 5’- CUGCCGCCUCCCGGGACGU-3’ (SEQ ID NO: 54) (antisense strand).
  • the oligonucleotide in the present disclosure comprises: 5’- UCCCGGGAGGCGGCAGUGU-3’ (SEQ ID NO: 55) (sense strand) and 5’- ACACUGCCGCCUCCCGGGA-3’ (SEQ ID NO: 56) (antisense strand).
  • the oligonucleotide in the present disclosure comprises: 5’- GGUGCAGAUAAUUAAUAAG-3’ (SEQ ID NO: 64) (sense strand) and 5’- CUUAUUAAUUAUCUGCACC-3’ (SEQ ID NO: 65) (antisense strand).
  • the oligonucleotide in the present disclosure comprises: 5’- GGUGCAGAUAAUUAAUAAG-3’ (SEQ ID NO: 64) (sense strand) and 5’- CUUAUUAAUUAUCUGCACC -3’ (SEQ ID NO: 65) (antisense strand); wherein the 1 st , 3 rd , 4 th , 7 th , 8 th , 9 th , 10 th , 11 th , 13 th , and 18 th nucleotides from 5’ to 3’of the sense strand is RNA and the 2 nd , 5 th , 6 th , 12 th , 14 th , 15 th , 16 th , 17 th , and 19 th nucleotide from 5’ to 3’of the sense strand comprise 2’-OMe at the sugar position.
  • the oligonucleotide in the present disclosure comprises: 5’- GGUGCAGAUAAUUAAUAAG-3’ (SEQ ID NO: 64) (sense strand) and 5’- CUUAUUAAUUAUCUGCACC -3’ (SEQ ID NO: 65) (antisense strand); wherein the 1 st , 2 nd , 3 rd , 4 th , 5 th , 6 th , 8 th , 9 th , 10 th , 11 th , 12 th , 13 th , 14 th , 15 th , 16 th , 18 th , and 19 th nucleotides from 5’ to 3’of the antisense strand is RNA and the 7 th and 17 th nucleotides from 5’ to 3’of the antisense strand comprise 2’-OMe at the sugar position.
  • the oligonucleotide in the present disclosure comprises: 5’- GGUGCAGAUAAUUAAUAAG-3’ (SEQ ID NO: 64) (sense strand) and 5’- CUUAUUAAUUAUCUGCACC -3’ (SEQ ID NO: 65) (antisense strand); wherein at least one of the backbone of the sense and/or antisense strands is linked to a phosphodiester bond, a phosphothioate bond, or a combination thereof.
  • the oligonucleotide in the present disclosure comprises: 5’- GGUGCAGAUAAUUAAUAAG-3’ (SEQ ID NO: 64) (sense strand) and 5’- CUUAUUAAUUAUCUGCACC -3’ (SEQ ID NO: 65) (antisense strand); wherein each of the backbone of the sense and antisense strands is linked by a phosphodiester bond.
  • the oligonucleotide in the present disclosure comprises: 5’- GGUGCAGAUAAUUAAUAAG-3’ (SEQ ID NO: 64) (sense strand) and 5’- CUUAUUAAUUAUCUGCACC -3’ (SEQ ID NO: 65) (antisense strand); wherein the 1 st , 2 nd , 3 rd , 4 th , 5 th , 6 th , 8 th , 12 th , 13 th , 14 th , 15 th , 16 th , 17 th , 18 th and 19 th nucleotides from 5’ to 3’of the sense strand comprise 2’-OMe at the sugar position and the 7 th , 9 th , 10 th and 11 th nucleotides from 5’ to 3’of the sense strand comprise 2’-F at the sugar position.
  • the oligonucleotide in the present disclosure comprises: 5’- GGUGCAGAUAAUUAAUAAG-3’ (SEQ ID NO: 64) (sense strand) and 5’- CUUAUUAAUUAUCUGCACC -3’ (SEQ ID NO: 65) (antisense strand); wherein the 1 st , 3 rd , 4 th , 5 th , 7 th , 8 th , 10 th , 11 th , 12 th , 13 th , 15 th , 17 th , 18 th , and 19 th nucleotides from 5’ to 3’of the antisense strand comprise 2’-OMe at the sugar position and the 2 nd , 6 th , 9 th , 14 th , and 16 th nucleotides from 5’ to 3’of the antisense strand comprise 2’-F at the sugar position.
  • the oligonucleotide in the present disclosure comprises: 5’- GGUGCAGAUAAUUAAUAAG-3’ (SEQ ID NO: 64) (sense strand) and 5’- CUUAUUAAUUAUCUGCACC -3’ (SEQ ID NO: 65) (antisense strand); wherein the nucleotides in the sense strand and/or the nucleotides in the antisense strand are linked by a phosphodiester bond, a phosphothioate bond, or any combination thereof.
  • the oligonucleotide in the present disclosure comprises: 5’- GGUGCAGAUAAUUAAUAAG-3’ (SEQ ID NO: 64) (sense strand) and 5’- CUUAUUAAUUAUCUGCACC -3’ (SEQ ID NO: 65) (antisense strand); wherein the 1 st , 2 nd and 3 rd nucleotides from 5’ to 3’of the sense strand, the 1 st , 2 nd and 3 rd nucleotides from 5’ to 3’of the antisense strand, and the 17 th , 18 th and 19 th nucleotides from 5’ to 3’of the antisense strand are linked by phosphorothioate bonds, and every other nucleotide of the sense and antisense strands are linked by phosphodiester bonds.
  • the oligonucleotide in the present disclosure comprises: 5’- AGCGGCUACAGCAGCCCCG-3’ (SEQ ID NO: 9) (sense strand) and 5’- CGGGGCUGCUGUAGCCGCU-3’ (SEQ ID NO: 10) (antisense strand); wherein the 1 st , 3 rd , 4 th , 7 th , 8 th , 9 th , 10 th , 11 th , 13 th , and 18 th nucleotides from 5’ to 3’ of the sense strand is RNA and the 2 nd , 5 th , 6 th , 12 th , 14 th , 15 th , 16 th , 17 th , and 19 th nucleotides from 5’ to 3’of the sense strand comprise 2’- OMe at the sugar position.
  • the oligonucleotide in the present disclosure comprises: 5’- AGCGGCUACAGCAGCCCCG-3’ (SEQ ID NO: 9) (sense strand) and 5’- CGGGGCUGCUGUAGCCGCU-3’ (SEQ ID NO: 10) (antisense strand); wherein the 1 st , 2 nd , 3 rd , 4 th , 5 th , 6 th , 8 th , 9 th , 10 th , 11 th , 12 th , 13 th , 14 th , 15 th ,16 th , 18 th , and 19 th nucleotides from 5’ to 3’of the antisense strand is RNA and the 7 th and 17 th nucleotides from 5’ to 3’ of the antisense strand comprise 2’-OMe at the sugar position.
  • the oligonucleotide in the present disclosure comprises: 5’- AGCGGCUACAGCAGCCCCG-3’ (SEQ ID NO: 9) (sense strand) and 5’- CGGGGCUGCUGUAGCCGCU-3’ (SEQ ID NO: 10) (antisense strand); wherein at least one of the backbone of the sense and/or antisense strands is linked to a phosphodiester bond, a phosphothioate bond, or a combination thereof.
  • the oligonucleotide in the present disclosure comprises: 5’- AGCGGCUACAGCAGCCCCG-3’ (SEQ ID NO: 9) (sense strand) and 5’- CGGGGCUGCUGUAGCCGCU-3’ (SEQ ID NO: 10) (antisense strand); wherein each of the backbone of the sense and antisense strands is linked by a phosphodiester bond.
  • the oligonucleotide in the present disclosure comprises: 5’- AGCGGCUACAGCAGCCCCG-3’ (SEQ ID NO: 9) (sense strand) and 5’- CGGGGCUGCUGUAGCCGCU-3’ (SEQ ID NO: 10) (antisense strand); wherein the 1 st , 2 nd , 3 rd , 4 th , 5 th , 6 th , 8 th , 12 th , 13 th , 14 th , 15 th , 16 th , 17 th , 18 th and 19 th nucleotides from 5’ to 3’of the sense strand comprise 2’-OMe at the sugar position and the 7 th , 9 th , 10 th and 11 th nucleotides from 5’ to 3’of the sense strand comprise 2’-F at the sugar position.
  • the oligonucleotide in the present disclosure comprises: 5’- AGCGGCUACAGCAGCCCCG-3’ (SEQ ID NO: 9) (sense strand) and 5’- CGGGGCUGCUGUAGCCGCU-3’ (SEQ ID NO: 10) (antisense strand); wherein the 1 st , 3 rd , 4 th , 5 th , 7 th , 8 th , 10 th , 11 th , 12 th , 13 th , 15 th , 17 th , 18 th , and 19 th nucleotides from 5’ to 3’of the antisense strand comprise 2’-OMe at the sugar position and the 2 nd , 6 th , 9 th , 14 th , and 16 th nucleotide from 5’ to 3’of the antisense strand comprise 2’-F at the sugar position.
  • the oligonucleotide in the present disclosure comprises: 5’- AGGAGGUGGCCAGGUGGAA-3’ (SEQ ID NO: 23) (sense strand) and 5’- UUCCACCUGGCCACCUCCU-3’ (SEQ ID NO: 24) (antisense strand); wherein the 1 st , 3 rd , 4 th , 7 th , 8 th , 9 th , 10 th , 11 th , 13 th , and 18 th nucleotides from 5’ to 3’of the sense strand is RNA and the 2 nd , 5 th , 6 th , 12 th , 14 th , 15 th , 16 th , 17 th , and 19 th nucleotides from 5’ to 3’of the sense strand comprise 2’- OMe at the sugar position.
  • the oligonucleotide in the present disclosure comprises: 5’- AGGAGGUGGCCAGGUGGAA-3’ (SEQ ID NO: 23) (sense strand) and 5’- UUCCACCUGGCCACCUCCU-3’ (SEQ ID NO: 24) (antisense strand); wherein the 1 st , 2 nd , 3 rd , 4 th , 5 th , 6 th , 8 th , 9 th , 10 th , 11 th , 12 th , 13 th , 14 th , 15 th , 16 th , 18 th , and 19 th nucleotides from 5’ to 3’of the antisense strand is RNA and the 7 th and 17 th nucleotides from 5’ to 3’of the antisense strand comprise 2’-OMe at the sugar position.
  • the oligonucleotide in the present disclosure comprises: 5’- AGGAGGUGGCCAGGUGGAA-3’ (SEQ ID NO: 23) (sense strand) and 5’- UUCCACCUGGCCACCUCCU-3’ (SEQ ID NO: 24) (antisense strand); wherein at least one of the backbone of the sense and/or antisense strands is linked to a phosphodiester bond, a phosphothioate bond, or a combination thereof.
  • the oligonucleotide in the present disclosure comprises: 5’- AGGAGGUGGCCAGGUGGAA-3’ (SEQ ID NO: 23) (sense strand) and 5’- UUCCACCUGGCCACCUCCU-3’ (SEQ ID NO: 24) (antisense strand); wherein each of the backbone of the sense and antisense strands is linked by a phosphodiester bond.
  • the oligonucleotide in the present disclosure comprises: 5’- AGGAGGUGGCCAGGUGGAA-3’ (SEQ ID NO: 23) (sense strand) and 5’- UUCCACCUGGCCACCUCCU-3’ (SEQ ID NO: 24) (antisense strand); wherein the 1 st , 2 nd , 3 rd , 4 th , 5 th , 6 th , 8 th , 12 th , 13 th , 14 th , 15 th , 16 th , 17 th , 18 th and 19 th nucleotides from 5’ to 3’of the sense strand comprise 2’-OMe at the sugar position and the 7 th , 9 th , 10 th and 11 th nucleotides from 5’ to 3’of the sense strand comprise 2’-F at the sugar position.
  • the oligonucleotide in the present disclosure comprises: 5’- AGGAGGUGGCCAGGUGGAA-3’ (SEQ ID NO: 23) (sense strand) and 5’- UUCCACCUGGCCACCUCCU-3’ (SEQ ID NO: 24) (antisense strand); wherein the 1 st , 3 rd , 4 th , 5 th , 7 th , 8 th , 10 th , 11 th , 12 th , 13 th , 15 th , 17 th , 18 th , and 19 th nucleotides from 5’ to 3’of the antisense strand comprise 2’-OMe at the sugar position and the 2 nd , 6 th , 9 th , 14 th , and 16 th nucleotides from 5’ to 3’ of the antisense strand comprise 2’-F at the sugar position.
  • the oligonucleotide in the present disclosure comprises: 5’- AGGAGGUGGCCAGGUGGAA-3’ (SEQ ID NO: 23) (sense strand) and 5’- UUCCACCUGGCCACCUCCU-3’ (SEQ ID NO: 24) (antisense strand); wherein at least one of the backbone of the sense and/or antisense strands is linked to a phosphodiester bond, a phosphothioate bond, or a combination thereof.
  • the oligonucleotide in the present disclosure comprises: 5’- AGGAGGUGGCCAGGUGGAA-3’ (SEQ ID NO: 23) (sense strand) and 5’- UUCCACCUGGCCACCUCCU-3’ (SEQ ID NO: 24) (antisense strand); wherein the 1 st , 2 nd and 3 rd nucleotides from 5’ to 3’of the sense strand, the 1 st , 2 nd and 3 rd nucleotides of the antisense strand, and the 17 th , 18 th and 19 th nucleotides from 5’ to 3’of the antisense strand are linked by phosphorothioate bonds, and every other nucleotide of the sense and antisense strands are linked by phosphodiester bonds.
  • the oligonucleotide in the present disclosure comprises: 5’- UGGCCAGGUGGAAGUAAAA-3’ (SEQ ID NO: 27) (sense strand) and 5’- UUUUACUUCCACCUGGCCA-3’ (SEQ ID NO: 28) (antisense strand); wherein the 1 st , 3 rd , 4 th , 7 th , 8 th , 9 th , 10 th , 11 th , 13 th , and 18 th nucleotides from 5’ to 3’of the sense strand is RNA and the 2 nd , 5 th , 6 th , 12 th , 14 th , 15 th , 16 th , 17 th , and 19 th nucleotides from 5’ to 3’ of the sense strand is 2’-OMe.
  • the oligonucleotide in the present disclosure comprises: 5’- UGGCCAGGUGGAAGUAAAA-3’ (SEQ ID NO: 27) (sense strand) and 5’- UUUUACUUCCACCUGGCCA-3’ (SEQ ID NO: 28) (antisense strand); wherein the 1 st , 2 nd , 3 rd , 4 th , 5 th , 6 th , 8 th , 9 th , 10 th , 11 th , 12 th , 13 th , 14 th , 15 th , 16 th , 18 th , and 19 th nucleotides from 5’ to 3’ of the antisense strand is RNA and the 7 th and 17 th nucleotides from 5’ to 3’ of the antisense strand comprise 2’-OMe at the sugar position.
  • the oligonucleotide in the present disclosure comprises: 5’- UGGCCAGGUGGAAGUAAAA-3’ (SEQ ID NO: 27) (sense strand) and 5’- UUUUACUUCCACCUGGCCA-3’ (SEQ ID NO: 28) (antisense strand); wherein at least one of the backbone of the sense and/or antisense strands is linked to a phosphodiester bond, a phosphothioate bond, or a combination thereof.
  • the oligonucleotide in the present disclosure comprises: 5’- UGGCCAGGUGGAAGUAAAA-3’ (SEQ ID NO: 27) (sense strand) and 5’- UUUUACUUCCACCUGGCCA-3’ (SEQ ID NO: 28) (antisense strand); wherein each of the backbone of the sense and antisense strands is linked by a phosphodiester bond.
  • the oligonucleotide in the present disclosure comprises: 5’- UGGCCAGGUGGAAGUAAAA-3’ (SEQ ID NO: 27) (sense strand) and 5’- UUUUACUUCCACCUGGCCA-3’ (SEQ ID NO: 28) (antisense strand); wherein the 1 st , 2 nd , 3 rd , 4 th , 5 th , 6 th , 8 th , 12 th , 13 th , 14 th , 15 th , 16 th , 17 th , 18 th , and 19 th nucleotides from 5’ to 3’of the sense strand comprises 2’ OMe at the sugar position and the 7 th , 9 th , 10 th , and 11 th nucleotides from 5’ to 3’of the sense strand comprises 2’ F at the sugar position.
  • the oligonucleotide in the present disclosure comprises: 5’- UGGCCAGGUGGAAGUAAAA-3’ (SEQ ID NO: 27) (sense strand) and 5’- UUUUACUUCCACCUGGCCA-3’ (SEQ ID NO: 28) (antisense strand); wherein the 1 st , 3 rd , 4 th , 5 th , 7 th , 8 th , 10 th , 11 th , 12 th , 13 th , 15 th , 17 th , 18 th , and 19 th nucleotides from 5’ to 3’of the antisense strand comprise 2’-OMe at the sugar position and the 2 nd , 6 th , 9 th , 14 th , and 16 th nucleotides from 5’ to 3’ of the antisense strand comprise 2’-F at the sugar position.
  • the oligonucleotide in the present disclosure comprises: 5’- UGGCCAGGUGGAAGUAAAA-3’ (SEQ ID NO: 27) (sense strand) and 5’- UUUUACUUCCACCUGGCCA-3’ (SEQ ID NO: 28) (antisense strand); wherein at least one of the backbone of the sense and/or antisense strands is linked to a phosphodiester bond, a phosphothioate bond, or a combination thereof.
  • the oligonucleotide in the present disclosure comprises: 5’- UGGCCAGGUGGAAGUAAAA-3’ (SEQ ID NO: 27) (sense strand) and 5’- UUUUACUUCCACCUGGCCA-3’ (SEQ ID NO: 28) (antisense strand); wherein the 1 st , 2 nd and 3 rd nucleotides from 5’ to 3’ of the sense strand, the 1 st , 2 nd and 3 rd nucleotides from 5’ to 3’of the antisense strand, and the 17 th , 18 th and 19 th nucleotides from 5’ to 3’of the antisense strand are linked by phosphorothioate bonds, and every other nucleotide of the sense and antisense strands are linked by phosphodiester bonds.
  • the oligonucleotide of the present disclosure is capable of inhibiting the expression of the human MAPT transcript in a cell which is expressing the human MAPT transcript. In some aspects, the oligonucleotide of the present disclosure is capable of inhibiting the expression of the monkey MAPT transcript in a cell which is expressing the monkey MAPT transcript. [0134] In some aspects, the sense and antisense strands in the oligonucleotide forms an RNA duplex are covalently linked by a single-stranded hairpin. In some aspects, the oligonucleotide comprises non-nucleotide materials.
  • the oligonucleotide further comprises an addition, deletion, substitution, or alteration of one or more nucleotides.
  • the present disclosure provides oligonucleotides wherein the sense and antisense RNA strands are stabilized against nuclease degradation.
  • the present disclosure provides an oligonucleotide that also comprises a 3′ overhang of 1 to 6 nucleotides or about 2 nucleotides.
  • the present disclosure provides an oligonucleotide that also comprises a 3′ overhang of 1 to 6 nucleotides or about 2 nucleotides with a sequence of: GCCACACUUGGACUGGACGTT (SEQ ID NO: 2), CUUAUUAAUUAUCUGCACCTT (SEQ ID NO: 6), AGAUCCAGCUUCUUAUUAATT (SEQ ID NO: 12), GACGUUGCUAAGAUCCAGCTT (SEQ ID NO: 38), GACGUGUUUGAUAUUAUCCTT (SEQ ID NO: 20), CCAGGGACGUGGGUGAUATT (SEQ ID NO: 26),CUUCGACUGGACUCUGUCCTT (SEQ ID NO: 42),GCUUGUGGGUUUCAAUCTT (SEQ ID NO: 44),CUGAUCACAAACCCUGCTT (SEQ ID NO: 32),UCCCGGGACGUGUUUGAUATT (SEQ ID NO: 48), orUGCCGCCUCCCGGGACGUGTT (SEQ ID NO: 2),
  • the sense RNA strand in the oligonucleotide comprises a first 3′ overhang
  • the antisense RNA strand in the oligonucleotide comprises a second 3′ overhang.
  • the first and second 3′ overhangs in the oligonucleotide separately comprise from 1 to about 6 nucleotides.
  • the first 3′ overhang in the oligonucleotide comprises a dinucleotide and the second 3′ overhang comprises a dinucleotide.
  • the present disclosure provides an oligonucleotide where the dinucleotide comprising the first and second 3′ overhangs is dithymidylic acid (TT) or diuridylic acid (uu). [0140] In some aspects, the present disclosure provides an oligonucleotide wherein the 3′ overhang is stabilized against nuclease degradation. [0141] In some aspects, the present disclosure provides an oligonucleotide that comprises at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten nucleoside analogues.
  • the present disclosure provides an oligonucleotide wherein the nucleoside analogue or analogues are one or more sugar modified nucleosides selected from the group consisting of Locked Nucleic Acid (LNA); 2'-O-alkyl-RNA; 2'-amino-DNA; 2'-fluoro- DNA; arabino nucleic acid (ANA); 2'-fluoro-ANA, hexitol nucleic acid (HNA), intercalating nucleic acid (INA), constrained ethyl nucleoside (cEt), 2'-O-methyl nucleic acid (2'-OMe), 2'-O- methoxyethyl nucleic acid (2'-MOE), and any combination thereof.
  • LNA Locked Nucleic Acid
  • ANA arabino nucleic acid
  • INA intercalating nucleic acid
  • cEt constrained ethyl nucleoside
  • 2'-OMe 2'-O-methyl nucleic
  • the present disclosure provides an oligonucleotide wherein the nucleoside analogue or analogues comprise a bicyclic sugar.
  • the bicyclic sugar comprises cEt, 2',4'-constrained 2′-O-methoxyethyl (cMOE), LNA, ⁇ -L-LNA, ⁇ -D-LNA, 2'-O,4'- C-ethylene-bridged nucleic acids (ENA), amino-LNA, oxy-LNA, or thio-LNA.
  • the present disclosure provides an oligonucleotide wherein the nucleoside analogue or analogues comprise an LNA.
  • the oligonucleotide comprises 5’-methylcytosine, pseudouridine, or 5-methoxyuridine. In some aspects, the oligonucleotide comprises one or more 5' methyl cytosine nucleobases. [0145] In some aspects, the present disclosure provides an oligonucleotide which reduces expression of MAPT mRNA in a cell by at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or about 100% compared to a cell not exposed to the oligonucleotide.
  • the present disclosure provides an oligonucleotide which reduces expression of MAPT protein in a cell by at least about 60%, at least about 70%, at least about 80%, at least about 90%, or at least about 95% compared to a cell not exposed to the oligonucleotide.
  • the present disclosure provides an oligonucleotide which comprises an internucleoside linkage selected from: a phosphodiester linkage, a phosphotriester linkage, a methylphosphonate linkage, a phosphoramidate linkage, a phosphorothioate linkage, and combinations thereof.
  • the internucleoside linkage comprises one or more stereo- defined, modified phosphate linkages.
  • the present disclosure provides a conjugate comprising oligonucleotides wherein the oligonucleotide is covalently attached to at least one non-nucleotide or non-polynucleotide moiety.
  • the present disclosure provides a conjugate comprising an oligonucleotide wherein the non-nucleotide or non-polynucleotide moiety comprises a protein, a fatty acid chain, a sugar residue, a glycoprotein, a polymer, or any combinations thereof.
  • the present disclosure provides a conjugate comprising an oligonucleotide wherein the non-nucleotide or non-polynucleotide moiety comprises a choleteryl moiety, a vinyl phosphonate moiety, or any combination thereof.
  • the present disclosure provides a conjugate comprising an oligonucleotide and a non-nucleotide moiety, wherein the oligonucleotide comprises, consists essentially of, or consists of: 5’-GGUGCAGAUAAUUAAUAAG-3’ (SEQ ID NO: 64) (sense strand) and/or 5’-CUUAUUAAUUAUCUGCACC-3’ (SEQ ID NO: 65) (antisense strand), wherein the 1 st , 3 rd , 4 th , 7 th , 8 th , 9 th , 10 th , 11 th , 13 th , and 18 th nucleotides from 5’ to 3’of the sense strand is RNA and the 2 nd , 5 th , 6 th , 12 th , 14 th , 15 th , 16 th , 17 th , and 19 th nucleotides from 5’ to
  • the present disclosure provides a conjugate comprising an oligonucleotide and a non-nucleotide moiety, wherein the oligonucleotide comprises, consists essentially of, or consists of: 5’-GGUGCAGAUAAUUAAUAAG-3’ (SEQ ID NO: 64) (sense strand) and/or 5’-CUUAUUAAUUAUCUGCACC-3’ (SEQ ID NO: 65) (antisense strand), wherein the 1 st , 3 rd , 4 th , 7 th , 8 th , 9 th , 10 th , 11 th , 13 th , and 18 th nucleotides from 5’ to 3’of the sense strand is RNA and the 2 nd , 5 th , 6 th , 12 th , 14 th , 15 th , 16 th , 17 th , and 19 th nucleotides from 5’ to
  • the present disclosure provides a conjugate comprising an oligonucleotide and a non-nucleotide moiety, wherein the oligonucleotide comprises, consists essentially of, or consists of: 5’-GGUGCAGAUAAUUAAUAAG-3’ (SEQ ID NO: 64) (sense strand) and/or 5’-CUUAUUAAUUAUCUGCACC-3’ (SEQ ID NO: 65) (antisense strand), wherein the 1 st , 2 nd , 3 rd , 4 th , 5 th , 6 th , 8 th , 12 th , 13 th , 14 th , 15 th , 16 th , 17 th , 18 th and 19 th nucleotides from 5’ to 3’of the sense strand comprise 2’-OMe at the sugar position and the 7 th , 9 th , 10 th and 11 th nucleotides from
  • the present disclosure provides a conjugate comprising an oligonucleotide and a non-nucleotide moiety, wherein the oligonucleotide comprises, consists essentially of, or consists of: 5’-GGUGCAGAUAAUUAAUAAG-3’ (SEQ ID NO: 64) (sense strand) and/or 5’-CUUAUUAAUUAUCUGCACC-3’ (SEQ ID NO: 65) (antisense strand), wherein the 1 st , 2 nd , 3 rd , 4 th , 5 th , 6 th , 8 th , 12 th , 13 th , 14 th , 15 th , 16 th , 17 th , 18 th and 19 th nucleotides from 5’ to 3’of the sense strand comprise 2’-OMe at the sugar position and the 7 th , 9 th , 10 th and 11 th nucleotides from
  • the present disclosure provides a conjugate comprising an oligonucleotide and a non-nucleotide moiety, wherein the oligonucleotide comprises, consists essentially of, or consists of: 5’-AGCGGCUACAGCAGCCCCG -3’ (SEQ ID NO: 9) (sense strand) and 5’-CGGGGCUGCUGUAGCCGCU-3’ (SEQ ID NO: 10) (antisense strand), wherein the 1 st , 3 rd , 4 th , 7 th , 8 th , 9 th , 10 th , 11 th , 13 th , and 18 th nucleotides from 5’ to 3’ of the sense strand is RNA and the 2 nd , 5 th , 6 th , 12 th , 14 th , 15 th , 16 th , 17 th , and 19 th nucleotides from 5’ to 3
  • the present disclosure provides a conjugate comprising an oligonucleotide and a non-nucleotide moiety, wherein the oligonucleotide comprises, consists essentially of, or consists of: 5’-AGCGGCUACAGCAGCCCCG -3’ (SEQ ID NO: 9) (sense strand) and 5’-CGGGGCUGCUGUAGCCGCU-3’ (SEQ ID NO: 10) (antisense strand), wherein the 1 st , 3 rd , 4 th , 7 th , 8 th , 9 th , 10 th , 11 th , 13 th , and 18 th nucleotides from 5’ to 3’ of the sense strand is RNA and the 2 nd , 5 th , 6 th , 12 th , 14 th , 15 th , 16 th , 17 th , and 19 th nucleotides from 5’ to 3
  • the present disclosure provides a conjugate comprising an oligonucleotide and a non-nucleotide moiety, wherein the oligonucleotide comprises, consists essentially of, or consists of: 5’-AGCGGCUACAGCAGCCCCG -3’ (SEQ ID NO: 9) (sense strand) and 5’-CGGGGCUGCUGUAGCCGCU-3’ (SEQ ID NO: 10) (antisense strand), wherein the 1 st , 2 nd , 3 rd , 4 th , 5 th , 6 th , 8 th , 12 th , 13 th , 14 th , 15 th , 16 th , 17 th , 18 th and 19 th nucleotides from 5’ to 3’of the sense strand comprise 2’-OMe at the sugar position and the 7 th , 9 th , 10 th and 11 th nucleotides from 5
  • the present disclosure provides a conjugate comprising an oligonucleotide and a non-nucleotide moiety, wherein the oligonucleotide comprises, consists essentially of, or consists of: 5’-AGCGGCUACAGCAGCCCCG -3’ (SEQ ID NO: 9) (sense strand) and 5’-CGGGGCUGCUGUAGCCGCU-3’ (SEQ ID NO: 10) (antisense strand), wherein the 1 st , 2 nd , 3 rd , 4 th , 5 th , 6 th , 8 th , 12 th , 13 th , 14 th , 15 th , 16 th , 17 th , 18 th and 19 th nucleotides from 5’ to 3’of the sense strand comprise 2’-OMe at the sugar position and the 7 th , 9 th , 10 th and 11 th nucleotides from 5
  • the present disclosure provides a conjugate comprising an oligonucleotide and a non-nucleotide moiety, wherein the nucleic acid sequence comprises, consists essentially of, or consists of: 5’-AGGAGGUGGCCAGGUGGAA-3’ (SEQ ID NO: 23) (sense strand) and 5’-UUCCACCUGGCCACCUCCU-3’ (SEQ ID NO: 24) (antisense strand), wherein the 1 st , 3 rd , 4 th , 7 th , 8 th , 9 th , 10 th , 11 th , 13 th , and 18 th nucleotides from 5’ to 3’of the sense strand is RNA and the 2 nd , 5 th , 6 th , 12 th , 14 th , 15 th , 16 th , 17 th , and 19 th nucleotides from 5’ to 3’of the sense strand is RNA and the
  • the present disclosure provides a conjugate comprising an oligonucleotide and a non-nucleotide moiety, wherein the oligonucleotide comprises, consists essentially of, or consists of: 5’-AGGAGGUGGCCAGGUGGAA-3’ (SEQ ID NO: 23) (sense strand) and 5’-UUCCACCUGGCCACCUCCU-3’ (SEQ ID NO: 24) (antisense strand), wherein the 1 st , 3 rd , 4 th , 7 th , 8 th , 9 th , 10 th , 11 th , 13 th , and 18 th nucleotides from 5’ to 3’of the sense strand is RNA and the 2 nd , 5 th , 6 th , 12 th , 14 th , 15 th , 16 th , 17 th , and 19 th nucleotides from 5’ to 3’
  • the present disclosure provides a conjugate comprising an oligonucleotide and a non-nucleotide moiety, wherein the oligonucleotide comprises, consists essentially of, or consists of: 5’-AGGAGGUGGCCAGGUGGAA-3’ (SEQ ID NO: 23) (sense strand) and 5’-UUCCACCUGGCCACCUCCU-3’ (SEQ ID NO: 24) (antisense strand), wherein the 1 st , 2 nd , 3 rd , 4 th , 5 th , 6 th , 8 th , 12 th , 13 th , 14 th , 15 th , 16 th , 17 th , 18 th and 19 th nucleotides from 5’ to 3’of the sense strand comprise 2’-OMe at the sugar position and the 7 th , 9 th , 10 th and 11 th nucleotides from 5’
  • the present disclosure provides a conjugate comprising an oligonucleotide and a non-nucleotide moiety, wherein the oligonucleotide comprises, consists essentially of, or consists of: 5’-AGGAGGUGGCCAGGUGGAA-3’ (SEQ ID NO: 23) (sense strand) and 5’-UUCCACCUGGCCACCUCCU-3’ (SEQ ID NO: 24) (antisense strand), wherein the 1 st , 2 nd , 3 rd , 4 th , 5 th , 6 th , 8 th , 12 th , 13 th , 14 th , 15 th , 16 th , 17 th , 18 th and 19 th nucleotides from 5’ to 3’of the sense strand comprise 2’-OMe at the sugar position and the 7 th , 9 th , 10 th and 11 th nucleotides from 5’
  • the present disclosure provides a conjugate comprising an oligonucleotide and a non-nucleotide moiety, wherein the oligonucleotide comprises, consists essentially of, or consists of: 5’-UGGCCAGGUGGAAGUAAAA-3’ (SEQ ID NO: 27) (sense strand) and 5’-UUUUACUUCCACCUGGCCA-3’ (SEQ ID NO: 28) (antisense strand); wherein the 1 st , 3 rd , 4 th , 7 th , 8 th , 9 th , 10 th , 11 th , 13 th , and 18 th nucleotides from 5’ to 3’of the sense strand is RNA and the 2 nd , 5 th , 6 th , 12 th , 14 th , 15 th , 16 th , 17 th , and 19 th nucleotides from 5’ to 3
  • the present disclosure provides a conjugate comprising an oligonucleotide and a non-nucleotide moiety, wherein the oligonucleotide comprises, consists essentially of, or consists of: 5’-UGGCCAGGUGGAAGUAAAA-3’ (SEQ ID NO: 27) (sense strand) and 5’-UUUUACUUCCACCUGGCCA-3’ (SEQ ID NO: 28) (antisense strand); wherein the 1 st , 3 rd , 4 th , 7 th , 8 th , 9 th , 10 th , 11 th , 13 th , and 18 th nucleotides from 5’ to 3’of the sense strand is RNA and the 2 nd , 5 th , 6 th , 12 th , 14 th , 15 th , 16 th , 17 th , and 19 th nucleotides from 5’ to 3
  • the present disclosure provides a conjugate comprising an oligonucleotide and a non-nucleotide moiety, wherein the oligonucleotide comprises, consists essentially of, or consists of: 5’-UGGCCAGGUGGAAGUAAAA-3’ (SEQ ID NO: 27) (sense strand) and 5’-UUUUACUUCCACCUGGCCA-3’ (SEQ ID NO: 28) (antisense strand); wherein the 1 st , 3 rd , 4 th , 7 th , 8 th , 9 th , 10 th , 11 th , 13 th , and 18 th nucleotides from 5’ to 3’of the sense strand is RNA and the 2 nd , 5 th , 6 th , 12 th , 14 th , 15 th , 16 th , 17 th , and 19 th nucleotides from 5’ to 3
  • the present disclosure provides a conjugate comprising an oligonucleotide and a non-nucleotide moiety, wherein the oligonucleotide comprises, consists essentially of, or consists of: 5’-UGGCCAGGUGGAAGUAAAA-3’ (SEQ ID NO: 27) (sense strand) and 5’-UUUUACUUCCACCUGGCCA-3’ (SEQ ID NO: 28) (antisense strand); wherein the 1 st , 2 nd , 3 rd , 4 th , 5 th , 6 th , 8 th , 12 th , 13 th , 14 th , 15 th , 16 th , 17 th , 18 th , and 19 th nucleotides from 5’ to 3’of the sense strand comprises 2’ OMe at the sugar position and the 7 th , 9 th , 10 th , and 11 th nucleo
  • the present disclosure provides a conjugate comprising an oligonucleotide and a non-nucleotide moiety, wherein the oligonucleotide comprises, consists essentially of, or consists of: 5’-UGGCCAGGUGGAAGUAAAA-3’ (SEQ ID NO: 27) (sense strand) and 5’-UUUUACUUCCACCUGGCCA-3’ (SEQ ID NO: 28) (antisense strand); wherein the 1 st , 2 nd , 3 rd , 4 th , 5 th , 6 th , 8 th , 12 th , 13 th , 14 th , 15 th , 16 th , 17 th , 18 th , and 19 th nucleotides from 5’ to 3’of the sense strand comprises 2’ OMe at the sugar position and the 7 th , 9 th , 10 th , and 11 th nucleo
  • the present disclosure provides a composition comprising an oligonucleotide or conjugate and a pharmaceutically acceptable carrier and/or a therapeutic agent.
  • the therapeutic agent is a Microtubule-associated protein tau antagonist.
  • the Microtubule-associated protein tau antagonist is an anti-Microtubule-associated protein tau antibody or fragment thereof.
  • the present disclosure provides an oligonucleotide, a conjugate, or a composition as a kit with instructions for use.
  • the present disclosure provides a method of inhibiting or reducing MAPT protein expression in a cell, the method comprising administering an oligonucleotide, a conjugate, or a composition to a cell expressing MAPT protein, wherein the MAPT protein expression in the cell is inhibited or reduced after the administration.
  • the present disclosure provides a method in which the oligonucleotide inhibits or reduces expression of MAPT mRNA in the cell after the administration.
  • the present disclosure provides a method wherein the expression of MAPT mRNA is reduced by at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or about 100% after the administration compared to a cell not exposed to the oligonucleotide.
  • the oligonucleotide reduces expression of MAPT protein in the cell after the administration by at least about 60%, at least about 70%, at least about 80%, or at least about 90% compared to a cell not exposed to the oligonucleotide.
  • the present disclosure provides methods of inhibiting or reducing MAPT protein expression in a cell where the cell is a neuron.
  • the present disclosure provides a method for treating a tauopathy in a subject in need thereof, comprising administering an effective amount of an oligonucleotide, a conjugate, or a composition to the subject.
  • the present disclosure provides methods of treating tauopathy in a subject wherein the tauopathy is selected from the group consisting of behavioral variant frontotemporal dementia, primary progressive aphasia, corticobasal syndrome, Richardson’s syndrome, Pick’s disease, Parkinsonism linked to chromosome 17, Primary age related tauopathy, Tangle-predominant dementia, Corticobasal degeneration, Progressive supranuclear palsy, Alzheimer’s disease, Parkinson’s disease, amyotrophic lateral sclerosis, globular glial tauopathy, Argyrophilic grain disease, Aging-related tau ⁇ astrogliopathy, and any combinations thereof.
  • the present disclosure provides methods of treating tauopathy in a subject that is human.
  • FIG.1A shows Western blot analyses of tau protein expression level after MAPT siRNA (100 nM) transfection for 48 h in differentiated SH-SY5Y.
  • the table below the Western blot shows the % inhibition of the total tau protein.
  • siMAPT 1, 2, 4, 5, 6, 8, 10, 11, 12, 14, 15, 21, 22, and 23 effective reduces effectively reduces the tau protein level.
  • FIG. 1B shows relative mRNA expression levels of tau after MAPT siRNA (100 nM) transfection in Tau-WT (3R2N and 4R2N) transfected SH-SY5Y cells and relative mRNA expression level of MAPT isoform (4R2N) after MAPT siRNA (100 nM) transfection in Tau-MUT (L266V, 4R2N isoform) transfected SH-SY5Y cells.
  • siMAPT 5, 6, 8, 9, 10, 12, 14, 15, 21, and 22 effectively reduces wild type tau mRNA.
  • siMAPT 1 and 17 effectively reduces Tau-MUT (L266V).
  • siMAPT 3, 11, and 23 effectively reduces both wild type tau and tau- MUT.
  • FIG. 2A shows Western blot analyses of tau protein expression levels following different concentration of MAPT siRNA (No.5) transfection for 48 h in SH-SY5Y. Beta ( ⁇ )-actin protein expression was used as a control. A graph of the % expression is shown, with its IC50.
  • the % tau protein expression is shown in the table for 1nM, 5nM, 10nM, 50nM, 100nM, 250nM, 500nM, and 1000nM of siRNA no. 5.
  • FIG. 2B shows Western blot analyses of tau protein expression levels following different concentration of MAPT siRNA (No.10) transfection for 48 h in SH-SY5Y. Beta ( ⁇ )-actin protein expression was used as a control. A graph of the % expression is shown, with its IC50. The % tau protein expression is shown in the table for 10nM, 100nM, and 1000nM of siRNA no. 10.
  • FIGs. 2C and 2D show relative mRNA expression levels of total tau following different concentration of MAPT siRNA (No.5) transfection (FIG.
  • FIGs.2E and 2F show relative mRNA expression levels of MAPT isoform (4R2N) after MAPT siRNA (No.5) transfection (FIG. 2E) and siRNA (No.10) transfection (FIG. 2F) for 48h in Tau-MUT (L226V, 4R2N isoform) transfected SH-SY5Y cells respectively.
  • FIGs.3A through 3F show relative mRNA expression levels of wild-type Tau, and 3R0N, 4R2N, and 4R1N Tau isoforms following different concentration of MAPT siRNA for 48 h in Tau-MUT ((L266V, 4R2N) and (P301S, 4R1N)) and Tau-WT (3R1N, 3R2N, 4R1N, 4R2N) and Tau-WT (3R0N) transfected SH-SY5Y cells.
  • FIG. 4A shows a schematic illustration of the experimental schedule. Mice were subjected to behavior test for 3days after ICV injection of A ⁇ Os (2 ⁇ g) (4-weeks). BIOK-Mapt siRNA (2 ⁇ g) was ICV injected 2-week post to A ⁇ Os.
  • FIG. 4C shows the results of a Y-maze test with vehicle control, A ⁇ Os and A ⁇ Os + Mapt siRNA (No.11) (left bar, middle bar, and right bar, respectively) for assessment of spatial working memory.
  • FIGs.4D through 4F show quantification of Western blot analyses of total tau (FIG. 4D), p-tau (pT181) (FIG.
  • FIG. 4E shows a summary of the quantified average Tau inhibition % for FIG.4D through 4F.
  • FIGs. 4H through 4J show the results of this test with bars representing vehicle control (Con), A ⁇ Os, and A ⁇ Os + Mapt siRNA (No.11) (left bar, middle bar, and right bar, respectively) conditions.
  • FIGs.5A through 5D show quantifications of Western blot analyses of total tau (t- tau) and hyperphosphorylated tau (p-tau) protein expression levels in each brain tissue such as hippocampus (FIG. 5A), prefrontal cortex (FIG. 5B), entorhinal cortex (FIG.5C), and striatum (FIG.5D) following ICV injection of BIOK-Mapt siRNA in P301S transgenic mice respectively.
  • FIG.6 shows siMAPT sense and antisense sequences, their sequence ID Nos, and their target exons.
  • FIG.7 shows the MAPT mRNA sequence.
  • FIG.8 shows the MAPT isoform mRNA sequences for 4R2N, 4R1N, 4R0N, 3R2N, 3R1N, and 3R0N.
  • FIGs. 9A through 9I show the sense strand and antisense strand for #3*_M1/M1/siMAPT-3*-On_chol (FIG. 9A), #5_M2/M2/siMAPT-5-On_chol (FIG. 9B), #14_M3/M3/siMAPT-14-On_chol (FIG. 9C), #17_M4/M4/siMAPT-17-On_chol (FIG.
  • FIGs. 9D #3*_M5/M5/siMAPT-3*-Am_chol (FIG. 9E), #5_M6/M6/siMAPT-5-Am_chol (FIG. 9F), #14_M7/M7/siMAPT-14-Am_chol (FIG. 9G), #17_M8/M8/siMAPT-17-Am_chol (FIG. 9H), and #17_M9/M9/siMAPT-17-On_chol_vinyl (FIG. 9I). Also shown is a cholesteryl and vinyl phosphonate moiety. [0185] FIGs.
  • FIGs. 10A through 10C show the relative expression of mTau MAPT RNA following administration with siRNA #1 in the hippocampus (FIG.10A), entorhinal cortex (FIG. 10B), and prefrontal cortex (FIG. 10C) (sub brain regions) of C57BL/6N Mice at 7, 21, and 28 days compared to a control group.
  • a table showing the inihibition percent at 7, 21, and 28 days is also shown for FIGs.10A through 10C. All data are mean ⁇ SD. Signifigance was set at ****P ⁇ 0.0001 compared to the WT (Vehicle) group. Signfigance was determined using a one-way ANOVA and post-hoc tukey’s test. [0186] FIGs.
  • FIGs. 10D through 10F show the relative expression of mTau MAPT RNA following administration with siRNA #17 in the hippocampus (FIG.10D), entorhinal cortex (FIG. 10E), and prefrontal cortex (FIG. 10F) (sub brain regions) of C57BL/6N Mice at 7, 21, and 28 days compared to a control group.
  • a table showing the inihibition percent at 7, 21, and 28 days is also shown for FIGs.10D through 10F. All data are mean ⁇ SD. Signifigance was set at ****P ⁇ 0.0001 compared to the WT (Vehicle) group. Signfigance was determined using a one-way ANOVA and post-hoc tukey’s test. [0187] FIGs.
  • FIGs. 11A through 11C show the relative expression of huTau MAPT RNA following administration with siRNA #1 in the hippocampus (FIG.11A), entorhinal cortex (FIG. 11B), and prefrontal cortex (FIG. 11C) (sub brain regions) of PS19 Mice at 7, 21, and 28 days compared to a control group.
  • a table showing the inihibition percent at 7, 21, and 28 days is also shown for FIGs. 11A through 11C. All data are mean ⁇ SD. Significance was set at ****P ⁇ 0.0001 compared to the WT (Vehicle) group. Significance was determined using a one-way ANOVA and post-hoc tukey’s test. [0188] FIGs.
  • FIGs.11D through 11F show the relative expression of huTau MAPT RNA following administration with siRNA #17 in the hippocampus (FIG.11D), entorhinal cortex (FIG. 11E), and prefrontal cortex (FIG. 11F) (sub brain regions) of PS19 Mice at 7, 21, and 28 days compared to a control group.
  • a table showing the inihibition percent at 7, 21, and 28 days is also shown for FIGs.11D through 11F. All data are mean ⁇ SD. Significance was set at ****P ⁇ 0.0001 compared to the WT (Vehicle) group. Significance was determined using a one-way ANOVA and post-hoc tukey’s test.
  • FIG.12A shows the schematic illustration of the experimental schedule for BDDS- MAPT siRNA efficacy test in PS19 mice. ICV injections took place on weekss 22, 25, 28, 31, 34, and 37. Administration of the nest building test occurred on weeks 26, 32, and 38. Administration of open field test occurred on weeks 27 and 33. Administration of the elevated plus maze test occurred on weeks 27, and 36. Administration of the Y-maze test occurred on week 39. Mice were sacrificed, tissue was harvested, and the brain was fixed at 40 weeks.
  • FIGs.12B through 12E show western blot analysis of human tau (FIG.12B), p-tau (pT181) (FIG.12C), p-tau (Ser 404) (FIG.
  • FIG. 12E p-tau expression levels in protein tissue lysates from tissue of PS19 ICV injected mice following MAPT siRNA #1 or #17 administration compared to WT and PS19 vehicle administration. All data are mean ⁇ SD. Significance was set at ****P ⁇ 0.0001 compared to the WT (Vehicle) group. Significance was set at ####P ⁇ 0.0001 compared to the PS19 (Vehicle) group. Significance was set at ⁇ P ⁇ 0.05, ⁇ P ⁇ 0.01 and ⁇ P ⁇ 0.0001 for indicated comparisons. Significance was determined using a one-way ANOVA and post-hoc tukey’s test.
  • FIG.13A shows immunohistochemistry (IHC) images of AT8 (Ser202/Thr205) in the hippocampus of wild-type (WT), PS19, PS19 + MAPT siRNA #1, and PS19 + MAPT siRNA #17 administered mice at 10 months of age.
  • FIG.13B shows a summary of the relative intensity of AT8 (Ser202/Thr205) in the hippocampus of wild-type (WT), PS19, PS19 + MAPT siRNA #1, and PS19 + MAPT siRNA #17 administered mice at 10 months of age.
  • FIG.13A shows immunohistochemistry (IHC) images of AT8 (Ser202/Thr205) in the hippocampus of wild-type (WT), PS19, PS19 + MAPT siRNA #1, and PS19 + MAPT siRNA #17 administered mice at 10 months of age.
  • FIG. 13C shows immunohistochemistry (IHC) images of AT8 (Ser202/Thr205) in the entorhinal cortex of wild- type (WT), PS19, PS19 + MAPT siRNA #1, and PS19 + MAPT siRNA #17 administered mice at 10 months of age.
  • FIG.13D shows a summary of the relative intensity of AT8 (Ser202/Thr205) in the entorhinal cortex of wild-type (WT), PS19, PS19 + MAPT siRNA #1, and PS19 + MAPT siRNA #17 administered mice at 10 months of age.
  • AT8 is contrasted against black background.
  • Scale bar 250 ⁇ m. All data are mean ⁇ SD.
  • FIG.14A shows the nest building score (measured using Deacon’s nesting score) for vehicle, PS19, PS19 + MAPT siRNA #1, and PS19 + MAPT siRNA #17 administered mice.
  • FIG.14B shows the untorn nestlet weight for vehicle, PS19, PS19 + MAPT siRNA #1, and PS19 + MAPT siRNA #17 administered mice.
  • FIG.14C shows the time in open arms for vehicle, PS19, PS19 + MAPT siRNA #1, and PS19 + MAPT siRNA #17 administered mice for the elevated plus maze test.
  • FIG.14D shows the time in closed arms for vehicle, PS19, PS19 + MAPT siRNA #1, and PS19 + MAPT siRNA #17 administered mice for the elevated plus maze test. All data are mean ⁇ SD. Significance was set at **P ⁇ 0.01 and ****P ⁇ 0.0001 compared to the WT (Vehicle) group. Significance was set at #P ⁇ 0.05 and ####P ⁇ 0.0001 compared to the PS19 (Vehicle) group.
  • FIG.15A and 15B show the results of an open field test for vehicle, PS19, PS19 + MAPT siRNA #1, and PS19 + MAPT siRNA #17 administered mice. Ambulatory distance is indicated on the left chart, and resting time is indicated of the right chart of FIG.15A. Rearing behavior (vertical activity) is shown on FIG.15B.
  • FIG.15C shows the results of a spatial working memory test for vehicle, PS19, PS19 + MAPT siRNA #1, and PS19 + MAPT siRNA #17 administered mice as determined by the Y-maze test. All data are mean ⁇ SD.
  • each modified siRNA M1, M2, M3, M4, M5, M6, M7, M8 and M9 was transfected twice with a 5- fold dilution between the two (10 nM and 50 nM).
  • each naked siRNA (siRNA #3*, siRNA #5, siRNA#14 and siRNA #17) was transfected at a concentration of 50 nM. All data are mean ⁇ SEM.
  • the siRNA reduces expression of MAPT protein in a cell following administration of the siRNA by at least about 60%
  • the MAPT levels are reduced by a range of 50% to 70%.
  • G,” “C,” “A,” “T,” and “U” each generally stand for a naturally-occurring nucleotide that contains guanine, cytosine, adenine, thymine, and uracil as a base, respectively.
  • nucleotide can refer to an alternative nucleotide, as further detailed below, or a surrogate replacement moiety.
  • nucleotide comprising inosine as its base can base pair with nucleotides containing adenine, cytosine, or uracil.
  • nucleotides containing uracil, guanine, or adenine can be replaced in the nucleotide sequences of dsRNA featured herein by a nucleotide containing, for example, inosine.
  • the nucleobase moiety is modified by changing the purine or pyrimidine into a modified purine or pyrimidine, such as substituted purine or substituted pyrimidine, such as an “alternative nucleobase” selected from isocytosine, pseudoisocytosine, 5- methyl cytosine, 5-thiozolo-cytosine, 5-propynyl-cytosine, 5-propynyl-uridine, 5-bromouridine 5- thiazolo-uridine, 2-thio-uridine, pseudouridine, 1-methylpseudouridine, 5-methoxyuridine, 2′- thio-thymine, inosine, diaminopurine, 6-aminopurine, 2-aminopurine, 2,6-diaminopurine, and 2- chloro-6-aminopurine.
  • a modified purine or pyrimidine such as substituted purine or substituted pyrimidine, such as an “alternative nucleobase” selected from
  • nucleobase moieties can be indicated by the letter code for each corresponding nucleobase, e.g. A, T, G, C, or U, wherein each letter can include alternative nucleobases of equivalent function.
  • a “sugar” or “sugar moiety,” includes naturally occurring sugars having a furanose ring.
  • a sugar also includes an “alternative sugar,” defined as a structure that is capable of replacing the furanose ring of a nucleoside.
  • alternative sugars are non-furanose (or 4′- substituted furanose) rings or ring systems or open systems.
  • Such structures include simple changes relative to the natural furanose ring, such as a six-membered ring, or can be more complicated as is the case with the non-ring system used in peptide nucleic acid.
  • Alternative sugars can include sugar surrogates wherein the furanose ring has been replaced with another ring system such as, for example, a morpholino or hexitol ring system.
  • Sugar moieties useful in the preparation of oligonucleotides having motifs include, without limitation, ⁇ -D-ribose, ⁇ -D-2′-deoxyribose, substituted sugars (such as 2′, 5′ and bis substituted sugars), 4′-S-sugars (such as 4′-S-ribose, 4′-S- 2′-deoxyribose and 4′-S-2′-substituted ribose), bicyclic alternative sugars (such as the 2′-O—CH2- 4′ or 2′-O—(CH 2 ) 2 -4′ bridged ribose derived bicyclic sugars) and sugar surrogates (such as when the ribose ring has been replaced with a morpholino or a hexitol ring system).
  • substituted sugars such as 2′, 5′ and bis substituted sugars
  • 4′-S-sugars such as 4′-S-ribose,
  • nucleotide refers to a glycoside comprising a sugar moiety, a base moiety and a covalently linked group (linkage group), such as a phosphate or phosphorothioate internucleotide linkage group, and covers both naturally occurring nucleotides, such as DNA or RNA, and non-naturally occurring nucleotides comprising modified sugar and/or base, which are also referred to as "nucleotide analogs" herein.
  • linkage group such as a phosphate or phosphorothioate internucleotide linkage group
  • nucleotide analogs refers to nucleotides having modified sugar moieties.
  • nucleotides having modified sugar moieties e.g., LNA
  • nucleotide analogs refers to nucleotides having modified nucleobase moieties.
  • nucleotides having modified nucleobase moieties include, but are not limited to, 5- methylcytosine, isocytosine, pseudoisocytosine, 5-bromouracil, 5-propynyluracil, 6-aminopurine, 2-aminopurine, inosine, diaminopurine, and 2-chloro-6-aminopurine.
  • nucleoside as used herein is used to refer to a glycoside comprising a sugar moiety and a base moiety, which can be covalently linked by the internucleotide linkages between the nucleosides of the siRNA.
  • nucleoside is often used to refer to a nucleic acid monomer or unit.
  • nucleoside can refer to the base alone, i.e., a nucleobase sequence comprising cytosine (DNA and RNA), guanine (DNA and RNA), adenine (DNA and RNA), thymine (DNA) and uracil (RNA), in which the presence of the sugar backbone and internucleotide linkages are implicit.
  • nucleotide can refer to a "nucleoside.”
  • nucleoside can be used, even when specifying the presence or nature of the linkages between the nucleosides.
  • alternative nucleoside refers to a nucleoside having an alternative sugar or an alternative nucleobase, such as those described herein.
  • an “alternative nucleotide,” as used herein, refers to a nucleotide having an alternative nucleoside or an alternative sugar, and an internucleoside linkage, which can include alternative nucleoside linkages.
  • oligonucleotide and polynucleotide are defined as it is generally understood by the skilled person as a molecule comprising two or more covalently linked nucleosides. Such covalently bound nucleosides can be referred to as nucleic acid molecules or oligomers. Oligonucleotides are commonly made in the laboratory by solid-phase chemical synthesis followed by purification.
  • Oligonucleotide refers to a short polynucleotide (e.g., of 100 or fewer linked nucleosides).
  • strand refers to an oligonucleotide comprising a chain of linked nucleosides.
  • a "strand comprising a nucleobase sequence” refers to an oligonucleotide comprising a chain of linked nucleosides that is described by the sequence referred to using the standard nucleobase nomenclature.
  • antisense refers to a nucleic acid comprising an oligonucleotide or polynucleotide that is sufficiently complementary to all or a portion of a gene, primary transcript, or processed mRNA, so as to interfere with expression of the endogenous gene (e.g., SCNA).
  • endogenous gene e.g., SCNA
  • “Complementary” polynucleotides are those that are capable of base pairing according to the standard Watson-Crick complementarity rules.
  • purines will base pair with pyrimidines to form a combination of guanine paired with cytosine (G:C) and adenine paired with either thymine (A:T) in the case of DNA, or adenine paired with uracil (A:U) in the case of RNA.
  • G:C guanine paired with cytosine
  • A:T thymine
  • A:U uracil
  • two polynucleotides can hybridize to each other even if they are not completely complementary to each other, provided that each has at least one region that is substantially complementary to the other.
  • antisense strand and "guide strand” refer to the strand of a dsRNA that includes a region that is substantially complementary to a target sequence, e.g., an MAPT mRNA.
  • RNAi RNA interference
  • the dsRNA reduces the expression of MAPT in a cell, e.g., a cell within a subject, such as a mammalian subject.
  • a cell e.g., a cell within a subject, such as a mammalian subject.
  • the majority of linked nucleosides of each strand of a dsRNA are ribonucleosides, but as described in detail herein, each or both strands can include one or more non-ribonucleosides, e.g., deoxyribonucleosides and/or alternative nucleosides.
  • siRNA and “short interfering RNA” (also known as “small interfering RNA”) refer to an RNA agent, such as a double-stranded agent, of about 10-50 nucleotides in length, the strands optionally having overhanging ends comprising, for example 1, 2, or 3 overhanging linked nucleosides, which is capable of directing or mediating RNA interference.
  • Naturally-occurring siRNAs are generated from longer dsRNA molecules (e.g., >25 linked nucleosides in length) by a cell's RNAi machinery (e.g., Dicer or a homolog thereof).
  • RNA agent having a stem-loop structure, comprising at least two regions of complementary sequence, the degree of complementarity and orientation of the regions being sufficient such that base pairing occurs between the regions, at least two of the regions being joined by a loop region which results from a lack of base pairing between nucleobases within the loop region.
  • nucleic acids or “nucleotides” is intended to encompass plural nucleic acids.
  • nucleic acids refers to a target sequence, e.g., pre-mRNAs, mRNAs, or DNAs in vivo or in vitro.
  • target sequence e.g., pre-mRNAs, mRNAs, or DNAs in vivo or in vitro.
  • the term refers to the nucleic acids or nucleotides in a target sequence
  • the nucleic acids or nucleotides can be naturally occurring sequences within a cell.
  • nucleic acids or “nucleotides” refer to a sequence in the siRNAs of the disclosure.
  • the nucleic acids or nucleotides are not naturally occurring, i.e., chemically synthesized, enzymatically produced, recombinantly produced, or any combination thereof.
  • the nucleic acids or nucleotides in the siRNAs are produced synthetically or recombinantly, but are not a naturally occurring sequence or a fragment thereof.
  • the nucleic acids or nucleotides in the siRNAs are not naturally occurring because they contain at least one nucleotide analog that is not naturally occurring in nature.
  • nucleic acid refers to a single nucleic acid segment, e.g., a DNA, an RNA, or an analog thereof, present in a polynucleotide.
  • Nucleic acid or “nucleoside” includes naturally occurring nucleic acids or non-naturally occurring nucleic acids.
  • nucleotide or “unit” and “monomer” are used interchangeably. It will be recognized that when referring to a sequence of nucleotides or monomers, what is referred to is the sequence of bases, such as A, T, G, C or U, and analogs thereof.
  • nucleotide length means the total number of the nucleotides (monomers) in a given sequence.
  • sequence of CUUAUUAAUUAUCUGCACC SEQ ID NO: 65
  • nucleotide length is therefore used herein interchangeably with “nucleotide number.”
  • nucleoside overhang refers to at least one unpaired nucleobase that protrudes from the duplex structure of a dsRNA. For example, when a 3'-end of one strand of a dsRNA extends beyond the 5'-end of the other strand, or vice versa, there is a nucleoside overhang.
  • a dsRNA can comprise an overhang of at least one nucleoside; alternatively, the overhang can comprise at least two nucleosides, at least three nucleosides, at least four nucleosides, at least five nucleosides or more.
  • a nucleoside overhang can comprise or consist of an alternative nucleoside, including a deoxynucleotide/nucleoside.
  • a nucleoside overhang can comprise or consist of one or more phosphorothioates bonds.
  • the overhang(s) can be on the sense strand, the antisense strand, or any combination thereof.
  • the nucleoside(s) of an overhang can be present on the 5'-end, 3'-end or both ends of either an antisense or sense strand of a dsRNA.
  • the overhang includes a self-complementary portion such that the overhang is capable of forming a hairpin structure that is stable under physiological conditions.
  • dsRNA a dsRNA that is blunt at both ends, i.e., no nucleoside overhang at either end of the molecule. Most often, such a molecule will be double stranded over its entire length.
  • cleavage region refers to a region that is located immediately adjacent to the cleavage site.
  • the cleavage site is the site on the target at which cleavage occurs.
  • the cleavage region comprises three bases on either end of, and immediately adjacent to, the cleavage site.
  • the cleavage region comprises two bases on either end of, and immediately adjacent to, the cleavage site.
  • the cleavage site specifically occurs at the site bound by nucleosides 10 and 11 of the antisense strand, and the cleavage region comprises nucleosides 11, 12, and 13.
  • a "coding region” or “coding sequence” is a portion of polynucleotide which consists of codons translatable into amino acids. Although a “stop codon” (TAG, TGA, or TAA) is typically not translated into an amino acid, it can be considered to be part of a coding region, but any flanking sequences, for example promoters, ribosome binding sites, transcriptional terminators, introns, untranslated regions (“UTRs”), and the like, are not part of a coding region.
  • non-coding region means a nucleotide sequence that is not a coding region. Examples of non-coding regions include, but are not limited to, promoters, ribosome binding sites, transcriptional terminators, introns, untranslated regions ("UTRs"), non- coding exons and the like.
  • exons can be wholly or part of the 5' untranslated region (5' UTR) or the 3' untranslated region (3' UTR) of each transcript.
  • the untranslated regions are important for efficient translation of the transcript and for controlling the rate of translation and half-life of the transcript.
  • region when used in the context of a nucleotide sequence refers to a section of that sequence.
  • region within a nucleotide sequence or “region within the complement of a nucleotide sequence” refers to a sequence shorter than the nucleotide sequence, but longer than at least 10 nucleotides located within the particular nucleotide sequence or the complement of the nucleotides sequence, respectively.
  • sequence or “subsequence” can also refer to a region of a nucleotide sequence.
  • downstream when referring to a nucleotide sequence, means that a nucleic acid or a nucleotide sequence is located 3' to a reference nucleotide sequence. In certain aspects, downstream nucleotide sequences relate to sequences that follow the starting point of transcription. For example, the translation initiation codon of a gene is located downstream of the start site of transcription.
  • upstream refers to a nucleotide sequence that is located 5' to a reference nucleotide sequence.
  • regulatory region refers to nucleotide sequences located upstream (5' non-coding sequences), within, or downstream (3' non-coding sequences) of a coding region, and which influence the transcription, RNA processing, stability, or translation of the associated coding region. Regulatory regions can include promoters, translation leader sequences, introns, polyadenylation recognition sequences, RNA processing sites, effector binding sites, UTRs, and stem-loop structures.
  • transcript can refer to a primary transcript that is synthesized by transcription of DNA and becomes a messenger RNA (mRNA) after processing, i.e., a precursor messenger RNA (pre-mRNA), and the processed mRNA itself.
  • mRNA messenger RNA
  • pre-mRNA precursor messenger RNA
  • transcript can be interchangeably used with "pre-mRNA” and "mRNA.” After DNA strands are transcribed to primary transcripts, the newly synthesized primary transcripts are modified in several ways to be converted to their mature, functional forms such as mRNA, tRNA, rRNA, lncRNA, miRNA and others. Thus, the term “transcript” can include exons, introns, 5' UTRs, and 3' UTRs. [0230]
  • expression refers to a process by which a polynucleotide produces a gene product, for example, a RNA or a polypeptide.
  • RNA messenger RNA
  • expression produces a "gene product.”
  • a gene product can be either a nucleic acid, e.g., a messenger RNA produced by transcription of a gene, or a polypeptide which is translated from a transcript.
  • Gene products described herein further include nucleic acids with post transcriptional modifications, e.g., polyadenylation or splicing, or polypeptides with post translational modifications, e.g., methylation, glycosylation, the addition of lipids, association with other protein subunits, or proteolytic cleavage.
  • nucleic acids refer to two or more sequences that are the same or have a specified percentage of nucleotides or amino acid residues that are the same, when compared and aligned (introducing gaps, if necessary) for maximum correspondence, not considering any conservative amino acid substitutions as part of the sequence identity.
  • the percent identity can be measured using sequence comparison software or algorithms or by visual inspection.
  • Various algorithms and software are known in the art that can be used to obtain alignments of amino acid or nucleotide sequences.
  • sequence alignment algorithm is the algorithm described in Karlin et al., 1990, Proc. Natl. Acad.
  • BLAST-2 Altschul et al., 1996, Methods in Enzymology, 266:460-480
  • ALIGN ALIGN-2
  • Megalign Megalign
  • the percent identity between two nucleotide sequences is determined using the GAP program in the GCG software package (e.g., using a NWSgapdna.CMP matrix and a gap weight of 40, 50, 60, 70, or 90 and a length weight of 1, 2, 3, 4, 5, or 6).
  • the GAP program in the GCG software package which incorporates the algorithm of Needleman and Wunsch (J.
  • Mol. Biol. (48):444-453 (1970)) can be used to determine the percent identity between two amino acid sequences (e.g., using either a BLOSUM 62 matrix or a PAM250 matrix, and a gap weight of 16, 14, 12, 10, 8, 6, or 4 and a length weight of 1, 2, 3, 4, 5).
  • the percent identity between nucleotide or amino acid sequences is determined using the algorithm of Myers and Miller (CABIOS, 4:11-17 (1989)).
  • the percent identity can be determined using the ALIGN program (version 2.0) and using a PAM120 with residue table, a gap length penalty of 12 and a gap penalty of 4.
  • One skilled in the art can determine appropriate parameters for maximal alignment by particular alignment software.
  • the default parameters of the alignment software are used.
  • the percentage identity "X" of a first nucleotide sequence to a second nucleotide sequence is calculated as 100 x (Y/Z), where Y is the number of amino acid residues scored as identical matches in the alignment of the first and second sequences (as aligned by visual inspection or a particular sequence alignment program) and Z is the total number of residues in the second sequence. If the length of a first sequence is longer than the second sequence, the percent identity of the first sequence to the second sequence will be higher than the percent identity of the second sequence to the first sequence.
  • Different regions within a single polynucleotide target sequence that align with a polynucleotide reference sequence can each have their own percent sequence identity. It is noted that the percent sequence identity value is rounded to the nearest tenth. For example, 80.11, 80.12, 80.13, and 80.14 are rounded down to 80.1, while 80.15, 80.16, 80.17, 80.18, and 80.19 are rounded up to 80.2. It also is noted that the length value will always be an integer.
  • the terms “homologous” and “homology” are interchangeable with the terms “identity” and “identical.”
  • the term “naturally occurring variant thereof” refers to variants of the MAPT polypeptide sequence or MAPT nucleic acid sequence (e.g., transcript) which exist naturally within the defined taxonomic group, such as mammalian, such as mouse, monkey, and human.
  • the term also can encompass any allelic variant of the MAPT -encoding genomic DNA which is found at Chromosomal position 17q21.31 by chromosomal translocation or duplication, and the RNA, such as mRNA derived therefrom.
  • Non- occurring variants can also include variants derived from alternative splicing of the MAPT mRNA.
  • the term when referenced to a specific polypeptide sequence, e.g., the term also includes naturally occurring forms of the protein, which can therefore be processed, e.g., by co- or post-translational modifications, such as signal peptide cleavage, proteolytic cleavage, glycosylation, etc.
  • the degree of "complementarity” is expressed as the percentage identity (or percentage homology) between the sequence of the siRNA (or region thereof) and the sequence of the target region (or the reverse complement of the target region) that best aligns therewith. The percentage is calculated by counting the number of aligned bases that are identical between the two sequences, dividing by the total number of contiguous monomers in the siRNA, and multiplying by 100.
  • complement indicates a sequence that is complementary to a reference sequence. It is well known that complementarity is the base principle of DNA replication and transcription as it is a property shared between two DNA or RNA sequences, such that when they are aligned antiparallel to each other, the nucleotide bases at each position in the sequences will be complementary, much like looking in the mirror and seeing the reverse of things.
  • the complement of a sequence of 5’"ATGC”3’ can be written as 3’"TACG”5’ or 5’"GCAT”3’.
  • the terms “reverse complement”, “reverse complementary” and “reverse complementarity” as used herein are interchangeable with the terms “complement”, “complementary” and “complementarity.” [0239]
  • the terms “corresponding to” and “corresponds to,” when referencing two separate nucleic acid or nucleotide sequences can be used to clarify regions of the sequences that correspond or are similar to each other based on homology and/or functionality, although the nucleotides of the specific sequences can be numbered differently.
  • different isoforms of a gene transcript can have similar or conserved portions of nucleotide sequences whose numbering can differ in the respective isoforms based on alternative splicing and/or other modifications.
  • different numbering systems can be employed when characterizing a nucleic acid or nucleotide sequence (e.g., a gene transcript and whether to begin numbering the sequence from the translation start codon or to include the 5'UTR).
  • nucleic acid or nucleotide sequence of different variants of a gene or gene transcript can vary.
  • nucleotide sequence of a MAPT transcript corresponding to nucleotides X to Y of SEQ ID NO: 57 refers to an MAPT transcript sequence (e.g., MAPT pre-mRNA or mRNA) that has an identical sequence or a similar sequence to nucleotides X to Y of SEQ ID NO: 57.
  • reference sequence refers to an MAPT transcript sequence (e.g., MAPT pre-mRNA or mRNA) that has an identical sequence or a similar sequence to nucleotides X to Y of SEQ ID NO: 57.
  • nucleotide analog and “corresponding nucleotide” are intended to indicate that the nucleobase in the nucleotide analog and the naturally occurring nucleotide have the same pairing, or hybridizing, ability.
  • the “corresponding nucleotide analog” contains a pentose unit (different from 2-deoxyribose) linked to an adenine.
  • “Potency” is normally expressed as an IC 50 or EC 50 value, in ⁇ M, nM or pM unless otherwise stated. Potency can also be expressed in terms of percent inhibition.
  • IC 50 is the median inhibitory concentration of a therapeutic molecule.
  • EC50 is the median effective concentration of a therapeutic molecule relative to a vehicle or control (e.g., saline).
  • IC 50 is the concentration of a therapeutic molecule that reduces a biological response, e.g., transcription of mRNA or protein expression, by 50% of the biological response that is achieved by the therapeutic molecule.
  • EC 50 is the concentration of a therapeutic molecule that produces 50% of the biological response, e.g., transcription of mRNA or protein expression.
  • IC 50 or EC 50 can be calculated by any number of means known in the art.
  • subject or “individual” or “animal” or “patient” or “mammal,” is meant any subject, particularly a mammalian subject, for whom diagnosis, prognosis, or therapy is desired.
  • Mammalian subjects include humans, domestic animals, farm animals, sports animals, and zoo animals including, e.g., humans, non-human primates, dogs, cats, guinea pigs, rabbits, rats, mice, horses, cattle, bears, and so on.
  • pharmaceutical composition refers to a preparation which is in such form as to permit the biological activity of the active ingredient to be effective, and which contains no additional components which are unacceptably toxic to a subject to which the composition would be administered.
  • composition can be sterile.
  • An "effective amount" of an siRNA as disclosed herein is an amount sufficient to carry out a specifically stated purpose. An “effective amount” can be determined empirically and in a routine manner, in relation to the stated purpose.
  • Terms such as “treating” or “treatment” or “to treat” or “alleviating” or “to alleviate” refer to both (1) therapeutic measures that cure, slow down, lessen symptoms of, and/or halt progression of a diagnosed pathologic condition or disorder and (2) prophylactic or preventative measures that prevent and/or slow the development of a targeted pathologic condition or disorder.
  • those in need of treatment include those already with the disorder; those prone to have the disorder; and those in whom the disorder is to be prevented.
  • a subject is successfully "treated” for a disease or condition disclosed elsewhere herein according to the methods provided herein if the patient shows, e.g., total, partial, or transient alleviation or elimination of symptoms associated with the disease or disorder.
  • the term “MAPT protein” refers to, as way of example not limitation, wild-type MAPT protein, MAPT protein variants, MAPT isoforms (3R-, 4R-, 0N3R, 3R0N, 1N3R, 3R1N, 2N3R, 3R2N, 0N4R, 4R0N, 1N4R, 4R1N, 2N4R, 4R2N), or any combination thereof.
  • MAPT isoforms can be defined by the amount of N regions and R regions they contain, so e.g., 0N3R and 3R0N are equivalent to each other, this is also true for 1N3R and 3R1N, 2N3R and 3R2N, 0N4R and 4R0N, 1N4R and 4R1N, and 2N4R and 4R2N. II.
  • oligonucleotides for use in modulating the function of nucleic acid molecules encoding mammalian Microtubule-associated protein tau, such as the MAPT nucleic acid, e.g., MAPT transcript, including MAPT pre-mRNA, and MAPT mRNA, or naturally occurring variants of such nucleic acid molecules encoding mammalian Microtubule-associated protein tau.
  • the oligonucleotides (e.g., siRNAs) that are capable of modulating the MAPT nucleic acid can bind to and modulate the expression of human MAPT, mouse MAPT, monkey MAPT, or any combination thereof.
  • the oligonucleotides are capable of modulating one or more MAPT isoform 3R-, MAPT isoform 4R-, MAPT isoform 0N3R, MAPT isoform 1N3R, MAPT isoform 2N3R, MAPT isoform 0N4R, MAPT isoform 1N4R, MAPT isoform 2N4R, or any combination thereof.
  • the oligonucleotides (e.g., siRNAs) that are capable of modulating the MAPT nucleic acid binds to human MAPT, mouse MAPT, and monkey MAPT.
  • the siRNA comprises a contiguous nucleotide sequence of from about 10 to about 30, such as 10–20, 16–20, or 15–25 nucleotides in length.
  • the siRNA of the disclosure comprises RNA (units). In some aspects, the siRNA comprises one or more DNA units. In one aspect, the siRNA according to the disclosure is a linear molecule or is synthesized as a linear molecule. In various aspects, the siRNA of the disclosure can consist entirely of the contiguous nucleotide region. Thus, in some aspects the siRNA is not substantially self-complementary. [0250] In one aspect, the siRNA of the disclosure can be in the form of any pharmaceutically acceptable salts.
  • the term "pharmaceutically acceptable salts" as used herein refers to derivatives of the siRNA of the disclosure wherein the siRNA is modified (e.g., addition of a cation) by making salts thereof. Such salts retain the desired biological activity of the siRNAs without imparting undesired toxicological effects.
  • the siRNA of the disclosure is in the form of a sodium salt.
  • the siRNA is in the form of a potassium salt. II.A.
  • the siRNA of the disclosure is capable of down-regulating (e.g., reducing or removing) expression of the MAPT mRNA or protein.
  • the siRNA of the disclosure can affect indirect inhibition of MAPT protein through the reduction in MAPT mRNA levels, typically in a mammalian cell, such as a human cell, such as a neuronal cell.
  • the present disclosure is directed to siRNAs that target one or more regions of the MAPT pre- mRNA.
  • Synonyms of MAPT are known and include TAU, TAU_HUMAN, MSTD, PPND, DDPAC, MAPTL, MTBT1, MTBT2, tau-40, FTDP-17, PPP1R103, FLJ31424, MGC138549, Neurofibrillary tangle protein, Tau-derived paired helical filament hexapeptide, protein phosphatase 1, regulatory subunit 103, G protein beta1/gamma2 subunit-interacting factor 1, Paired helical filament-tau, PHF-tau, and Tau-PHF6.
  • the mRNA sequence for Homo sapiens microtubule associated protein tau (MAPT), transcript variant 1, can be found under publicly available Accession Number NM_016835.5, and the reference sequence for Homo sapiens microtubule associated protein tau (MAPT) can be found under publicly available Accession Number NG_007398.2.
  • the sequence for MAPT protein can be found under publicly available Accession Numbers: P10636 (primary accession) and, P18518, Q14799, Q15549, Q15550, Q15551, Q1RMF6, Q53YB1, Q5CZI7, Q5XWF0, Q6QT54, Q9UDJ3, Q9UMH0, and Q9UQ96 (secondary accessions), each of which is incorporated by reference herein in its entirety.
  • Natural variants of the MAPT gene product are known.
  • natural variants of MAPT protein can contain one or more amino acid substitutions.
  • the siRNAs of the present disclosure can be designed to reduce or inhibit expression of the natural variants of the MAPT protein.
  • siRNAs of the disclosure can be used to reduce or inhibit the expression of a SNP or alternatively spliced MAPT transcript containing one or more mutations and consequently reduce the formation of a mutated MAPT protein.
  • MAPT protein mutants include, but are not limited to a MAPT protein comprising one or more mutations selected from: A2S, P4T, R5C, R5L, R5H, E9K, M11L, G16V, T17M, L20F, G21V, R23S, G27E, T30K, T30I, G55R, D34Y, G37V, A41T, P59L, P78A, G86S, A91T, A91V, T95M, A103G, A74G, A45G, G78S, G49S, G107S, G107D, G78D, G49D, G107V, G49V, G78V, I108V, I79V, I50V, S113T, S55T, S84T, E115D, E57D, E86D, E117K, E59K, E88K, A60G, A89G, A118G, A143S, R155H, E183K, P126S, P201
  • MAPT protein mutants include, but are not limited to a MAPT protein comprising one or more mutations selected from: L266V and P301S, and any combination thereof.
  • the siRNA of the disclosure can be designed to reduce or inhibit expression of any mutants of MAPT proteins.
  • Alternative exon splicing of the MAPT gene transcript can produce six common isoforms of the MAPT protein, which include 0N4R, 1N4R, 2N4R, 0N3R, 1N3R, and 2N3R (also represented by 4R- and 3R-). Neurodegenerative conditions are associated with over- and under- expression of these isoforms, with specific neurodegenerative conditions being associated with specific isoforms.
  • target nucleic acid sequence of the siRNAs is MAPT mRNA.
  • the "target nucleic acid” comprises an exon region of an MAPT protein-encoding nucleic acids or naturally occurring variants thereof, and RNA nucleic acids derived therefrom.
  • the "target nucleic acid” can be a cDNA or a synthetic oligonucleotide derived from the above DNA or RNA nucleic acid targets.
  • the mRNA encoding MAPT protein is shown as SEQ ID NO: 57. See FIG 7.
  • the MAPT mRNA sequences of the six most common MAPT isoforms are shown as SEQ ID NOs: 58-63.
  • the amino acid sequence for MAPT protein can be found under publicly available Accession Number P10636. Specifically 0N3R can be found under P10636-2, 0N4R can be found under P10636-6, 1N3R can be found under P10636-4, 1N4R can be found under P10636-7 (, 2N3R can be found under P10636-5, and 2N4R can be found under P10636-8.
  • the ASO of the disclosure is capable of hybridizing to the target nucleic acid (e.g., MAPT transcript) under physiological condition, i.e., in vivo condition. In some aspects, the ASO of the disclosure is capable of hybridizing to the target nucleic acid (e.g., MAPT transcript) in vitro.
  • the ASO of the disclosure is capable of hybridizing to the target nucleic acid (e.g., MAPT transcript) in vitro under stringent conditions.
  • Stringency conditions for hybridization in vitro are dependent on, inter alia, productive cell uptake, RNA accessibility, temperature, free energy of association, salt concentration, and time (see, e.g., Stanley T Crooks, Antisense Drug Technology: Principles, Strategies and Applications, 2 nd Edition, CRC Press (2007)).
  • conditions of high to moderate stringency are used for in vitro hybridization to enable hybridization between substantially similar nucleic acids, but not between dissimilar nucleic acids.
  • stringent hybridization conditions include hybridization in 5X saline-sodium citrate (SSC) buffer (0.75 M sodium chloride/0.075 M sodium citrate) for 1 hour at 40° C, followed by washing the sample 10 times in 1X SSC at 40° C and 5 times in 1X SSC buffer at room temperature.
  • SSC 5X saline-sodium citrate
  • In vivo hybridization conditions consist of intracellular conditions (e.g., physiological pH and intracellular ionic conditions) that govern the hybridization of antisense oligonucleotides with target sequences.
  • In vivo conditions can be mimicked in vitro by relatively low stringency conditions.
  • hybridization can be carried out in vitro in 2X SSC (0.3 M sodium chloride/0.03 M sodium citrate), 0.1% SDS at 37°C.
  • oligonucleotide Sequences [0257]
  • the oligonucleotides, e.g., siRNA, of the disclosure comprise a specific contiguous nucleotide sequence which corresponds to the complement of a region of MAPT transcript, e.g., a nucleotide sequence corresponding to SEQ ID NO: 57.
  • the disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are capable of hybridizing to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: i. 5’-AACGUCCAGUCCAAGUGUGGC-3’ (SEQ ID NO: 1) (sense strand) or 5’-GCCACACUUGGACUGGACGTT-3’ (SEQ ID NO: 2) (antisense strand); ii.
  • MTT Microtubule-associated protein tau
  • 5’-AGCAACGUCCAGUCCAAG-3’ (SEQ ID NO: 3) (sense strand) or 5’-CUUGGACUGGACGUUGCU-3’ (SEQ ID NO: 4) (antisense strand); iii. 5’-AAGGUGCAGAUAAUUAAUAAG-3’ (SEQ ID NO: 5) (sense strand) or 5’-CUUAUUAAUUAUCUGCACCTT -3’ (SEQ ID NO: 6) (antisense strand); iv.
  • 5’-UGAGAACCUGAAGCACCAG -3’ (SEQ ID NO: 7) (sense strand) or 5’-CUGGUGCUUCAGGUUCUCA-3’ (SEQ ID NO: 8) (antisense strand); v. 5’-AGCGGCUACAGCAGCCCCG -3’ (SEQ ID NO: 9) (sense strand) or 5’-CGGGGCUGCUGUAGCCGCU-3’ (SEQ ID NO: 10) (antisense strand); vi. 5’-AAUUAAUAAGAAGCUGGAUCU-3’ (SEQ ID NO: 11) (sense strand) or 5’-AGAUCCAGCUUCUUAUUAATT-3’ (SEQ ID NO: 12) (antisense strand); vii.
  • 5’-GAGCCCAAGAAGGUGGCAG-3’ (SEQ ID NO: 13) (sense strand) or 5’-CUGCCACCUUCUUGGGCUC-3’ (SEQ ID NO: 14) (antisense strand); viii. 5’-ACCCGGGAGCCCAAGAAG-3’ (SEQ ID NO: 15) (sense strand) or 5’-CUUCUUGGGCUCCCGGGU-3’ (SEQ ID NO: 16) (antisense strand); ix. 5’-AGAAGCUGGAUCUUAGCAA-3’ (SEQ ID NO: 17) (sense strand) or 5’-UUGCUAAGAUCCAGCUUCU-3’ (SEQ ID NO: 18) (antisense strand); x.
  • 5’-AAGCAGGGUUUGUGAUCAG-3’ (SEQ ID NO: 31) (sense strand) or 5’-CUGAUCACAAACCCUGCTT-3’ (SEQ ID NO: 32) (antisense strand); or xvii. 5’-CUCGGGACUUCAAAAUCAG-3’ (SEQ ID NO: 33) (sense strand) or 5’-CUGAUUUUGAAGUCCCGAG-3’ (SEQ ID NO: 34) (antisense strand).
  • the disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are capable of hybridizing to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: i. 5’-AGGGCAGCCUGUGGGAGAA-3’ (SEQ ID NO: 35) (sense strand) or 5’-UUCUCCCACAGGCUGCCCU-3’ (SEQ ID NO: 36) (antisense strand); or ii.
  • MTT Microtubule-associated protein tau
  • oligonucleotides e.g., siRNA
  • the oligonucleotides comprise a specific contiguous nucleotide sequence which corresponds to the complement of a region of MAPT transcript, e.g., a nucleotide sequence corresponding to SEQ ID NO: 57.
  • the disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are capable of hybridizing to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: i. 5’-AAGCUGGAUCUUAGCAACGUC-3’ (SEQ ID NO: 37) (sense strand) or 5’-GACGUUGCUAAGAUCCAGCTT-3’ (SEQ ID NO: 38) (antisense strand); ii.
  • MTT Microtubule-associated protein tau
  • 5’-AUCAAACACGUCCCGGGAG-3’ (SEQ ID NO: 49) (sense strand) or 5’-CUCCCGGGACGUGUUUGAU-3’ (SEQ ID NO: 50) (antisense strand); viii. 5’-AACACGUCCCGGGAGGCGGCA-3’ (SEQ ID NO: 51) (sense strand) or 5’-UGCCGCCUCCCGGGACGUGTT-3’ (SEQ ID NO: 52) (antisense strand); ix. 5’-ACGUCCCGGGAGGCGGCAG-3’ (SEQ ID NO: 53) (sense strand) or 5’-CUGCCGCCUCCCGGGACGU-3’ (SEQ ID NO: 54) (antisense strand); x.
  • the contiguous nucleotide sequence is complementary to the nucleic acid sequence.
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- AACGUCCAGUCCAAGUGUGGC-3’ (SEQ ID NO: 1) (sense strand) or 5’-GCCACACUUGGACUGGACGTT-3’ (SEQ ID NO: 2) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- AGCAACGUCCAGUCCAAG-3’ (SEQ ID NO: 3) (sense strand) or 5’- CUUGGACUGGACGUUGCU-3’ (SEQ ID NO: 4) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- AAGGUGCAGAUAAUUAAUAAG-3’ (SEQ ID NO: 5) (sense strand) or 5’-CUUAUUAAUUAUCUGCACCTT -3’ (SEQ ID NO: 6) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- UGAGAACCUGAAGCACCAG -3’ (SEQ ID NO: 7) (sense strand)or 5’- CUGGUGCUUCAGGUUCUCA-3’ (SEQ ID NO: 8) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- AGCGGCUACAGCAGCCCCG -3’ (SEQ ID NO: 9) (sense strand) or 5’-CGGGGCUGCUGUAGCCGCU-3’ (SEQ ID NO: 10) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- AAUUAAUAAGAAGCUGGAUCU-3’ (SEQ ID NO: 11) (sense strand) or 5’-AGAUCCAGCUUCUUAUUAATT-3’ (SEQ ID NO: 12) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- GAGCCCAAGAAGGUGGCAG-3’ (SEQ ID NO: 13) (sense strand) or 5’-CUGCCACCUUCUUGGGCUC-3’ (SEQ ID NO: 14) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- ACCCGGGAGCCCAAGAAG-3’ (SEQ ID NO: 15) (sense strand) or 5’-CUUCUUGGGCUCCCGGGU-3’ (SEQ ID NO: 16) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- AGAAGCUGGAUCUUAGCAA-3’ (SEQ ID NO: 17) (sense strand) or 5’-UUGCUAAGAUCCAGCUUCU-3’ (SEQ ID NO: 18) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- AAGGAUAAUAUCAAACACGUC-3’ (SEQ ID NO: 19) (sense strand) or 5’-GACGUGUUUGAUAUUAUCCTT-3’ (SEQ ID NO: 20) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- AUAAUAUCAAACACGUCCC-3’ (SEQ ID NO: 21) (sense strand) or 5’-GGGACGUGUUUGAUAUUAU-3’ (SEQ ID NO: 22) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- AGGAGGUGGCCAGGUGGAA-3’ (SEQ ID NO: 23) (sense strand) or 5’-UUCCACCUGGCCACCUCCU-3’ (SEQ ID NO: 24) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-AAUAUCACCCACGUCCCUGG-3’ (SEQ ID NO: 25) (sense strand) or 5’-CCAGGGACGUGGGUGAUATT-3’ (SEQ ID NO: 26) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- UGGCCAGGUGGAAGUAAAA-3’ (SEQ ID NO: 27) (sense strand) or 5’-UUUUACUUCCACCUGGCCA-3’ (SEQ ID NO: 28) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- CCAAAGCCAAGACAGACCA-3’ (SEQ ID NO: 29) (sense strand) or 5’-UGGUCUGUCUUGGCUUUGG-3’ (SEQ ID NO: 30) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- AAGCAGGGUUUGUGAUCAG-3’ (SEQ ID NO: 31) (sense strand) or 5’-CUGAUCACAAACCCUGCTT-3’ (SEQ ID NO: 32) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- CUCGGGACUUCAAAAUCAG-3’ (SEQ ID NO: 33) (sense strand) or 5’-CUGAUUUUGAAGUCCCGAG-3’ (SEQ ID NO: 34) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- AGGGCAGCCUGUGGGAGAA-3’ (SEQ ID NO: 35) (sense strand) or 5’-UUCUCCCACAGGCUGCCCU-3’ (SEQ ID NO: 36) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- AAGCUGGAUCUUAGCAACGUC-3’ (SEQ ID NO: 37) (sense strand) or 5’-GACGUUGCUAAGAUCCAGCTT-3’ (SEQ ID NO: 38) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-AGUGUGCAAAUAGUCUACAA-3’ (SEQ ID NO: 39) (sense strand) or 5’-UUGUAGACUAUUUGCACACU-3’ (SEQ ID NO: 40) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- AAGGACAGAGUCCAGUCGAAG-3’ (SEQ ID NO: 41) (sense strand) or 5’-CUUCGACUGGACUCUGUCCTT-3’ (SEQ ID NO: 42) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- AAGAUUGAAACCCACAAGC-3’ (SEQ ID NO: 43) (sense strand) or 5’-GCUUGUGGGUUUCAAUCTT-3’ (SEQ ID NO: 44) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- GAAACCCACAAGCUGACCU-3’ (SEQ ID NO: 45) (sense strand) or 5’-AGGUCAGCUUGUGGGUUUC-3’ (SEQ ID NO: 46) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- AAUAUCAAACACGUCCCGGGA-3’ (SEQ ID NO: 47) (sense strand) or 5’-UCCCGGGACGUGUUUGAUATT-3’ (SEQ ID NO: 48) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- AUCAAACACGUCCCGGGAG-3’ (SEQ ID NO: 49) (sense strand) or 5’-CUCCCGGGACGUGUUUGAU-3’ (SEQ ID NO: 50) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- AACACGUCCCGGGAGGCGGCA-3’ (SEQ ID NO: 51) (sense strand) or 5’-UGCCGCCUCCCGGGACGUGTT-3’ (SEQ ID NO: 52) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- ACGUCCCGGGAGGCGGCAG-3’ (SEQ ID NO: 53) (sense strand) or 5’-CUGCCGCCUCCCGGGACGU-3’ (SEQ ID NO: 54) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’- UCCCGGGAGGCGGCAGUGU-3’ (SEQ ID NO: 55) (sense strand) or 5’-ACACUGCCGCCUCCCGGGA-3’ (SEQ ID NO: 56) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-GGUGCAGAUAAUUAAUAAG-3’ (SEQ ID NO: 64) (sense strand) or 5’-CUUAUUAAUUAUCUGCACC-3’ (SEQ ID NO: 65) (antisense strand).
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-GGUGCAGAUAAUUAAUAAG-3’ (SEQ ID NO: 64) (sense strand) or 5’-CUUAUUAAUUAUCUGCACC -3’ (SEQ ID NO: 65) (antisense strand); wherein the 1 st , 3 rd , 4 th , 7 th , 8 th , 9 th , 10 th , 11 th , 13 th , and 18 th nucleotides from 5’ to 3’of the sense strand is RNA and the 2 nd , 5 th , 6 th , 12 th , 14 th , 15 th , 16
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-GGUGCAGAUAAUUAAUAAG-3’ (SEQ ID NO: 64) (sense strand) or 5’-CUUAUUAAUUAUCUGCACC -3’ (SEQ ID NO: 65) (antisense strand); wherein at least one of the backbone of the sense and/or antisense strands is linked to a phosphodiester bond, a phosphothioate bond, or a combination thereof.
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-GGUGCAGAUAAUUAAUAAG-3’ (SEQ ID NO: 64) (sense strand) or 5’-CUUAUUAAUUAUCUGCACC -3’ (SEQ ID NO: 65) (antisense strand); wherein each of the backbone of the sense and antisense strands is linked by a phosphodiester bond.
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-GGUGCAGAUAAUUAAUAAG-3’ (SEQ ID NO: 64) (sense strand) or 5’-CUUAUUAAUUAUCUGCACC -3’ (SEQ ID NO: 65) (antisense strand); wherein the 1 st , 2 nd , 3 rd , 4 th , 5 th , 6 th , 8 th , 12 th , 13 th , 14 th , 15 th , 16 th , 17 th , 18 th and 19 th nucleotides from 5’ to 3’of the sense strand comprise 2’-OMe at the sugar position and the 7
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-GGUGCAGAUAAUUAAUAAG-3’ (SEQ ID NO: 64) (sense strand) or 5’-CUUAUUAAUUAUCUGCACC -3’ (SEQ ID NO: 65) (antisense strand); wherein the 1 st , 3 rd , 4 th , 5 th , 7 th , 8 th , 10 th , 11 th , 12 th , 13 th , 15 th , 17 th , 18 th , and 19 th nucleotides from 5’ to 3’of the antisense strand comprise 2’-OMe at the sugar position and the 2 nd
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-GGUGCAGAUAAUUAAUAAG-3’ (SEQ ID NO: 64) (sense strand) or 5’-CUUAUUAAUUAUCUGCACC -3’ (SEQ ID NO: 65) (antisense strand); wherein the nucleotides in the sense strand and/or the nucleotides in the antisense strand are linked by a phosphodiester bond, a phosphothioate bond, or any combination thereof.
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-AGCGGCUACAGCAGCCCCG-3’ (SEQ ID NO: 9) (sense strand) or 5’-CGGGGCUGCUGUAGCCGCU-3’ (SEQ ID NO: 10) (antisense strand); wherein the 1 st , 2 nd , 3 rd , 4 th , 5 th , 6 th , 8 th , 9 th , 10 th , 11 th , 12 th , 13 th , 14 th , 15 th , 16 th , 18 th , and 19 th nucleotides from 5’ to 3’of the antisense strand is
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-AGCGGCUACAGCAGCCCCG-3’ (SEQ ID NO: 9) (sense strand) or 5’-CGGGGCUGCUGUAGCCGCU-3’ (SEQ ID NO: 10) (antisense strand); wherein at least one of the backbone of the sense and/or antisense strands is linked to a phosphodiester bond, a phosphothioate bond, or a combination thereof.
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-AGCGGCUACAGCAGCCCCG-3’ (SEQ ID NO: 9) (sense strand) or 5’-CGGGGCUGCUGUAGCCGCU-3’ (SEQ ID NO: 10) (antisense strand); and wherein each of the backbone of the sense and antisense strands is linked by a phosphodiester bond.
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-AGCGGCUACAGCAGCCCCG-3’ (SEQ ID NO: 9) (sense strand) or 5’-CGGGGCUGCUGUAGCCGCU-3’ (SEQ ID NO: 10) (antisense strand); wherein the 1 st , 3 rd , 4 th , 5 th , 7 th , 8 th , 10 th , 11 th , 12 th , 13 th , 15 th , 17 th , 18 th , and 19 th nucleotides from 5’ to 3’of the antisense strand comprise 2’-OMe at the sugar position and the 2 nd
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-AGCGGCUACAGCAGCCCCG-3’ (SEQ ID NO: 9) (sense strand) or 5’-CGGGGCUGCUGUAGCCGCU-3’ (SEQ ID NO: 10) (antisense strand); wherein at least one of the backbone of the sense and/or antisense strands is linked to a phosphodiester bond, a phosphothioate bond, or a combination thereof.
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-AGCGGCUACAGCAGCCCCG-3’ (SEQ ID NO: 9) (sense strand) or 5’-CGGGGCUGCUGUAGCCGCU-3’ (SEQ ID NO: 10) (antisense strand); wherein the 1 st , 2 nd and 3 rd nucleotides from 5’ to 3’of the sense strand, the 1 st , 2 nd and 3 rd nucleotides from 5’ to 3’of the antisense nucleotides, and the 17 th , 18 th and 19 th nucleotides from 5’ to 3’of the antisense strand are linked by phospho
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-AGGAGGUGGCCAGGUGGAA-3’ (SEQ ID NO: 23) (sense strand) or 5’-UUCCACCUGGCCACCUCCU-3’ (SEQ ID NO: 24) (antisense strand); wherein the 1 st , 3 rd , 4 th , 7 th , 8 th , 9 th , 10 th , 11 th , 13 th , and 18 th nucleotides from 5’ to 3’of the sense strand is RNA and the 2 nd , 5 th , 6 th , 12 th , 14 th , 15 th , 16 th
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-AGGAGGUGGCCAGGUGGAA-3’ (SEQ ID NO: 23) (sense strand) or 5’-UUCCACCUGGCCACCUCCU-3’ (SEQ ID NO: 24) (antisense strand); wherein the 1 st , 2 nd , 3 rd , 4 th , 5 th , 6 th , 8 th , 9 th , 10 th , 11 th , 12 th , 13 th , 14 th , 15 th , 16 th , 18 th , and 19 th nucleotides from 5’ to 3’of the antisense strand is
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-AGGAGGUGGCCAGGUGGAA-3’ (SEQ ID NO: 23) (sense strand) or 5’-UUCCACCUGGCCACCUCCU-3’ (SEQ ID NO: 24) (antisense strand); wherein the 1 st , 2 nd , 3 rd , 4 th , 5 th , 6 th , 8 th , 12 th , 13 th , 14 th , 15 th , 16 th , 17 th , 18 th and 19 th nucleotides from 5’ to 3’of the sense strand comprise 2’-OMe at the sugar position and the 7 th
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-AGGAGGUGGCCAGGUGGAA-3’ (SEQ ID NO: 23) (sense strand) or 5’-UUCCACCUGGCCACCUCCU-3’ (SEQ ID NO: 24) (antisense strand); wherein the 1 st , 3 rd , 4 th , 5 th , 7 th , 8 th , 10 th , 11 th , 12 th , 13 th , 15 th , 17 th , 18 th , and 19 th nucleotides from 5’ to 3’of the antisense strand comprise 2’-OMe at the sugar position and the 2 nd
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-AGGAGGUGGCCAGGUGGAA-3’ (SEQ ID NO: 23) (sense strand) or 5’-UUCCACCUGGCCACCUCCU-3’ (SEQ ID NO: 24) (antisense strand); wherein at least one of the backbone of the sense and/or antisense strands is linked to a phosphodiester bond, a phosphothioate bond, or a combination thereof.
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-UGGCCAGGUGGAAGUAAAA-3’ (SEQ ID NO: 27) (sense strand) or 5’-UUUUACUUCCACCUGGCCA-3’ (SEQ ID NO: 28) (antisense strand); wherein the 1 st , 3 rd , 4 th , 7 th , 8 th , 9 th , 10 th , 11 th , 13 th , and 18 th nucleotides from 5’ to 3’of the sense strand is RNA and the 2 nd , 5 th , 6 th , 12 th , 14 th , 15 th , 16
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-UGGCCAGGUGGAAGUAAAA-3’ (SEQ ID NO: 27) (sense strand) or 5’-UUUUACUUCCACCUGGCCA-3’ (SEQ ID NO: 28) (antisense strand); wherein the 1 st , 2 nd , 3 rd , 4 th , 5 th , 6 th , 8 th , 9 th , 10 th , 11 th , 12 th , 13 th , 14 th , 15 th , 16 th , 18 th , and 19 th nucleotides from 5’ to 3’ of the antisense strand
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-UGGCCAGGUGGAAGUAAAA-3’ (SEQ ID NO: 27) (sense strand) or 5’-UUUUACUUCCACCUGGCCA-3’ (SEQ ID NO: 28) (antisense strand); wherein at least one of the backbone of the sense and/or antisense strands is linked to a phosphodiester bond, a phosphothioate bond, or a combination thereof.
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-UGGCCAGGUGGAAGUAAAA-3’ (SEQ ID NO: 27) (sense strand) or 5’-UUUUACUUCCACCUGGCCA-3’ (SEQ ID NO: 28) (antisense strand); wherein each of the backbone of the sense and antisense strands is linked by a phosphodiester bond.
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-UGGCCAGGUGGAAGUAAAA-3’ (SEQ ID NO: 27) (sense strand) or 5’-UUUUACUUCCACCUGGCCA-3’ (SEQ ID NO: 28) (antisense strand); wherein at least one of the backbone of the sense and/or antisense strands is linked to a phosphodiester bond, a phosphothioate bond, or a combination thereof.
  • MTT Microtubule-associated protein tau
  • the present disclosure provides an oligonucleotide comprising a contiguous nucleotide sequence of 10 to 30 nucleotides in length that are complementary to a nucleic acid sequence within a Microtubule-associated protein tau (MAPT) transcript, wherein the contiguous nucleotide sequence comprises: 5’-UGGCCAGGUGGAAGUAAAA-3’ (SEQ ID NO: 27) (sense strand) or 5’-UUUUACUUCCACCUGGCCA-3’ (SEQ ID NO: 28) (antisense strand); wherein the 1 st , 2 nd and 3 rd nucleotides from 5’ to 3’ of the sense strand, the 1 st , 2 nd and 3 rd nucleotides from 5’ to 3’of the antisense strand, and the 17 th , 18 th and 19 th nucleotides from 5’ to 3’of the antisense strand are linked by phosphoroth
  • the oligonucleotide e.g. siRNA
  • the oligonucleotide is double stranded and comprises: i. 5’-AACGUCCAGUCCAAGUGUGGC-3’ (SEQ ID NO: 1) (sense strand) and/or 5’-GCCACACUUGGACUGGACGTT-3’ (SEQ ID NO: 2) (antisense strand); ii. 5’-AGCAACGUCCAGUCCAAG-3’ (SEQ ID NO: 3) (sense strand) and/or 5’-CUUGGACUGGACGUUGCU-3’ (SEQ ID NO: 4) (antisense strand); iii.
  • 5’-AGCGGCUACAGCAGCCCCG -3’ (SEQ ID NO: 9) (sense strand) and/or 5’-CGGGGCUGCUGUAGCCGCU-3’ (SEQ ID NO: 10) (antisense strand); vi. 5’-AAUUAAUAAGAAGCUGGAUCU-3’ (SEQ ID NO: 11) (sense strand) and/or 5’-AGAUCCAGCUUCUUAUUAATT-3’ (SEQ ID NO: 12) (antisense strand); vii.
  • 5’-GAGCCCAAGAAGGUGGCAG-3’ (SEQ ID NO: 13) (sense strand) and/or 5’-CUGCCACCUUCUUGGGCUC-3’ (SEQ ID NO: 14) (antisense strand); viii. 5’-ACCCGGGAGCCCAAGAAG-3’ (SEQ ID NO: 15) (sense strand) and/or 5’-CUUCUUGGGCUCCCGGGU-3’ (SEQ ID NO: 16) (antisense strand); ix.
  • 5’-AAUAUCACCCACGUCCCUGG-3’ (SEQ ID NO: 25) (sense strand) and/or 5’-CCAGGGACGUGGGUGAUATT-3’ (SEQ ID NO: 26) (antisense strand); xiv. 5’-UGGCCAGGUGGAAGUAAAA-3’ (SEQ ID NO: 27) (sense strand) and/or 5’-UUUUACUUCCACCUGGCCA-3’ (SEQ ID NO: 28) (antisense strand); xv.
  • 5’-CCAAAGCCAAGACAGACCA-3’ (SEQ ID NO: 29) (sense strand) and/or 5’-UGGUCUGUCUUGGCUUUGG-3’ (SEQ ID NO: 30) (antisense strand); xvi. 5’-AAGCAGGGUUUGUGAUCAG-3’ (SEQ ID NO: 31) (sense strand) and/or 5’-CUGAUCACAAACCCUGCTT-3’ (SEQ ID NO: 32) (antisense strand); or xvii.
  • the oligonucleotide e.g. siRNA
  • the oligonucleotide is double stranded and comprises: i. 5’-AGGGCAGCCUGUGGGAGAA-3’ (SEQ ID NO: 35) (sense strand) and/or 5’-UUCUCCCACAGGCUGCCCU-3’ (SEQ ID NO: 36) (antisense strand) or ii.
  • the oligonucleotide e.g. siRNA
  • the oligonucleotide is double stranded and comprises: i. 5’-AAGCUGGAUCUUAGCAACGUC-3’ (SEQ ID NO: 37) (sense strand) and/or 5’-GACGUUGCUAAGAUCCAGCTT-3’ (SEQ ID NO: 38) (antisense strand); ii.
  • 5’-AAGAUUGAAACCCACAAGC-3’ (SEQ ID NO: 43) (sense strand) and/or 5’-GCUUGUGGGUUUCAAUCTT-3’ (SEQ ID NO: 44) (antisense strand); v. 5’-GAAACCCACAAGCUGACCU-3’ (SEQ ID NO: 45) (sense strand) and/or 5’-AGGUCAGCUUGUGGGUUUC-3’ (SEQ ID NO: 46) (antisense strand); vi.
  • 5’-AACACGUCCCGGGAGGCGGCA-3’ (SEQ ID NO: 51) (sense strand) and/or 5’-UGCCGCCUCCCGGGACGUGTT-3’ (SEQ ID NO: 52) (antisense strand); ix. 5’-ACGUCCCGGGAGGCGGCAG-3’ (SEQ ID NO: 53) (sense strand) and/or 5’-CUGCCGCCUCCCGGGACGU-3’ (SEQ ID NO: 54) (antisense strand); or x.
  • the oligonucleotide comprises: 5’- AACGUCCAGUCCAAGUGUGGC-3’ (SEQ ID NO: 1) (sense strand) and 5’- GCCACACUUGGACUGGACGTT-3’ (SEQ ID NO: 2) (antisense strand).
  • the oligonucleotide e.g., siRNA
  • the oligonucleotide comprises: 5’- AGCAACGUCCAGUCCAAG-3’ (SEQ ID NO: 3) (sense strand) and 5’-CUUGGACUGGACGUUGCU- 3’ (SEQ ID NO: 4) (antisense strand).
  • the oligonucleotide, e.g., siRNA comprises: 5’- AAGGUGCAGAUAAUUAAUAAG-3’ (SEQ ID NO: 5) (sense strand) and 5’- CUUAUUAAUUAUCUGCACCTT-3’ (SEQ ID NO: 6) (antisense strand).
  • the oligonucleotide comprises: 5’- UGAGAACCUGAAGCACCAG-3’ (SEQ ID NO: 7) (sense strand) and 5’- CUGGUGCUUCAGGUUCUCA-3’ (SEQ ID NO: 8) (antisense strand).
  • the oligonucleotide, e.g., siRNA comprises: 5’- AGCGGCUACAGCAGCCCCG-3’ (SEQ ID NO: 9) (sense strand) and 5’- CGGGGCUGCUGUAGCCGCU-3’ (SEQ ID NO: 10) (antisense strand).
  • the oligonucleotide e.g., siRNA
  • the oligonucleotide comprises: 5’- AAUUAAUAAGAAGCUGGAUCU-3’ (SEQ ID NO: 11) (sense strand) and 5’- AGAUCCAGCUUCUUAUUAATT-3’ (SEQ ID NO: 12) (antisense strand).
  • the oligonucleotide, e.g., siRNA comprises: 5’- GAGCCCAAGAAGGUGGCAG-3’ (SEQ ID NO: 13) (sense strand) and 5’- CUGCCACCUUCUUGGGCUC-3’ (SEQ ID NO: 14) (antisense strand).
  • the oligonucleotide e.g., siRNA
  • the oligonucleotide comprises: 5’- ACCCGGGAGCCCAAGAAG-3’ (SEQ ID NO: 15) (sense strand) and 5’-CUUCUUGGGCUCCCGGGU- 3’ (SEQ ID NO: 16) (antisense strand).
  • the oligonucleotide, e.g., siRNA comprises: 5’- AGAAGCUGGAUCUUAGCAA-3’ (SEQ ID NO: 17) (sense strand) and 5’- UUGCUAAGAUCCAGCUUCU-3’ (SEQ ID NO: 18) (antisense strand).
  • the oligonucleotide e.g., siRNA
  • the oligonucleotide comprises: 5’- AAGGAUAAUAUCAAACACGUC-3’ (SEQ ID NO: 19) (sense strand) and 5’- GACGUGUUUGAUAUUAUCCTT-3’ (SEQ ID NO: 20) (antisense strand).
  • the oligonucleotide, e.g., siRNA comprises: 5’- AUAAUAUCAAACACGUCCC-3’ (SEQ ID NO: 21) (sense strand) and 5’- GGGACGUGUUUGAUAUUAU-3’ (SEQ ID NO: 22) (antisense strand).
  • the oligonucleotide e.g., siRNA
  • the oligonucleotide comprises: 5’- AGGAGGUGGCCAGGUGGAA-3’ (SEQ ID NO: 23) (sense strand) and 5’- UUCCACCUGGCCACCUCCU-3’ (SEQ ID NO: 24) (antisense strand).
  • the oligonucleotide, e.g., siRNA comprises: 5’- AAUAUCACCCACGUCCCUGG-3’ (SEQ ID NO: 25) (sense strand) and 5’- CCAGGGACGUGGGUGAUATT-3’ (SEQ ID NO: 26) (antisense strand).
  • the oligonucleotide e.g., siRNA
  • the oligonucleotide comprises: 5’- UGGCCAGGUGGAAGUAAAA-3’ (SEQ ID NO: 27) (sense strand) and 5’- UUUUACUUCCACCUGGCCA-3’ (SEQ ID NO: 28) (antisense strand).
  • the oligonucleotide, e.g., siRNA comprises: 5’- CCAAAGCCAAGACAGACCA-3’ (SEQ ID NO: 29) (sense strand) and 5’- UGGUCUGUCUUGGCUUUGG-3’ (SEQ ID NO: 30) (antisense strand).
  • the oligonucleotide e.g., siRNA
  • the oligonucleotide comprises: 5’- AAGCAGGGUUUGUGAUCAG-3’ (SEQ ID NO: 31) (sense strand) and 5’- CUGAUCACAAACCCUGCTT-3’ (SEQ ID NO: 32) (antisense strand).
  • the oligonucleotide, e.g., siRNA comprises: 5’- CUCGGGACUUCAAAAUCAG-3’ (SEQ ID NO: 33) (sense strand) and 5’- CUGAUUUUGAAGUCCCGAG-3’ (SEQ ID NO: 34) (antisense strand).
  • the oligonucleotide e.g., siRNA
  • the oligonucleotide comprises: 5’- AGGGCAGCCUGUGGGAGAA-3’ (SEQ ID NO: 35) (sense strand) and 5’- UUCUCCCACAGGCUGCCCU-3’ (SEQ ID NO: 36) (antisense strand).
  • the oligonucleotide, e.g., siRNA comprises: 5’- AAGCUGGAUCUUAGCAACGUC-3’ (SEQ ID NO: 37) (sense strand) and 5’- GACGUUGCUAAGAUCCAGCTT-3’ (SEQ ID NO: 38) (antisense strand).
  • the oligonucleotide e.g., siRNA
  • the oligonucleotide comprises: 5’- AGUGUGCAAAUAGUCUACAA-3’ (SEQ ID NO: 39) (sense strand) and 5’- UUGUAGACUAUUUGCACACU-3’ (SEQ ID NO: 40) (antisense strand).
  • the oligonucleotide, e.g., siRNA comprises: 5’- AAGGACAGAGUCCAGUCGAAG-3’ (SEQ ID NO: 41) (sense strand) and 5’- CUUCGACUGGACUCUGUCCTT-3’ (SEQ ID NO: 42) (antisense strand).
  • the oligonucleotide comprises: 5’- AAGAUUGAAACCCACAAGC-3’ (SEQ ID NO: 43) (sense strand) and 5’- GCUUGUGGGUUUCAAUCTT-3’ (SEQ ID NO: 44) (antisense strand).
  • the oligonucleotide, e.g., siRNA comprises: 5’- GAAACCCACAAGCUGACCU-3’ (SEQ ID NO: 45) (sense strand) and 5’- AGGUCAGCUUGUGGGUUUC-3’ (SEQ ID NO: 46) (antisense strand).
  • the oligonucleotide e.g., siRNA
  • the oligonucleotide comprises: 5’- AAUAUCAAACACGUCCCGGGA-3’ (SEQ ID NO: 47) (sense strand) and 5’- UCCCGGGACGUGUUUGAUATT-3’ (SEQ ID NO: 48) (antisense strand).
  • the oligonucleotide, e.g., siRNA comprises: 5’- AUCAAACACGUCCCGGGAG-3’ (SEQ ID NO: 49) (sense strand) and 5’- CUCCCGGGACGUGUUUGAU-3’ (SEQ ID NO: 50) (antisense strand).
  • the oligonucleotide e.g., siRNA
  • the oligonucleotide comprises: 5’- AACACGUCCCGGGAGGCGGCA-3’ (SEQ ID NO: 51) (sense strand) and 5’- UGCCGCCUCCCGGGACGUGTT-3’ (SEQ ID NO: 52) (antisense strand).
  • the oligonucleotide, e.g., siRNA comprises: 5’- ACGUCCCGGGAGGCGGCAG-3’ (SEQ ID NO: 53) (sense strand) and 5’- CUGCCGCCUCCCGGGACGU-3’ (SEQ ID NO: 54) (antisense strand).
  • the oligonucleotide e.g., siRNA
  • the oligonucleotide comprises: 5’- UCCCGGGAGGCGGCAGUGU-3’ (SEQ ID NO: 55) (sense strand) and 5’- ACACUGCCGCCUCCCGGGA-3’ (SEQ ID NO: 56) (antisense strand).
  • the oligonucleotide, e.g., siRNA comprises: 5’- GGUGCAGAUAAUUAAUAAG-3’ (SEQ ID NO: 64) (sense strand) and 5’- CUUAUUAAUUAUCUGCACC-3’ (SEQ ID NO: 65) (antisense strand).
  • the oligonucleotide e.g., siRNA
  • the oligonucleotide e.g., siRNA
  • the oligonucleotide e.g, siRNA
  • the oligonucleotide e.g., siRNA
  • the oligonucleotide e.g., siRNA
  • the oligonucleotide e.g., siRNA
  • the oligonucleotide e.g., siRNA
  • the oligonucleotide e.g., siRNA
  • the oligonucleotide e.g., siRNA
  • the oligonucleotide e.g., siRNA
  • the oligonucleotide e.g., siRNA
  • the oligonucleotide e.g., siRNA
  • the oligonucleotide e.g., siRNA
  • the oligonucleotide e.g., siRNA
  • the oligonucleotide e.g., siRNA
  • the oligonucleotide e.g., siRNA
  • the oligonucleotide e.g., siRNA
  • the oligonucleotide e.g., siRNA
  • the oligonucleotide e.g., siRNA
  • the oligonucleotide e.g., siRNA
  • the oligonucleotide e.g., siRNA
  • the oligonucleotide e.g., siRNA
  • the oligonucleotide e.g., siRNA
  • the oligonucleotide e.g., siRNA
  • the oligonucleotide e.g., siRNA
  • the oligonucleotide e.g., siRNA
  • the oligonucleotide e.g., siRNA
  • siRNA Nos.1, 2, 3, 4, 5, 6, 8, 9, 10, 11, 12, 14, 15, 17, 20, 21, 22, and 23 reduced the expression level of total tau protein. In some aspects siRNA Nos.1, 2, 4, 5, 6, 8, 10, 11, 12, 14, 15, 21, 22, and 23 effectively reduced the expression level of total tau protein. [0387] In some aspects, siRNA Nos.7, 13, 16, 17, 20, 25, 26, 27, and 28 failed to reduce the expression level of total tau protein. [0388] In some aspects, siRNA Nos.1 and 17 effectively reduced the expression level of mutant MAPT (L266V). [0389] In some aspects, siRNA Nos.3, 11, and 23 effectively reduced the expression level of wild-type & mutant MAPT.
  • siRNA Nos.5, 6, 8, 9, 10, 12, 14, 15, 21, and 22 effectively reduced expression levels of wild-type MAPT (4R2N).
  • siRNA Nos.1 and 17 effectively reduced the expression level of 4R- MAPT isoforms.
  • siRNA Nos.20 and 23 effectively reduced the expression level of wild-type, 4R-, and 3R- MAPT isoforms.
  • siRNA Nos.3 and 11 effectively reduced the expression level of wild-type and 4R- MAPT isoforms.
  • siRNA No.5 effectively reduced the expression level of wild-type and 3R- MAPT isoforms.
  • siRNA Nos. 1, 2, 4, 5, 6, 8, 10, 11, 12, 14, 15, 21, 22 and 23 effectively reduced the expression level of total tau protein and will be effective for diseases characterized by the aggregation and deposition of tau. (e.g., Alzheimer’s disease, Parkinson’s disease, and amyotrophic lateral sclerosis).
  • siRNA Nos.1 and 17 specifically reduced 4R-tau isoforms thus, will provide therapeutic effects in treating 4R-tauopathies (e.g., Progressive supranuclear palsy, Corticobasal degeneration, Globular glial tauopathy, Argyrophilic grain disease, and Aging-related tau ⁇ astrogliopathy).
  • siRNA Nos.20 and 23 reduced all forms of tau, thus will provide therapeutic effects in treating (3R-)+(4R-) tauopathies (e.g., Primary age related tauopathy, Tangle-predominant dementia, Frontotemporal dementia, and parkinsonism linked to chromosome 17)
  • siRNA Nos.3 and 11 reduced wild-type (except 3R-tau isoforms) and 4R-tau isoforms, thus will provide therapeutic effects in treating 3R/4R imbalance tauopathies induced by abnormally increased 4R-tau isoforms.
  • siRNA No. 17 efficiently reduced tau hyperphosphorylation and restored recognition decline in P301S transgenic mice.
  • siRNA Nos.1, 2, 3, 4, 5, 6, 8, 9, 10, 11, 12, 14, 15, 17, 20, 21, 22, and 23 exhibited knockdown effects.
  • siRNA Nos.7, 13, 16, 18, 19, 24, 25, 26, 27, and 28 did not exhibit knockdown effects.
  • the disclosure provides an siRNA which comprises a contiguous nucleotide sequence of a total of from 10-30 nucleotides, such as 10-15 nucleotides, 10-20 nucleotides, or 10-25 nucleotides in length wherein the contiguous nucleotide sequence has at least about 85%, at least about 90%, at least about 95%, at least about 98%, or at least about 99% sequence identity to a region within the complement of a mammalian MAPT transcript, such as SEQ ID NO: 57 or SEQ ID NOS: 58-63.
  • the siRNAs of the disclosure bind to the target nucleic acid sequence (e.g., MAPT transcript) and are capable of inhibiting or reducing expression of the MAPT transcript by at least about 10%, at least about 20% , at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or about 100% in a tissue (e.g., a brain region) of a mouse expressing a human MAPT gene (e.g., P301L) compared to the control.
  • a tissue e.g., a brain region
  • a mouse expressing a human MAPT gene e.g., P301L
  • the siRNAs are capable of reducing expression of MAPT protein by at least about 10%, at least about 20% , at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or about 100% in a tissue (e.g., a brain region) of a mouse expressing a human MAPT gene (e.g., P301L) compared to the control.
  • a tissue e.g., a brain region
  • a human MAPT gene e.g., P301L
  • the siRNA of the disclosure bind to the target nucleic acid sequence (e.g., MAPT transcript) and are capable of inhibiting or reducing expression of the MAPT transcript by at least about 10%, at least about 20% , at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or about 100% in a tissue (e.g., a brain region) of a cyno expressing the wild-type MAPT gene when administered in vivo compared to the control.
  • a tissue e.g., a brain region
  • the siRNAs are capable of reducing expression of MAPT protein by at least about 10%, at least about 20% , at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or about 100% in a tissue (e.g., a brain region) of a cyno expressing the wild-type MAPT gene when administered in vivo compared to the control.
  • the siRNAs of the disclosure bind to the MAPT transcript and inhibit or reduce expression of the MAPT mRNA by at least about 10% or about 20% compared to the normal (i.e.
  • the siRNA reduces expression of MAPT protein in a cell following administration of the siRNA by at least 60%, at least 70%, at least 80%, or at least 90% compared to a cell not exposed to the siRNA (i.e., control).
  • the siRNA reduces expression of MAPT protein in a cell following administration of the siRNA by at least about 60%, at least about 70%, at least about 80%, or at least about 90% compared to a cell not exposed to the siRNA (i.e., control).
  • the siRNA of the disclosure has at least one property selected from: (1) reduces expression of MAPT mRNA in a cell, e.g., HeLa cell, compared to a control cell that has not been exposed to the siRNA; (2) does not significantly reduce calcium oscillations in a cell; (3) does not significantly reduce tubulin intensity in a cell; (4) reduces expression of Microtubule-associated protein tau protein in a cell; and (5) any combinations thereof compared to a control cell that has not been exposed to the siRNA.
  • a cell e.g., HeLa cell
  • the siRNA can tolerate 1, 2, 3, or 4 (or more) mismatches, when hybridizing to the target sequence and still sufficiently bind to the target to show the desired effect, i.e., down-regulation of the target mRNA and/or protein. Mismatches can, for example, be compensated by increased length of the siRNA nucleotide sequence and/or an increased number of nucleotide analogs, which are disclosed elsewhere herein.
  • the siRNA of the disclosure comprises no more than 3 mismatches when hybridizing to the target sequence. In other aspects, the contiguous nucleotide sequence comprises no more than 2 mismatches when hybridizing to the target sequence.
  • the contiguous nucleotide sequence comprises no more than 1 mismatch when hybridizing to the target sequence.
  • the nucleotide sequence of the ASO can comprise additional 5' or 3' nucleotides, such as, independently, 1, 2, 3, 4 or 5 additional nucleotides 5' and/or 3', which are non-complementary to the target sequence.
  • the siRNA of the disclosure can, in some aspects, comprise a contiguous nucleotide sequence which is flanked 5' and/or 3' by additional nucleotides.
  • the additional 5' and/or 3' nucleotides are naturally occurring nucleotides, such as DNA or RNA. II.C.
  • the siRNAs can comprise a contiguous nucleotide sequence of a total of 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 contiguous nucleotides in length. [0413] In some aspects, the siRNAs comprise a contiguous nucleotide sequence of a total of about 10–22, such as 10-21 or 12–18, such as 13–17 or 12–16, such as 13, 14, 15, 16, 17, 18, 19, 20, or 21 contiguous nucleotides in length. [0414] In some aspects, the siRNAs comprise a contiguous nucleotide sequence of a total of 16, 17, 18, 19, or 20 (16 to 20) contiguous nucleotides in length.
  • the siRNA comprises a sense strand and an antisense strand, each of the strands comprises 19 contiguous nucleotides in length.
  • the siRNA according to the disclosure consists of no more than 22 nucleotides, such as no more than 21 or 20 nucleotides, such as no more than 18 nucleotides, such as 15, 16 or 17 nucleotides.
  • the siRNA of the disclosure comprises less than 22 nucleotides. It should be understood that when a range is given for an siRNA, or contiguous nucleotide sequence length, the range includes the lower and upper lengths provided in the range, for example from (or between) 10–30, includes both 10 and 30. II.D.
  • the siRNAs comprise one or more non-naturally occurring nucleotide analogs.
  • "Nucleotide analogs” or “nucleotide analogues” as used herein are variants of natural nucleotides, such as DNA or RNA nucleotides, by virtue of modifications in the sugar and/or base moieties. Analogs could in principle be merely "silent” or “equivalent” to the natural nucleotides in the context of the oligonucleotide, i.e. have no functional effect on the way the oligonucleotide works to inhibit target gene expression.
  • Such "equivalent" analogs can nevertheless be useful if, for example, they are easier or cheaper to manufacture, or are more stable to storage or manufacturing conditions, or represent a tag or label. In some aspects, however, the analogs will have a functional effect on the way in which the siRNA works to inhibit expression; for example by producing increased binding affinity to the target and/or increased resistance to intracellular nucleases and/or increased ease of transport into the cell.
  • nucleoside analogs are described by e.g. Freier & Altmann; Nucl. Acid Res., 1997, 25, 4429-4443 and Uhlmann; Curr. Opinion in Drug Development, 2000, 3(2), 293-213, and in Scheme 1.
  • the oligonucleotide comprises a sense strand comprising 5’ RMRRMMRRRRRMRMMMMRM3’, wherein R is RNA and M is 2’-OMe and/or an antisense strand comprising 5’ RRMRRRRRRRRRMRRRR3’, wherein R is RNA and M is 2’-OMe.
  • the oligonucleotide comprises a 5’ methyl cytosine.
  • the oligonucleotide comprises a sense strand comprising 5’ MMMMMMFMFFFMMMMMM3’, wherein M is 2’-OMe and F is 2’-F and/or an antisense strand comprising 5’ MMMFMFMMMMFMMFMMMF3’, wherein M is 2’-OMe and F is 2’-F.
  • the oligonucleotide comprises a 5’ methyl cytosine. II.D.1.
  • nucleobase includes the purine (e.g., adenine and guanine) and pyrimidine (e.g., uracil, thymine and cytosine) moiety present in nucleosides and nucleotides which form hydrogen bonds in nucleic acid hybridization.
  • pyrimidine e.g., uracil, thymine and cytosine
  • nucleobase also encompasses modified nucleobases which may differ from naturally occurring nucleobases, but are functional during nucleic acid hybridization.
  • nucleobase moiety is modified by modifying or replacing the nucleobase.
  • nucleobase refers to both naturally occurring nucleobases such as adenine, guanine, cytosine, thymine, uracil, xanthine and hypoxanthine, as well as non-naturally occurring variants. Such variants are for example described in Hirao et al., (2012) Accounts of Chemical Research vol 45 page 2055 and Bergstrom (2009) Current Protocols in Nucleic Acid Chemistry Suppl.371.4.1.
  • the nucleobase moiety is modified by changing the purine or pyrimidine into a modified purine or pyrimidine, such as substituted purine or substituted pyrimidine, such as a nucleobase selected from isocytosine, pseudoisocytosine, 5-methyl cytosine, 5-thiozolo-cytosine, 5-propynyl-cytosine, 5-propynyl-uracil, 5-bromouracil 5-thiazolo-uracil, 2- thio-uracil, 2’thio-thymine, inosine, diaminopurine, 6-aminopurine, 2-aminopurine, 2,6- diaminopurine and 2-chloro-6-aminopurine.
  • a nucleobase selected from isocytosine, pseudoisocytosine, 5-methyl cytosine, 5-thiozolo-cytosine, 5-propynyl-cytosine, 5-propynyl-uracil, 5-brom
  • the nucleobase moieties may be indicated by the letter code for each corresponding nucleobase, e.g., A, T, G, C or U, wherein each letter may optionally include modified nucleobases of equivalent function.
  • the nucleobase moieties are selected from A, T, G, C, and 5-methyl cytosine.
  • 5-methyl cytosine LNA nucleosides may be used.
  • the siRNA of the disclosure can comprise one or more nucleosides which have a modified sugar moiety, i.e.
  • modifications include those where the ribose ring structure is modified, e.g.
  • HNA hexose ring
  • LNA ribose ring
  • UPA unlinked ribose ring which typically lacks a bond between the C2' and C3' carbons
  • Other sugar modified nucleosides include, for example, bicyclohexose nucleic acids (WO2011/017521) or tricyclic nucleic acids (WO2013/154798).
  • Modified nucleosides also include nucleosides where the sugar moiety is replaced with a non-sugar moiety, for example in the case of peptide nucleic acids (PNA), or morpholino nucleic acids.
  • Sugar modifications also include modifications made via altering the substituent groups on the ribose ring to groups other than hydrogen, or the 2’-OH group naturally found in RNA nucleosides. Substituents may, for example be introduced at the 2’, 3’, 4’ or 5’ positions.
  • Nucleosides with modified sugar moieties also include 2’ modified nucleosides, such as 2’ substituted nucleosides.
  • the sugar modification comprises an affinity enhancing sugar modification, e.g., LNA.
  • An affinity enhancing sugar modification increases the binding affinity of the ASOs to the target RNA sequence (e.g., intron1/exon2 junction of MAPT mRNA).
  • an siRNA comprising a sugar modification disclosed herein has a binding affinity to a target RNA sequence that is enhanced by at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100% compared to a control (e.g., an siRNA without such sugar modification).
  • a 2’ sugar modified nucleoside is a nucleoside which has a substituent other than H or –OH at the 2’ position (2’ substituted nucleoside) or comprises a 2’ linked biradical, and includes 2’ substituted nucleosides and LNA (2’ – 4’ biradical bridged) nucleosides.
  • the 2’ modified sugar may provide enhanced binding affinity and/or increased nuclease resistance to the oligonucleotide.
  • Examples of 2’ substituted modified nucleosides are 2’-O-alkyl-RNA, 2’- O-methyl-RNA, 2’-alkoxy-RNA, 2’-O-methoxyethyl-RNA (MOE), 2’-amino-DNA, 2’-Fluoro- RNA, and 2’-F-ANA nucleoside.
  • MOE methoxyethyl-RNA
  • 2’-amino-DNA 2’-Fluoro- RNA
  • 2’-F-ANA nucleoside examples of 2’ substituted modified nucleosides.
  • LNA nucleosides are modified nucleosides which comprise a linker group (referred to as a biradical or a bridge) between C2’ and C4’ of the ribose sugar ring of a nucleotide. These nucleosides are also termed bridged nucleic acid or bicyclic nucleic acid (BNA) in the literature.
  • linker group referred to as a biradical or a bridge
  • –X-Y- designates –O-CH 2 - or –O-CH(CH 3 )-.
  • Z is selected from -O-, -S-, and -N(R a )-, and R a and, when present R b , each is independently selected from hydrogen, optionally substituted C1-6-alkyl, optionally substituted C 2-6 -alkenyl, optionally substituted C 2-6 -alkynyl, hydroxy, optionally substituted C 1-6 - alkoxy, C 2-6 -alkoxyalkyl, C 2-6 -alkenyloxy, carboxy, C 1-6 -alkoxycarbonyl, C 1-6 -alkylcarbonyl, formyl, aryl, aryloxy-carbonyl, aryloxy, arylcarbonyl, heteroaryl, heteroaryloxy-carbonyl, heteroaryloxy, heteroarylcarbonyl, amino, mono- and
  • R 1 , R 2 , R 3 , R 5 and R 5* are independently selected from the group consisting of: hydrogen, optionally substituted C1-6-alkyl, optionally substituted C2-6-alkenyl, optionally substituted C 2-6 -alkynyl, hydroxy, C 1-6 -alkoxy, C 2-6 -alkoxyalkyl, C 2-6 -alkenyloxy, carboxy, C 1-6 -alkoxycarbonyl, C 1-6 -alkylcarbonyl, formyl, aryl, aryloxy-carbonyl, aryloxy, arylcarbonyl, heteroaryl, heteroaryloxy-carbonyl, heteroaryloxy, heteroarylcarbonyl, amino, mono- and di(C1-6-alkyl)amino, carbamoyl, mono- and di(C1-6-alkyl)-amino-carbonyl, amino-C1- 6 -alkyl-aminocarbonyl, amino-C1-
  • R 1 , R 2 , R 3 , R 5 and R 5* are independently selected from C1-6 alkyl, such as methyl, and hydrogen. [0437] In some aspects R 1 , R 2 , R 3 , R 5 and R 5* are all hydrogen. [0438] In some aspects R 1 , R 2 , R 3 , are all hydrogen, and either R 5 and R 5* is also hydrogen and the other of R 5 and R 5* is other than hydrogen, such as C 1-6 alkyl such as methyl. [0439] In some aspects, R a is either hydrogen or methyl. In some aspects, when present, R b is either hydrogen or methyl. [0440] In some aspects, one or both of R a and R b is hydrogen.
  • one of R a and R b is hydrogen and the other is other than hydrogen.
  • one of R a and R b is methyl and the other is hydrogen.
  • both of R a and R b are methyl.
  • the biradical –X-Y- is –O-CH 2 -, W is O, and all of R 1 , R 2 , R 3 , R 5 and R 5* are all hydrogen.
  • LNA nucleosides are disclosed in WO99/014226, WO00/66604, WO98/039352 and WO2004/046160 which are all hereby incorporated by reference, and include what are commonly known as beta-D-oxy LNA and alpha-L-oxy LNA nucleosides.
  • the biradical –X-Y- is –S-CH 2 -
  • W is O
  • all of R 1 , R 2 , R 3 , R 5 and R 5* are all hydrogen.
  • Such thio LNA nucleosides are disclosed in WO99/014226 and WO2004/046160.
  • the biradical –X-Y- is –NH-CH 2 -, W is O, and all of R 1 , R 2 , R 3 , R 5 and R 5* are all hydrogen.
  • Such amino LNA nucleosides are disclosed in WO99/014226 and WO2004/046160.
  • the biradical –X-Y- is –O-CH 2 -CH 2 - or –O-CH 2 -CH 2 - CH 2 -, W is O, and all of R 1 , R 2 , R 3 , R 5 and R 5* are all hydrogen.
  • LNA nucleosides are disclosed in WO00/047599 and Morita et al., Bioorganic & Med.Chem. Lett. 12 73-76, which are hereby incorporated by reference, and include what are commonly known as 2’-O-4’C-ethylene bridged nucleic acids (ENA).
  • the biradical –X-Y- is –O-CH2-
  • W is O
  • all of R 1 , R 2 , R 3 , and one of R 5 and R 5* are hydrogen
  • the other of R 5 and R 5* is other than hydrogen such as C1-6 alkyl, such as methyl.
  • Such 5’ substituted LNA nucleosides are disclosed in WO2007/134181.
  • the biradical –X-Y- is –O-CR a R b -, wherein one or both of R a and R b are other than hydrogen, such as methyl, W is O, and all of R 1 , R 2 , R 3 , and one of R 5 and R 5* are hydrogen, and the other of R 5 and R 5* is other than hydrogen such as C 1-6 alkyl, such as methyl.
  • R a and R b are other than hydrogen, such as methyl
  • W is O
  • all of R 1 , R 2 , R 3 , and one of R 5 and R 5* are hydrogen
  • the other of R 5 and R 5* is other than hydrogen such as C 1-6 alkyl, such as methyl.
  • Such bis modified LNA nucleosides are disclosed in WO2010/077578.
  • the biradical –X-Y- designate the bivalent linker group –O- CH(CH 2 OCH 3 )- (2’ O-methoxyethyl bicyclic nucleic acid - Seth at al., 2010, J. Org. Chem. Vol 75(5) pp.1569-81). In some aspects, the biradical –X-Y- designate the bivalent linker group –O- CH(CH2CH3)- (2’O-ethyl bicyclic nucleic acid - Seth at al., 2010, J. Org. Chem. Vol 75(5) pp. 1569-81).
  • the biradical –X-Y- is –O-CHR a -, W is O, and all of R 1 , R 2 , R 3 , R 5 and R 5* are all hydrogen.
  • Such 6’ substituted LNA nucleosides are disclosed in WO10036698 and WO07090071.
  • the biradical –X-Y- is –O-CH(CH2OCH3)-, W is O, and all of R 1 , R 2 , R 3 , R 5 and R 5* are all hydrogen.
  • Such LNA nucleosides are also known as cyclic MOEs in the art (cMOE) and are disclosed in WO07090071.
  • the biradical –X-Y- designates the bivalent linker group –O- CH(CH 3 )-. – in either the R- or S- configuration.
  • the biradical –X-Y- together designate the bivalent linker group –O-CH 2 -O-CH 2 - (Seth et al., 2010, J. Org. Chem).
  • the biradical –X-Y- is –O-CH(CH3)-, W is O, and all of R 1 , R 2 , R 3 , R 5 and R 5* are all hydrogen.
  • methyl LNA nucleosides are also known as cET nucleosides in the art, and may be either (S)cET or (R)cET stereoisomers, as disclosed in WO07090071 (beta-D) and WO2010/036698 (alpha-L)).
  • the biradical –X-Y- is –O-CR a R b -, wherein in neither R a or R b is hydrogen, W is O, and all of R 1 , R 2 , R 3 , R 5 and R 5* are all hydrogen.
  • R a and R b are both methyl.
  • Such 6’ di-substituted LNA nucleosides are disclosed in WO 2009006478.
  • the biradical –X-Y- is –S-CHR a -
  • W is O
  • all of R 1 , R 2 , R 3 , R 5 and R 5* are all hydrogen.
  • substituted thio LNA nucleosides are disclosed in WO11156202.
  • R a is methyl.
  • Such vinyl carbo LNA nucleosides are disclosed in WO08154401 and WO09067647.
  • the biradical –X-Y- is –N(-OR a )-, W is O, and all of R 1 , R 2 , R 3 , R 5 and R 5* are all hydrogen.
  • R a is C1-6 alkyl such as methyl.
  • LNA nucleosides are also known as N substituted LNAs and are disclosed in WO2008/150729.
  • the biradical –X-Y- together designate the bivalent linker group –O-NR a -CH3- (Seth et al., 2010, J. Org. Chem).
  • the biradical –X-Y- is –N(R a )-, W is O, and all of R 1 , R 2 , R 3 , R 5 and R 5* are all hydrogen.
  • R a is C 1-6 alkyl such as methyl.
  • R 5 and R 5* is hydrogen and, when substituted the other of R 5 and R 5* is C1-6 alkyl such as methyl.
  • R 1 , R 2 , R 3 may all be hydrogen, and the biradical –X-Y- may be selected from –O-CH2- or –O-CH(CR a )-, such as –O-CH(CH3)-.
  • the biradical is –CR a R b -O-CR a R b -, such as CH 2 -O-CH 2 -, W is O and all of R 1 , R 2 , R 3 , R 5 and R 5* are all hydrogen.
  • R a is C1-6 alkyl such as methyl.
  • LNA nucleosides are also known as conformationally restricted nucleotides (CRNs) and are disclosed in WO2013036868.
  • the biradical is –O-CR a R b -O-CR a R b -, such as O-CH2-O-CH2-, W is O and all of R 1 , R 2 , R 3 , R 5 and R 5* are all hydrogen.
  • R a is C1-6 alkyl such as methyl.
  • LNA nucleosides are also known as COC nucleotides and are disclosed in Mitsuoka et al., Nucleic Acids Research 200937(4), 1225-1238. [0460] It will be recognized than, unless specified, the LNA nucleosides may be in the beta-D or alpha-L stereoisoform. [0461] Certain examples of LNA nucleosides are presented in Scheme 1. Scheme 1 [0462] As illustrated in the examples, in some aspects of the disclosure the LNA nucleosides in the oligonucleotides are beta-D-oxy-LNA nucleosides. II.E.
  • Nuclease mediated degradation refers to an oligonucleotide capable of mediating degradation of a complementary nucleotide sequence when forming a duplex with such a sequence.
  • the oligonucleotide may function via nuclease mediated degradation of the target nucleic acid, where the oligonucleotides of the disclosure are capable of recruiting a nuclease, particularly and endonuclease, preferably endoribonuclease (RNase), such as RNase H.
  • RNase endoribonuclease
  • oligonucleotide designs which operate via nuclease mediated mechanisms are oligonucleotides which typically comprise a region of at least 5 or 6 DNA nucleosides and are flanked on one side or both sides by affinity enhancing nucleosides, for example gapmers.
  • II.F. RNase H Activity and Recruitment [0465] The RNase H activity of an oligonucleotide refers to its ability to recruit RNase H when in a duplex with a complementary RNA molecule and induce cleavage and subsequent degradation of the complementary RNA molecule.
  • WO01/23613 provides in vitro methods for determining RNase H activity, which may be used to determine the ability to recruit RNase H.
  • an oligonucleotide is deemed capable of recruiting RNase H if it, when provided with a complementary target nucleic acid sequence, has an initial rate, as measured in pmol/l/min, of at least 5%, such as at least 10% or more than 20% of the of the initial rate determined when using a oligonucleotide having the same base sequence as the modified oligonucleotide being tested, but containing only DNA monomers, with phosphorothioate linkages between all monomers in the oligonucleotide, and using the methodology provided by Example 91 - 95 of WO01/23613.
  • an oligonucleotide is deemed essentially incapable of recruiting RNase H if, when provided with the complementary target nucleic acid, the RNase H initial rate, as measured in pmol/l/min, is less than 20%, such as less than 10%, such as less than 5% of the initial rate determined when using a oligonucleotide having the same base sequence as the oligonucleotide being tested, but containing only DNA monomers, with no 2’ substitutions, with phosphorothioate linkages between all monomers in the oligonucleotide, and using the methodology provided by Example 91 - 95 of WO01/23613. II.G.
  • Internucleotide Linkages [0467] The monomers of the siRNA described herein are coupled together via linkage groups. Suitably, each monomer is linked to the 3' adjacent monomer via a linkage group. [0468] The person having ordinary skill in the art would understand that, in the context of the present disclosure, the 5' monomer at the end of an siRNA does not comprise a 5' linkage group, although it may or may not comprise a 5' terminal group. [0469]
  • linkage group and “internucleotide linkage” are intended to mean a group capable of covalently coupling together two nucleotides. Specific and preferred examples include phosphate groups and phosphorothioate groups.
  • nucleotides of the siRNA of the disclosure or contiguous nucleotides sequence thereof are coupled together via linkage groups.
  • each nucleotide is linked to the 3' adjacent nucleotide via a linkage group.
  • Suitable internucleotide linkages include those listed within WO2007/031091, for example the internucleotide linkages listed on the first paragraph of page 34 of WO2007/031091 (hereby incorporated by reference in its entirety).
  • internucleotide linkages examples include phosphodiester linkage, a phosphotriester linkage, a methylphosphonate linkage, a phosphoramidate linkage, a phosphorothioate linkage, and combinations thereof.
  • Suitable sulphur (S) containing internucleotide linkages as provided herein may be preferred.
  • Phosphorothioate internucleotide linkages are also preferred, particularly for the gap region (B) of gapmers.
  • Phosphorothioate linkages can also be used for the flanking regions (A and C, and for linking A or C to D, and within region D, as appropriate).
  • Regions A, B and C can, however, comprise internucleotide linkages other than phosphorothioate, such as phosphodiester linkages, particularly, for instance when the use of nucleotide analogs protects the internucleotide linkages within regions A and C from endo- nuclease degradation – such as when regions A and C comprise LNA nucleotides.
  • the internucleotide linkages in the siRNA can be phosphodiester, phosphorothioate or boranophosphate so as to allow RNase H cleavage of targeted RNA. Phosphorothioate is preferred for improved nuclease resistance and other reasons, such as ease of manufacture.
  • the internucleotide linkages comprise one or more stereo-defined internucleotide linkages (e.g., such as stereo-defined modified phosphate linkages, e.g., phosphodiester, phosphorothioate, or boranophosphate linkages with a defined stereochemical structure).
  • stereo-defined internucleotide linkage is used interchangeably with "chirally controlled internucleotide linkage” and refers to a internucleotide linkage in which the stereochemical designation of the phosphorus atom is controlled such that a specific amount of R p or S p of the internucleotide linkage is present within an siRNA strand.
  • the stereochemical designation of a chiral linkage can be defined (controlled) by, for example, asymmetric synthesis.
  • An siRNA having at least one stereo-defined internucleotide linkage can be called as a stereo-defined siRNA, which includes both a fully stereo- defined siRNA and a partially stereo-defined ASO.
  • the nucleotides and/or nucleotide analogs are linked to each other by means of phosphorothioate groups.
  • the oligonucleotide comprises a sense strand comprising 5’ RMRRMMRRRRRMRMMMMRM3’, wherein R is RNA and M is 2’-OMe and/or an antisense strand comprising 5’ RRMRRRRRRRRRMRRRR3’, wherein R is RNA and M is 2’-OMe.
  • the oligonucleotide comprises a phosphothioate bond between the first and second nucleotides and the second and third nucleotides of the sense strand and/or the first and second nucleotides, the second and third nucleotides, the 17 th and 18 th nucleotides, and the 18 th and 19 th nucletoides of the antisense strand.
  • the oligonucleotide comprises a phosphothiate bond.
  • the oligonucleotide comprises 5’ methyl cytosine.
  • the oligonucleotide comprises a sense strand comprising 5’ MMMMMMFMFFFMMMMMM3’, wherein M is 2’-OMe and F is 2’-F and/or an antisense strand comprising 5’ MMMFMFMMMMFMMFMMMF3’, wherein M is 2’-OMe and F is 2’-F.
  • the oligonucleotide comprises a 5’ methyl cytosine.
  • the oligonucleotide comprises a phosphothioate bond between the first and second nucleotides and the second and third nucleotides of the sense strand and/or the first and second nucleotides, the 17 th and 18 th nucleotides, and the 18 th and 19 th nucletoides of the antisense strand.
  • the oligonucleotide comprises a phosphothiate bond.
  • the oligonucleotide comprises 5’ methyl cytosine.
  • phosphodiester linkages such as one or two linkages
  • nucleotide analog units typically in region A and or C
  • all remaining linkage groups are either phosphodiester or phosphorothioate, or a mixture thereof.
  • all the internucleotide linkage groups are phosphorothioate.
  • linkages are phosphorothioate linkages
  • alternative linkages such as those disclosed herein can be used, for example phosphate (phosphodiester) linkages can be used, particularly for linkages between nucleotide analogs, such as LNA, units.
  • phosphate (phosphodiester) linkages can be used, particularly for linkages between nucleotide analogs, such as LNA, units.
  • C residues are annotated as 5- 'methyl modified cytosine
  • one or more of the Cs present in the siRNA can be unmodified C residues. II.H.
  • conjugate refers to an oligonucleotide which is covalently linked to a non-nucleotide moiety (conjugate moiety or region C or third region).
  • Conjugation of the oligonucleotide of the disclosure to one or more non-nucleotide moieties may improve the pharmacology of the oligonucleotide, e.g. by affecting the activity, cellular distribution, cellular uptake or stability of the oligonucleotide.
  • the conjugate moiety modify or enhance the pharmacokinetic properties of the oligonucleotide by improving cellular distribution, bioavailability, metabolism, excretion, permeability, and/or cellular uptake of the oligonucleotide.
  • the conjugate may target the oligonucleotide to a specific organ, tissue or cell type and thereby enhance the effectiveness of the oligonucleotide in that organ, tissue or cell type.
  • the conjugate may serve to reduce activity of the oligonucleotide in non-target cell types, tissues or organs, e.g., off target activity or activity in non- target cell types, tissues or organs.
  • WO 93/07883 and WO2013/033230 provides suitable conjugate moieties.
  • Further suitable conjugate moieties are those capable of binding to the asialoglycoprotein receptor (ASGPr).
  • ASGPr asialoglycoprotein receptor
  • tri-valent N-acetylgalactosamine conjugate moieties are suitable for binding to the ASGPr, see for example WO 2014/076196, WO 2014/207232 and WO 2014/179620.
  • Oligonucleotide conjugates and their synthesis has also been reported in comprehensive reviews by Manoharan in Antisense Drug Technology, Principles, Strategies, and Applications, S.T.
  • the non-nucleotide moiety is selected from the group consisting of carbohydrates, cell surface receptor ligands, drug substances, hormones, lipophilic substances, polymers, proteins, peptides, toxins (e.g. bacterial toxins), vitamins, viral proteins (e.g. capsids), and combinations thereof.
  • the present disclosure provides a conjugate comprising an oligonucleotide wherein the non-nucleotide or non-polynucleotide moiety comprises a protein, a fatty acid chain, a sugar residue, a glycoprotein, a polymer, or any combinations thereof.
  • the present disclosure provides a conjugate comprising an oligonucleotide wherein the non-nucleotide or non-polynucleotide moiety comprises a choleteryl moiety, a vinyl phosphonate moiety, or any combination thereof.
  • the present disclosure provides a conjugate comprising an oligonucleotide and a non-nucleotide moiety, wherein the oligonucleotide comprises, consists essentially of, or consists of: 5’-GGUGCAGAUAAUUAAUAAG-3’ (SEQ ID NO: 64) (sense strand) and/or 5’-CUUAUUAAUUAUCUGCACC-3’ (SEQ ID NO: 65) (antisense strand), wherein the 1 st , 3 rd , 4 th , 7 th , 8 th , 9 th , 10 th , 11 th , 13 th , and 18 th nucleotides from 5’ to 3’of the sense strand is RNA and the 2 nd , 5 th , 6 th , 12 th , 14 th , 15 th , 16 th , 17 th , and 19 th nucleotides from 5’ to
  • the present disclosure provides a conjugate comprising an oligonucleotide and a non-nucleotide moiety, wherein the oligonucleotide comprises, consists essentially of, or consists of: 5’-GGUGCAGAUAAUUAAUAAG-3’ (SEQ ID NO: 64) (sense strand) and/or 5’-CUUAUUAAUUAUCUGCACC-3’ (SEQ ID NO: 65) (antisense strand), wherein the 1 st , 3 rd , 4 th , 7 th , 8 th , 9 th , 10 th , 11 th , 13 th , and 18 th nucleotides from 5’ to 3’of the sense strand is RNA and the 2 nd , 5 th , 6 th , 12 th , 14 th , 15 th , 16 th , 17 th , and 19 th nucleotides from 5’ to
  • the present disclosure provides a conjugate comprising an oligonucleotide and a non-nucleotide moiety, wherein the oligonucleotide comprises, consists essentially of, or consists of: 5’-GGUGCAGAUAAUUAAUAAG-3’ (SEQ ID NO: 64) (sense strand) and/or 5’-CUUAUUAAUUAUCUGCACC-3’ (SEQ ID NO: 65) (antisense strand), wherein the 1 st , 2 nd , 3 rd , 4 th , 5 th , 6 th , 8 th , 12 th , 13 th , 14 th , 15 th , 16 th , 17 th , 18 th and 19 th nucleotides from 5’ to 3’of the sense strand comprise 2’-OMe at the sugar position and the 7 th , 9 th , 10 th and 11 th nucleotides from
  • the present disclosure provides a conjugate comprising an oligonucleotide and a non-nucleotide moiety, wherein the oligonucleotide comprises, consists essentially of, or consists of: 5’-GGUGCAGAUAAUUAAUAAG-3’ (SEQ ID NO: 64) (sense strand) and/or 5’-CUUAUUAAUUAUCUGCACC-3’ (SEQ ID NO: 65) (antisense strand), wherein the 1 st , 2 nd , 3 rd , 4 th , 5 th , 6 th , 8 th , 12 th , 13 th , 14 th , 15 th , 16 th , 17 th , 18 th and 19 th nucleotides from 5’ to 3’of the sense strand comprise 2’-OMe at the sugar position and the 7 th , 9 th , 10 th and 11 th nucleotides from
  • the present disclosure provides a conjugate comprising an oligonucleotide and a non-nucleotide moiety, wherein the oligonucleotide comprises, consists essentially of, or consists of: 5’-AGCGGCUACAGCAGCCCCG -3’ (SEQ ID NO: 9) (sense strand) and 5’-CGGGGCUGCUGUAGCCGCU-3’ (SEQ ID NO: 10) (antisense strand), wherein the 1 st , 3 rd , 4 th , 7 th , 8 th , 9 th , 10 th , 11 th , 13 th , and 18 th nucleotides from 5’ to 3’ of the sense strand is RNA and the 2 nd , 5 th , 6 th , 12 th , 14 th , 15 th , 16 th , 17 th , and 19 th nucleotides from 5’ to 3
  • the present disclosure provides a conjugate comprising an oligonucleotide and a non-nucleotide moiety, wherein the oligonucleotide comprises, consists essentially of, or consists of: 5’-AGCGGCUACAGCAGCCCCG -3’ (SEQ ID NO: 9) (sense strand) and 5’-CGGGGCUGCUGUAGCCGCU-3’ (SEQ ID NO: 10) (antisense strand), wherein the 1 st , 3 rd , 4 th , 7 th , 8 th , 9 th , 10 th , 11 th , 13 th , and 18 th nucleotides from 5’ to 3’ of the sense strand is RNA and the 2 nd , 5 th , 6 th , 12 th , 14 th , 15 th , 16 th , 17 th , and 19 th nucleotides from 5’ to 3
  • the present disclosure provides a conjugate comprising an oligonucleotide and a non-nucleotide moiety, wherein the oligonucleotide comprises, consists essentially of, or consists of: 5’-AGCGGCUACAGCAGCCCCG -3’ (SEQ ID NO: 9) (sense strand) and 5’-CGGGGCUGCUGUAGCCGCU-3’ (SEQ ID NO: 10) (antisense strand), wherein the 1 st , 2 nd , 3 rd , 4 th , 5 th , 6 th , 8 th , 12 th , 13 th , 14 th , 15 th , 16 th , 17 th , 18 th and 19 th nucleotides from 5’ to 3’of the sense strand comprise 2’-OMe at the sugar position and the 7 th , 9 th , 10 th and 11 th nucleotides from 5
  • the present disclosure provides a conjugate comprising an oligonucleotide and a non-nucleotide moiety, wherein the oligonucleotide comprises, consists essentially of, or consists of: 5’-AGCGGCUACAGCAGCCCCG -3’ (SEQ ID NO: 9) (sense strand) and 5’-CGGGGCUGCUGUAGCCGCU-3’ (SEQ ID NO: 10) (antisense strand), wherein the 1 st , 2 nd , 3 rd , 4 th , 5 th , 6 th , 8 th , 12 th , 13 th , 14 th , 15 th , 16 th , 17 th , 18 th and 19 th nucleotides from 5’ to 3’of the sense strand comprise 2’-OMe at the sugar position and the 7 th , 9 th , 10 th and 11 th nucleotides from 5
  • the present disclosure provides a conjugate comprising an oligonucleotide and a non-nucleotide moiety, wherein the oligonucleotide comprises, consists essentially of, or consists of: 5’-AGGAGGUGGCCAGGUGGAA-3’ (SEQ ID NO: 23) (sense strand) and 5’-UUCCACCUGGCCACCUCCU-3’ (SEQ ID NO: 24) (antisense strand), wherein the 1 st , 3 rd , 4 th , 7 th , 8 th , 9 th , 10 th , 11 th , 13 th , and 18 th nucleotides from 5’ to 3’of the sense strand is RNA and the 2 nd , 5 th , 6 th , 12 th , 14 th , 15 th , 16 th , 17 th , and 19 th nucleotides from 5’ to 3’
  • the present disclosure provides a conjugate comprising an oligonucleotide and a non-nucleotide moiety, wherein the oligonucleotide comprises, consists essentially of, or consists of: 5’-AGGAGGUGGCCAGGUGGAA-3’ (SEQ ID NO: 23) (sense strand) and 5’-UUCCACCUGGCCACCUCCU-3’ (SEQ ID NO: 24) (antisense strand), wherein the 1 st , 3 rd , 4 th , 7 th , 8 th , 9 th , 10 th , 11 th , 13 th , and 18 th nucleotides from 5’ to 3’of the sense strand is RNA and the 2 nd , 5 th , 6 th , 12 th , 14 th , 15 th , 16 th , 17 th , and 19 th nucleotides from 5’ to 3’
  • the present disclosure provides a conjugate comprising an oligonucleotide and a non-nucleotide moiety, wherein the oligonucleotide comprises, consists essentially of, or consists of: 5’-AGGAGGUGGCCAGGUGGAA-3’ (SEQ ID NO: 23) (sense strand) and 5’-UUCCACCUGGCCACCUCCU-3’ (SEQ ID NO: 24) (antisense strand), wherein the 1 st , 2 nd , 3 rd , 4 th , 5 th , 6 th , 8 th , 12 th , 13 th , 14 th , 15 th , 16 th , 17 th , 18 th and 19 th nucleotides from 5’ to 3’of the sense strand comprise 2’-OMe at the sugar position and the 7 th , 9 th , 10 th and 11 th nucleotides from 5’
  • the present disclosure provides a conjugate comprising an oligonucleotide and a non-nucleotide moiety, wherein the oligonucleotide comprises, consists essentially of, or consists of: 5’-AGGAGGUGGCCAGGUGGAA-3’ (SEQ ID NO: 23) (sense strand) and 5’-UUCCACCUGGCCACCUCCU-3’ (SEQ ID NO: 24) (antisense strand), wherein the 1 st , 2 nd , 3 rd , 4 th , 5 th , 6 th , 8 th , 12 th , 13 th , 14 th , 15 th , 16 th , 17 th , 18 th and 19 th nucleotides from 5’ to 3’of the sense strand comprise 2’-OMe at the sugar position and the 7 th , 9 th , 10 th and 11 th nucleotides from 5’
  • the present disclosure provides a conjugate comprising an oligonucleotide and a non-nucleotide moiety, wherein the oligonucleotide comprises, consists essentially of, or consists of: 5’-UGGCCAGGUGGAAGUAAAA-3’ (SEQ ID NO: 27) (sense strand) and 5’-UUUUACUUCCACCUGGCCA-3’ (SEQ ID NO: 28) (antisense strand); wherein the 1 st , 3 rd , 4 th , 7 th , 8 th , 9 th , 10 th , 11 th , 13 th , and 18 th nucleotides from 5’ to 3’of the sense strand is RNA and the 2 nd , 5 th , 6 th , 12 th , 14 th , 15 th , 16 th , 17 th , and 19 th nucleotides from 5’ to 3
  • the present disclosure provides a conjugate comprising an oligonucleotide and a non-nucleotide moiety, wherein the oligonucleotide comprises, consists essentially of, or consists of: 5’-UGGCCAGGUGGAAGUAAAA-3’ (SEQ ID NO: 27) (sense strand) and 5’-UUUUACUUCCACCUGGCCA-3’ (SEQ ID NO: 28) (antisense strand); wherein the 1 st , 3 rd , 4 th , 7 th , 8 th , 9 th , 10 th , 11 th , 13 th , and 18 th nucleotides from 5’ to 3’of the sense strand is RNA and the 2 nd , 5 th , 6 th , 12 th , 14 th , 15 th , 16 th , 17 th , and 19 th nucleotides from 5’ to 3
  • the present disclosure provides a conjugate comprising an oligonucleotide and a non-nucleotide moiety, wherein the oligonucleotide comprises, consists essentially of, or consists of: 5’-UGGCCAGGUGGAAGUAAAA-3’ (SEQ ID NO: 27) (sense strand) and 5’-UUUUACUUCCACCUGGCCA-3’ (SEQ ID NO: 28) (antisense strand); wherein the 1 st , 2 nd , 3 rd , 4 th , 5 th , 6 th , 8 th , 12 th , 13 th , 14 th , 15 th , 16 th , 17 th , 18 th , and 19 th nucleotides from 5’ to 3’of the sense strand comprises 2’ OMe at the sugar position and the 7 th , 9 th , 10 th , and 11 th nucleo
  • the conjugate comprises an oligonucleotide and a non nucleotide moiety, wherein the oligonucleotide comprises a sense strand comprising 5’ RMRRMMRRRRRMRMMMMRM3’, wherein R is RNA and M is 2’-OMe and/or an antisense strand comprising 5’ RRMRRRRRRRRRMRRRR3’, wherein R is RNA and M is 2’-OMe.
  • the oligonucleotide comprises a phosphothioate bond between the first and second nucleotides and the second and third nucleotides of the sense strand and/or the first and second nucleotides, the 17 th and 18 th nucleotides, and the 18 th and 19 th nucletoides of the antisense strand.
  • the oligonucleotide in the conjugate comprises a phosphothiate bond.
  • the oligonucleotide comprises 5’ methyl cytosine.
  • the non nucleotide moiety in the conjugate is a cholesteryl moiety linked to the sense or antisense strand.
  • the non nucleotide moiety comprises a vinyl phosphonate moiety.
  • the conjugate comprises an oligonucleotide and a non nucleotide moiety, wherein the oligonucleotide comprises a sense strand comprising 5’ MMMMMMFMFFFMMMMMM3’, wherein M is 2’-OMe and F is 2’-F and/or an antisense strand comprising 5’ MMMFMFMMMMFMMFMMMF3’, wherein M is 2’-OMe and F is 2’-F.
  • the oligonucleotide comprises a 5’ methyl cytosine.
  • the non nucleotide moiety comprises a vinyl phosphonate moiety.
  • the oligonucleotide, e.g., siRNA, of the disclosure can be used in pharmaceutical formulations and compositions. Suitably, such compositions comprise a pharmaceutically acceptable diluent, carrier, salt or adjuvant.
  • the oligonucleotide, e.g., siRNA, of the disclosure can be included in a unit formulation such as in a pharmaceutically acceptable carrier or diluent in an amount sufficient to deliver to a patient a therapeutically effective amount without causing serious side effects in the treated patient.
  • the formulated drug may comprise pharmaceutically acceptable binding agents and adjuvants.
  • Capsules, tablets, or pills can contain for example the following compounds: microcrystalline cellulose, gum or gelatin as binders; starch or lactose as excipients; stearates as lubricants; various sweetening or flavoring agents.
  • the dosage unit may contain a liquid carrier like fatty oils.
  • coatings of sugar or enteric agents may be part of the dosage unit.
  • the oligonucleotide formulations can also be emulsions of the active pharmaceutical ingredients and a lipid forming a micellular emulsion.
  • compositions of the present disclosure can be administered in a number of ways depending upon whether local or systemic treatment is desired and upon the area to be treated.
  • Administration can be (a) oral (b) pulmonary, e.g., by inhalation or insufflation of powders or aerosols, including by nebulizer; intratracheal, intranasal, (c) topical including epidermal, transdermal, ophthalmic and to mucous membranes including vaginal and rectal delivery; or (d) parenteral including intravenous, intraarterial, subcutaneous, intraperitoneal or intramuscular injection or infusion; or intracranial, e.g., intrathecal, intra-cerebroventricular, or intraventricular, administration.
  • the oligonucleotide e.g., siRNA
  • the oligonucleotide e.g., siRNA
  • Pharmaceutical compositions and formulations for topical administration can include transdermal patches, ointments, lotions, creams, gels, drops, sprays, suppositories, liquids and powders. Conventional pharmaceutical carriers, aqueous, powder or oily bases, thickeners and the like may be necessary or desirable.
  • topical formulations include those in which the oligonucleotide, e.g., siRNA, of the disclosure are in admixture with a topical delivery agent such as lipids, liposomes, fatty acids, fatty acid esters, steroids, chelating agents and surfactants.
  • a topical delivery agent such as lipids, liposomes, fatty acids, fatty acid esters, steroids, chelating agents and surfactants.
  • compositions and formulations for oral administration include but are not limited to powders or granules, microparticulates, nanoparticulates, suspensions or solutions in water or non-aqueous media, capsules, gel capsules, sachets, tablets or minitablets.
  • compositions and formulations for parenteral, intrathecal, intra-cerebroventricular, or intraventricular administration can include sterile aqueous solutions which can also contain buffers, diluents and other suitable additives such as, but not limited to, penetration enhancers, carrier compounds and other pharmaceutically acceptable carriers or excipients.
  • sterile aqueous solutions which can also contain buffers, diluents and other suitable additives such as, but not limited to, penetration enhancers, carrier compounds and other pharmaceutically acceptable carriers or excipients.
  • Pharmaceutical compositions of the present disclosure include, but are not limited to, solutions, emulsions, and liposome-containing formulations. These compositions may be generated from a variety of components that include, but are not limited to, preformed liquids, self- emulsifying solids and self-emulsifying semisolids.
  • Delivery of drug to the target tissue can be enhanced by carrier-mediated delivery including, but not limited to, cationic liposomes, cyclodextrins, porphyrin derivatives, branched chain dendrimers, polyethylenimine polymers, nanoparticles and microspheres (Dass CR. J Pharm Pharmacol 2002; 54(1):3-27).
  • carrier-mediated delivery including, but not limited to, cationic liposomes, cyclodextrins, porphyrin derivatives, branched chain dendrimers, polyethylenimine polymers, nanoparticles and microspheres.
  • the formulations are prepared by uniformly and intimately bringing into association the active ingredients with liquid carriers or finely divided solid carriers or both, and then, if necessary, shaping the product.
  • the formulation can include a sterile diluent, buffers, regulators of tonicity and antibacterials.
  • the active oligonucleotides e.g., siRNAs
  • the carriers can be physiological saline or phosphate buffered saline.
  • WO2007/031091 (A2), published March 22, 2007, further provides suitable pharmaceutically acceptable diluent, carrier and adjuvants - which are hereby incorporated by reference.
  • IV. Diagnostics [0517] This disclosure further provides a diagnostic method useful during diagnosis of MAPT related diseases, e.g., a tauopathy.
  • Non-limiting examples of tauopathy include, but are not limited to, behavioral variant frontotemporal dementia, primary progressive aphasia, corticobasal syndrome, Richardson’s syndrome, Pick’s disease, Parkinsonism linked to chromosome 17, Primary age related tauopathy, Tangle-predominant dementia, Corticobasal degeneration, Progressive supranuclear palsy, Alzheimer’s disease, Parkinson’s disease, amyotrophic lateral sclerosis, Argyrophilic grain disease, Aging-related tau ⁇ astrogliopathy, and globular glial tauopathy.
  • the oligonucleotides, e.g., siRNAs, of the disclosure can be used to measure expression of MAPT transcript in a tissue or body fluid from an individual and comparing the measured expression level with a standard MAPT transcript expression level in normal tissue or body fluid, whereby an increase in the expression level compared to the standard is indicative of a disorder treatable by an oligonucleotide, e.g., siRNA, of the disclosure.
  • the oligonucleotides, e.g., siRNAs, of the disclosure can be used to assay MAPT transcript levels in a biological sample using any methods known to those of skill in the art. (Touboul et. al., Anticancer Res.
  • kits comprising ASOs
  • This disclosure further provides kits that comprise an oligonucleotide, e.g., siRNA, of the disclosure described herein and that can be used to perform the methods described herein.
  • kits comprises at least one oligonucleotide, e.g., siRNA, in one or more containers.
  • the kits contain all of the components necessary and/or sufficient to perform a detection assay, including all controls, directions for performing assays, and any necessary software for analysis and presentation of results.
  • the disclosed oligonucleotide, e.g., siRNA can be readily incorporated into one of the established kit formats which are well known in the art. VI. Methods of Using [0522]
  • the oligonucleotides, e.g., siRNAs, of the disclosure can be utilized for therapeutics and prophylaxis.
  • MAPT is a 352-441 amino acid protein preferentially expressed in neurons at pre- synaptic terminals where it is thought to play a role in regulating synaptic transmission.
  • MAPT is natively unfolded, but can form a “paperclip” like structure. Preclusion of this structure via post- translational modifications, truncations, and mutations can cause abnormal aggregation of the protein.
  • P301L One mutation that has been extensively studied is P301L. This mutation is associated with misfolding and hyperphosphorylation of the protein leading to the formation of neurofibrillary tangles.
  • tauopathies consist of aggregated, insoluble accumulations of misfolded MAPT proteins and are a characteristic feature of some members of a group of neurodegenerative diseases collectively known as tauopathies, notably Dementia and Alzheimer’s Disease.
  • Other tauopathies are characterized by Pick bodies, astrocytic plaques, tufted astrocytes, and astrocytic inclusions.
  • MAPT can form pathological aggregates in neurons known as Neurofibrillary Tangles, which are characteristic of both Alzheimer’s Disease and Dementia.
  • the present oligonucleotides e.g., siRNAs
  • the oligonucleotides, e.g., siRNAs, of the disclosure reduce the number of neurofibrillary tangles, Pick bodies, astrocytic plaques, tufted astrocytes, and astrocytic inclusions, or prevent formation of neurofibrillary tangle, Pick bodies, astrocytic plaques, tufted astrocytes, and astrocytic inclusions, .
  • oligodendrocytes Reports of either undetectable or low levels of MAPT mRNA expression in oligodendrocytes suggest that some pathological form of MAPT is propagated from neurons, where it is highly expressed, to oligodendrocytes.
  • the oligonucleotides, e.g., siRNAs, of the disclosure reduce or prevent propagation of MAPT, e.g., pathological form of MAPT, from neurons.
  • the oligonucleotides can be used in research, e.g., to specifically inhibit the synthesis of MAPT protein (typically by degrading or inhibiting the mRNA and thereby prevent protein formation) in cells and experimental animals thereby facilitating functional analysis of the target or an appraisal of its usefulness as a target for therapeutic intervention.
  • methods of down-regulating the expression of MAPT mRNA and/or MAPT protein in cells or tissues comprising contacting the cells or tissues, in vitro or in vivo, with an effective amount of one or more of the oligonucleotides, e.g., siRNAs, conjugates or compositions of the disclosure.
  • an animal or a human, suspected of having a disease or disorder, which can be treated by modulating the expression of MAPT transcript and/or MAPT protein is treated by administering oligonucleotide, e.g., siRNA, compounds in accordance with this disclosure.
  • oligonucleotide e.g., siRNA
  • methods of treating a mammal, such as treating a human, suspected of having or being prone to a disease or condition, associated with expression of MAPT transcript and/or MAPT protein by administering a therapeutically or prophylactically effective amount of one or more of the oligonucleotides, e.g., siRNAs, or compositions of the disclosure.
  • the oligonucleotide e.g., siRNA, a conjugate or a pharmaceutical composition according to the disclosure is typically administered in an effective amount.
  • the oligonucleotide, e.g., siRNA, or conjugate of the disclosure is used in therapy.
  • the disclosure further provides for an oligonucleotide, e.g., siRNA, according to the disclosure, for use in treating one or more of the diseases referred to herein, such as a disease selected from the group consisting of behavioral variant frontotemporal dementia, primary progressive aphasia, corticobasal syndrome, Richardson’s syndrome, Pick’s disease, Parkinsonism linked to chromosome 17, Primary age related tauopathy, Tangle-predominant dementia, Corticobasal degeneration, Progressive supranuclear palsy, globular glial tauopathy, Alzheimer’s disease, Parkinson’s disease, amyotrophic lateral sclerosis, Argyrophilic grain disease, Aging- related tau ⁇ astrogliopathy, and any combinations thereof.
  • a disease selected from the group consisting of behavioral variant frontotemporal dementia, primary progressive aphasia, corticobasal syndrome, Richardson’s syndrome, Pick’s disease, Parkinsonism linked to chromosome 17, Primary age related tauopathy, Tang
  • the disclosure further provides for a method for treating tauopathies, the method comprising administering an effective amount of one or more oligonucleotides, e.g., siRNAs, conjugates, or pharmaceutical compositions thereof to an animal in need thereof (such as a patient in need thereof).
  • the disease, disorder, or condition is associated with overexpression of MAPT gene transcript and/or MAPT protein.
  • the disclosure also provides for methods of inhibiting (e.g., by reducing) the expression of MAPT gene transcript and/or MAPT protein in a cell or a tissue, the method comprising contacting the cell or tissue, in vitro or in vivo, with an effective amount of one or more oligonucleotides, e.g., siRNAs, conjugates, or pharmaceutical compositions thereof, of the disclosure to affect degradation of expression of MAPT gene transcript thereby reducing MAPT protein.
  • oligonucleotides e.g., siRNAs, conjugates, or pharmaceutical compositions thereof
  • the oligonucleotides are used to reduce the expression of MAPT mRNA in one or more sections of brain, e.g., hippocampus, brainstem, striatum, or any combinations thereof.
  • the oligonucleotides reduce the expression of MAPT mRNA, e.g., in brain stem and/or striatum, less than 70%, less than 60%, less than 50%, less than 40%, less than 30%, less than 20%, less than 10%, or less than 5% compared to the MAPT mRNA expression after administration of or exposure to a vehicle (no ASO), at day 3, day 5, day 7, day 10, day 14, day 15, day 20, day 21, or day 25.
  • a vehicle no ASO
  • the expression of MAPT mRNA is maintained below 70%, below 60%, below 50%, below 40%, below 30%, below 20%, below 10%, or below 5% compared to the MAPT mRNA expression after administration of or exposure to a vehicle (no oligonucleotide) until day 28, day 30, day 32, day 35, day 40, day 42, day 45, day 49, day 50, day 56, day 60, day 63, day 70, or day 75.
  • a vehicle no oligonucleotide
  • the oligonucleotides, e.g., siRNAs, of the present disclosure reduces MAPT mRNA and/or MAPT protein expression in medulla, caudate putamen, pons cerebellum, lumbar spinal cord, frontal cortex, and/or any combinations thereof.
  • the disclosure also provides for the use of the oligonucleotide, e.g., siRNA, or conjugate of the disclosure as described for the manufacture of a medicament.
  • the disclosure also provides for a composition comprising the oligonucleotide, e.g., siRNA, or conjugate thereof for use in treating a disorder as referred to herein, or for a method of the treatment of as a disorder as referred to herein.
  • the present disclosure also provides oligonucleotides, e.g., siRNAs, or conjugates for use in therapy.
  • the present disclosure additionally provides oligonucleotides, e.g., siRNAs, or conjugates for use in the treatment of tauopathies.
  • the disclosure further provides for a method for inhibiting MAPT protein in a cell which is expressing MAPT comprising administering an oligonucleotide, e.g., siRNA, or a conjugate according to the disclosure to the cell so as to affect the inhibition of MAPT protein in the cell.
  • the disclosure includes a method of reducing, ameliorating, preventing, or treating neuronal hyperexcitability in a subject in need thereof comprising administering an oligonucleotide, e.g., siRNA, or a conjugate according to the disclosure.
  • the disclosure also provides for a method for treating a disorder as referred to herein the method comprising administering an oligonucleotide, e.g., siRNA, or a conjugate according to the disclosure as herein described and/or a pharmaceutical composition according to the disclosure to a patient in need thereof.
  • an oligonucleotide e.g., siRNA
  • the oligonucleotides, e.g., siRNAs, and other compositions according to the disclosure can be used for the treatment of conditions associated with over expression or expression of mutated version of MAPT protein.
  • the disclosure provides for the oligonucleotide, e.g., siRNA, or the conjugate according to disclosure, for use as a medicament, such as for the treatment of tauopathies.
  • the tauopathy is a disease selected from the group consisting of behavioral variant frontotemporal dementia, primary progressive aphasia, corticobasal syndrome, Richardson’s syndrome, Pick’s disease, Parkinsonism linked to chromosome 17, Primary age related tauopathy, Tangle-predominant dementia, Corticobasal degeneration, Progressive supranuclear palsy, and globular glial tauopathy, Alzheimer’s disease, Parkinson’s disease, amyotrophic lateral sclerosis, Argyrophilic grain disease, Aging-related tau ⁇ astrogliopathy, and any combinations thereof.
  • the disclosure further provides use of an oligonucleotide, e.g., siRNA, of the disclosure in the manufacture of a medicament for the treatment of a disease, disorder or condition as referred to herein.
  • an oligonucleotide e.g., siRNA
  • the oligonucleotide, e.g., siRNA, or conjugate of the disclosure is used for the manufacture of a medicament for the treatment of a tauopathy.
  • one aspect of the disclosure is directed to a method of treating a mammal suffering from or susceptible to conditions associated with abnormal levels of MAPT i.e., a tauopathy), comprising administering to the mammal a therapeutically effective amount of an oligonucleotide, e.g., siRNA, targeted to MAPT transcript that comprises one or more LNA units.
  • an oligonucleotide e.g., siRNA
  • the oligonucleotide, e.g., siRNA, a conjugate or a pharmaceutical composition according to the disclosure is typically administered in an effective amount.
  • the disease or disorder as referred to herein, can, in some aspects be associated with a mutation in the MAPT gene or a gene whose protein product is associated with or interacts with MAPT protein.
  • the target MAPT is a mutated form or a non human species of the MAPT sequence.
  • An interesting aspect of the disclosure is directed to the use of an oligonucleotide, e.g., siRNA, (compound) as defined herein or a conjugate as defined herein for the preparation of a medicament for the treatment of a disease, disorder or condition as referred to herein.
  • the methods of the disclosure can be employed for treatment or prophylaxis against diseases caused by abnormal levels of MAPT protein.
  • diseases caused by abnormal levels of MAPT protein are tauopathies.
  • tauopathies include behavioral variant frontotemporal dementia, primary progressive aphasia, corticobasal syndrome, Richardson’s syndrome, Pick’s disease, Parkinsonism linked to chromosome 17, Primary age related tauopathy, Tangle-predominant dementia, Corticobasal degeneration, Progressive supranuclear palsy, globular glial tauopathy, Alzheimer’s disease, Parkinson’s disease, amyotrophic lateral sclerosis, Argyrophilic grain disease, and Aging-related tau ⁇ astrogliopathy.
  • the disclosure is furthermore directed to a method for treating abnormal levels of MAPT protein, the method comprising administering an oligonucleotide, e.g., siRNA, of the disclosure, or a conjugate of the disclosure or a pharmaceutical composition of the disclosure to a patient in need thereof.
  • an oligonucleotide e.g., siRNA
  • the disclosure also relates to an oligonucleotide, e.g., siRNA, a composition or a conjugate as defined herein for use as a medicament.
  • the disclosure further relates to use of a compound, composition, or a conjugate as defined herein for the manufacture of a medicament for the treatment of abnormal levels of MAPT protein or expression of mutant forms of MAPT protein (such as allelic variants, such as those associated with one of the diseases referred to herein).
  • a patient who is in need of treatment is a patient suffering from or likely to suffer from the disease or disorder.
  • VII. Vectors and Delivery Systems [0548]
  • the oligonucleotides, e.g., siRNAs, of the present disclosure can be administered, e.g., to a subject suffering from a disease or condition associated with abnormal (e.g., increased) inflammasome activity, using any relevant delivery system known in the art.
  • the delivery system is a vector.
  • the present disclosure provides a vector comprising an oligonucleotide, e.g., siRNA, of the present disclosure.
  • the vector is viral vector.
  • the viral vector is an adenoviral vector or an adenoassociated viral vector.
  • the viral vector is an AAV that has a serotype of AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, or any combination thereof.
  • the adenoviral vector is a third generation adenoviral vector.
  • ADEASYTM is by far the most popular method for creating adenoviral vector constructs.
  • the system consists of two types of plasmids: shuttle (or transfer) vectors and adenoviral vectors.
  • the transgene of interest is cloned into the shuttle vector, verified, and linearized with the restriction enzyme PmeI.
  • This construct is then transformed into ADEASIER-1 cells, which are BJ5183 E. coli cells containing PADEASYTM.
  • PADEASYTM is a ⁇ 33Kb adenoviral plasmid containing the adenoviral genes necessary for virus production.
  • the shuttle vector and the adenoviral plasmid have matching left and right homology arms which facilitate homologous recombination of the transgene into the adenoviral plasmid.
  • Recombinant adenoviral plasmids are then verified for size and proper restriction digest patterns to determine that the transgene has been inserted into the adenoviral plasmid, and that other patterns of recombination have not occurred. Once verified, the recombinant plasmid is linearized with PacI to create a linear dsDNA construct flanked by ITRs. 293 or 911 cells are transfected with the linearized construct, and virus can be harvested about 7-10 days later.
  • the viral vector is a retroviral vector, e.g., a lentiviral vector (e.g., a third or fourth generation lentiviral vector).
  • Lentiviral vectors are usually created in a transient transfection system in which a cell line is transfected with three separate plasmid expression systems. These include the transfer vector plasmid (portions of the HIV provirus), the packaging plasmid or construct, and a plasmid with the heterologous envelop gene (env) of a different virus.
  • the three plasmid components of the vector are put into a packaging cell which is then inserted into the HIV shell.
  • the virus portions of the vector contain insert sequences so that the virus cannot replicate inside the cell system.
  • Current third generation lentiviral vectors encode only three of the nine HIV-1 proteins (Gag, Pol, Rev), which are expressed from separate plasmids to avoid recombination-mediated generation of a replication-competent virus.
  • fourth generation lentiviral vectors the retroviral genome has been further reduced (see, e.g., TAKARA® LENTI-XTM fourth- generation packaging systems).
  • Any AAV vector known in the art can be used in the methods disclosed herein.
  • the AAV vector can comprise a known vector or can comprise a variant, fragment, or fusion thereof.
  • the AAV vector is selected from the group consisting of AAV type 1 (AAV1), AAV2, AAV3A, AVV3B, AAV4, AAV5, AAV6, AAV7, AAV8, AVV9, AVV10, AVV11, AVV12, AVV13, AAVrh.74, avian AAV, bovine AAV, canine AAV, equine AAV, goat AVV, primate AAV, non-primate AAV, bovine AAV, shrimp AVV, snake AVV, and any combination thereof.
  • the AAV vector is derived from an AAV vector selected from the group consisting of AAV1, AAV2, AAV3A, AVV3B, AAV4, AAV5, AAV6, AAV7, AAV8, AVV9, AVV10, AVV11, AVV12, AVV13, AAVrh.74, avian AAV, bovine AAV, canine AAV, equine AAV, goat AVV, primate AAV, non-primate AAV, ovine AAV, shrimp AVV, snake AVV, and any combination thereof.
  • the AAV vector is a chimeric vector derived from at least two AAV vectors selected from the group consisting of AAV1, AAV2, AAV3A, AVV3B, AAV4, AAV5, AAV6, AAV7, AAV8, AVV9, AVV10, AVV11, AVV12, AVV13, AAVrh.74, avian AAV, bovine AAV, canine AAV, equine AAV, goat AVV, primate AAV, non-primate AAV, ovine AAV, shrimp AVV, snake AVV, and any combination thereof.
  • the AAV vector comprises regions of at least two different AAV vectors known in the art.
  • the AAV vector comprises an inverted terminal repeat from a first AAV (e.g., AAV1, AAV2, AAV3A, AVV3B, AAV4, AAV5, AAV6, AAV7, AAV8, AVV9, AVV10, AVV11, AVV12, AVV13, AAVrh.74, avian AAV, bovine AAV, canine AAV, equine AAV, goat AVV, primate AAV, non-primate AAV, ovine AAV, shrimp AVV, snake AVV, or any derivative thereof) and a second inverted terminal repeat from a second AAV (e.g., AAV1, AAV2, AAV3A, AVV3B, AAV4, AAV5, AAV6, AAV7, AAV8, AVV9, AVV10, AVV11, AVV12, AVV13, AAVrh.74, avian AAV, bovine AAV, canine AAV, equine A
  • the AVV vector comprises a portion of an AAV vector selected from the group consisting of AAV1, AAV2, AAV3A, AVV3B, AAV4, AAV5, AAV6, AAV7, AAV8, AVV9, AVV10, AVV11, AVV12, AVV13, AAVrh.74, avian AAV, bovine AAV, canine AAV, equine AAV, goat AVV, primate AAV, non-primate AAV, ovine AAV, shrimp AVV, snake AVV, and any combination thereof.
  • the AAV vector comprises AAV2.
  • the AVV vector comprises a splice acceptor site.
  • the AVV vector comprises a promoter. Any promoter known in the art can be used in the AAV vector of the present disclosure.
  • the promoter is an RNA Pol III promoter.
  • the RNA Pol III promoter is selected from the group consisting of the U6 promoter, the H1 promoter, the 7SK promoter, the 5S promoter, the adenovirus 2 (Ad2) VAI promoter, and any combination thereof.
  • the promoter is a cytomegalovirus immediate-early gene (CMV) promoter, an EF1a promoter, an SV40 promoter, a PGK1 promoter, a Ubc promoter, a human beta actin promoter, a CAG promoter, a TRE promoter, a UAS promoter, a Ac5 promoter, a polyhedrin promoter, a CaMKIIa promoter, a GAL1 promoter, a GAL10 promoter, a TEF promoter, a GDS promoter, a ADH1 promoter, a CaMV35S promoter, or a Ubi promoter.
  • the promoter comprises the U6 promoter.
  • the AAV vector comprises a constitutively active promoter (constitutive promoter).
  • the constitutive promoter is selected from the group consisting of hypoxanthine phosphoribosyl transferase (HPRT), adenosine deaminase, pyruvate kinase, beta-actin promoter, cytomegalovirus (CMV), simian virus (e.g., SV40), papilloma virus, adenovirus, human immunodeficiency virus (HIV), Rous sarcoma virus, a retrovirus long terminal repeat (LTR), Murine stem cell virus (MSCV) and the thymidine kinase promoter of herpes simplex virus.
  • HPRT hypoxanthine phosphoribosyl transferase
  • CMV cytomegalovirus
  • simian virus e.g., SV40
  • papilloma virus adenovirus
  • the promoter is an inducible promoter.
  • the inducible promoter is a tissue specific promoter.
  • the tissue specific promoter drives transcription of the coding region of the AVV vector in a neuron, a glial cell, or in both a neuron and a glial cell.
  • the AVV vector comprises one or more enhancers.
  • the one or more enhancer are present in the AAV alone or together with a promoter disclosed herein.
  • the AAV vector comprises a 3'UTR poly(A) tail sequence.
  • the 3'UTR poly(A) tail sequence is selected from the group consisting of bGH poly(A), actin poly(A), hemoglobin poly(A), and any combination thereof. In some aspects, the 3'UTR poly(A) tail sequence comprises bGH poly(A). [0561] In some aspects, an oligonucleotide, e.g., siRNA, disclosed herein is administered with a delivery agent.
  • Non-limiting examples of delivery agents that can be used include an exosome, a lipidoid, a liposome, a lipoplex, a lipid nanoparticle, an extracellular vesicle, a synthetic vesicle, a polymeric compound, a peptide, a protein, a cell, a nanoparticle mimic, a nanotube, a micelle, a viral vector, or a conjugate.
  • the present disclosure also provides a composition comprising an oligonucleotide, e.g., siRNA, of the present disclosure and a delivery agent.
  • the delivery agent comprises a carrier unit, e.g., that can self-assemble into micelles or be incorporated into micelles.
  • the delivery agent comprises a cationic carrier unit comprising [WP]-L1-[CC]-L2-[AM] (formula I) or [WP]-L1-[AM]-L2-[CC] (formula II) wherein WP is a water-soluble biopolymer moiety; CC is a positively charged (i.e., cationic) carrier moiety; AM is an adjuvant moiety; and, L1 and L2 are independently optional linkers, and wherein when mixed with a nucleic acid at an ionic ratio of about 1:1, the cationic carrier unit forms a micelle.
  • the oligonucleotide, e.g., siRNA, and the cationic carrier unit are capable of associating with each other (e.g., via a covalent bond or a non-valent bond) to form a micelle when mixed together.
  • composition comprising an oligonucleotide, e.g., siRNA, of the present disclosure interacts with the cationic carrier unit via an ionic bond.
  • the water-soluble biopolymer moiety comprises poly(alkylene glycols), poly(oxyethylated polyol), poly(olefinic alcohol), poly(vinylpyrrolidone), poly(hydroxyalkylmethacrylamide), poly(hydroxyalkylmethacrylate), poly(saccharides), poly( ⁇ - hydroxy acid), poly(vinyl alcohol), polyglycerol, polyphosphazene, polyoxazolines (“POZ”) poly(N-acryloylmorpholine), or any combinations thereof.
  • the water-soluble polymer comprises polyethylene glycol (“PEG”), polyglycerol, or poly(propylene glycol) (“PPG").
  • the water-soluble biopolymer moiety comprises: , (formula III), wherein n is 1-1000.
  • the n is at least about 110, at least about 111, at least about 112, at least about 113, at least about 114, at least about 115, at least about 116, at least about 117, at least about 118, at least about 119, at least about 120, at least about 121, at least about 122, at least about 123, at least about 124, at least about 125, at least about 126, at least about 127, at least about 128, at least about 129, at least about 130, at least about 131, at least about 132, at least about 133, at least about 134, at least about 135, at least about 136, at least about 137, at least about 138, at least about 139, at least about 140, or at least about 141.
  • the n is about 80 to about 90, about 90 to about 100, about 100 to about 110, about 110 to about 120, about 120 to about 130, about 140 to about 150, about 150 to about 160.
  • the water-soluble biopolymer moiety is linear, branched, or dendritic.
  • the cationic carrier moiety comprises one or more basic amino acids.
  • the cationic carrier moiety comprises at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least 11, at least 12, at least 13, at least 14, at last 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, at least 25, at least 26, at least 27, at least 28, at least 29, at least 30, at least 31, at least 32, at least 33, at least 34, at least 35, at least 36, at least 37, at least 38, at least 39, at least 40, at least 41, at least 42, at least 43, at least 44, at least 45, at least 46, at least 47, at least 48, at least 49, or at least 50 basic amino acids.
  • the cationic carrier moiety comprises about 30 to about 50 basic amino acids.
  • the basic amino acid comprises arginine, lysine, histidine, or any combination thereof.
  • the cationic carrier moiety comprises about 40 lysine monomers.
  • the adjuvant moiety is capable of modulating an immune response, an inflammatory response, and/or a tissue microenvironment.
  • the adjuvant moiety comprises an imidazole derivative, an amino acid, a vitamin, or any combination thereof.
  • the adjuvant moiety comprises: , (formula IV), wherein each of G1 and G2 is H, an aromatic ring, or 1-10 alkyl, or G1 and G2 together form an aromatic ring, and wherein n is 1-10.
  • the adjuvant moiety comprises nitroimidazole.
  • the adjuvant moiety comprises metronidazole, tinidazole, nimorazole, dimetridazole, pretomanid, ornidazole, megazol, azanidazole, benznidazole, or any combination thereof.
  • the adjuvant moiety comprises an amino acid.
  • the adjuvant moiety comprises (formula V), wherein Ar is wherein each of Z1 and Z2 is H or OH.
  • the adjuvant moiety comprises a vitamin.
  • the vitamin comprises a cyclic ring or cyclic hetero atom ring and a carboxyl group or hydroxyl group.
  • the vitamin comprises: (formula VI), wherein each of Y1 and Y2 is C, N, O, or S, and wherein n is 1 or 2.
  • the vitamin is selected from the group consisting of vitamin A, vitamin B1, vitamin B2, vitamin B3, vitamin B6, vitamin B7, vitamin B9, vitamin B12, vitamin C, vitamin D2, vitamin D3, vitamin E, vitamin M, vitamin H, and any combination thereof.
  • the vitamin is vitamin B3.
  • the adjuvant moiety comprises at least about two, at least about three, at least about four, at least about five, at least about six, at least about seven, at least about eight, at least about nine, at least about ten, at least about 11, at least about 12, at least about 13, at least about 14, at least about 15, at least about 16, at least about 17, at least about 18, at least about 19, or at least about 20 vitamin B3.
  • the adjuvant moiety comprises about 10 vitamin B3.
  • the composition comprises a water-soluble biopolymer moiety with about 120 to about 130 PEG units, a cationic carrier moiety comprising a poly-lysine with about 30 to about 40 lysines, and an adjuvant moiety with about 5 to about 10 vitamin B3.
  • the composition comprises (i) a water-soluble biopolymer moiety with about 100 to about 200 PEG units, (ii) about 30 to about 40 lysines with an amine group (e.g., about 32 lysines), (iii) about 15 to 20 lysines, each having a thiol group (e.g., about 16 lysines, each with a thiol group), and (iv) about 30 to 40 lysines fused to vitamin B3 (e.g., about 32 lysines, each fused to vitamin B3).
  • an amine group e.g., about 32 lysines
  • a thiol group e.g., about 16 lysines, each with a thiol group
  • vitamin B3 e.g., about 32 lysines, each fused to vitamin B3
  • the composition further comprises a targeting moiety, e.g., a LAT1 targeting ligand, e.g., phenyl alanine, linked to the water soluble polymer.
  • a targeting moiety e.g., a LAT1 targeting ligand, e.g., phenyl alanine
  • the thiol groups in the composition form disulfide bonds.
  • the composition comprises (1) a micelle comprising (i) about 100 to about 200 PEG units, (ii) about 30 to about 40 lysines with an amine group (e.g., about 32 lysines), (iii) about 15 to 20 lysines, each having a thiol group (e.g., about 16 lysines, each with a thiol group), and (iv) about 30 to 40 lysines fused to vitamin B3 (e.g., about 32 lysines, each fused to vitamin B3), and (2) an oligonucleotide, e.g., siRNA, wherein the oligonucleotide, e.g., siRNA, is encapsulated within the micelle.
  • an oligonucleotide e.g., siRNA
  • the composition further comprises a targeting moiety, e.g., a LAT1 targeting ligand, e.g., phenyl alanine, linked to the PEG units.
  • a targeting moiety e.g., a LAT1 targeting ligand, e.g., phenyl alanine
  • the thiol groups in the micelle form disulfide bonds.
  • the present disclosure also provides a micelle comprising an oligonucleotide, e.g., siRNA, of the present disclosure, wherein the oligonucleotide, e.g., siRNA, and the delivery agent are associated with each other.
  • the association is a covalent bond, a non-covalent bond, or an ionic bond.
  • the positive charge of the cationic carrier moiety of the cationic carrier unit is sufficient to form a micelle when mixed with the oligonucleotide, e.g., siRNA, disclosed herein in a solution, wherein the overall ionic ratio of the positive charges of the cationic carrier moiety of the cationic carrier unit and the negative charges of the oligonucleotide, e.g., siRNA, (or vector comprising the inhibitor) in the solution is about 1: 1.
  • the cationic carrier unit is capable of protecting the oligonucleotide, e.g., siRNA, of the present disclosure from enzymatic degradation. See PCT Publication No.
  • WO2020/261227 which is herein incorporated by reference in its entirety.
  • the practice of the present disclosure will employ, unless otherwise indicated, conventional techniques of cell biology, cell culture, molecular biology, transgenic biology, microbiology, recombinant DNA, and immunology, which are within the skill of the art. Such techniques are explained fully in the literature. See, for example, Sambrook et al., ed. (1989) Molecular Cloning A Laboratory Manual (2nd ed.; Cold Spring Harbor Laboratory Press); Sambrook et al., ed. (1992) Molecular Cloning: A Laboratory Manual, (Cold Springs Harbor Laboratory, NY); D. N.
  • EXAMPLE 1 [0582] Various siRNAs (FIG. 6) at 100nM were transfected into SH-SY5Y cells. Cells were transfected using the reagent in vivo-jetPEI ® -PC (101000171) from polyplus.
  • neuroblastoma cell line SH-SY5Y was cultured in 1:1 mixture of Ham’s F12 and Dulbecco’s modified Eagle’s medium (DMEM; Hyclone) with 10% fetal bovine serum (FBS; Hyclone) and 1% penicillin/streptomycin (Gibco) for 3-4 days before experimentation and then sub-cultured for differentiation in 12-well culture plates (Corning Costar, Corning, NY, USA) at 1.5x10 5 cells/well. A day after plating, the media was removed and replaced with SH-SY5Y media with 3% FBS containing 10 ⁇ M all-trans-Retinoic acid (ATRA) and cells were then grown for 3 days.
  • DMEM Dulbecco’s modified Eagle’s medium
  • FBS fetal bovine serum
  • Gibco penicillin/streptomycin
  • the media was replaced with SH-SY5Y media with 3% FBS containing 80 nM 12-O-tetradecanoyl-phorbol-13-acetate (TPA) and cells were grown for an additional 3 days.
  • TPA 12-O-tetradecanoyl-phorbol-13-acetate
  • the media was replaced with SH-SY5Y media with 3% FBS.
  • SH-SY5Y human cells can be differentiated to a more mature neuron-like phenotype that is characterized by neuronal markers.
  • the cells were incubated for 48 hours. Microtubule associated protein tau protein expression was measured by Western Blot.
  • cell lysis was performed in a RIPA buffer (iNtRON Biotechnology, Seongnam, Korea) with protease inhibitor cocktail (Roche, Basel, Switzerland), phosphatase inhibitor cocktail 2 (Sigma-Aldrich, P5726), and phosphatase inhibitor cocktail 3 (Sigma-Aldrich, P0044).
  • the mixture was centrifuged at 4 °C for 15 min at 13,000 rpm.
  • the concentration of the supernatant was measured using DC protein assay reagents (Bio-Rad, Hercules, CA, USA).
  • Proteins were then separated in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS- PAGE) and transferred to polyvinylidene fluoride (PVDF) membranes (Millipore, Burling-ton, MA, USA). The membranes were incubated with the appropriate primary antibodies. Species- specific horseradish peroxidase (HRP)-conjugated secondary antibodies were used to detect the bound antibodies. The protein bands of interest were analyzed using chemiluminescence detection.
  • SDS- PAGE sodium dodecyl sulfate-polyacrylamide gel electrophoresis
  • PVDF polyvinylidene fluoride
  • HRP horseradish peroxidase
  • the antibodies used are as follows: ⁇ -actin (sc-47778) from Santa Cruz Biotechnology; Tau monoclonal antibody (TAU-5) (AHB0042), p-Tau (Thr181) (MN1050), and p-Tau (Ser202/ Thr205, AT8) (MN1020) from Invitrogen; Tau monoclonal antibody (TAU-46) (4019S) and p- Tau (Ser202) from cell signaling technology. [0584] Signals on western blots were normalized to those of ACTB (Actin Beta). The relative tau protein expression levels between the non-transfection control and the candidate siRNAs (FIG.6) transfection were compared.
  • Candidate siRNAs exhibiting an inhibitory effect of more than 30% compared to the control were selected for further analysis. Because only endogenous tau levels were measured in the intitial screen, the knockdown effect could not be efficiently confirmed. As such, candidate siRNAs exhibiting a 10% or greater knockdown effect in first screening step were re-tested under the condition of tau overexpression. Candidate siRNAs (No.3, 9, 17, and 20) (FIG.6) were confirmed to exhbit an efficient inhibitory effect of more than 30% compared to the control. The results are shown in FIG 1A. Table 1 shows the inhibition % effect if each siRNA on total tau protein expression.
  • siMAPT Nos.7, 13, 16, 18, 19, 24, 25, 26, 27, and 28 did not have knockdown effects.
  • siMAPT Nos. 3, 4, 5, 8, and 9 targeted exon 9.
  • siMAPT Nos.1, 2, 3, 6, 10, 11, 12, 7, 13, 24, 25, 26, 27, and 28 targeted exon 10.
  • siMAPT Nos. 14 and 13 targeted exon 11.
  • siMAPT Nos. 20, 21, 22, 23, 18 and 19 targeted exon 13.
  • siMAPT No.3* was precicted to behave similarly to siMAPT No.3.
  • FIG.3A shows the knockdown efficacy by siRNA No. 1 (FIG.3A), siRNA No.17 (FIG.3B), siRNA No.20 (FIG. 3C), siRNA No.23 (FIG. 3D), siRNA No.3 (FIG.3E), and siRNA No.11 (FIG.3F) of wild-type Tau, 3R0N isoform, 4R1N isoform, and 4R2N isoform (FIG.8).
  • EXAMPLE 5 [0593] In order to show that MAPT siRNA can restore memory deficit in AD acute mouse model, mice were subjected to behavior test for 3 days after ICV injection of A ⁇ Os (2 ⁇ g) (4- weeks).
  • FIG. 4G shows that the average Tau inhibition % for FIG. 4D-4F were 23.25% ⁇ 32.33, 53.61% ⁇ 10.48, and 51.54% ⁇ 6.21 respectively.
  • FIGs. 5A to 5D shows the Western blot analysis of total tau (t-tau) and hyperphosphorylated tau (p-tau) protein expression levels in each brain tissue such as hippocampus (FIG. 5A), prefrontal cortex (FIG. 5B), entorhinal cortex (FIG. 5C), and striatum (FIG. 5D) following ICV injection of BIOK-Mapt siRNA in P301S transgenic mice.
  • FIG. 5E shows a Y- maze test with vehicle control and BIOK-Mapt siRNA (Nos.
  • siRNA No. 17 (FIG.6) ability to specifically reduce the expression of 4R- MAPT isoforms efficiently restores recognition decline in the AD acute mouse model.
  • siRNA No. 17 (FIG. 6) efficiently reduces tau hyperphosphorylation, but not total tau protein expression.
  • siRNA No. 17’s (FIG. 6) recovery effect on recognition decline is likely due to its ability to efficiently reduce tau hyperphosphorylation, which is a characteristic of tauopathies, without a similar reduction in tota tau protein expression.
  • siRNA No.11 (FIG. 6) also reduces tau hyperphosphorylation, but reduces total tau protein expression at a similar rate.
  • siRNA No. 11 did not cause a recovery effect on recognition decline, and this is likely because of the loss of functional tau proteins.
  • siRNA #1 is more effective than siRNA #17 at reducing mTau mRNA levels in the hippocampus, entorhinal cortex, and prefrontal cortex of C57BL/6N mice.
  • EXAMPLE 8 Evaluating the Effectiveness of MAPT siRNA #1 and #17 in Reducing huTau mRNA in Brain Subregions of PS19 Mice [0602]
  • the PS19 (P301S Tg) mouse is a tauopathy pathogenic model that expresses the 1N4R transgene (P301S mutation) of human tau.
  • PS19 mice (The Jackson Laboratory) overexpressing mutant human MAPT P301S were used for Examples 8-12. Genotyping was performed by ear snipping, DNA isolation, and polymerase chain reaction (PCR) with the primers of MAPT.
  • the C57BL/6N mice used in this study were obtained from Koatech. Mice had free access to a regular diet (S-01010; felako) and purified water. Mice were maintained on a 12/12 h dark/light cycle (lights on at 7:00 a.m.) in a climate-controlled vivarium (22 °C). Before the experiment, mice were handled daily for three days. The daily behavioral observation took place between 10 a.m. and 5 p.m.
  • the stereotaxic injection was delivered into the right lateral ventricle (AP: - 0.2 mm, ML: -0.8 mm, DV: -2.5 mm, relative to the bregma) at a rate of 1 ⁇ l/min for a total dose of 2 ⁇ g/4 ⁇ l. After the injection, the needle was left in place for an additional 5 minutes before being slowly retracted. [0605] The results of Example 8 are shown in FIGs. 11A-11F.
  • huTau mRNA is decreased by 30 ⁇ 40%, as measured by qRT-PCR results at 7, 21, and 28 days, in the hippocampus (FIG.11A), entorhinal cortex (FIG.11B), and prefrontal cortex (FIG.11C) (sub brain regions) regions of PS19 mice 4 weeks after a single injection of siRNA #1.
  • the results show that huTau mRNA is decreased by 60 ⁇ 80%, as measured by qRT-PCR results at 7, 21, and 28 days, in the hippocampus (FIG.11D), entorhinal cortex (FIG.11E), and prefrontal cortex (FIG. 11F) (sub brain regions) regions of PS19 mice 4 weeks after a single injection of siRNA #17.
  • siRNA #17 is more effective than siRNA #1 at reducing huTau mRNA (4R-tau isoform) levels in the hippocampus, entorhinal cortex, and prefrontal cortex of PS19 mice.
  • MAPT siRNA #1 and #17 Impact on Tau Protein Expressions in PS19 Mice are as follows: in vivo-jetPEI ® -PC (101000171) from polyplus.
  • the antibodies used in Example 9 are as follows: ⁇ -actin (sc-47778), Tau13 (sc- 21796) from Santa Cruz Biotechnology; p-Tau (Thr181) (MN1050) and p-Tau (Ser202/ Thr205, AT8) (MN1020) from Invitrogen; Tau monoclonal antibody p-Tau (Ser396) (9632) from cell signaling technology.
  • ⁇ -actin sc-47778)
  • Tau13 sc- 21796
  • MN1050 p-Tau
  • Ser202/ Thr205, AT8 MN1020
  • Tau monoclonal antibody p-Tau Tau monoclonal antibody p-Tau (Ser396) (9632) from cell signaling technology.
  • MAPT siRNA #17 reversed hyper-phosphorylated tau (tau pathology markers) in aged PS19 mice more effectively than MAPT siRNA #1 (FIGs. 12B-12E).
  • MAPT siRNA #17 which specifically targets against human tau in the 9 months old PS19 tauopathy mouse model, showed a marked decrease and even reversal of tau hyperphosphorylation (Tau-13 (FIG. 12B), Thr181 (FIG. 12C), Ser404 (FIG. 12D), Ser396 (FIG. 12E)) pathology across the brain regions despite the age of the PS19 mice.
  • MAPT siRNA #1 and #17 Effect on Tau Pathology in the Hippocampus and Entorhinal Cortex of PS19 Mice
  • PBS cold phosphate buffer solution
  • the entorhinal cortex a lateral cut was made to separate it from the adjoining regions, carefully preserving its structure.
  • the prefrontal cortex dissection involved a frontal sectioning (approximately 2 mm from the anterior tip of the brain).
  • Brain sections were prepared for free-floating immunohistochemical staining. The sections were blocked with 5% normal donkey serum and then incubated for 48 h at 4 °C with the primary antibody to Mouse Anti-AT-8 (1:200, Thermo Fisher Scientific, MN1020). The sections were then incubated for 2 h at room temperature with a corresponding Donkey Anti-Mouse Alexa fluor 488-conjugated secondary antibody (1:400, Invitrogen, A-21202).
  • An identifiable but flat nest more than 90% of the Nestlet is torn and the material 4 is gathered into a nest within a quarter of the cage floor area, but the nest is flat, with walls higher than mouse body height (of a mouse curled up on its side for less than 50% of its circumference.
  • 5 A (near) perfect nest more than 90% of the Nestlet is torn and the nest is a crater, w ith walls higher than mouse body height for more than 50% of its circumference.
  • the elevated plus maze (EPM) test was conducted at the age of 27 and 36 weeks to test levels of anxiety in PS19 mice. The EPM test consisted of a black acrylic platform, 30 cm in diameter and 6 cm wide, elevated 50 cm above the ground.
  • the maze was divided into four equal quadrants, two of which were enclosed with black acrylic walls 20 cm in height. There were walls on both the inner and outer sides of the platform to provide the ‘closed’ areas of the maze. The ‘open’ arms had no walls. The maze was surrounded by curtains to prevent any distractions. Videos were taken using a camera that was positioned above the maze. Mouse behavior was analyzed using the EthoVision software. [0615] Typical behavioral deficits caused by tauopathy include decreased nesting activity and reduced anxiety. [0616] The results of Example 11 are shown in FIGs.14A-14D. In the nest building test (FIGs.
  • MAPT siRNA #17 alleviated cognitive decline and acute behavioral deficits in aged PS19 mice more effectively than MAPT siRNA #1 as measured by the nest building test and the elevated plus maze test.
  • EXAMPLE 12 Therapeutic Effects of MAPT siRNA #1 and #17 on Hyperactivity and Spatial Memory in PS19 Mice Measured by Open Field and Y-Maze Tests [0617] The Y-maze test was used to evaluate spatial working memory and was conducted when the mice were 39 weeks old. The test was conducted in black plastic arms of a Y-shaped maze. A mouse was placed in the center and was allowed to freely explore the arms for 5 min. The experiment was recorded with EthoVision software 11.5.
  • the alternation percentage was calculated by dividing the number of three consecutive arm entries (triads) by the number of possible triads ⁇ 100 (Total arm entries - 2).
  • the open field test was conducted when the mice were 27 and 33 weeks old to test the level of hyperactivity in the mice. The mice were brought to the experimental room for at least 1 hr of acclimation to the experimental room conditions prior to testing.
  • Activity chambers Med Associates Inc, St Albans, VT; 27 x 27 x 20.3 cm
  • Mice were placed in the center of the chamber and their behavior as recorded for 30 min in 5-minute segments.
  • Example 12 Quantitative analysis was performed on the following three dependent measures: distance, resting time, and rearing counts. Animals were tested at low-stress conditions where the light was lowered to approximately 10-30 lux of red light. [0619] Typical behavioral impairment caused by tauopathy include distinct hyperactivity and decline of working memory [0620] The results of Example 12 are shown in FIGs. 15A-15C. In the open field test, increased hyperactivity was observed in PS19 mice compared to WT mice as measured by distance traveled, resting time, and rearing count (FIGs. 15A-15B). However, this hyperactivity was significantly reduced following treatment with either MAPT siRNA #1 or #17 (FIGs.15A-15B).
  • FIGs. 9A-9I The chemically modified MAPT siRNAs are shown in FIGs. 9A-9I. Specifically, the sense strand and antisense strand for #3*_M1/M1/siMAPT-3*-On_chol (FIG. 9A), #5_M2/M2/siMAPT-5-On_chol (FIG. 9B), #14_M3/M3/siMAPT-14-On_chol (FIG.

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Abstract

La présente invention concerne un ARNsi qui cible un transcrit MAPT dans une cellule, conduisant à une expression réduite de la protéine MAPT. La réduction de l'expression de la protéine MAPT est bénéfique pour le traitement de certains troubles médicaux, par exemple, une tauopathie.
PCT/IB2024/053120 2023-03-30 2024-03-29 Arnsi ciblant la protéine tau associée aux microtubules et leurs utilisations Ceased WO2024201423A2 (fr)

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