Modified multisegment antisense oligonucleotides for use

Abstract

Modified multisegment antisense oligonucleotides and methods for reducing target mRNA, DNA, and protein expression are provided. Such methods, oligonucleotides, and compositions are useful for treating, preventing, or ameliorating diseases or disorders.

Description

[Technical Field]

[0001] Related Applications This application claims priority to and the benefit of PCT Application No. PCT / CN2023 / 000034, filed January 10, 2023, and PCT Application No. PCT / CN2023 / 106924, filed July 12, 2023, the contents of each of which are incorporated herein in their entirety. [Background technology]

[0002] Antisense technology has emerged as an effective means for reducing the expression of specific gene products, and therefore may prove to be uniquely useful in many therapeutic applications.Antisense therapy differs from nucleoside therapy in that it can directly target the transcripts of targets and antigens of interest and therapeutic value.Antisense technology has emerged as an effective means for reducing the expression of certain gene products, and therefore may prove to be uniquely useful in many therapeutic, diagnostic and research applications for the regulation of diseases of interest.

[0003] Many of the antisense oligonucleotides (ASOs) currently in clinical use and testing have minimal efficacy in reducing target levels and / or raised safety concerns. Therefore, there is a need in the art for improved targeted antisense oligonucleotides. Provided herein are compositions and methods that address this need. Summary of the Invention

[0004] The present disclosure provides modified multi-segment antisense oligonucleotide structures that are complementary to nucleotide sequences in gene transcripts. The modified multi-segment antisense oligonucleotides of the present disclosure comprise, in 5' to 3' order, at least a 5' wing segment, a first gap segment (G1), a first separator segment (S1), a second gap segment (G2), and a 3' wing segment (W2), each segment connected by an internucleoside linkage, and at least one nucleoside of each of W1, W2, and S1 is modified. In some embodiments, the modified multi-segment antisense oligonucleotide can have an additional separator segment flanked by additional gap segments.

[0005] These modified multi-segment antisense oligonucleotides contain a separated gap region in combination with strategically positioned binding enhancers. The target of these modified multi-segment antisense oligonucleotides can be any target, for example, a mammalian target or a viral target. In some embodiments, the modified multisegment antisense oligonucleotides are directed against HBV, DM1 protein kinase (DMPK), alpha-1 antitrypsin (AAT), transthyretin (TTR), proprotein convertase subtilisin / kexin type 9 (PCSK9), apolipoprotein B (ApoB), apolipoprotein C-III (ApoCIII), TNF-alpha, SERPINA8 (AGT), complement factor B (CFB), diacylglycerol O-acyltransferase 2 (DGAT2), patatin-like phospholipase domain containing 3 (PNPLA3), 5'-aminolevulinate synthase 1 (5'-aminolevulinate synthase 1), and / or 5'-aminolevulinate synthase 2 (5'-aminolevulinate synthase 1). 1, ALAS1), or hydroxyacid oxidase 1. In some embodiments, the modified multisegment antisense oligonucleotide targets Mat1α, (pro)renin receptor / (P)RR, COVID-19 5′UTR, Acc1, Acc2, ACE2, androgen receptor, ApoB, ASGR1, ASO, ATXN2, B1AR, B2AR, C9ORF72, caspase 2, CD19, CD4, chikungunya virus, CLPro, complement component 5, COVID19, COX2, CTGF, DGAT2, DMD, DMPK, DNM2, DUX4, E2 gene, EGFR, envelope, EphA2, epithelial sodium channel alpha subunit, exon 100 dystrophin, exon 101 dystrophin, exon 102 dystrophin, exon 103 dystrophin,Exon 104 dystrophin, Exon 105 dystrophin, Exon 106 dystrophin, Exon 107 dystrophin, Exon 108 dystrophin, Exon 109 dystrophin, Exon 110 dystrophin, Exon 111 dystrophin, Exon 112 dystrophin, Exon 113 dystrophin, Exon 114 dystrophin, Exon 115 dystrophin, Exon 116 dystrophin, Exon 117 dystrophin, Exon 118 dystrophin, Exon 119 dystrophin, Exon 120 dystrophin dystrophin, exon 121 dystrophin, exon 122 dystrophin, exon 123 dystrophin, exon 124 dystrophin, exon 125 dystrophin, exon 126 dystrophin, exon 127 dystrophin, exon 128 dystrophin, exon 129 dystrophin, exon 130 dystrophin, exon 131 dystrophin, exon 132 dystrophin, exon 133 dystrophin, exon 134 dystrophin, exon 135 dystrophin, exon 136 dystrophin, exon 1 37 dystrophin, exon 138 dystrophin, exon 139 dystrophin, exon 140 dystrophin, exon 141 dystrophin, exon 142 dystrophin, exon 143 dystrophin, exon 144 dystrophin, exon 145 dystrophin, exon 146 dystrophin, exon 147 dystrophin, exon 148 dystrophin, exon 149 dystrophin, exon 150 dystrophin, exon 151 dystrophin, exon 152 dystrophin, exon 44 dystrophin, exon exon 53 dystrophin, exon 54 dystrophin, exon 55 dystrophin, exon 56 dystrophin, exon 57 dystrophin, exon 58 dystrophin, exon 59 dystrophin, exon 60 dystrophin, exon 61 dystrophin, exon 62 dystrophin, exon 63 dystrophin, exon 64 dystrophin, exon 65 dystrophin, exon 66 dystrophin, exon 67 dystrophin, exon 68 dystrophin, exon 69 dystrophin, exon 70 dystrophin,Exon 71 dystrophin, Exon 72 dystrophin, Exon 73 dystrophin, Exon 74 dystrophin, Exon 75 dystrophin, Exon 76 dystrophin, Exon 77 dystrophin, Exon 78 dystrophin, Exon 79 dystrophin, Exon 80 dystrophin, Exon 81 dystrophin, Exon 82 dystrophin, Exon 83 dystrophin, Exon 84 dystrophin, Exon 85 dystrophin, Exon 86 dystrophin, Exon 87 dystrophin, Exon 88 dystrophin, Exon 89 dystrophin, Exon 90 dystrophin, Exon 91 dystrophin, Exon 92 dystrophin, Exon 93 dystrophin, Exon 94 dystrophin, Exon 95 dystrophin, Exon 96 dystrophin, Exon 97 dystrophin, Exon 98 dystrophin, Exon 99 dystrophin Trophin, Fabp3, Factor VII, Factor XI, FAK, FGFR4, FOXP3, FUS, FXII, GFAP, GFP, glycogen synthase, H1N1, HBV, heat shock protein 47, heat shock protein 48, heat shock protein 49, heat shock protein 50, heat shock protein 51, heat shock protein 52, heat shock protein 53, heat shock protein 54, heat shock protein 55, heat shock protein 56, heat shock protein 57, heat shock protein 58, heat shock protein 59, helicase, HIV, HOXB13, HPRT, HPV, Hsd11β1, HTT, hydroxyacid oxidase 1, hydroxysteroid 17β-dehydrogenase 13, hypoxia-inducible factor 2α, kinesin spindle protein and vascular endothelial growth factor, KRAS, leader, leucine-rich repeat kinase 2 (Leucine-rich repeat kinase 2, LRRK2), MERS-CoV, MGMT, miR-16, miR-21, MMP-2, MMP-9, MTL-CEBPA, MuRF1, Mycobacterium tuberculosis, COVID-19 N, rabies N 123, rabies N 749, rabies N 903, rabies N1082, rabies N53, rabies N8, chikungunya virus ns1 gene,Targeting chikungunya virus ns2 gene, chikungunya virus ns3 gene, chikungunya virus ns4 gene, ORF1 b, ORF1 a, HIV P24, rabies P330, P53, rabies P721, rabies P91, PCSK9, PDGF, PDL1, COVID-19 PLP, PNPLA3, polo-like kinase 1, protein kinase N3, RAF1, RAF-1, RDRP, RSV, SMN2, SNCA, COVID-19 spike, STAT3, TAU, TGFB1, TGFB1 and Cox2, TMPRSS2, TMPRSS6, TNFα, transthyretin, VEGF, VEGFR2, VER2, xanthine dehydrogenase, or YAP1. They are useful for treating both acute and chronic disease states, including, but not limited to, where the oligonucleotide target is an oncogenic target, an inflammatory target, a metabolic disease target, a cardiovascular target, a liver disease target, an infectious disease target, a neurological disease target, a neuromuscular disease target, an ophthalmic disease target, a renal disease target, a respiratory disease target, a hematological disease target, a wound healing target, a transplant target, an autoimmune disease target, a neuropsychiatric target, etc.

[0006] The present disclosure is based, at least in part, on the findings that a discontinuous gap segment adjacent to the 5' wing segment and the 3' wing segment in the modified multisegment antisense oligonucleotide of the present disclosure provides improved activity over conventional gapmers (antisense oligonucleotides composed of a central DNA segment flanked by wing segments). One or more separator segments positioned directly between the two gap segments, thus providing discontinuity between the gap segments, surprisingly provides improved activity of the modified multisegment antisense oligonucleotide. The inventors unexpectedly discovered that positioning a separator segment at a specific position in the multisegment antisense oligonucleotide of the present disclosure provides improved activity. Without being bound by theory or mechanism, one or more separator segments adjacent to the gap segments provide improved physicochemical properties (e.g., solubility) and a favorable secondary structure for alternative interactions with nucleases (e.g., endonucleases such as RNase H) when duplexed with a target nucleic acid, thereby increasing the ability of the modified multisegment antisense oligonucleotide of the present disclosure to target and / or degrade the target mRNA.

[0007] Furthermore, certain types of nucleoside modifications in the separator segment provide unexpected improvements in activity. Without being bound by theory or mechanism, the inventors have observed that certain modifications of nucleosides in the separator segment, including 2'-O-methoxyethyl and 2'-O-methyl modifications, increase the ability of the modified multisegment antisense oligonucleotides of the present disclosure to target and / or degrade target mRNAs.

[0008] Without being bound by theory or mechanism, certain combinations of nucleoside modifications and separator segment positioning within modified multisegment antisense oligonucleotides improve interaction with RNase H endonucleases without impairing hybridization to the target sequence.

[0009] Substitution of nucleosides in conventional gapmers (i.e., those that are not multisegmented) with sugar-modified nucleosides (e.g., 2'-O-MOE) results in enhanced binding affinity with the target sequence. This type of enhancement has been explored in conventional gapmer design to form a more stable duplex with the target sequence, thereby improving gapmer activity. Duplex stability can be determined by quantifying the melting temperature (Tm). A higher melting temperature traditionally indicates enhanced binding to the target sequence, which would be expected to result in increased nuclease recruitment and functional activity (e.g., silencing of target expression).

[0010] Contrary to current teachings in the art and unlike conventional gapmers that are not multi-segmented, the present inventors have unexpectedly discovered that introducing a specific nucleoside (e.g., 2'-O-MOE) as a separator at a specific position in a contiguous sequence of deoxynucleosides unexpectedly reduces Tm, instead of increasing it. More surprisingly, modified multi-segment antisense oligonucleotides with such a Tm reduction correlated with improved biological activity and potency compared to the corresponding conventional gapmer design. Without being bound by theory or mechanism, certain combinations of nucleoside modifications and the positioning of separator segments within the full-length multi-segment antisense oligonucleotide may result in changes in secondary structure that are advantageous for enhanced interaction with nucleases (e.g., endonucleases such as RNase H) and improved functional activity.

[0011] The present disclosure is based, at least in part, on research findings showing that certain nucleoside modifications at certain positions in the 5' wing segment and / or 3' wing segment reduce in vivo toxicity. The inventors unexpectedly discovered that certain modified nucleosides at certain positions in the 3' wing segment (e.g., LNA) reduced in vivo toxicity. Without being bound by theory or mechanism, certain combinations of nucleoside modifications and their positioning within the 5' wing segment and / or 3' wing segment contribute to the complementarity of modified multisegment antisense oligonucleotides while reducing interactions with off-target sequences. The modified multisegment antisense oligonucleotides described herein have one or more of increased in vivo activity, reduced off-target binding, reduced toxicity, and a wider safety margin, which are advantageous for therapeutic use.

[0012] Provided herein are methods, modified multi-segment antisense oligonucleotides, and compositions for modulating expression of target mRNAs and proteins.

[0013] Also provided are methods, modified multi-segment antisense oligonucleotides, and compositions useful for preventing, treating, and ameliorating diseases, disorders, and conditions. [Brief explanation of the drawings]

[0014] [Figure 1A] 1 shows a series of graphs depicting inhibition of PCSK9 mRNA levels in Hepa1-6 cells following treatment with modified multi-segment antisense oligonucleotides of the present disclosure. [Figure 1B] 1 shows a series of graphs depicting inhibition of PCSK9 mRNA levels in Hepa1-6 cells following treatment with modified multi-segment antisense oligonucleotides of the present disclosure. [Figure 1C]1 shows a series of graphs depicting inhibition of PCSK9 mRNA levels in Hepa1-6 cells following treatment with modified multi-segment antisense oligonucleotides of the present disclosure. [Figure 1D] 1 shows a series of graphs depicting inhibition of PCSK9 mRNA levels in Hepa1-6 cells following treatment with modified multi-segment antisense oligonucleotides of the present disclosure. [Figure 1E] 1 shows a series of graphs depicting inhibition of PCSK9 mRNA levels in Hepa1-6 cells following treatment with modified multi-segment antisense oligonucleotides of the present disclosure. [Figure 2A] 1 shows a series of graphs depicting inhibition of PCSK9 mRNA levels in Hepa1-6 cells following treatment with modified multi-segment antisense oligonucleotides of the present disclosure. [Figure 2B] 1 shows a series of graphs depicting inhibition of PCSK9 mRNA levels in Hepa1-6 cells following treatment with modified multi-segment antisense oligonucleotides of the present disclosure. [Figure 2C] 1 shows a series of graphs depicting inhibition of PCSK9 mRNA levels in Hepa1-6 cells following treatment with modified multi-segment antisense oligonucleotides of the present disclosure. [Figure 2D] 1 shows a series of graphs depicting inhibition of PCSK9 mRNA levels in Hepa1-6 cells following treatment with modified multi-segment antisense oligonucleotides of the present disclosure. [Figure 2E] 1 shows a series of graphs depicting inhibition of PCSK9 mRNA levels in Hepa1-6 cells following treatment with modified multi-segment antisense oligonucleotides of the present disclosure. [Figure 2F] 1 shows a series of graphs depicting inhibition of PCSK9 mRNA levels in Hepa1-6 cells following treatment with modified multi-segment antisense oligonucleotides of the present disclosure. [Figure 3A]1 shows a series of graphs depicting inhibition of DMPK mRNA levels in Sol8 cells following treatment with modified multisegment antisense oligonucleotides of the present disclosure. [Figure 3B] 1 shows a series of graphs depicting inhibition of DMPK mRNA levels in Sol8 cells following treatment with modified multisegment antisense oligonucleotides of the present disclosure. [Figure 3C] 1 shows a series of graphs depicting inhibition of DMPK mRNA levels in Sol8 cells following treatment with modified multisegment antisense oligonucleotides of the present disclosure. [Figure 3D] 1 shows a series of graphs depicting inhibition of DMPK mRNA levels in Sol8 cells following treatment with modified multisegment antisense oligonucleotides of the present disclosure. [Figure 3E] 1 shows a series of graphs depicting inhibition of DMPK mRNA levels in Sol8 cells following treatment with modified multisegment antisense oligonucleotides of the present disclosure. [Figure 4A] 1 shows a series of graphs depicting inhibition of DMPK mRNA levels in Sol8 cells following treatment with modified multisegment antisense oligonucleotides of the present disclosure. [Figure 4B] 1 shows a series of graphs depicting inhibition of DMPK mRNA levels in Sol8 cells following treatment with modified multisegment antisense oligonucleotides of the present disclosure. [Figure 4C] 1 shows a series of graphs depicting inhibition of DMPK mRNA levels in Sol8 cells following treatment with modified multisegment antisense oligonucleotides of the present disclosure. [Figure 4D] 1 shows a series of graphs depicting inhibition of DMPK mRNA levels in Sol8 cells following treatment with modified multisegment antisense oligonucleotides of the present disclosure. [Figure 4E] 1 shows a series of graphs depicting inhibition of DMPK mRNA levels in Sol8 cells following treatment with modified multisegment antisense oligonucleotides of the present disclosure. [Figure 4F] 1 shows a series of graphs depicting inhibition of DMPK mRNA levels in Sol8 cells following treatment with modified multisegment antisense oligonucleotides of the present disclosure. [Figure 5A] 1 shows a series of graphs depicting inhibition of TTR mRNA levels in AML-12 cells following treatment with modified multisegment antisense oligonucleotides of the present disclosure. [Figure 5B] 1 shows a series of graphs depicting inhibition of TTR mRNA levels in AML-12 cells following treatment with modified multisegment antisense oligonucleotides of the present disclosure. [Figure 5C] 1 shows a series of graphs depicting inhibition of TTR mRNA levels in AML-12 cells following treatment with modified multisegment antisense oligonucleotides of the present disclosure. [Figure 5D] 1 shows a series of graphs depicting inhibition of TTR mRNA levels in AML-12 cells following treatment with modified multisegment antisense oligonucleotides of the present disclosure. [Figure 6A] 1 shows a series of graphs depicting inhibition of TTR mRNA levels in AML-12 cells following treatment with modified multisegment antisense oligonucleotides of the present disclosure. [Figure 6B] 1 shows a series of graphs depicting inhibition of TTR mRNA levels in AML-12 cells following treatment with modified multisegment antisense oligonucleotides of the present disclosure. [Figure 6C] 1 shows a series of graphs depicting inhibition of TTR mRNA levels in AML-12 cells following treatment with modified multisegment antisense oligonucleotides of the present disclosure. [Figure 6D] 1 shows a series of graphs depicting inhibition of TTR mRNA levels in AML-12 cells following treatment with modified multisegment antisense oligonucleotides of the present disclosure. [Figure 7A]1 shows a series of graphs depicting inhibition of AAT mRNA levels in AML-12 cells following treatment with modified multi-segment antisense oligonucleotides of the present disclosure. [Figure 7B] 1 shows a series of graphs depicting inhibition of AAT mRNA levels in AML-12 cells following treatment with modified multi-segment antisense oligonucleotides of the present disclosure. [Figure 7C] 1 shows a series of graphs depicting inhibition of AAT mRNA levels in AML-12 cells following treatment with modified multi-segment antisense oligonucleotides of the present disclosure. [Figure 7D] 1 shows a series of graphs depicting inhibition of AAT mRNA levels in AML-12 cells following treatment with modified multi-segment antisense oligonucleotides of the present disclosure. [Figure 8] 1 shows a graph depicting the effect of modified multi-segment antisense oligonucleotides of the present disclosure on plasma PCSK9 protein levels. [Figure 9] 1 shows a graph depicting percent inhibition of plasma PCSK9 protein levels after treatment with modified multi-segment antisense oligonucleotides of the present disclosure. [Figure 10] 1 shows a graph depicting the effect of modified multi-segment antisense oligonucleotides of the present disclosure on plasma TTR protein levels. [Figure 11] 1 shows a graph depicting the percent inhibition of plasma TTR protein levels after treatment with modified multi-segment antisense oligonucleotides of the present disclosure. [Figure 12A] 1 shows a graph depicting the effect of modified multi-segment antisense oligonucleotides of the present disclosure on hepatitis B virus (HBV) surface antigen (HBsAg) levels in genotype C HBV transgenic mice. [Figure 12B]1 shows a graph depicting the effect of modified multi-segment antisense oligonucleotides of the present disclosure on hepatitis B virus (HBV) surface antigen (HBsAg) levels in genotype C HBV transgenic mice. [Figure 13] 1 shows a graph depicting the effect of modified multi-segment antisense oligonucleotides of the present disclosure on HBsAg levels in pcDNA3.1-preS2-GTD HDI-HBV mice. [Figure 14] 1 shows a graph depicting the effect of modified multi-segment antisense oligonucleotides of the present disclosure on HBsAg levels in pcDNA3.1-preS2-GTD HDI-HBV mice. [Figure 15] 1 shows a graph depicting the effect of modified multi-segment antisense oligonucleotides of the present disclosure on HBsAg levels in pcDNA3.1-preS2-GTD HDI-HBV mice. [Figure 16A] 1 shows a graph depicting the effect of modified multi-segment antisense oligonucleotides of the present disclosure on HBsAg levels in pcDNA3.1-preS2-GTD HDI-HBV mice. [Figure 16B] 1 shows a graph depicting the effect of modified multi-segment antisense oligonucleotides of the present disclosure on HBsAg levels in pcDNA3.1-preS2-GTD HDI-HBV mice. [Figure 17] 1 shows a graph depicting the effect of modified multi-segment antisense oligonucleotides of the present disclosure on HBsAg levels in pcDNA3.1-preS2-GTD HDI-HBV mice. [Figure 18] 1 shows a graph depicting the effect of modified multisegment antisense oligonucleotides of the present disclosure on HBsAg levels in pAAV-1.2HBV-GTA HDI-HBV mice. [Figure 19]1 shows a graph depicting the effect of modified multi-segment antisense oligonucleotides of the present disclosure on HBsAg levels in pcDNA3.1-preS2-GTD HDI-HBV mice. [Figure 20] A 20% precast UREA-TBE PAGE gel of RNase H cleavage products is shown. Lane 1: AUS1233, lane 2: AUS1493, lane 3: AUS1493, lane 4: AUS1013, lane 5: AUS1014, lane 6: AUS1015, lane 7: AUS1212, lane 8: AUS1654, lane 9: AUS1655, lane 10: molecular weight marker. [Figure 21-1] A table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure is provided and is incorporated in their entirety into the detailed description. The modifications at each position of the modified multi-segment antisense oligonucleotide sequence are read using the legend in Table 2.2. AUS1010 to AUS1714 (SEQ ID NO: 11 to SEQ ID NO: 666) contain examples of modified multi-segment antisense oligonucleotides of the present disclosure. [Figure 21-2] A table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure is provided and is incorporated in their entirety into the detailed description. The modifications at each position of the modified multi-segment antisense oligonucleotide sequence are read using the legend in Table 2.2. AUS1010 to AUS1714 (SEQ ID NO: 11 to SEQ ID NO: 666) contain examples of modified multi-segment antisense oligonucleotides of the present disclosure. [Figure 21-3] A table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure is provided and is incorporated in their entirety into the detailed description. The modifications at each position of the modified multi-segment antisense oligonucleotide sequence are read using the legend in Table 2.2. AUS1010 to AUS1714 (SEQ ID NO: 11 to SEQ ID NO: 666) contain examples of modified multi-segment antisense oligonucleotides of the present disclosure. [Figure 21-4]A table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure is provided and is incorporated in their entirety into the detailed description. The modifications at each position of the modified multi-segment antisense oligonucleotide sequence are read using the legend in Table 2.2. AUS1010 to AUS1714 (SEQ ID NO: 11 to SEQ ID NO: 666) contain examples of modified multi-segment antisense oligonucleotides of the present disclosure. [Figure 21-5] A table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure is provided and is incorporated in their entirety into the detailed description. The modifications at each position of the modified multi-segment antisense oligonucleotide sequence are read using the legend in Table 2.2. AUS1010 to AUS1714 (SEQ ID NO: 11 to SEQ ID NO: 666) contain examples of modified multi-segment antisense oligonucleotides of the present disclosure. [Figure 21-6] A table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure is provided and is incorporated in their entirety into the detailed description. The modifications at each position of the modified multi-segment antisense oligonucleotide sequence are read using the legend in Table 2.2. AUS1010 to AUS1714 (SEQ ID NO: 11 to SEQ ID NO: 666) contain examples of modified multi-segment antisense oligonucleotides of the present disclosure. [Figure 21-7] A table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure is provided and is incorporated in their entirety into the detailed description. The modifications at each position of the modified multi-segment antisense oligonucleotide sequence are read using the legend in Table 2.2. AUS1010 to AUS1714 (SEQ ID NO: 11 to SEQ ID NO: 666) contain examples of modified multi-segment antisense oligonucleotides of the present disclosure. [Figure 21-8]A table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure is provided and is incorporated in their entirety into the detailed description. The modifications at each position of the modified multi-segment antisense oligonucleotide sequence are read using the legend in Table 2.2. AUS1010 to AUS1714 (SEQ ID NO: 11 to SEQ ID NO: 666) contain examples of modified multi-segment antisense oligonucleotides of the present disclosure. [Figure 21-9] A table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure is provided and is incorporated in their entirety into the detailed description. The modifications at each position of the modified multi-segment antisense oligonucleotide sequence are read using the legend in Table 2.2. AUS1010 to AUS1714 (SEQ ID NO: 11 to SEQ ID NO: 666) contain examples of modified multi-segment antisense oligonucleotides of the present disclosure. [Figure 21-10] A table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure is provided and is incorporated in their entirety into the detailed description. The modifications at each position of the modified multi-segment antisense oligonucleotide sequence are read using the legend in Table 2.2. AUS1010 to AUS1714 (SEQ ID NO: 11 to SEQ ID NO: 666) contain examples of modified multi-segment antisense oligonucleotides of the present disclosure. [Figure 21-11] A table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure is provided and is incorporated in their entirety into the detailed description. The modifications at each position of the modified multi-segment antisense oligonucleotide sequence are read using the legend in Table 2.2. AUS1010 to AUS1714 (SEQ ID NO: 11 to SEQ ID NO: 666) contain examples of modified multi-segment antisense oligonucleotides of the present disclosure. [Figure 22-1]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-2]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-3]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-4]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-5]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-6]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-7]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-8]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-9]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-10]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-11]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-12]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-13]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-14]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-15]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-16]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-17]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-18]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-19]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-20]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-21]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-22]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-23]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-24]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-25]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-26]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-27]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-28]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-29]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-30]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-31]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-32]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-33]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-34]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-35]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-36]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-37]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-38]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-39]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-40]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-41]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-42]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-43]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-44]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-45]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-46]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-47]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-48]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-49]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-50]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-51]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-52]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-53]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-54]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-55]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-56]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-57]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-58]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-59]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-60]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-61]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-62]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-63]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-64]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-65]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-66]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-67]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-68]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-69]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-70]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-71]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-72]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-73]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-74]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-75]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-76]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-77]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-78]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-79]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-80]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-81]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-82]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-83]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-84]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-85]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-86]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-87]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-88]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-89]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-90]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-91]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-92]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-93]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-94]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-95]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-96]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-97]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-98]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-99]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-100]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-101]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-102]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-103]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-104]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-105]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-106]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-107]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-108]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-109]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-110]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-111]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-112]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-113]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-114]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-115]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-116]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-117]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-118]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-119]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. [Figure 22-120]22 shows a table of exemplary modified multi-segment antisense oligonucleotides of the present disclosure.In Figure 22, lowercase font represents 2'-deoxynucleoside, and uppercase font represents nucleoside that has any one of the modifications shown in the section entitled "exemplary modifications," such as 2'-O-methoxyethylnucleoside, 2'-O-methoxyethylcytidine, 2'-O-methylnucleoside, 2'-O-methylnucleoside, 2'-OHnucleoside, GNA, LNA, LNA with cytosine base, 2'-fluoro 2'-deoxynucleoside, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof. In Figure 22, lowercase "c" represents 2'-deoxycytidine or 2'-deoxy5-methylcytidine, and uppercase "C" represents cytidine or 5-methylcytidine with or without the sugar modifications described above. SEQ ID NOs: 5,000 to 30,983 are exemplary modified multisegment antisense oligonucleotides. DETAILED DESCRIPTION OF THE INVENTION

[0015] It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the invention as claimed. As used herein, the use of the singular includes the plural unless specifically stated otherwise. As used herein, the use of "or" means "and / or" unless specifically stated otherwise. Furthermore, the use of the term "including" and other forms, such as "includes" and "included," is not limiting. Also, terms such as "element" or "component" encompass both elements and components comprising a single unit and elements and components comprising two or more subunits, unless specifically stated otherwise.

[0016] The section headings used herein are for organizational purposes only and should not be construed as limiting the subject matter described. All documents or portions of documents cited in this application, including, but not limited to, patents, patent applications, articles, books, and treatises, are expressly incorporated herein by reference in their entirety as well as for the portion of the document discussed herein.

[0017] definition Unless specific definitions are provided, the nomenclature utilized in connection with, and the procedures and techniques of, analytical chemistry, synthetic organic chemistry, and pharmaceutical and medicinal chemistry described herein are well known and commonly used in the art. Standard techniques may be used for chemical synthesis and chemical analysis. Where permitted, all patents, applications, published applications, and other publications, GENBANK accession numbers, and associated sequence information available through databases such as the National Center for Biotechnology Information (NCBI), and other data referred to throughout this disclosure are incorporated by reference in their entirety as well as for portions of the documents discussed herein.

[0018] Unless otherwise indicated, the following terms have the following meanings.

[0019] "2'-O-Methoxyethyl" (2'-MOE and 2'-O(CH2)2-OCH3) refers to an O-methoxy-ethyl modification at the 2' position of the furanose ring. A 2'-O-methoxyethyl modified sugar is a modified sugar.

[0020] "2'-MOE nucleoside" (also 2'-O-methoxyethyl nucleoside) means a nucleoside that includes a 2'-MOE modified sugar moiety.

[0021] "2'-substituted nucleoside" means a nucleoside that includes a substituent other than H or OH at the 2' position of the furanosyl ring. In certain embodiments, 2'-substituted nucleosides include nucleosides with bicyclic sugar modifications.

[0022] "3' target site" refers to the region of a target nucleic acid that is complementary to the 3' end of a particular antisense oligonucleotide.

[0023] "5' target site" refers to the region of a target nucleic acid that is complementary to the 5' end of a particular antisense oligonucleotide.

[0024] "5-methylcytosine" means a cytosine modified with a methyl group attached to position 5. 5-methylcytosine is a modified nucleobase.

[0025] "About" means within ±7% of a value. For example, when it is stated that "an oligonucleotide affected at least about 70% inhibition of a target," it is implied that the target level is inhibited within a range of 63% to 77%.

[0026] "Acceptable safety profile" means a pattern of side effects that is within clinically acceptable limits.

[0027] "Active pharmaceutical agent" means the substance in a pharmaceutical composition that provides a therapeutic benefit when administered to an individual.

[0028] "Active target region" means a target region to which one or more active antisense oligonucleotides are targeted. "Active antisense oligonucleotide" means an antisense oligonucleotide that reduces target nucleic acid levels or protein levels.

[0029] "Administered simultaneously" refers to the simultaneous administration of two agents in any manner such that the pharmacological effects of both are apparent in the patient at the same time. Simultaneous administration does not require that both agents be administered in a single pharmaceutical composition, in the same dosage form, or by the same route of administration. The effects of both agents need not be simultaneous. The effects need only overlap over a period of time, not necessarily be coextensive.

[0030] "Animal" refers to humans or non-human animals, including, but not limited to, mice, rats, rabbits, dogs, cats, pigs, primates, and non-human primates, including, but not limited to, monkeys and chimpanzees.

[0031] "Antibody" refers to a molecule characterized by reacting specifically with an antigen in some way, with the antibody and antigen each being defined in terms of the other. An antibody may be a complete antibody molecule or a subunit of heavy chains, light chains, F ab Area, and F c The term "region" may refer to any fragment or region thereof, such as a region.

[0032] "Antisense activity" means any detectable or measurable activity resulting from hybridization of an antisense oligonucleotide to its target nucleic acid. In certain embodiments, antisense activity is a reduction in the amount or expression of a target nucleic acid or a protein encoded by such a target nucleic acid.

[0033] "Antisense oligonucleotide" or "ASO" refers to an oligomeric oligonucleotide capable of undergoing hybridization to a target nucleic acid through hydrogen bonding. Examples of antisense oligonucleotides include single-stranded oligonucleotides having a nucleobase sequence that allows hybridization to a corresponding region or segment of a target nucleic acid.

[0034] "Antisense inhibition" refers to a reduction in target nucleic acid levels in the presence of an antisense oligonucleotide complementary to the target nucleic acid compared to target nucleic acid levels in the absence of the antisense oligonucleotide.

[0035] "Antisense mechanisms" are all mechanisms involving hybridization of an oligonucleotide with a target nucleic acid, the result or effect of which is either target degradation or target occupancy, including, for example, the simultaneous cessation of cellular machinery involved in transcription or splicing.

[0036] "Area under the curve" or "AUC" is the integral of the concentration of a drug in plasma as a function of time. The AUC is calculated over the time for which data are available, e.g., until the drug is no longer detectable (AUC 0-t ), the area under the curve extrapolated from time 0 to infinity (AUC 0-￥ ), or within a specific shortened time frame, e.g., 24 hours after administration (AUC 0-24 ) can be determined.

[0037] "Base complementarity" refers to the capacity for precise base pairing (i.e., hybridization) of nucleobases of an antisense oligonucleotide with corresponding nucleobases in a target nucleic acid, mediated by Watson-Crick, Hoogsteen, or reversed Hoogsteen hydrogen bonding between corresponding nucleobases.

[0038] "Bicyclic sugar" means a furanose ring modified by bridging two non-geminal carbon atoms. A bicyclic sugar is a modified sugar.

[0039] "Body weight" refers to the total body weight of an animal, including all tissues, including adipose tissue.

[0040] "Cap structure" or "terminal cap moiety" means a chemical modification incorporated at either end of an antisense compound.

[0041] "cEt (constrained ethyl)" or "constrained ethyl" means a bicyclic sugar moiety containing a bridge connecting the 4'-carbon and the 2'-carbon, wherein the bridge has the formula: 4'-CH(CH3)-O-2'.

[0042] "Constrained ethyl nucleoside" (also cEt nucleoside) means a nucleoside that includes a bicyclic sugar moiety containing a 4'-CH(CH3)-O-2' bridge.

[0043] A "chemically distinct region" refers to a region of an antisense compound that is chemically distinct in some way from another region of the same antisense compound. For example, a region having 2'-O-methoxyethyl nucleotides is chemically distinct from a region having nucleotides that do not have 2'-O-methoxyethyl modifications.

[0044] "Co-administration" means the administration of two or more pharmaceutical agents to an individual. The two or more pharmaceutical agents can be in a single pharmaceutical composition or in separate pharmaceutical compositions. Each of the two or more pharmaceutical agents can be administered through the same or different routes of administration. Co-administration includes parallel or sequential administration.

[0045] "Complementarity" means the capacity for pairing between nucleobases of a first nucleic acid and a second nucleic acid.

[0046] "Diluent" means an ingredient in a composition that lacks pharmacological activity, but is pharmaceutically necessary or desirable. For example, in an injected drug, the diluent may be a liquid, such as saline.

[0047] "Dosage unit" means the form in which a pharmaceutical agent is provided, such as a pill, tablet, or other dosage unit known in the art.

[0048] "Dose" refers to a specified amount of a pharmaceutical agent provided in a single administration or over a specified period of time. In certain embodiments, a dose may be administered in two or more boluses, tablets, or injections. For example, in certain embodiments where subcutaneous administration is desired, the desired dose requires a volume that does not easily fit into a single injection. In such embodiments, two or more injections may be used to achieve the desired dose. In certain embodiments, a dose may be administered in two or more injections to minimize injection site reactions in individuals. In other embodiments, a pharmaceutical agent is administered over an extended period of time or continuously. A dose may be described as the amount of pharmaceutical agent per hour, day, week, or month.

[0049] A "dosing regimen" is a combination of doses designed to achieve one or more desired effects.

[0050] "Duration" refers to the period of time that an activity or event lasts. In certain embodiments, the treatment period is the period during which a dose of a pharmaceutical agent is administered.

[0051] An "effective amount," in the context of modulating an activity or treating or preventing a condition, means that an amount of active ingredient is administered to a subject in need of such modulation, treatment, or prevention, either in a single dose or as part of a series, that is effective to modulate the effect, or treat or prevent, or ameliorate the condition. The effective amount will vary depending on the health and physical condition of the subject being treated, the taxonomic group of the subject being treated, the formulation of the composition, an evaluation of the medical condition, and other relevant factors.

[0052] "Efficacy" means the ability to produce a desired effect.

[0053] "Expression" includes all functions by which a gene's coded information is converted into structures present and operating in a cell, including, but not limited to, the products of transcription and translation.

[0054] The term "fragment," when applied to polynucleotides, is understood to mean a nucleotide sequence that is reduced in length compared to a reference nucleic acid or nucleotide sequence and that comprises, consists essentially of, and / or consists of a nucleotide sequence of contiguous nucleotides that is identical or nearly identical (e.g., 60%, 70%, 80%, 90%, 92%, 95%, 98%, or 99% identical) to the reference nucleic acid or nucleotide sequence. Such nucleic acid fragments according to the invention may, where appropriate, be included in the larger polynucleotide of which they are a component. In some embodiments, such fragments may comprise, consist essentially of, and / or consist of an oligonucleotide having a length of at least about 8, 10, 12, 15, 20, 25, 30, 35, 40, 45, 50, 75, 100, 150, 200, or more contiguous nucleotides of a nucleic acid or nucleotide sequence according to the invention.

[0055] "Fully complementary" or "100% complementary" means that each nucleobase of a first nucleic acid has a complementary nucleobase in a second nucleic acid. In certain embodiments, the first nucleic acid is an antisense oligonucleotide and the target nucleic acid is the second nucleic acid.

[0056] A "fully modified motif" refers to an antisense oligonucleotide that contains a contiguous sequence of nucleosides, essentially where each nucleoside is a sugar-modified nucleoside having a uniform modification.

[0057] "Gapmer" means an antisense oligonucleotide in which an internal region having a contiguous sequence of linked deoxynucleosides that supports the recruitment of a nuclease (e.g., an RNase, e.g., RNase H) is positioned between external regions having one or more nucleosides, wherein the nucleosides comprising the internal region may be chemically different from the nucleosides comprising the external regions. The internal region may be referred to as the "gap," and each of the 5' and 3' external regions may be referred to as a "wing."

[0058] A "gap" is an internal segment of a modified multisegment antisense oligonucleotide or gapmer, comprising one or more linked deoxynucleosides, and is positioned directly or indirectly between the 5' wing (W1) and the 3' wing (W2). A gap may be referred to interchangeably as a "gap," "gap region," or "gap segment."

[0059] "Hybridization" refers to the annealing of complementary nucleic acid molecules. In certain embodiments, complementary nucleic acid molecules include, but are not limited to, antisense oligonucleotides and nucleic acid targets. In certain embodiments, complementary nucleic acid molecules include, but are not limited to, antisense oligonucleotides and nucleic acid targets.

[0060] "Directly adjacent" means that there are no intervening elements between the immediately adjacent elements.

[0061] "Individual" means a human or non-human animal selected for treatment or therapy.

[0062] "Individual compliance" means adherence by an individual to a recommended or prescribed therapy.

[0063] "Induce," "inhibit," "enhance," "elevate," "increase," "reduce," and the like generally refer to a quantitative difference between two states. Such terms can refer to a statistically significant difference between two states. Such terms apply, for example, to levels of expression and levels of activity. As used herein, the terms "inhibit" or "reduce," or grammatical variations thereof, refer to a decrease or reduction in a specified level or activity by at least about 5%, about 10%, about 15%, about 25%, about 35%, about 40%, about 50%, about 60%, about 75%, about 80%, about 90%, about 95%, or more. In some embodiments, inhibition or reduction results in little or essentially no detectable activity (at most, an insignificant amount, e.g., less than about 10% or even 5%).

[0064] "Inhibiting expression or activity" refers to a reduction, blocking, or the like of expression or activity, and does not necessarily indicate a complete elimination of expression or activity.

[0065] "Injection site reaction" means inflammation or abnormal redness of the skin at the site of injection in an individual.

[0066] "Intraperitoneal administration" means administration through infusion or injection into the peritoneum.

[0067] "Intravenous administration" means administration into a vein.

[0068] "Extended" antisense oligonucleotides have one or more additional amino acids compared to the antisense oligonucleotides disclosed herein.

[0069] [ka] It has a nucleoside.

[0070] "Locked nucleic acid" or "LNA" or "LNA nucleoside" refers to a nucleic acid monomer having a bridge connecting two carbon atoms between the 4' and 2' positions of the nucleoside sugar unit, thereby forming a bicyclic sugar. Examples of such bicyclic sugars include, but are not limited to, A) α-L-methyleneoxy (4'-CH2-O-2') LNA, (B) β-D-methyleneoxy (4'-CH2-O-2')-LNA, (C) ethyleneoxy (4'-(CH2)2-O-2') LNA, (D) aminooxy (4'-CH2-O-NI-2') LNA, and oxyamino (4'-CH2-NI-O-2') LNA, as depicted below.

[0071] As used herein, LNA oligonucleotides include, but are not limited to, oligonucleotides having at least one bridge between the 4' and 2' sugar positions, each of which independently represents -[C(R 1 )(R 2 )]n-, -C(R1 )=C(R 2 )-, -C(R 1 )=N-, -C(=NR 1 )-, -C(=O)-, -C(=S)-, -O-, -Si(R 1 )2-, -S(=O) x - and -N(R 1 )-, wherein x is 0, 1, or 2; n is 1, 2, 3, or 4; and each R 1 and R 2 are independently H, a protecting group, hydroxyl, C1-C 12 Alkyl, substituted C1-C 12 Alkyl, C2-C 12 Alkenyl, substituted C2-C 12 Alkenyl, C2-C 12 Alkynyl, substituted C2-C 12 Alkynyl, C5-C 20 Aryl, substituted C5-C 20 Aryl, heterocyclic radical, substituted heterocyclic radical, heteroaryl, substituted heteroaryl, C5-C7 alicyclic radical, substituted C5-C7 alicyclic radical, halogen, OJ 1 , N.J. I J 2 , S.J. 1 , N3, COOJ 1 , acyl (C(=O)-H), substituted acyl, CN, sulfonyl (S(=O)2-J 1 ), or sulfoxyl (S(=O)-J 1 ) and each J 1 and J 2 are independently H, C1 to C 12 Alkyl, substituted C1-C 12 Alkyl, C2-C 12 Alkenyl, substituted C2-C 12 Alkenyl, C2-C 12 Alkynyl, substituted C2-C 12 Alkynyl, C5-C 20 Aryl, substituted C5-C 20 Aryl, acyl (C(=O)-H), substituted acyl, heterocyclic radical, substituted heterocyclic radical, C1-C 12 Aminoalkyl, substituted C1-C12 It is an aminoalkyl, or a protecting group.

[0072] Examples of 4'-2' bridging groups encompassed within the definition of LNA include those of the formula: --[C(R 1 )(R 2 )] n -, -[C(R 1 )(R 2 )] n -O-, -C(R 1 )(R 2 )-N(R 1 )-O-, or -C(R 1 )(R 2 )-ON(R 1 )-. Additionally, other bridging groups encompassed within the definition of LNA include, but are not limited to, 4'-CH2-2', 4'-(CH2)2-2', 4'-(CH2)3-2', 4'-CH2-O-2', 4'-(CH2)2-O-2', 4'-CH2-ON(R 1 )-2', and 4'-CH2-N(R 1 )-O-2'-bridge, wherein each R 1 and R 2 are independently H, a protecting group, or C1-C 12 It is alkyl.

[0073] Also included within the definition of LNA according to the present invention are LNAs in which the 2'-hydroxyl group of the ribosyl sugar ring is connected to the 4' carbon atom of the sugar ring, thereby forming a methyleneoxy (4'-CH2-O-2') bridge to form a bicyclic sugar moiety. The bridge can also be a methylene (-CH2-) group connecting the 2' oxygen atom and the 4' carbon atom, for which the term methyleneoxy (4'-CH2-O-2') LNA is used. Furthermore, for bicyclic sugar moieties with an ethylene bridging group at this position, the term ethyleneoxy (4'-CH2CH2-O-2') LNA is used. The isomer of methyleneoxy (4'-CH2-O-2') LNA, A-L-methyleneoxy (4'-CH2-O-2'), is also encompassed within the definition of LNA as used herein.

[0074] "Maximum plasma concentration" or "Cmax " refers to the peak concentration of a drug in plasma after a dose of the drug has been given to a subject. Methods for measuring drug concentrations are known to those skilled in the art and include liquid chromatography and tandem mass spectrometry, among others.

[0075] A "mismatch" or "non-complementary nucleobase" refers to an instance where a nucleobase of a first nucleic acid is unable to pair with the corresponding nucleobase of a second or target nucleic acid.

[0076] A "modified internucleoside linkage" refers to a substitution or any change from a naturally occurring internucleoside bond (ie, a phosphodiester internucleoside bond).

[0077] "Modified nucleobase" means any nucleobase other than adenine, cytosine, guanine, thymidine, or uracil. "Unmodified nucleobase" means the purine bases adenine (A) and guanine (G), and the pyrimidine bases thymine (T), cytosine (C), and uracil (U).

[0078] "Modified nucleoside" means a nucleoside having, independently, a modified sugar moiety and / or a modified nucleobase.

[0079] "Modified nucleotide" means a nucleotide having, independently, a modified sugar moiety, modified internucleoside linkage, or modified nucleobase.

[0080] "Modified oligonucleotide" means an oligonucleotide containing at least one modified internucleoside linkage, modified sugar, and / or modified nucleobase.

[0081] "Modified sugar" means a substitution and / or any change from a naturally occurring sugar moiety.

[0082] "Monomer" refers to a single unit of an oligomer. Monomers include, but are not limited to, nucleosides and nucleotides, whether naturally occurring or modified.

[0083] "Motif" refers to the pattern of unmodified and modified nucleosides in an antisense oligonucleotide. "Natural sugar moiety" refers to the sugar moiety found in DNA (2'-H) or RNA (2'-OH). "Naturally occurring internucleoside linkage" refers to a 3' to 5' phosphodiester linkage.

[0084] "Modified multi-segment antisense oligonucleotide" refers to an antisense oligonucleotide having at least five structural segments, including a 5' wing (W1), a 3' wing (W2), at least two gaps (at least G1 and G2), and at least one separator (at least S1). The separator is flanked by two gap regions. The wings are at the ends of the modified multi-segment antisense oligonucleotide. From 5' to 3', exemplary modified multi-segment antisense oligonucleotides have the formula of Formula I or Formula II. The modified multi-segment antisense oligonucleotides of the present disclosure further comprise at least one modified nucleoside in the 5' wing, at least one nucleoside modification in the separator, and at least one modified nucleoside in the 3' wing. "Modified multi-segment antisense oligonucleotides" may be referred to interchangeably as "multi-segment modified antisense oligonucleotides" or "multi-segment oligonucleotides."

[0085] "Non-complementary nucleobases" refers to a pair of nucleobases that do not form hydrogen bonds with one another or otherwise support hybridization.

[0086] "Nucleic acid" refers to a molecule composed of monomeric nucleotides. Nucleic acids include, but are not limited to, ribonucleic acid (RNA), deoxyribonucleic acid (DNA), single-stranded nucleic acids, double-stranded nucleic acids, small interfering ribonucleic acids (siRNA), and microRNAs (miRNAs).

[0087] "Nucleobase" means a nitrogenous heterocyclic base moiety capable of pairing with a base of another nucleic acid.

[0088] " Nucleobase complementarity " refers to a nucleobase that can base pair with another nucleobase. For example, in DNA, adenine (A) is complementary to thymine (T). For example, in RNA, adenine (A) is complementary to uracil (U). In certain embodiments, complementary nucleobase refers to the nucleobase of an antisense oligonucleotide that can base pair with the nucleobase of its target nucleic acid. For example, if the nucleobase at a specific position of an antisense oligonucleotide can hydrogen bond with the nucleobase at a specific position of a target nucleic acid, the hydrogen bond position between the oligonucleotide and the target nucleic acid is considered to be complementary in this nucleobase pair.

[0089] "Nucleobase sequence" means the order of contiguous nucleobases independent of any sugar, linkage, and / or nucleobase modifications.

[0090] "Nucleoside mimic" includes structures used to replace sugars or sugars and bases, but not necessarily linkages, at one or more positions in an oligomeric compound, such as morpholino, cyclohexenyl, cyclohexyl, tetrahydropyranyl, bicyclo, or tricyclo sugar mimics, e.g., nucleoside mimics having non-furanose sugar units. Nucleotide mimics include structures used to replace nucleosides and linkages at one or more positions in an oligomeric compound, such as peptide nucleic acids or morpholinos (morpholinos linked by -N(H)-C(=O)-O- or other non-phosphodiester linkages). Sugar surrogate overlaps slightly with the broader term nucleoside mimic, but is intended to refer only to the replacement of the sugar unit (furanose ring). The tetrahydropyranyl ring provided herein represents an example of a sugar surrogate in which the furanose sugar group is replaced with a tetrahydropyranyl ring system. "Mimetic" refers to groups substituted on the sugar, nucleobase, and / or internucleoside linkage. Generally, a mimetic is used in place of the sugar or sugar-internucleoside linkage combination, and the nucleobase is maintained for hybridization to a selected target.

[0091] "Off-target effect" refers to an unwanted or adverse biological effect associated with the modulation of RNA or protein expression of a gene other than the intended target nucleic acid.

[0092] "Oligomeric compound" means a polymer of linked monomeric subunits that is capable of hybridizing to at least one region of a nucleic acid molecule.

[0093] "Oligonucleotide" means a polymer of nucleosides linked through internucleoside linkages, each of which may be modified or unmodified, independently of the other.

[0094] "Parenteral administration" means administration by injection (e.g., bolus injection) or infusion. Parenteral administration includes subcutaneous, intravenous, intramuscular, intraarterial, intraperitoneal, or intracranial administration, e.g., intrathecal or intraventricular.

[0095] "Peptide" means a molecule formed by linking at least two amino acids by an amide bond. As used herein, "peptide" refers to, but is not limited to, polypeptides and proteins.

[0096] "Pharmaceutically acceptable carrier" means a medium or diluent that does not interfere with the structure of the oligonucleotide. Certain such carriers allow the pharmaceutical composition to be formulated as, for example, tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions, and troches for oral ingestion by a subject.

[0097] "Pharmaceutically acceptable derivatives" include pharmaceutically acceptable salts, conjugates, prodrugs, or isomers of the oligonucleotides described herein.

[0098] "Pharmaceutically acceptable salt" means a physiologically and pharmaceutically acceptable salt of an antisense oligonucleotide, i.e., a salt that retains the desired biological activity of the parent oligonucleotide and does not impart undesired toxicological effects thereto.

[0099] "Pharmaceutical product" means a substance that provides a therapeutic benefit when administered to an individual.

[0100] "Pharmaceutical composition" refers to a mixture of substances suitable for administration to a subject. For example, a pharmaceutical composition may comprise an antisense oligonucleotide and a sterile aqueous solution. In certain embodiments, the pharmaceutical composition exhibits activity in a free uptake assay in certain cell lines.

[0101] "Phosphorothioate linkage" means an internucleoside linkage in which the phosphodiester bond has been modified by replacing one of the non-bridging oxygen atoms with a sulfur atom. A phosphorothioate linkage is a modified internucleoside linkage.

[0102] "Prevention" or "preventing" refers to delaying or forestalling the onset or progression of a condition or disease for a period of hours to days, preferably weeks to months.

[0103] "Prodrug" means a therapeutic agent that is prepared in an inactive form that is converted to an active form (i.e., a drug) within the body or cells thereof by the action of endogenous enzymes or other chemicals and / or conditions.

[0104] A "prophylactically effective amount" refers to an amount of a pharmaceutical agent that provides a prophylactic or preventative benefit to an animal.

[0105] "Recommended therapy" means a therapeutic regimen recommended by a medical professional for the treatment, amelioration, or prevention of a disease.

[0106] "Ribonucleotide" means a nucleotide having a hydroxy group at the 2' position of the sugar moiety of the nucleotide. Ribonucleotides can be modified with any of a variety of substituents.

[0107] "Salts" means physiologically and pharmaceutically acceptable salts of antisense oligonucleotides, i.e., salts that retain the desired biological activity of the parent oligonucleotide and do not impart undesired toxicological effects thereto.

[0108] A "segment" may be referred to interchangeably as a "region" or a "portion."

[0109] The "separator" in the modified multisegment antisense oligonucleotides of the present disclosure is positioned directly between and separates the two gap regions, and is positioned between the two gap regions in the modified multisegment antisense oligonucleotides. The separator can have one or more nucleosides, which are chemically different from the nucleosides comprising the gap. The separator segment comprises nucleosides modified to confer properties such as enhanced inhibitory activity, increased binding affinity to the target nucleic acid, reduced in vivo toxicity, or resistance to degradation by in vivo nucleases. The modified multisegment antisense oligonucleotides can comprise one or more separator segments. Exemplary modified multisegment antisense oligonucleotides of the present disclosure can comprise one, two, three, four, five, or six separator segments. In some embodiments, the modified multisegment antisense oligonucleotide comprises one separator segment. In some embodiments, the modified multisegment antisense oligonucleotide comprises two separator segments.

[0110] "Shortened" or "truncated" versions of the antisense oligonucleotides taught herein have one, two, or more nucleosides missing.

[0111] "Side effects" refer to physiological responses resulting from treatment other than the desired effect. In certain embodiments, side effects include, but are not limited to, injection site reactions, liver function test abnormalities, renal function abnormalities, hepatotoxicity, nephrotoxicity, central nervous system abnormalities, and myopathy. For example, elevated aminotransferase levels in serum may indicate hepatotoxicity or liver function abnormalities. For example, increased bilirubin may indicate hepatotoxicity or liver function abnormalities.

[0112] As used herein, "significant" means measurable or observable; for example, a significant result, such as a significant improvement or a significant reduction, generally refers to a measurable or observable result, such as a measurable or observable improvement or reduction.

[0113] As used herein, a "site" is defined as a unique nucleobase position within a target nucleic acid.

[0114] "Specifically hybridizable" refers to an antisense oligonucleotide that has a sufficient degree of complementarity between the antisense oligonucleotide and the target nucleic acid to induce the desired effect, but that exhibits minimal or no effect on non-target nucleic acids under conditions where specific binding is desired, i.e., physiological conditions in in vivo assays and therapeutic treatments. "Stringent hybridization conditions" or "stringent conditions" refer to conditions under which an oligomeric oligonucleotide hybridizes to its target sequence but only to a minimal number of other sequences.

[0115] "Target nucleic acid," "target RNA," "target RNA transcript," and "nucleic acid target" all refer to a nucleic acid that can be targeted by an antisense oligonucleotide.

[0116] "Target region" means a portion of a target nucleic acid to which one or more antisense oligonucleotides are targeted.

[0117] "Target segment" means the sequence of nucleotides of a target nucleic acid to which an antisense oligonucleotide is targeted.

[0118] "Therapeutically effective amount" means an amount of a pharmaceutical agent that provides a therapeutic benefit to an individual.

[0119] "Treatment" refers to the administration of a composition to bring about an alteration or improvement of a disease or condition.

[0120] "Unmodified" nucleobase means the purine bases adenine (A) and guanine (G), and the pyrimidine bases thymine (T), cytosine (C), and uracil (U).

[0121] "Unmodified nucleotide" means a nucleotide composed of naturally occurring nucleobases, sugar moieties, and internucleoside linkages. In certain embodiments, an unmodified nucleotide is an RNA nucleotide (i.e., a 3-D-ribonucleoside) or a DNA nucleotide (i.e., a 3-D-deoxyribonucleoside).

[0122] A "validated target segment" is defined as at least an 8 nucleobase portion (ie, 8 contiguous nucleobases) of a target region to which an active oligomeric oligonucleotide is targeted.

[0123] "Wings" are terminal segments of gapmers or modified multisegment antisense oligonucleotides that have been modified to confer properties to the oligonucleotide, such as enhanced inhibitory activity, enhanced biological activity, increased binding affinity for a target nucleic acid, reduced in vivo toxicity, or resistance to in vivo nuclease degradation. Wings may also be referred to as "wings," "wing regions," or "wing segments." As used herein, wings comprise at least two linked nucleosides, a subset of which may comprise one or more deoxynucleosides, although the entire wing cannot be composed entirely of deoxynucleosides. In some embodiments, wings comprise 2 to 8 linked nucleosides. In some embodiments, wings comprise 2 to 6 linked nucleosides.

[0124] The modified multi-segment antisense oligonucleotides of the present disclosure comprise a 5' wing segment (W1) located at the 5' end of the modified multi-segment antisense oligonucleotide, wherein the 3' terminal residue of W1 is not a deoxynucleoside.The modified multi-segment antisense oligonucleotides of the present disclosure also comprise a 3' wing segment (W2) located at the 3' end of the modified multi-segment antisense oligonucleotide, wherein the 5' terminal residue of W2 is not a deoxynucleoside.

[0125] The 5' wing (W1) begins at the 5' end of the modified multisegment antisense oligonucleotide, extends in the 5' to 3' direction, and is directly linked to the deoxynucleoside of the first gap, thus representing the 3' end of W1 and the 5' end of the first gap (G1).

[0126] The 3' wing (W2) begins at the 3' end of the modified multisegment antisense oligonucleotide, extends in the 3' to 5' direction, and is directly linked to the deoxynucleosides of the gap, thus representing the 3' and 5' ends of the final gap of W2.

[0127] Exemplary Modified Multisegment Antisense Oligonucleotides The present disclosure provides at least the following exemplary modified multi-segment antisense oligonucleotides: The modified multi-segment antisense oligonucleotides of the present disclosure are intended to be complementary to transcripts of genes, which in some embodiments can be one or more of HBV, DM1 protein kinase (DMPK), alpha-1 antitrypsin (AAT), transthyretin (TTR), proprotein convertase subtilisin / kexin type 9 (PCSK9), apolipoprotein B (ApoB), apolipoprotein C-III (ApoCIII), TNF-alpha, SERPINA8 (AGT), complement factor B (CFB), diacylglycerol O-acyltransferase 2 (DGAT2), patatin-like phospholipase domain-containing 3 (PNPLA3), 5'-aminolevulinic acid synthase 1 (ALAS1), or hydroxyacid oxidase 1.

[0128] In some embodiments, the modified multi-segment antisense oligonucleotides of the present disclosure may be complementary to a transcript of a gene, the gene being Mat1α, (pro)renin receptor / (P)RR, COVID-19 5′UTR, Acc1, Acc2, ACE2, androgen receptor, ApoB, ASGR1, ASO, ATXN2, B1AR, B2AR, C9ORF72, caspase 2, CD19, CD4, chikungunya virus, CLPro, complement component 5, COVID19, COX2, CTGF, DGAT2, DMD, DMPK, DNM2, DUX4, E2 gene, EGFR, envelope, EphA2, epithelial sodium channel alpha subunit, exon 100 dystrophin, exon 101 dystrophin , exon 102 dystrophin, exon 103 dystrophin, exon 104 dystrophin, exon 105 dystrophin, exon 106 dystrophin, exon 107 dystrophin, exon 108 dystrophin, exon 109 dystrophin, exon 110 dystrophin, exon 111 dystrophin, exon 112 dystrophin, exon 113 dystrophin, exon 114 dystrophin, exon 115 dystrophin, exon 116 dystrophin , exon 117 dystrophin, exon 118 dystrophin, exon 119 dystrophin, exon 120 dystrophin, exon 121 dystrophin, exon 122 dystrophin, exon 123 dystrophin, exon 124 dystrophin, exon 125 dystrophin, exon 126 dystrophin, exon 127 dystrophin, exon 128 dystrophin, exon 129 dystrophin, exon 130 dystrophin, exon 131 dystrophin , exon 132 dystrophin, exon 133 dystrophin, exon 134 dystrophin, exon 135 dystrophin, exon 136 dystrophin, exon 137 dystrophin, exon 138 dystrophin, exon 139 dystrophin, exon 140 dystrophin, exon 141 dystrophin, exon 142 dystrophin, exon 143 dystrophin, exon 144 dystrophin, exon 145 dystrophin, exon 146 dystrophin,Exon 147 dystrophin, Exon 148 dystrophin, Exon 149 dystrophin, Exon 150 dystrophin, Exon 151 dystrophin, Exon 152 dystrophin, Exon 44 dystrophin, Exon 53 dystrophin, Exon 54 dystrophin, Exon 55 dystrophin, Exon 56 dystrophin, Exon 57 dystrophin, Exon 58 dystrophin, Exon 59 dystrophin, Exon 60 dystrophin, Exon 61 dystrophin, Exon 62 dystrophin, Exon 63 dystrophin, exon 64 dystrophin, exon 65 dystrophin, exon 66 dystrophin, exon 67 dystrophin, exon 68 dystrophin, exon 69 dystrophin, exon 70 dystrophin, exon 71 dystrophin, exon 72 dystrophin, exon 73 dystrophin, exon 74 dystrophin, exon 75 dystrophin, exon 76 dystrophin, exon 77 dystrophin, exon 78 dystrophin, exon 79 dystrophin, exon 80 dystrophin, Exon 81 dystrophin, Exon 82 dystrophin, Exon 83 dystrophin, Exon 84 dystrophin, Exon 85 dystrophin, Exon 86 dystrophin, Exon 87 dystrophin, Exon 88 dystrophin, Exon 89 dystrophin, Exon 90 dystrophin, Exon 91 dystrophin, Exon 92 dystrophin, Exon 93 dystrophin, Exon 94 dystrophin, Exon 95 dystrophin, Exon 96 dystrophin, Exon 97 dystrophin, Exon 98 dystrophin Fin, exon 99 dystrophin, Fabp3, Factor VII, Factor XI, FAK, FGFR4, FOXP3, FUS, FXII, GFAP, GFP, glycogen synthase, H1N1, HBV, heat shock protein 47, heat shock protein 48, heat shock protein 49, heat shock protein 50, heat shock protein 51, heat shock protein 52, heat shock protein 53, heat shock protein 54, heat shock protein 55, heat shock protein 56, heat shock protein 57, heat shock protein 58,Heat shock protein 59, helicase, HIV, HOXB13, HPRT, HPV, Hsd11β1, HTT, hydroxyacid oxidase 1, hydroxysteroid 17β-dehydrogenase 13, hypoxia-inducible factor 2α, kinesin spindle protein and vascular endothelial growth factor, KRAS, leader, leucine-rich repeat kinase 2 (LRRK2), MERS-CoV, MGMT, miR-16, miR-21, MMP-2, MMP-9, MTL-CEBPA, MuRF1, Mycobacterium tuberculosis, COVID-19 N, rabies N 123, rabies N 749, rabies N 903, rabies N1082, rabies N53, rabies N8, chikungunya virus ns1 gene, chikungunya virus ns2 gene, chikungunya virus ns3 gene, chikungunya virus ns4 gene, ORF1 b, ORF1 a, HIV P24, rabies P330, P53, rabies P721, rabies P91, PCSK9, PDGF, PDL1, COVID-19 PLP, PNPLA3, polo-like kinase 1, protein kinase N3, RAF1, RAF-1, RDRP, RSV, SMN2, SNCA, COVID-19 spike, STAT3, TAU, TGFB1, TGFB1 and Cox2, TMPRSS2, TMPRSS6, TNFα, transthyretin, VEGF, VEGFR2, VER2, xanthine dehydrogenase, and YAP1.

[0129] Therefore, in some embodiments, the modified multi-segment antisense oligonucleotides provided herein have one or more of the following characteristics: higher biological activity, lower off-target binding and toxicity, and a wider safety margin, which may be advantageous for therapeutic use.Furthermore, in some embodiments, the modified multi-segment antisense oligonucleotides disclosed herein have higher inhibition of target genes compared with conventional gapmers (not multi-segmented) with the same template sequence.

[0130] Thus, in one aspect, the present disclosure provides a method for producing a 5' to 3' amino acid sequence comprising: 1. A modified multi-segment antisense oligonucleotide comprising: 5'W1-G1-S1-G2-W2 3' (Formula I), During the ceremony, W1 is the 5' wing segment; W2 is the 3' wing segment; G1 is the first gap segment; S1 is the first separator segment; G2 is the second gap segment; - is an internucleoside linkage, Modified multi-segment antisense oligonucleotides are provided in which at least one nucleoside of each of W1, W2, and S1 is modified.

[0131] In some embodiments, the modified multi-segment antisense oligonucleotide is of Formula I, wherein W1 comprises 2 to 8 linked nucleosides, G1 comprises 2 to 7 linked deoxynucleosides, S1 comprises 1 to 2 nucleosides, G2 comprises 2 to 7 linked deoxynucleosides, and W2 comprises 2 to 8 linked nucleosides.

[0132] In some embodiments, the modified multi-segment antisense oligonucleotide is of Formula I, wherein W1 comprises 2 to 8 linked nucleosides, G1 comprises 2 to 7 linked deoxynucleosides, S1 comprises 1 nucleoside, G2 comprises 2 to 7 linked deoxynucleosides, and W2 comprises 2 to 8 linked nucleosides.

[0133] In some embodiments, the modified multi-segment antisense oligonucleotide is of Formula I, wherein W1 comprises 2 to 8 linked nucleosides, G1 comprises 2 to 7 linked deoxynucleosides, S1 comprises 2 nucleosides, G2 comprises 2 to 7 linked deoxynucleosides, and W2 comprises 2 to 8 linked nucleosides.

[0134] In some embodiments, the modified multi-segment antisense oligonucleotide is of Formula I, wherein W1 comprises 2 to 6 linked nucleosides, G1 comprises 2 to 7 linked deoxynucleosides, S1 comprises 1 nucleoside, G2 comprises 2 to 7 linked deoxynucleosides, and W2 comprises 2 to 6 linked nucleosides.

[0135] In some embodiments, the modified multi-segment antisense oligonucleotide is of Formula I, wherein W1 comprises 2 to 8 linked nucleosides, G1 comprises 2 to 7 linked deoxynucleosides, S1 comprises 1 nucleoside, G2 comprises 2 to 7 linked deoxynucleosides, and W2 comprises 2 to 8 linked nucleosides.

[0136] In some embodiments, the modified multi-segment antisense oligonucleotide is of Formula I, wherein W1 comprises 4 to 6 linked nucleosides, G1 comprises 1 to 6 linked deoxynucleosides, S1 comprises 1 linked nucleoside, G2 comprises 1 to 6 linked deoxynucleosides, and W2 comprises 4 to 6 linked nucleosides.

[0137] In some embodiments, the modified multi-segment antisense oligonucleotide is of Formula I, wherein W1 comprises 4 to 6 linked nucleosides, G1 comprises 5 linked deoxynucleosides, S1 comprises 1 linked nucleoside, G2 comprises 5 linked deoxynucleosides, and W2 comprises 4 to 6 linked nucleosides.

[0138] In some embodiments, the modified multi-segment antisense oligonucleotide is of Formula I, wherein W1 comprises 4 linked nucleosides, G1 comprises 5 linked deoxynucleosides, S1 comprises 1 linked nucleoside, G2 comprises 5 linked deoxynucleosides, and W2 comprises 5 linked nucleosides.

[0139] In some embodiments, the modified multi-segment antisense oligonucleotide is of Formula I, wherein W1 comprises 5 linked nucleosides, G1 comprises 4 linked deoxynucleosides, S1 comprises 1 linked nucleoside, G2 comprises 5 linked deoxynucleosides, and W2 comprises 5 linked nucleosides.

[0140] In some embodiments, the modified multisegment antisense oligonucleotide is of Formula I, wherein the W1-G1-S1-G2-W2 motif is 2-4-1-5-2, 3-4-1-4-2, 3-4-1-5-3, 3-4-1-5-4, 3-4-1-5-8, 3-5-1-4-2, 3-5-1-5-2, 3-5-1-5-3, 3 -6-1-5-5, 3-6-1-6-4, 4-4-1-5-3, 4-4-1-5-4, 4-5-1-4-13, 4-5-1-4-6, 4-5-1-5-1, 4-5-1-5-10, 4-5-1-5-11, 4-5-1-5-13, 4-5-1-5-3, 4-5-1-5-4, 4-5-1-5-5, 4-5-1-5-6, 4-5-1- 5-7, 4-5-1-5-8, 4-5-1-5-9, 4-5-1-6-4, 4-6-1-4-5, 5-1-1-8-5, 5-2-1-7-5, 5-3-1-6-5, 5-4-1-4-6, 5-4-1-5-25, 5-4-1-5-4, 5-4-1-5-5, 5-4-1-6-4, 5-4-1-7-3, 5-4-1-8-2, 5-4- 2-4-5, 5-5-2-5-5, 5-5-2-6-5, 5-6-2-6-5, 5-5-1-4-25, 5-5-1-4-5, 5-6-1-3-5, 5-7-1-2-5, 5-8-1-1-5, 6-3-1-3-7, 6-3-1-4-6, 6-4-1-4-5, or 6-7-1-1-5, where each number represents the number of nucleosides. That is, for example, the modified multi-segment antisense oligonucleotide of formula I (W1-G1-S1-G2-W2) is 2-4-1-5-2, and has 2 nucleosides in W1, 4 nucleosides in G1, 1 nucleoside in S1, 5 nucleosides in G2, and 2 nucleosides in W2.

[0141] In some embodiments, the modified multi-segment antisense oligonucleotide is of Formula I and the W1-G1-S1-G2-W2 motif is 4-5-1-5-5. In some embodiments, the modified multi-segment antisense oligonucleotide is of Formula I and the W1-G1-S1-G2-W2 motif is 5-4-1-5-5. In some embodiments, the modified multi-segment antisense oligonucleotide is of Formula I and the W1-G1-S1-G2-W2 motif is 5-5-1-5-5. In some embodiments, the modified multi-segment antisense oligonucleotide is of Formula I and the W1-G1-S1-G2-W2 motif is 5-5-1-4-5. In some embodiments, the modified multi-segment antisense oligonucleotide is of Formula I and the W1-G1-S1-G2-W2 motif is 6-5-1-5-5.

[0142] In some embodiments, the modified multi-segment antisense oligonucleotide comprises, 5' to 3', 5'W1-G1-S1-G2-S2-G3-W2 3' (Formula II), During the ceremony, S2 is a second separator segment; G3 is the third gap segment, - is an internucleoside linkage, At least one nucleoside of each of W1, S1, S2, and W2 is a modified nucleoside.

[0143] In some embodiments, the modified multi-segment antisense oligonucleotide is of Formula II, wherein W1 comprises 2 to 8 linked nucleosides, G1 comprises 2 to 7 linked deoxynucleosides, S1 comprises 1 linked nucleoside, G2 comprises 1 to 2 linked deoxynucleosides, S2 comprises 1 linked nucleoside, G3 comprises 2 to 7 linked deoxynucleosides, and W2 comprises 2 to 8 linked nucleosides.

[0144] In some embodiments, the modified multi-segment antisense oligonucleotide is of Formula II, wherein W1 comprises 2 to 8 linked nucleosides, G1 comprises 2 to 7 linked deoxynucleosides, S1 comprises 1 linked nucleoside, G2 comprises 1 linked deoxynucleoside, S2 comprises 1 linked nucleoside, G3 comprises 2 to 7 linked deoxynucleosides, and W2 comprises 2 to 8 linked nucleosides.

[0145] In some embodiments, the modified multi-segment antisense oligonucleotide is of Formula II, wherein W1 comprises 2 to 8 linked nucleosides, G1 comprises 2 to 7 linked deoxynucleosides, S1 comprises 1 linked nucleoside, G2 comprises 2 linked deoxynucleosides, S2 comprises 1 linked nucleoside, G3 comprises 2 to 7 linked deoxynucleosides, and W2 comprises 2 to 8 linked nucleosides.

[0146] In some embodiments, the modified multi-segment antisense oligonucleotide is of Formula II, wherein W1 comprises 2 to 6 linked nucleosides, G1 comprises 2 to 7 linked deoxynucleosides, S1 comprises 1 linked nucleoside, G2 comprises 1 to 2 linked deoxynucleosides, S2 comprises 1 linked nucleoside, G3 comprises 2 to 7 linked deoxynucleosides, and W2 comprises 2 to 6 linked nucleosides.

[0147] In some embodiments, the modified multi-segment antisense oligonucleotide is of Formula II, wherein W1 comprises 2 to 6 linked nucleosides, G1 comprises 2 to 7 linked deoxynucleosides, S1 comprises 1 linked nucleoside, G2 comprises 1 linked deoxynucleoside, S2 comprises 1 linked nucleoside, G3 comprises 2 to 7 linked deoxynucleosides, and W2 comprises 2 to 6 linked nucleosides.

[0148] In some embodiments, the modified multi-segment antisense oligonucleotide is of Formula II, wherein W1 comprises 2 to 6 linked nucleosides, G1 comprises 2 to 7 linked deoxynucleosides, S1 comprises 1 linked nucleoside, G2 comprises 2 linked deoxynucleosides, S2 comprises 1 linked nucleoside, G3 comprises 2 to 7 linked deoxynucleosides, and W2 comprises 2 to 6 linked nucleosides.

[0149] In some embodiments, the modified multisegment antisense oligonucleotide is of Formula I, wherein the W1-G1-S1-G2-S2-G3-W2 motif is 5-1-1-3-1-3-6, 5-1-1-3-1-4-5, 5-1-1-6-1-1-5, 5-2-1-2-1-2-7, 5-2-1-2-1-4-5, 5-2-1-4-1-2-5, 5-3-1-1-1-4-5, 5-3-1-2-1-3-5, 5-3-1-4-1-1-5, 5-4-1-1-1-3-5, 5-4-1-2-1-1-6, 5-4-1-2-1-2-5, 5-4-1-3-1-1-5, 6-3-1-1-1-1-7, 6-3-1-3-1-1-5, 6-4-1-1-1-2-5, or 7-2-1-2-1-2-5. That is, for example, a modified multisegment antisense oligonucleotide of Formula II (W1-G1-S1-G2-S2-G3-W2), which is 5-1-1-3-1-3-6, has five nucleosides in W1, one nucleoside in G1, one nucleoside in S1, three nucleosides in G2, one nucleoside in S1, three nucleosides in G2, one nucleoside in S2, three nucleosides in G3, and two nucleosides in W2.

[0150] In some embodiments, the modified multi-segment antisense oligonucleotide comprises, 5' to 3', 5'W1-G1-S1-G2-S2-G3-S3-G4-W2 3' (Formula III), During the ceremony, S3 is the third separator segment; G4 is the fourth gap segment, - is an internucleoside linkage, At least one nucleoside of each of W1, S1, S2, S3, and W2 is a modified nucleoside.

[0151] In some embodiments, the modified multi-segment antisense oligonucleotide comprises, 5' to 3', 5'W1-G1-S1-G2-S2-G3-S3-G4-S4-G5-W2 3' (Formula IV), During the ceremony, S4 is the fourth separator segment; G5 is the fifth gap segment, - is an internucleoside linkage, and at least one nucleoside of each of W1, S1, S2, S3, S4, and W2 is a modified nucleoside. In some embodiments, the modified multi-segment antisense oligonucleotide comprises, from 5' to 3', 5'W1-G1-S1-G2-S2-G3-S3-G4-S4-G5-S5-G6-W2 3' (Formula V), During the ceremony, S5 is the fifth separator segment; G6 is the sixth gap segment, - is an internucleoside linkage and at least one nucleoside of each of W1, S1, S2, S3, S4, S5, and W2 is a modified nucleoside.

[0152] In some embodiments, the modified multi-segment antisense oligonucleotide comprises, 5' to 3', 5'W1-G1-S1-G2-S2-G3-S3-G4-S4-G5-S5-G6-S6-G7-W2 3' (Formula VI), During the ceremony, S6 is a sixth separator segment; G7 is the seventh gap segment, - is an internucleoside linkage and at least one nucleoside of each of W1, S1, S2, S3, S4, S5, S6, and W2 is a modified nucleoside.

[0153] In some embodiments, W1 comprises 2 to 25 linked nucleosides. In some embodiments, W1 comprises one or more linked deoxynucleosides.

[0154] In some embodiments, W2 comprises 2 to 35 linked nucleosides. In some embodiments, W2 comprises one or more linked deoxynucleosides.

[0155] In some embodiments, any one or more of G1, G2, G3, G4, G5, G6, and / or G7 comprises 1 to 10 linked deoxynucleosides.

[0156] In some embodiments, any one or more of S1, S2, S3, S4, S5, and / or S6 comprises 1, 2, 3, 4, or 5 linked nucleosides.

[0157] In some embodiments, the modified multi-segmented antisense oligonucleotide is 12 to 50 nucleobases in length, 13 to 50 nucleobases in length, or 18 to 50 nucleobases in length. In some embodiments, the modified multi-segmented antisense oligonucleotide is about 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, or 28 nucleobases in length. In some embodiments, the modified multi-segmented antisense oligonucleotide is at least 16 nucleobases in length. In some embodiments, the modified multi-segmented antisense oligonucleotide is at least 17 nucleobases in length. In some embodiments, the modified multi-segmented antisense oligonucleotide is at least 18 nucleobases in length. In some embodiments, the modified multi-segmented antisense oligonucleotide is at least 19 nucleobases in length. In some embodiments, the modified multi-segmented antisense oligonucleotide is 20 nucleobases in length.

[0158] In some embodiments, the modified multi-segmented antisense oligonucleotide is of Formula I and G1-S1-G2 is 6 to 14 nucleobases in length. In some embodiments, the modified multi-segmented antisense oligonucleotide is of Formula I and G1-S1-G2 is 8 to 12 nucleobases in length. In some embodiments, the modified multi-segmented antisense oligonucleotide is of Formula II and G1-S1-G2-S2-G3 is 8 to 13 nucleobases in length. In some embodiments, the modified multi-segmented antisense oligonucleotide is of Formula II and G1-S1-G2-S2-G3 is 8 to 12 nucleobases in length.

[0159] The following embodiments relate to any of Formulas I-VI. In some embodiments, G1, G2, G3, G4, G5, G6, and G7 can comprise nucleosides containing 2'-deoxynucleoside modifications. In some embodiments, any one or more of G1, G2, G3, G4, G5, G6, and G7 comprise nucleosides containing a 2'-deoxy 5-methylcytidine sugar modification. In some embodiments, S1, S2, S3, S4, S5, and / or S6 comprise nucleosides containing a 2'-O-methoxyethyl sugar modification. In some embodiments, S1, S2, S3, S4, S5, and / or S6 comprise nucleosides containing 5-methylcytidine. In some embodiments, S1, S2, S3, S4, S5, and / or S6 comprise nucleosides containing a 2'-O-methyl sugar modification. In some embodiments, S1, S2, S3, S4, S5, and / or S6 comprise nucleosides comprising a 2'-OH sugar modification. In some embodiments, S1, S2, S3, S4, S5, and / or S6 comprise nucleosides comprising a 2'-fluoro sugar modification. In some embodiments, S1, S2, S3, S4, S5, and / or S6 comprise nucleosides comprising a 2'-fluoro-arabinonucleic acid (2'-fluoro-ANA) sugar modification. In some embodiments, S1, S2, S3, S4, S5, and / or S6 comprise glycol nucleic acid (GNA). In some embodiments, S1, S2, S3, S4, S5, and / or S6 comprise bridged nucleic acids (e.g., LNA, cET, cMOE). In some embodiments, S1, S2, S3, S4, S5, and / or S6 comprise LNA. In some embodiments, W1 comprises a nucleoside comprising a 2'-deoxy sugar modification. In some embodiments, W1 comprises a nucleoside comprising a 2'-O-methoxyethyl sugar modification. In some embodiments, W1 comprises a 2'-O-methoxyethyl 5-methylcytidine at position 2 of a 20-mer sequence (a sequence having 20 nucleobases). In some embodiments, W1 comprises a nucleoside comprising a 2'-O-methyl sugar modification.In some embodiments, W1 comprises a 2'-O-methyl 5-methylcytidine at position 2 of the 20-mer. In some embodiments, W1 comprises a nucleoside comprising a 2'-fluoro sugar modification. In some embodiments, W1 comprises a nucleoside comprising a 2'-fluoro-arabinonucleic acid (2'-fluoro-ANA) modification. In some embodiments, W1 comprises a glycol nucleic acid (GNA). In some embodiments, W1 comprises a modified nucleoside, wherein the modified nucleoside is a bridged nucleic acid (e.g., LNA, cET, cMOE). In some embodiments, W1 comprises a modified nucleoside, wherein the modified nucleoside is a locked nucleic acid (LNA). In some embodiments, W2 comprises a nucleoside comprising a 2'-deoxy sugar modification. In some embodiments, W2 comprises a nucleoside comprising a 2'-O-methoxyethyl sugar modification (e.g., the 2'-O-methoxyethyl sugar modification is at position 15, 16, 17, 18, 19, and / or 20 of the 20-mer). In some embodiments, W2 comprises a 2'-O-methoxyethyl 5-methylcytidine. In some embodiments, W2 comprises a nucleoside comprising a 2'-O-methyl sugar modification (e.g., the 2'-O-methyl sugar modification is at position 15, 16, 17, 18, 19, and / or 20 of the 20-mer). In some embodiments, W2 comprises a nucleoside comprising a 2'-fluoro sugar modification. In some embodiments, W2 comprises a nucleoside comprising a 2'-fluoro-arabinonucleic acid (2'-fluoro-ANA) modification. In some embodiments, W2 comprises a modified nucleoside, wherein the modified nucleoside is a glycol nucleic acid (GNA). In some embodiments, W2 comprises a modified nucleoside, and the modified nucleoside is a bridged nucleic acid (e.g., LNA, cET, cMOE). In some embodiments, W2 comprises a modified nucleoside, and the modified nucleoside is a locked nucleic acid (LNA) (e.g., the LNA is at positions 16, 17, 18, 19, and / or 20 of the 20-mer).

[0160] Exemplary Modifications The modified multi-segment antisense oligonucleotide of the present disclosure can comprise linked nucleosides, such as linked deoxynucleosides, linked ribonucleosides, and linked deoxyribonucleosides.Thus, the modified multi-segment antisense oligonucleotide of the present disclosure can comprise deoxyribonucleotides, ribonucleotides, or a mixture of both.In some embodiments, the modified multi-segment antisense oligonucleotide is a modified multi-segment antisense oligonucleotide, for example, the modified multi-segment antisense oligonucleotide comprises one or more sugar modifications, one or more modified internucleoside linkages, and / or one or more base modifications.The characteristics of the modified multi-segment antisense oligonucleotide of the present disclosure will be discussed in more detail below.

[0161] In some embodiments, the modified multi-segment antisense oligonucleotide is conjugated to a moiety or conjugate and is referred to as a conjugated modified multi-segment antisense oligonucleotide. The characteristics of the conjugated modified multi-segment antisense oligonucleotide of the present disclosure are discussed in more detail below. The conjugated modified multi-segment antisense oligonucleotide may be modified or unmodified.

[0162] In some embodiments, the modified multi-segment antisense oligonucleotides of the present disclosure comprise a modified multi-segment antisense oligonucleotide comprising any one of SEQ ID NOs: 11-1453 or 5000-30983, or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 sequence modifications thereto. As used herein, the term "sequence modification" refers to sequence identity (e.g., an A to T modification).

[0163] In some embodiments, the modified multi-segment antisense oligonucleotides of the present disclosure comprise a modified multi-segment antisense oligonucleotide comprising any one of SEQ ID NOs: 11-1453 or 5000-30983, or at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% sequence identity thereto.

[0164] In some embodiments, a vaccine comprises only one unique modified multi-segment antisense oligonucleotide, although for clarity, a vaccine may comprise copies of the same unique modified multi-segment antisense oligonucleotide. In some embodiments, a vaccine comprises a plurality of different modified multi-segment antisense oligonucleotides, for example, 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 or more different modified multi-segment antisense oligonucleotides of the present disclosure.

[0165] Characteristics of modified multisegment antisense oligonucleotides As contemplated herein, modified multi-segment antisense oligonucleotides include substitutions or changes to internucleoside linkages, sugar moieties, and nucleic acid bases.Modified multi-segment antisense oligonucleotides confer desirable properties, such as enhanced cellular uptake, enhanced affinity for nucleic acid targets, increased stability in the presence of nucleases, or increased inhibitory activity.Chemically modified nucleosides can also be used to increase the binding affinity to their target nucleic acids.

[0166] In some embodiments, the modified multi-segment antisense oligonucleotides of the present disclosure comprise one or more modified internucleoside linkages. In some embodiments, the modified multi-segment antisense oligonucleotides of the present disclosure comprise one or more modified internucleoside linkages, wherein the modified internucleoside linkages are phosphorodiamidate linkages between morpholino nucleoside mimics. In some embodiments, the modified multi-segment antisense oligonucleotides of the present disclosure comprise one or more modified internucleoside linkages, wherein the modified internucleoside linkages are phosphorothioate internucleoside linkages. In some embodiments, the phosphorothioate modifications comprise a mixture of Sp and Rp stereoisomers. In some embodiments, the phosphorothioate modifications comprise Sp stereoisomers. In some embodiments, the phosphorothioate modifications comprise Rp stereoisomers.

[0167] In some embodiments, the modified multi-segment antisense oligonucleotides of the present disclosure comprise one or more sugar modifications. In some embodiments, the modified multi-segment antisense oligonucleotides of the present disclosure comprise one or more linked nucleosides that comprise a 2' sugar modification (a sugar modification at the C2' position). In exemplary embodiments, the modified multisegment antisense oligonucleotides of the present disclosure comprise one or more modifications to the 2'-deoxynucleoside, such as a 2'-O-methyl modification, a 2'-O-ethyl modification, a 2'-O-(2-methoxyethyl) modification, a 2'-fluoro modification, a 2'-amino modification, a 2'-O-propyl modification, a 2'-O-butyl modification, a 2'-O-cyclopropylmethyl modification, a 2'-O-(2-hydroxyethyl) modification, a 2'-O-[2-(methylamino)-2-oxyethyl] modification, a 2'-O-2-[2-(N,N-dimethylamino)ethoxy]ethyl] modification, a 2'-O-(2-dimethylaminoethyl) modification, a 2'-O-(3-aminopropyl) modification, a 2'-O-(2-aminopropyl) modification, a 2'-O- and / or 2'-fluoro-arabino (2'-fluoro-ANA) modifications.

[0168] In some embodiments, modified multi-segment antisense oligonucleotides comprise one or more bicyclic sugar-modified nucleosides. In some embodiments, modified multi-segment antisense oligonucleotides of the present disclosure comprise one or more bridged nucleic acids, wherein the 2'-oxygen is linked to the 4' carbon of ribose through a bridging carbon to form a bridged nucleic acid (BNA), for example, linked through a 2',4'-methylene bridge to form a locked nucleic acid (LNA), or are, for example, 2',4'-constrained 2'-O-ethyl modified ((R)-cET or (S)-cET), or are, for example, 2',4'-constrained 2'-O-methoxyethyl modified ((R)-cMOE or (S)-cMOE).

[0169] In some embodiments, the modified multi-segment antisense oligonucleotide comprises one or more carbocyclic LNAs (cLNAs), in which the 2'-oxygen atom in LNA is replaced with a carbon atom. In some exemplary embodiments, the modified multi-segment antisense oligonucleotides of the present disclosure comprise one or more methanocarbabicyclo[3.1.0]hexane sugars, either North C2'-exo (N-MC) or South C3'-exo (S-MC), or 2'-F-NMC. In some embodiments, the modified multi-segment antisense oligonucleotide comprises (R)-Me-cLNA, or (S)-Me-cLNA, or F-LNA, or methylene-cLNA, as shown below.

[0170] [ka]

[0171] [ka]

[0172] In some embodiments, the modified multi-segment antisense oligonucleotide comprises one or more of the following: α-L-LNA, bcDNA, or tcDNA.

[0173] [ka]

[0174] In some embodiments, the modified multi-segment antisense oligonucleotides of the present disclosure comprise one or more morpholino modifications in which a morpholino group replaces the ribose sugar. In some embodiments, the modified multi-segment antisense oligonucleotides of the present disclosure comprise a phosphorodiamidate morpholino modification in which the sugar-phosphate backbone is replaced with a phosphorodiamidate morpholino moiety. In some embodiments, the modified multi-segment antisense oligonucleotides of the present disclosure comprise one or more peptide nucleic acid (PNA) modifications in which the sugar-phosphate or sugar-phosphorothioate backbone is replaced with a peptide backbone. In some embodiments, the modified multi-segment antisense oligonucleotides of the present disclosure comprise one or more glycol nucleic acid (GNA) modifications in which a propylene glycol group replaces the ribose sugar, sometimes referred to as glycerol nucleic acid. In some embodiments, the modified multi-segment antisense oligonucleotides of the present disclosure comprise one or more phosphate analogs, E vinylphosphonate (E-VP), shown below, and an amide backbone linkage.

[0175] [ka]

[0176] In some embodiments, the modified multi-segment antisense oligonucleotide of the present disclosure comprises one or more base modifications.In some embodiments, the modified multi-segment antisense oligonucleotide of the present disclosure comprises one or more nucleosides that are 5-methylcytidine.In some embodiments, the modified multi-segment antisense oligonucleotide of the present disclosure comprises one or more nucleosides that are N1-methyl-pseudouridine.

[0177] In some embodiments, the modified multi-segment antisense oligonucleotides of the present disclosure comprise one or more modifications to the 3' and / or 5' ends of the oligonucleotide, for example, 3'- and / or 5'-amino modifications.

[0178] In some embodiments, the modified multi-segmented antisense oligonucleotides of the present disclosure comprise segmented oligonucleotides, creating one or more internal regions (gaps) with multiple nucleotides that support RNase H cleavage, positioned between external regions (5' and 3' wing segments) with multiple nucleotides that are chemically distinct from the nucleosides of the internal regions. In oligonucleotides that exhibit complementarity to targets with segmented motifs, the gap segments can hybridize with the target RNA sequence for endonuclease cleavage, while the wing segments contain modified nucleosides. In some embodiments, the gap regions are distinguished by the type of sugar moiety that comprises each distinct region. The types of sugar moieties used to demarcate the region of the gap can, in some embodiments, include β-D-ribonucleosides, β-D-deoxyribonucleosides, 2'-modified nucleosides (such 2'-modified nucleosides can include 2'-MOE and 2'-O-CH3, among others, as described in the previous paragraph), and bicyclic sugar-modified nucleosides (such bicyclic sugar-modified nucleosides can include those with a constrained ethyl). In some embodiments, the nucleosides in the wings can include modified sugar moieties, including, for example, 2'-MOE and a bicyclic sugar moiety, e.g., a constrained ethyl or LNA. In some embodiments, the wings can include a combination of several modified and unmodified sugar moieties described in the previous paragraph. In some embodiments, the wings may comprise various combinations of bicyclic sugar moieties, such as 2'-MOE nucleosides, constrained ethyl nucleosides, or LNA nucleosides, and 2'-deoxynucleosides. A single internal region (gap) may be present or may be further segmented with separator segments, thus resulting in discontinuous gap segments, which in some embodiments provide improved activity (e.g., reduction of HBsAg, HBeAg, and / or HBcAg levels in serum or other bodily fluids or tissues) over conventional oligonucleotides with continuous gaps. One or more separator segments positioned directly between gap segments may be beneficial.

[0179] Exemplary modified multi-segment antisense oligonucleotides of the present disclosure include, but are not limited to, those illustrated in Table 2.1, Table 1A, Table 1B, Table 1C, FIG. 21, and FIG.

[0180] Table 2.2 provides a description of the modifications at each of the residues shown in Table 2.1, Table 1A, and Table 1B. In some embodiments, the modified multisegment antisense oligonucleotide is selected from the group consisting of SEQ ID NOs: 668-677, 679-682, 684-688, 690-692, 698-1107, 1109-1120, 1122-1125, 1127-1138, 1140, 1145-1156, 1158-1161, 1163-1174, 1176-1179, 1181-1192, 1194-1197, 1199-1210, 1212-1215, 1217-1228, 1230-1233, 1235-1246 , 1248 to 1251, 1253 to 1264, 1266 to 1269, 1271 to 1282, 1284 to 1287, 1289 to 1300, 1302 to 1305, 1307 to 1318, 1320 to 1323, 1325 to 1336, 1338 to 1341, 1343 to 1354, 1356 to 1359, 1361 to 1372, 1374 to 1377, 1379 to 1390, 1392 to 1395, 1397 to 1408, 1410 to 1413, 1415 to 1426, and 1428 to 1431.

[0181] [Table 1-1]

[0182] [Table 1-2]

[0183] [Table 1-3]

[0184] [Table 1-4]

[0185] [Table 1-5]

[0186] [Table 1-6]

[0187] [Table 1-7]

[0188] [Table 1-8]

[0189] [Table 1-9]

[0190] [Table 1-10]

[0191] [Table 2]

[0192] Conjugated Oligonucleotides The oligonucleotides of the present disclosure may be further covalently linked to one or more conjugate groups / moieties. In some embodiments, such groups enhance the activity, cellular distribution, and / or cellular uptake of the resulting vaccine. Exemplary conjugate groups include, but are not limited to, carbohydrate moieties (e.g., N-acetylgalactosamine-containing carbohydrate chains or galactose-containing carbohydrate chains) and lipid moieties (e.g., cholesterol and phospholipids). Other conjugate groups include, but are not limited to, phospholipids, biotin or other affinity tags (e.g., streptavidin), polymers (e.g., polyethylene glycol (PEG), polylysine), cell-penetrating peptides (e.g., Tat, penetratin), enzymes (e.g., horseradish peroxidase, alkaline phosphatase), antibodies or antibody fragments (e.g., anti-HER2, anti-EGFR), proteins or peptides (e.g., growth factors, cytokines), metal nanoparticles (e.g., gold nanoparticles), small molecule drugs (e.g., doxorubicin, paclitaxel), phenazine, folate, phenanthridine, anthraquinone, acridine, fluorescein, rhodamine, coumarin, and dyes.

[0193] The oligonucleotides of the present disclosure can also be modified to have one or more stabilizing groups, typically attached to one or both ends of the oligonucleotide, to enhance properties such as, for example, nuclease stability. Stabilizing groups include cap structures. These terminal modifications can protect oligonucleotides with terminal nucleic acids from exonuclease degradation and can aid in intracellular delivery and / or localization. The cap can be present at the 5' end (5' cap), the 3' end (3' cap), or both ends. Cap structures are well known in the art and include, for example, reverse deoxyabasic caps.

[0194] The conjugated oligonucleotides of the present disclosure may be modified as described in the previous section.

[0195] A. 5' Wing Segment, General (W1) In certain embodiments, at least one of the nucleosides in the 5' wing segment comprises a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, each of the nucleosides in the 5' wing segment comprises a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, the 5' wing segment comprises a nucleoside having a 2'-deoxy sugar. In certain embodiments, the 5' wing segment comprises a nucleoside having a 2'-deoxy sugar, and the other nucleoside in the 5' wing segment comprises a 2'-O(CH2)2-OCH3 sugar.

[0196] In certain embodiments, one, two, three, or four of the nucleosides in the 5' wing segment comprise a sugar modification described herein. In certain embodiments, one, two, three, or four of the nucleosides in the 5' wing segment comprise a bicyclic sugar. In certain embodiments, one, two, three, or four of the nucleosides in the 5' wing segment comprise a constrained ethyl sugar.

[0197] In certain embodiments, one, two, three, or four of the nucleosides in the 5' wing segment comprise a locked nucleic acid. In certain embodiments, one, two, three, or four of the nucleosides in the 5' wing segment comprise a 4'-CH2-O-2' sugar. In certain embodiments, two of the nucleosides in the 5' wing segment comprise a locked nucleic acid. In certain embodiments, two of the nucleosides in the 5' wing segment comprise a 4'-CH2-O-2' sugar.

[0198] B1. 5' wing segment (W1) with four linked nucleosides In certain embodiments, a 5' wing segment consists of four linked nucleosides. In certain embodiments, a 5' wing segment consists of four linked nucleosides, each of the four linked nucleosides comprising a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, a 5' wing segment consists of four linked nucleosides, three of the four linked nucleosides comprising a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, a 5' wing segment consists of four linked nucleosides, two of the four linked nucleosides comprising a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, a 5' wing segment consists of four linked nucleosides, one of the four linked nucleosides comprising a nucleoside having a 2'-O(CH2)2-OCH3 sugar.

[0199] B2. 5' wing segment with four linked nucleosides, 2'-deoxy combination (W1) In certain embodiments, a 5' wing segment is comprised of four linked nucleosides, one of which comprises a nucleoside having a 2'-deoxy sugar. In certain embodiments, a 5' wing segment is comprised of four linked nucleosides, two of which comprise a nucleoside having a 2'-deoxy sugar. In certain embodiments, a 5' wing segment is comprised of four linked nucleosides, one of which comprises a nucleoside having a 2'-deoxy sugar, and each of the other three linked nucleosides comprises a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, a 5' wing segment is comprised of four linked nucleosides, one of the four linked nucleosides comprises a nucleoside having a 2'-deoxy sugar, one of the four linked nucleosides comprises a locked nucleic acid, and each of the other two linked nucleosides comprises a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, a 5' wing segment is comprised of four linked nucleosides, one of the four linked nucleosides comprises a nucleoside having a 2'-deoxy sugar, two of the four linked nucleosides comprise a locked nucleic acid, and the other linked nucleoside comprises a nucleoside having a 2'-O(CH2)2-OCH3 sugar.

[0200] B3. 5' wing segment with four linked nucleosides, 2'-LNA combination (W1) In certain embodiments, a 5' wing segment is comprised of four linked nucleosides, one of the four linked nucleosides comprises a locked nucleic acid. In certain embodiments, a 5' wing segment is comprised of four linked nucleosides, two of the four linked nucleosides comprise a locked nucleic acid. In certain embodiments, a 5' wing segment is comprised of four linked nucleosides, one of the four linked nucleosides comprises a 4'-CH2-O-2' sugar. In certain embodiments, a 5' wing segment is comprised of four linked nucleosides, two of the four linked nucleosides comprise a 4'-CH2-O-2' sugar.

[0201] In certain embodiments, a 5' wing segment is comprised of four linked nucleosides, one of which comprises a locked nucleic acid, and each of the other three linked nucleosides comprises a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, a 5' wing segment is comprised of four linked nucleosides, two of which comprise a locked nucleic acid, and each of the other two linked nucleosides comprises a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, a 5' wing segment is comprised of four linked nucleosides, one of which comprises a 4'-CH2-O-2' sugar, and each of the other three linked nucleosides comprises a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, the 5' wing segment consists of four linked nucleosides, two of which include a 4'-CH2-O-2' sugar and each of the other two linked nucleosides includes a nucleoside having a 2'-O(CH2)2-OCH3 sugar.

[0202] C1. 5' wing segment (W1) with five linked nucleosides In certain embodiments, a 5' wing segment consists of five linked nucleosides. In certain embodiments, a 5' wing segment consists of five linked nucleosides, each of the five linked nucleosides comprising a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, a 5' wing segment consists of five linked nucleosides, four of the five linked nucleosides comprising a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, a 5' wing segment consists of five linked nucleosides, three of the five linked nucleosides comprising a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, a 5' wing segment consists of five linked nucleosides, two of the five linked nucleosides comprising a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, the 5' wing segment consists of five linked nucleosides, one of the five linked nucleosides comprising a nucleoside having a 2'-O(CH2)2-OCH3 sugar.

[0203] C2. 5' wing segment with five linked nucleosides, 2'-deoxy combination (W1) In certain embodiments, a 5' wing segment is comprised of five linked nucleosides, one of which includes a nucleoside having a 2'-deoxy sugar. In certain embodiments, a 5' wing segment is comprised of five linked nucleosides, two of which include a nucleoside having a 2'-deoxy sugar. In certain embodiments, a 5' wing segment is comprised of five linked nucleosides, one of which includes a nucleoside having a 2'-deoxy sugar, and each of the other four linked nucleosides includes a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, a 5' wing segment is comprised of five linked nucleosides, one of the five linked nucleosides comprises a nucleoside having a 2'-deoxy sugar, one of the five linked nucleosides comprises a locked nucleic acid, and each of the other three linked nucleosides comprises a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, a 5' wing segment is comprised of five linked nucleosides, one of the five linked nucleosides comprises a nucleoside having a 2'-deoxy sugar, two of the five linked nucleosides comprise a locked nucleic acid, and the other two linked nucleosides comprise a nucleoside having a 2'-O(CH2)2-OCH3 sugar.

[0204] C3. 5' wing segment with five linked nucleosides, 2'-LNA combination (W1) In certain embodiments, the 5' wing segment is comprised of five linked nucleosides, one of the five linked nucleosides comprises a locked nucleic acid. In certain embodiments, the 5' wing segment is comprised of five linked nucleosides, two of the five linked nucleosides comprise a locked nucleic acid. In certain embodiments, the 5' wing segment is comprised of five linked nucleosides, one of the five linked nucleosides comprises a 4'-CH2-O-2' sugar. In certain embodiments, the 5' wing segment is comprised of five linked nucleosides, two of the five linked nucleosides comprise a 4'-CH2-O-2' sugar.

[0205] In certain embodiments, a 5' wing segment is comprised of five linked nucleosides, one of which comprises a locked nucleic acid, and each of the other four linked nucleosides comprises a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, a 5' wing segment is comprised of five linked nucleosides, two of which comprise a locked nucleic acid, and each of the other three linked nucleosides comprises a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, a 5' wing segment is comprised of five linked nucleosides, one of which comprises a 4'-CH2-O-2' sugar, and each of the other four linked nucleosides comprises a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, the 5' wing segment consists of five linked nucleosides, two of the five linked nucleosides include a 4'-CH2-O-2' sugar, and each of the other three linked nucleosides includes a nucleoside having a 2'-O(CH2)2-OCH3 sugar.

[0206] D.3' Wing Segment, General (W2) In certain embodiments, at least one of the nucleosides in the 3' wing segment comprises a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, each of the nucleosides in the 3' wing segment comprises a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, the 3' wing segment comprises a nucleoside having a 2'-deoxy sugar. In certain embodiments, the 3' wing segment comprises a nucleoside having a 2'-deoxy sugar, and the other nucleoside in the 3' wing segment comprises a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, the 3' wing segment further comprises two nucleosides, each having a 2'-deoxy sugar.

[0207] In certain embodiments, one, two, three, or four of the nucleosides in the 3' wing segment comprise a sugar modification described herein. In certain embodiments, one, two, three, or four of the nucleosides in the 3' wing segment comprise a bicyclic sugar. In certain embodiments, one, two, three, or four of the nucleosides in the 3' wing segment comprise a constrained ethyl sugar.

[0208] In certain embodiments, one, two, three, or four of the nucleosides in the 3' wing segment comprise a locked nucleic acid. In certain embodiments, one, two, three, or four of the nucleosides in the 3' wing segment comprise a 4'-CH2-O-2' sugar. In certain embodiments, two of the nucleosides in the 3' wing segment comprise a locked nucleic acid. In certain embodiments, two of the nucleosides in the 3' wing segment comprise a 4'-CH2-O-2' sugar. In certain embodiments, three of the nucleosides in the 3' wing segment comprise a locked nucleic acid. In certain embodiments, three of the nucleosides in the 3' wing segment comprise a 4'-CH2-O-2' sugar.

[0209] E1. 3' wing segment (W2) with four linked nucleosides In certain embodiments, a 3' wing segment consists of four linked nucleosides. In certain embodiments, a 3' wing segment consists of four linked nucleosides, each of the four linked nucleosides comprising a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, a 3' wing segment consists of four linked nucleosides, three of the four linked nucleosides comprising a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, a 3' wing segment consists of four linked nucleosides, two of the four linked nucleosides comprising a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, a 3' wing segment consists of four linked nucleosides, one of the four linked nucleosides comprising a nucleoside having a 2'-O(CH2)2-OCH3 sugar.

[0210] E2. 3' wing segment with four linked nucleosides, 2'-deoxy combination (W2) In certain embodiments, a 3' wing segment is comprised of four linked nucleosides, one of which comprises a nucleoside having a 2'-deoxy sugar. In certain embodiments, a 3' wing segment is comprised of four linked nucleosides, two of which comprise a nucleoside having a 2'-deoxy sugar. In certain embodiments, a 3' wing segment is comprised of four linked nucleosides, one of which comprises a nucleoside having a 2'-deoxy sugar, and each of the other three linked nucleosides comprises a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, a 3' wing segment is comprised of four linked nucleosides, one of the four linked nucleosides comprises a nucleoside having a 2'-deoxy sugar, one of the four linked nucleosides comprises a locked nucleic acid, and each of the other two linked nucleosides comprises a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, a 3' wing segment is comprised of four linked nucleosides, one of the four linked nucleosides comprises a nucleoside having a 2'-deoxy sugar, two of the four linked nucleosides comprise a locked nucleic acid, and the other linked nucleoside comprises a nucleoside having a 2'-O(CH2)2-OCH3 sugar.

[0211] E3. 3' wing segment with four linked nucleosides, 2'-LNA combination (W2) In certain embodiments, the 3' wing segment is comprised of four linked nucleosides, one of the four linked nucleosides comprises a locked nucleic acid. In certain embodiments, the 3' wing segment is comprised of four linked nucleosides, two of the four linked nucleosides comprise a locked nucleic acid. In certain embodiments, the 3' wing segment is comprised of four linked nucleosides, one of the four linked nucleosides comprises a 4'-CH2-O-2' sugar. In certain embodiments, the 3' wing segment is comprised of four linked nucleosides, two of the four linked nucleosides comprise a 4'-CH2-O-2' sugar.

[0212] In certain embodiments, a 3' wing segment is comprised of four linked nucleosides, one of which comprises a locked nucleic acid, and each of the other three linked nucleosides comprises a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, a 3' wing segment is comprised of four linked nucleosides, two of which comprise a locked nucleic acid, and each of the other two linked nucleosides comprises a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, a 3' wing segment is comprised of four linked nucleosides, three of which comprise a locked nucleic acid, and one linked nucleoside comprises a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, a 3' wing segment consists of four linked nucleosides, one of which includes a 4'-CH2-O-2' sugar, and each of the other three linked nucleosides includes a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, a 3' wing segment consists of four linked nucleosides, two of which include a 4'-CH2-O-2' sugar, and each of the other two linked nucleosides includes a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, a 3' wing segment consists of four linked nucleosides, three of which include a 4'-CH2-O-2' sugar, and the other linked nucleoside includes a nucleoside having a 2'-O(CH2)2-OCH3 sugar.

[0213] F1. 3' wing segment (W2) with five linked nucleosides In certain embodiments, a 3' wing segment consists of five linked nucleosides. In certain embodiments, a 3' wing segment consists of five linked nucleosides, each of the five linked nucleosides comprising a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, a 3' wing segment consists of five linked nucleosides, four of the five linked nucleosides comprising a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, a 3' wing segment consists of five linked nucleosides, three of the five linked nucleosides comprising a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, a 3' wing segment consists of five linked nucleosides, two of the five linked nucleosides comprising a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, the 3' wing segment consists of five linked nucleosides, one of the five linked nucleosides comprising a nucleoside having a 2'-O(CH2)2-OCH3 sugar.

[0214] F2. 3' wing segment with five linked nucleosides, 2'-deoxy combination (W2) In certain embodiments, a 3' wing segment is comprised of five linked nucleosides, one of which includes a nucleoside having a 2'-deoxy sugar. In certain embodiments, a 3' wing segment is comprised of five linked nucleosides, two of which include a nucleoside having a 2'-deoxy sugar. In certain embodiments, a 3' wing segment is comprised of five linked nucleosides, one of which includes a nucleoside having a 2'-deoxy sugar, and each of the other four linked nucleosides includes a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, a 3' wing segment is comprised of five linked nucleosides, one of the five linked nucleosides comprises a nucleoside having a 2'-deoxy sugar, one of the five linked nucleosides comprises a locked nucleic acid, and each of the other three linked nucleosides comprises a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, a 3' wing segment is comprised of five linked nucleosides, one of the five linked nucleosides comprises a nucleoside having a 2'-deoxy sugar, two of the five linked nucleosides comprise a locked nucleic acid, and the other two linked nucleosides comprise a nucleoside having a 2'-O(CH2)2-OCH3 sugar.

[0215] F3. 3' wing segment with five linked nucleosides, 2'-LNA combination (W2) In certain embodiments, the 3' wing segment is comprised of five linked nucleosides, one of the five linked nucleosides comprises a locked nucleic acid. In certain embodiments, the 3' wing segment is comprised of five linked nucleosides, two of the five linked nucleosides comprise a locked nucleic acid. In certain embodiments, the 3' wing segment is comprised of five linked nucleosides, one of the five linked nucleosides comprises a 4'-CH2-O-2' sugar. In certain embodiments, the 3' wing segment is comprised of five linked nucleosides, two of the five linked nucleosides comprise a 4'-CH2-O-2' sugar.

[0216] In certain embodiments, the 3' wing segment is comprised of five linked nucleosides, one of which comprises a locked nucleic acid, and each of the other four linked nucleosides comprises a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, the 3' wing segment is comprised of five linked nucleosides, two of which comprise a locked nucleic acid, and each of the other three linked nucleosides comprises a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, the 3' wing segment is comprised of five linked nucleosides, three of which comprise a locked nucleic acid, and each of the other two linked nucleosides comprises a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, a 3' wing segment consists of five linked nucleosides, one of which includes a 4'-CH2-O-2' sugar and each of the other four linked nucleosides includes a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, a 3' wing segment consists of five linked nucleosides, two of which include a 4'-CH2-O-2' sugar and each of the other three linked nucleosides includes a nucleoside having a 2'-O(CH2)2-OCH3 sugar.

[0217] G1. 3' wing segment (W2) with six linked nucleosides In certain embodiments, a 3' wing segment consists of six linked nucleosides. In certain embodiments, a 3' wing segment consists of six linked nucleosides, each of the six linked nucleosides comprising a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, a 3' wing segment consists of six linked nucleosides, five of the six linked nucleosides comprising a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, a 3' wing segment consists of six linked nucleosides, four of the six linked nucleosides comprising a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, a 3' wing segment consists of six linked nucleosides, three of the six linked nucleosides comprising a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, a 3' wing segment is comprised of six linked nucleosides, two of which include a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, a 3' wing segment is comprised of six linked nucleosides, one of which includes a nucleoside having a 2'-O(CH2)2-OCH3 sugar.

[0218] G2. 3' wing segment with six linked nucleosides, 2'-deoxy combination (W2) In certain embodiments, a 3' wing segment is comprised of six linked nucleosides, one of which includes a nucleoside having a 2'-deoxy sugar. In certain embodiments, a 3' wing segment is comprised of six linked nucleosides, two of which include a nucleoside having a 2'-deoxy sugar. In certain embodiments, a 3' wing segment is comprised of six linked nucleosides, one of which includes a nucleoside having a 2'-deoxy sugar, and each of the other five linked nucleosides includes a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, a 3' wing segment consists of six linked nucleosides, one of the six linked nucleosides comprises a nucleoside having a 2'-deoxy sugar, one of the six linked nucleosides comprises a locked nucleic acid, and each of the other four linked nucleosides comprises a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, a 3' wing segment consists of six linked nucleosides, one of the five linked nucleosides comprises a nucleoside having a 2'-deoxy sugar, two of the six linked nucleosides comprise a locked nucleic acid, and the other three linked nucleosides comprise a nucleoside having a 2'-O(CH2)2-OCH3 sugar.

[0219] G3. 3' wing segment with six linked nucleosides, 2'-LNA combination (W2) In certain embodiments, the 3' wing segment is comprised of six linked nucleosides, one of the six linked nucleosides comprising a locked nucleic acid. In certain embodiments, the 3' wing segment is comprised of six linked nucleosides, two of the six linked nucleosides comprising a locked nucleic acid. In certain embodiments, the 3' wing segment is comprised of six linked nucleosides, one of the six linked nucleosides comprising a 4'-CH2-O-2' sugar. In certain embodiments, the 3' wing segment is comprised of six linked nucleosides, two of the six linked nucleosides comprising a 4'-CH2-O-2' sugar.

[0220] In certain embodiments, the 3' wing segment is comprised of six linked nucleosides, one of which comprises a locked nucleic acid, and each of the other five linked nucleosides comprises a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, the 3' wing segment is comprised of six linked nucleosides, two of which comprise a locked nucleic acid, and each of the other four linked nucleosides comprises a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, the 3' wing segment is comprised of six linked nucleosides, three of which comprise a locked nucleic acid, and each of the other three linked nucleosides comprises a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, a 3' wing segment consists of six linked nucleosides, one of which includes a 4'-CH2-O-2' sugar, and each of the other five linked nucleosides includes a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, a 3' wing segment consists of six linked nucleosides, two of which include a 4'-CH2-O-2' sugar, and each of the other four linked nucleosides includes a nucleoside having a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, a 3' wing segment consists of six linked nucleosides, three of which include a 4'-CH2-O-2' sugar, and each of the other three linked nucleosides includes a nucleoside having a 2'-O(CH2)2-OCH3 sugar.

[0221] H-0. Gap segment In some embodiments, the modified multi-segment antisense oligonucleotide comprises or consists of at least one gap segment. In some embodiments, the modified multi-segment antisense oligonucleotide comprises or consists of at least two gap segments. In some embodiments, the modified multi-segment antisense oligonucleotide comprises or consists of at least three gap segments. In some embodiments, the modified multi-segment antisense oligonucleotide comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 gap segments. The gap segments are sequentially referred to as the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, or tenth gap segments, respectively, with the first gap segment being closest to the 5' end of the modified multi-segment antisense oligonucleotide and the last gap segment being closest to the 3' end of the modified multi-segment antisense oligonucleotide. In some embodiments, the modified multi-segment antisense oligonucleotide comprises or consists of two gap segments. In some embodiments, the modified multi-segment antisense oligonucleotide comprises or consists of three gap segments. In some embodiments, the modified multi-segment antisense oligonucleotide comprises or consists of four gap segments. In some embodiments, the modified multi-segment antisense oligonucleotide comprises or consists of five gap segments. In some embodiments, the modified multi-segment antisense oligonucleotide comprises or consists of six gap segments. In some embodiments, the modified multi-segment antisense oligonucleotide comprises or consists of seven gap segments.

[0222] In some embodiments, a gap segment comprises or consists of 1 to 20 linked nucleosides. In some embodiments, a gap segment comprises or consists of 2 to 7 linked nucleosides. In some embodiments, a gap segment consists of 1 nucleoside. In some embodiments, a gap segment consists of 2 linked nucleosides. In some embodiments, a gap segment consists of 3 linked nucleosides. In some embodiments, a gap segment consists of 4 linked nucleosides. In some embodiments, a gap segment consists of 5 linked nucleosides. In some embodiments, a gap segment consists of 6 linked nucleosides. In some embodiments, a gap segment consists of 7 linked nucleosides. In some embodiments, a gap segment consists of 8 linked nucleosides. In some embodiments, a gap segment consists of 9 linked nucleosides. In some embodiments, a gap segment consists of 10 linked nucleosides.

[0223] H-1. First gap segment (G1) In certain embodiments, the first gap segment comprises or consists of 1, 2, 3, 4, 5, 6, 7, or 8 linked nucleosides. In certain embodiments, the first gap segment comprises or consists of 2 to 7 linked nucleosides. In certain embodiments, the first gap segment comprises or consists of 4 or 5 linked nucleosides. In certain embodiments, the first gap segment comprises or consists of 4 linked nucleosides. In certain embodiments, the first gap segment comprises or consists of 5 linked nucleosides.

[0224] In certain embodiments, the first gap segment comprises or consists of 1, 2, 3, 4, 5, 6, 7, or 8 linked nucleosides, each having a 2'-deoxy sugar. In certain embodiments, the first gap segment comprises or consists of 2 to 7 linked nucleosides, each having a 2'-deoxy sugar. In certain embodiments, the first gap segment comprises or consists of 4 or 5 linked nucleosides, each having a 2'-deoxy sugar. In certain embodiments, the first gap segment comprises or consists of 4 linked nucleosides, each having a 2'-deoxy sugar. In certain embodiments, the first gap segment comprises or consists of 5 linked nucleosides, each having a 2'-deoxy sugar.

[0225] H-2. Second gap segment (G2) In certain embodiments, the second gap segment comprises or consists of 1, 2, 3, 4, 5, 6, 7, or 8 linked nucleosides. In certain embodiments, the second gap segment comprises or consists of 2 to 7 linked nucleosides. In certain embodiments, the second gap segment comprises or consists of 1 or 2 linked nucleosides. In certain embodiments, the second gap segment comprises or consists of 4 or 5 linked nucleosides. In certain embodiments, the second gap segment comprises 1 nucleoside. In certain embodiments, the second gap segment comprises or consists of 2 linked nucleosides. In certain embodiments, the second gap segment comprises or consists of 4 linked nucleosides. In certain embodiments, the second gap segment comprises or consists of 5 linked nucleosides.

[0226] In certain embodiments, the second gap segment comprises or consists of 1, 2, 3, 4, 5, 6, 7, or 8 linked nucleosides, each having a 2'-deoxy sugar. In certain embodiments, the second gap segment comprises or consists of 2 to 7 linked nucleosides, each having a 2'-deoxy sugar. In certain embodiments, the second gap segment comprises or consists of 1 or 2 linked nucleosides, each having a 2'-deoxy sugar. In certain embodiments, the second gap segment comprises one nucleoside having a 2'-deoxy sugar. In certain embodiments, the second gap segment comprises or consists of two linked nucleosides, each having a 2'-deoxy sugar. In certain embodiments, the second gap segment comprises or consists of 4 linked nucleosides, each having a 2'-deoxy sugar. In certain embodiments, the second gap segment comprises or consists of 5 linked nucleosides, each having a 2'-deoxy sugar, hi certain embodiments, the second gap segment comprises or consists of 6 linked nucleosides, each having a 2'-deoxy sugar.

[0227] H-3. Third gap segment (G3) In certain embodiments, the third gap segment comprises or consists of 1, 2, 3, 4, 5, 6, 7, or 8 linked nucleosides. In certain embodiments, the third gap segment comprises or consists of 2 to 7 linked nucleosides. In certain embodiments, the third gap segment comprises or consists of 4 or 5 linked nucleosides. In certain embodiments, the third gap segment comprises or consists of 4 linked nucleosides. In certain embodiments, the third gap segment comprises or consists of 5 linked nucleosides.

[0228] In certain embodiments, the third gap segment comprises or consists of 1, 2, 3, 4, 5, 6, 7, or 8 linked nucleosides, each having a 2'-deoxy sugar. In certain embodiments, the third gap segment comprises or consists of 2 to 7 linked nucleosides, each having a 2'-deoxy sugar. In certain embodiments, the third gap segment comprises or consists of 4 or 5 linked nucleosides, each having a 2'-deoxy sugar. In certain embodiments, the third gap segment comprises or consists of 4 linked nucleosides, each having a 2'-deoxy sugar. In certain embodiments, the third gap segment comprises or consists of 5 linked nucleosides, each having a 2'-deoxy sugar.

[0229] J. Separator segment In some embodiments, the separator segment comprises 1, 2, 3, 4, or 5 linked nucleosides. In some embodiments, the separator segment comprises 1 nucleoside. In some embodiments, the separator segment comprises 2 nucleosides.

[0230] In some embodiments, the nucleoside comprises a modification. Exemplary modifications include any one of the modifications set forth in the section entitled Exemplary Modifications, including, but not limited to, 2'-O-methoxyethyl nucleosides, 2'-O-methoxyethyl cytidine, 2'-O-methyl nucleosides, 2'-O-methyl nucleosides, 2'-OH nucleosides, GNA, LNA, LNA with a cytosine base, 2'-fluoro 2'-deoxynucleosides, 2'-fluoro 2'-deoxycytidine, 2'-F-arabinonucleic acid (2'-F-ANA), or 2'-F-arabinonucleic acid (2'-F-ANA), or combinations thereof.

[0231] In certain embodiments, the separator segment consists of one nucleoside containing a 2'-OCH3 sugar. In certain embodiments, the separator segment consists of one nucleoside containing a 2'-O(CH2)2-OCH3 sugar.

[0232] In certain embodiments, the separator segment is located at position 7, 8, 9, 10, 11, 12, 13, or 14 of a modified multisegment antisense oligonucleotide that is 16 residues in length. In certain embodiments, the separator segment is located at position 7, 8, 9, 10, 11, 12, 13, or 14 of a modified multisegment antisense oligonucleotide that is 17 residues in length. In certain embodiments, the separator segment is located at position 7, 8, 9, 10, 11, 12, 13, or 14 of a modified multisegment antisense oligonucleotide that is 18 residues in length. In certain embodiments, the separator segment is located at position 7, 8, 9, 10, 11, 12, 13, or 14 of a modified multisegment antisense oligonucleotide that is 19 residues in length. In certain embodiments, the separator segment is located at position 7, 8, 9, 10, 11, 12, 13, or 14 of a modified multisegment antisense oligonucleotide that is 20 residues in length. In certain embodiments, the separator segment is at position 7, 8, 9, 10, 11, 12, 13, or 14 of a modified multisegment antisense oligonucleotide that is 21 residues in length. In certain embodiments, the separator segment is at position 7, 8, 9, 10, 11, 12, 13, or 14 of a modified multisegment antisense oligonucleotide that is 22 residues in length.

[0233] In certain embodiments, the separator segment is at position 9, 10, 11, or 12 of a modified multisegment antisense oligonucleotide that is 20 residues in length.

[0234] In certain embodiments, the separator segment is at position 10 of a modified multisegment antisense oligonucleotide that is 20 residues long.

[0235] In certain embodiments, the separator segment is located at position 10 of a 20-residue long modified multisegment antisense oligonucleotide and consists of a single nucleoside containing a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, the separator segment is located at position 7, or 8, or 9, or 10, or 11, or 12, or 13, or 14 of a 20-residue long modified multisegment antisense oligonucleotide. In certain embodiments, the separator segment is located at position 9, or 10, or 11 of a 20-residue long modified multisegment antisense oligonucleotide. In certain embodiments, the separator segment is located at position 10 of a 20-residue long modified multisegment antisense oligonucleotide.

[0236] In certain embodiments, the separator segment consists of one nucleoside containing a 2'-O(CH2)2-OCH3 sugar. In certain embodiments, the separator segment consists of one nucleoside containing a 2'-OCH3 sugar. In certain embodiments, the separator segment is located at position 7, or 8, or 9, or 10, or 11, or 12, or 13, or 14 of a modified multisegment antisense oligonucleotide that is 20 residues long. In certain embodiments, the separator segment is located at position 9, or 10, or 11 of a modified multisegment antisense oligonucleotide that is 20 residues long. In certain embodiments, the separator segment is located at position 10 of a modified multisegment antisense oligonucleotide that is 20 residues long.

[0237] K. Oligonucleotide Backbone In certain embodiments, modified multi-segment antisense oligonucleotide is single-stranded.In certain embodiments, at least one internucleoside linkage is modified internucleoside linkage.In certain embodiments, each internucleoside linkage is phosphorothioate internucleoside linkage.In some embodiments, the modified multi-segment antisense oligonucleotide of the present disclosure comprises one or more modified internucleoside linkages, and modified internucleoside linkage is phosphorodiamidate linkage between morpholino nucleoside mimics.

[0238] L. Modified Nucleobases In certain embodiments, the modified multi-segment antisense oligonucleotide comprises at least one modified nucleobase.In certain embodiments, the modified multi-segment antisense oligonucleotide comprises 1, 2, or 3 modified nucleobases.In certain embodiments, the modified nucleobase is 5-methylcytosine.

[0239] In certain embodiments, one or more modified nucleosides in a wing segment have a modified sugar. In certain embodiments, the modified sugar is a bicyclic sugar. In certain embodiments, the modified nucleoside is an LNA nucleoside. In certain embodiments, the modified nucleoside is a 2'-substituted nucleoside. In certain embodiments, the 2'-substituted nucleoside comprises a nucleoside having a bicyclic sugar modification. In certain embodiments, the modified nucleoside is a 2'-MOE nucleoside. In certain embodiments, the modified nucleoside is a constrained ethyl (cEt) nucleoside. In certain embodiments, each modified nucleoside in each wing segment is independently a 2'-MOE nucleoside or a nucleoside having a bicyclic sugar modification, e.g., a constrained ethyl (cEt) nucleoside or an LNA nucleoside.

[0240] 4-5-1-5-5 modified multisegment antisense oligonucleotides In certain embodiments, the modified multisegment antisense oligonucleotide comprises a 5' wing segment consisting of four linked nucleosides, a first gap segment consisting of five linked nucleosides having 2'-deoxy sugars, a separator segment at position 10 of the 20-residue long modified multisegment antisense oligonucleotide, a second gap segment consisting of five linked nucleosides having 2'-deoxy sugars, and a 3' wing segment consisting of five linked nucleosides. In exemplary embodiments, the modified multisegment antisense oligonucleotide of this paragraph further comprises that the cytosines at positions 2, 13, and 20 of the 20-residue long modified multisegment antisense oligonucleotide are each 5-methylcytosine. In exemplary embodiments of the modified multisegment antisense oligonucleotide of this paragraph, each cytosine in the modified multisegment antisense oligonucleotide is a 5-methylcytosine. In exemplary embodiments, the modified multisegment antisense oligonucleotide of this paragraph further comprises internucleoside linkages, each of which is a phosphorothioate internucleoside linkage. In an exemplary embodiment, the modified multi-segment antisense oligonucleotide of this paragraph further comprises that the cytosines at positions 2, 13, and 20 of the modified multi-segment antisense oligonucleotide that is 20 residues in length are each 5-methylcytosine, and further comprises internucleoside linkages that are each phosphorothioate internucleoside linkages.

[0241] In certain embodiments, a modified multi-segment antisense oligonucleotide comprises a 5' wing segment described in Sections B1, B2, or B3, a first gap segment consisting of five linked nucleosides having 2'-deoxy sugars, a separator segment at position 10 of the 20-residue long modified multi-segment antisense oligonucleotide, a second gap segment consisting of five linked nucleosides having 2'-deoxy sugars, and a 3' wing segment described in Sections F1, F2, or F3. In exemplary embodiments, the modified multi-segment antisense oligonucleotide of this paragraph further comprises that the cytosines at positions 2, 13, and 20 of the 20-residue long modified multi-segment antisense oligonucleotide are each 5-methylcytosine. In exemplary embodiments, the modified multi-segment antisense oligonucleotide of this paragraph further comprises internucleoside linkages that are each phosphorothioate internucleoside linkages. In an exemplary embodiment, the modified multi-segment antisense oligonucleotide of this paragraph further comprises that the cytosines at positions 2, 13, and 20 of the modified multi-segment antisense oligonucleotide that is 20 residues in length are each 5-methylcytosine, and further comprises internucleoside linkages that are each phosphorothioate internucleoside linkages.

[0242] Exemplary annotations of positions and segments of AUS 1493 or SEQ ID NO: 456 are shown below.

[0243] [Table 3]

[0244] In certain embodiments, the oligonucleotide or composition comprises a salt of a modified multi-segment antisense oligonucleotide.

[0245] In certain embodiments, the oligonucleotide or composition further comprises a pharmaceutically acceptable carrier or diluent.

[0246] In certain embodiments, the compound or modified multi-segment antisense oligonucleotide is single-stranded.

[0247] In certain embodiments, at least one nucleoside of the modified multi-segment antisense oligonucleotide comprises a modified sugar. In certain embodiments, at least one modified sugar comprises a 2'-O-methoxyethyl group (2'-O(CH2)2-OCH3). In certain embodiments, the modified sugar comprises a 2'-O-CH3 group. In certain embodiments, the modified nucleobase is 2'-O-methoxyethyl 5-methylcytidine.

[0248] In certain embodiments, at least one modified sugar is a bicyclic sugar. In certain embodiments, at least one modified sugar bicyclic sugar comprises a 4'-(CH2)-O-2' bridge, and n is 1 or 2. In certain embodiments, the bicyclic sugar comprises a 4'-CH2-O-2' bridge. In certain embodiments, the bicyclic sugar comprises a 4'-CH(CH3)-O-2' bridge.

[0249] method The present disclosure provides methods of treating a subject having a disease or disorder involving a target.

[0250] In some embodiments, the disease or disorder is a DMPK-related disease or disorder. In some embodiments, the disease or disorder is an AAT-related disease or disorder. In some embodiments, the disease or disorder is a TTR-related disease or disorder. In some embodiments, the disease or disorder is a PCSK9-related disease or disorder. In some embodiments, the disease or disorder is an ApoB-related disease or disorder. In some embodiments, the disease or disorder is a TNF-alpha-related disease or disorder. In some embodiments, the disease or disorder is an ApoCIII-alpha-related disease or disorder. In some embodiments, the disease or disorder is a DMPK-related disease or disorder. In some embodiments, the disease or disorder is an AGT-related disease or disorder. In some embodiments, the disease or disorder is a CFB-related disease or disorder. In some embodiments, the disease or disorder is a DGAT2-related disease or disorder. In some embodiments, the disease or disorder is a PNPLA3-related disease or disorder. In some embodiments, the disease or disorder is an ALAS1-related disease or disorder. In some embodiments, the disease or disorder is a hydroxyacid oxidase 1-related disease or disorder.

[0251] In some embodiments, the disease or disorder is selected from the group consisting of non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), COVID19, skin aging conditions, prostate cancer, atherosclerosis, chronic kidney disease, chikungunya virus, amyotrophic lateral sclerosis, progressive malignant solid tumors, non-arteritic anterior ischemic optic neuropathy, HIV, immunoglobulin A nephropathy, inflammation and injury, human Tenon's capsule fibroblasts, anti-fibrosis, cystic fibrosis, Duchenne muscular dystrophy (DMD), myotonic dystrophy 1, centronuclear myopathy, facioscapulohumeral muscular dystrophy, and fibrosis. dystrophy, FSHD), pancreatic cancer, solid tumors, cystic fibrosis, antithrombosis, coagulation disorders, thrombosis disorders, Alexander disease, Pompe disease, H1N1 influenza, HBV, liver fibrosis, HPV, Huntington's disease, clear cell renal cell carcinoma, pancreatic cancer, Parkinson's disease, MERS-CoV, inflammatory bowel disease, Alport syndrome, advanced liver cancer, muscle atrophy and myotonic dystrophy, mycobacterium tuberculosis, rabies, hypercholesterolemia, pulmonary fibrosis, primary or secondary liver cancer, breast cancer, RSV, spinal muscular atrophy, multiple system atrophy, cancer, Alzheimer's disease and frontotemporal degeneration, thalassemia / low-risk myelodysplastic syndromes, autoimmunity, hATTR, hereditary transthyretin (ATTRv) amyloidosis, or gout.

[0252] In some embodiments, the disease or disorder is associated with Mat1α, (pro)renin receptor / (P)RR, COVID-19 5′UTR, Acc1, Acc2, ACE2, androgen receptor, ApoB, ASGR1, ASO, ATXN2, B1AR, B2AR, C9ORF72, caspase 2, CD19, CD4, chikungunya virus, CLPro, complement component 5, COVID19, COX2, CTGF, DGAT2, DMD, DMPK, DNM2, DUX4, E2 gene, EGFR, envelope, EphA2, epithelial sodium channel alpha subunit, exon 100 dystrophin, exon 101 dystrophin, exon 102 dystrophin. dystrophin, exon 103 dystrophin, exon 104 dystrophin, exon 105 dystrophin, exon 106 dystrophin, exon 107 dystrophin, exon 108 dystrophin, exon 109 dystrophin, exon 110 dystrophin, exon 111 dystrophin, exon 112 dystrophin, exon 113 dystrophin, exon 114 dystrophin, exon 115 dystrophin, exon 116 dystrophin, exon 117 dystrophin, exon 118 Dystrophin, exon 119 dystrophin, exon 120 dystrophin, exon 121 dystrophin, exon 122 dystrophin, exon 123 dystrophin, exon 124 dystrophin, exon 125 dystrophin, exon 126 dystrophin, exon 127 dystrophin, exon 128 dystrophin, exon 129 dystrophin, exon 130 dystrophin, exon 131 dystrophin, exon 132 dystrophin, exon 133 dystrophin, exon Exon 134 dystrophin, exon 135 dystrophin, exon 136 dystrophin, exon 137 dystrophin, exon 138 dystrophin, exon 139 dystrophin, exon 140 dystrophin, exon 141 dystrophin, exon 142 dystrophin, exon 143 dystrophin, exon 144 dystrophin, exon 145 dystrophin, exon 146 dystrophin, exon 147 dystrophin, exon 148 dystrophin, exon 149 dystrophin,Exon 150 dystrophin, exon 151 dystrophin, exon 152 dystrophin, exon 44 dystrophin, exon 53 dystrophin, exon 54 dystrophin, exon 55 dystrophin, exon 56 dystrophin, exon 57 dystrophin, exon 58 dystrophin, exon 59 dystrophin, exon 60 dystrophin, exon 61 dystrophin, exon 62 dystrophin, exon 63 dystrophin, exon 64 dystrophin, exon 65 dystrophin, exon 66 dystrophin dystrophin, exon 67 dystrophin, exon 68 dystrophin, exon 69 dystrophin, exon 70 dystrophin, exon 71 dystrophin, exon 72 dystrophin, exon 73 dystrophin, exon 74 dystrophin, exon 75 dystrophin, exon 76 dystrophin, exon 77 dystrophin, exon 78 dystrophin, exon 79 dystrophin, exon 80 dystrophin, exon 81 dystrophin, exon 82 dystrophin, exon 83 dystrophin, exon 84 Dystrophin, exon 85 dystrophin, exon 86 dystrophin, exon 87 dystrophin, exon 88 dystrophin, exon 89 dystrophin, exon 90 dystrophin, exon 91 dystrophin, exon 92 dystrophin, exon 93 dystrophin, exon 94 dystrophin, exon 95 dystrophin, exon 96 dystrophin, exon 97 dystrophin, exon 98 dystrophin, exon 99 dystrophin, Fabp3, Factor VII, Factor XI, FAK, FGFR4, FO XP3, FUS, FXII, GFAP, GFP, glycogen synthase, H1N1, HBV, heat shock protein 47, heat shock protein 48, heat shock protein 49, heat shock protein 50, heat shock protein 51, heat shock protein 52, heat shock protein 53, heat shock protein 54, heat shock protein 55, heat shock protein 56, heat shock protein 57, heat shock protein 58, heat shock protein 59, helicase, HIV, HOXB13, HPRT, HPV, Hsd11β1, HTT,Hydroxyacid oxidase 1, hydroxysteroid 17β-dehydrogenase 13, hypoxia-inducible factor 2α, kinesin spindle protein and vascular endothelial growth factor, KRAS, leader, leucine-rich repeat kinase 2 (LRRK2), MERS-CoV, MGMT, miR-16, miR-21, MMP-2, MMP-9, MTL-CEBPA, MuRF1, Mycobacterium tuberculosis, COVID-19 N, rabies N 123, rabies N 749, rabies N 903, rabies N1082, rabies N53, rabies N8, chikungunya virus ns1 gene, chikungunya virus ns2 gene, chikungunya virus ns3 gene, chikungunya virus ns4 gene, ORF1 b, ORF1 a, HIV P24, rabies P330, P53, rabies P721, rabies P91, PCSK9, PDGF, PDL1, COVID-19 PLP, PNPLA3, polo-like kinase 1, protein kinase N3, RAF1, RAF-1, RDRP, RSV, SMN2, SNCA, COVID-19 spike, STAT3, TAU, TGFB1, TGFB1 and Cox2, TMPRSS2, TMPRSS6, TNFα, transthyretin, VEGF, VEGFR2, VER2, xanthine dehydrogenase, or YAP1-related diseases or disorders.

[0253] In certain embodiments, the subject is a human.

[0254] In certain embodiments, the subject is a non-human primate, eg, a monkey, eg, a cynomolgus monkey.

[0255] In certain embodiments, the subject is a rodent, such as a mouse or a rat.

[0256] In some embodiments, the modified multi-segment antisense oligonucleotides of the present disclosure have increased in vivo activity (compared to an unmodified sequence having the same nucleobase sequence).

[0257] In some embodiments, the modified multi-segment antisense oligonucleotides of the present disclosure provide reduced off-target binding (compared to an unmodified sequence having the same nucleobase sequence).

[0258] In some embodiments, the use of modified multi-segment antisense oligonucleotides of the present disclosure results in reduced toxicity (compared to sequences having the same nucleobase sequence but that are not modified).

[0259] In certain embodiments, the modified multi-segment antisense oligonucleotide or pharmaceutical composition is designated as the first drug.In certain embodiments, the method comprises administering the first drug and one or more second drugs.In certain embodiments, the first drug and one or more second drugs are administered simultaneously.In certain embodiments, the first drug and one or more second drugs are administered sequentially or simultaneously.In certain embodiments, the first drug and one or more second drugs are not administered simultaneously.

[0260] The modified multi-segment antisense oligonucleotides of the present disclosure and pharmaceutical compositions comprising them can be administered to a subject by any suitable route of administration. Examples of routes of administration include parenteral, for example, intravenous, intradermal, subcutaneous, oral (e.g., inhalation), transdermal (topical), intraocular, intramuscular, intraperitoneal (into a body cavity), and transmucosal administration.

[0261] In certain embodiments, administering comprises parenteral administration, hi certain embodiments, administering comprises subcutaneous administration, hi certain embodiments, administering comprises intravenous injection or infusion.

[0262] In some embodiments, ECs in cells treated with modified multisegment antisense oligonucleotides 50 was measured as a surrogate for the efficacy of the modified multisegment antisense oligonucleotide. In some embodiments, EC 50 In some embodiments, the EC50 is less than 250nM, less than 200nM, less than 150nM, less than 100nM, less than 90nM, less than 80nM, less than 70nM, less than 65nM, less than 60nM, less than 55nM, less than 50nM, less than 49nM, less than 47nM, less than 46nM, less than 45nM, less than 44nM, less than 43nM, less than 42nM, less than 41nM, less than 40nM, less than 39nM, less than 38nM, less than 37nM, less than 36nM, less than 35nM, less than 34nM, less than 33nM, less than 32nM, less than 31nM, less than 30nM, less than 29nM, less than 28nM, less than 27nM, less than 26nM , less than 25 nM, less than 24 nM, less than 23 nM, less than 22 nM, less than 21 nM, less than 20 nM, less than 19 nM, less than 18 nM, less than 17 nM, less than 16 nM, less than 15 nM, less than 14 nM, less than 13 nM, less than 12 nM, less than 11 nM, less than 10 nM, less than 9 nM, less than 8 nM, less than 7 nM, less than 6 nM, less than 5 nM, less than 4 nM, less than 3 nM, less than 2 nM, less than 1 nM, less than 0.9 nM, less than 0.8 nM, less than 0.7 nM, less than 0.6 nM, less than 0.5 nM, less than 0.4 nM, less than 0.3 nM, less than 0.2 nM, or less than 0.1 nM.

[0263] In some embodiments, ECs in cells treated with modified multisegment antisense oligonucleotides 50 / EC in cells treated with a reference oligonucleotide (e.g., a modified antisense oligonucleotide having the same template nucleobase sequence without a separator segment) 50 The ratio of EC is calculated as a measure of efficacy in reducing target protein expression levels. In some embodiments, the ratio is from about 0.05 to about 250. In some embodiments, the ratio is from about 0.05 to about 100. In some embodiments, the EC 50The ratios are: less than 250, less than 150, less than 100, less than 90, less than 80, less than 70, less than 65, less than 60, less than 55, less than 50, less than 49, less than 47, less than 46, less than 45, less than 44, less than 43, less than 42, less than 41, less than 40, less than 39, less than 38, less than 37, less than 36, less than 35, less than 34, less than 33, less than 32, less than 31, less than 30, less than 29, less than 28, less than 27, less than 26, less than 25 less than, 24, less than 23, less than 22, less than 21, less than 20, less than 19, less than 18, less than 17, less than 16, less than 15, less than 14, less than 13, less than 12, less than 11, less than 10, less than 9, less than 8, less than 7, less than 6, less than 5, less than 4, less than 3, less than 2, less than 1, less than 0.9, less than 0.8, less than 0.7, less than 0.6, less than 0.5, less than 0.4, less than 0.3, less than 0.2, or less than 0.1.

[0264] In some embodiments, the MTT CC25 value (nM) is used as a measure of cytotoxicity. In some embodiments, the MTT CC25 of cells treated with the modified multisegment antisense oligonucleotides described herein is about 10 nM to about 250 nM. In some embodiments, MTT CC25 is greater than 10 nM, greater than 15 nM, greater than 20 nM, greater than 25 nM, greater than 30 nM, greater than 35 nM, greater than 40 nM, greater than 45 nM, greater than 50 nM, greater than 55 nM, greater than 60 nM, greater than 65 nM, greater than 70 nM, greater than 75 nM, greater than 80 nM, greater than 85 nM, greater than 90 nM, greater than 95 nM, greater than 100 nM, greater than 110 nM, greater than 120 nM, greater than 130 nM, greater than 140 nM, greater than 150 nM, greater than 160 nM, greater than 170 nM, greater than 180 nM, greater than 190 nM, greater than 200 nM, greater than 210 nM, greater than 220 nM, greater than 230 nM, greater than 240 nM, or greater than 250 nM.

[0265] In some embodiments, CCK8 CC30 (nM) is used as a measure of cytotoxicity. In some embodiments, the CCK8 CC30 of cells treated with the modified multisegment antisense oligonucleotides described herein is about 10 nM to about 250 nM. In some embodiments, CCK8 CC30 is greater than 10 nM, greater than 15 nM, greater than 20 nM, greater than 25 nM, greater than 30 nM, greater than 35 nM, greater than 40 nM, greater than 45 nM, greater than 50 nM, greater than 55 nM, greater than 60 nM, greater than 65 nM, greater than 70 nM, greater than 75 nM, greater than 80 nM, greater than 85 nM, greater than 90 nM, greater than 95 nM, greater than 100 nM, greater than 110 nM, greater than 120 nM, greater than 130 nM, greater than 140 nM, greater than 150 nM, greater than 160 nM, greater than 170 nM, greater than 180 nM, greater than 190 nM, greater than 200 nM, greater than 210 nM, greater than 220 nM, greater than 230 nM, greater than 240 nM, or greater than 250 nM.

[0266] In some embodiments, the ratio of MTT CC25 or CCK8 CC30 of cells treated with a modified multisegment antisense oligonucleotide to MTT CC25 or CCK8 CC30 of cells treated with a reference oligonucleotide (e.g., a modified antisense oligonucleotide having the same template nucleobase sequence without a separator segment) is calculated as a measure of cytotoxicity (relative cytotoxicity). In some embodiments, the ratio of MTT CC25 or CCK8 CC30 of cells treated with a modified multisegment antisense oligonucleotide to MTT CC25 or CCK8 CC30 of cells treated with a reference oligonucleotide (e.g., a modified antisense oligonucleotide having the same template nucleobase sequence without a separator segment) is calculated as a measure of cytotoxicity (relative cytotoxicity). The CC30 ratio is less than 250, less than 150, less than 100, less than 90, less than 80, less than 70, less than 65, less than 60, less than 55, less than 50, less than 49, less than 47, less than 46, less than 45, less than 44, less than 43, less than 42, less than 41, less than 40, less than 39, less than 38, less than 37, less than 36, less than 35, less than 34, less than 33, less than 32, less than 31, less than 30, less than 29, less than 28, less than 27, less than 26 , less than 25, less than 24, less than 23, less than 22, less than 21, less than 20, less than 19, less than 18, less than 17, less than 16, less than 15, less than 14, less than 13, less than 12, less than 11, less than 10, less than 9, less than 8, less than 7, less than 6, less than 5, less than 4, less than 3, less than 2, less than 1, less than 0.9, less than 0.8, less than 0.7, less than 0.6, less than 0.5, less than 0.4, less than 0.3, less than 0.2, or less than 0.1.

[0267] In some embodiments, the "C / E ratio" is used as a measure of the effectiveness of reducing target protein expression levels relative to cytotoxicity. In some embodiments, the C / E ratio is calculated by taking the ratio of the MTT CC25 (nM) of the modified multi-segment antisense oligonucleotide to the MTT CC25 (nM) of a reference oligonucleotide (e.g., a modified antisense oligonucleotide having the same template nucleobase sequence but without a separator segment), and calculating it as the EC of the modified multi-segment antisense oligonucleotide. 50The C / E ratio is calculated by taking the ratio of CCK8 CC25 (nM) of the modified multisegment antisense oligonucleotide to the CCK8 CC25 (nM) of the reference oligonucleotide (e.g., a modified antisense oligonucleotide having the same template nucleobase sequence without a separator segment) and dividing it by the EC50 of the modified multisegment antisense oligonucleotide. 50 The C / E ratio is calculated by dividing the EC50 of a reference oligonucleotide (e.g., a modified antisense oligonucleotide having the same template nucleobase sequence but without a separator segment) by the EC50 of the reference oligonucleotide. In some embodiments, the C / E ratio is from about 0.01 to about 50. In some embodiments, the C / E ratio is greater than 1, greater than 1.1, greater than 1.2, greater than 1.3, greater than 1.4, greater than 1.5, greater than 1.6, greater than 1.7, greater than 1.8, greater than 1.9, greater than 2.0, greater than 2.1, greater than 2.2, greater than 2.3, greater than 2.4, greater than 2.5, greater than 2.6, greater than 2.7, greater than 2.8, greater than 2.9, greater than 3.0, greater than 10, greater than 11, greater than 12, greater than 13, greater than 14, greater than 15, greater than 16, greater than 17, greater than 18, greater than 19, greater than 20, greater than 21, greater than 22, greater than 23, greater than 24, greater than 25, greater than 30, greater than 35, greater than 40, greater than 45, greater than 50, greater than 55, greater than 65, greater than 70, greater than 80, greater than 90, or greater than 100.

[0268] In some embodiments, the melting temperature of a modified multi-segment antisense oligonucleotide is used as a measure of stability. In some embodiments, the melting temperature of a modified multi-segment antisense oligonucleotide described herein is at least 0.01 times lower than that of a reference oligonucleotide (e.g., a modified antisense oligonucleotide having the same template nucleobase sequence but without a separator segment). In some embodiments, the melting temperature is about 0.01 times, about 0.02 times, about 0.03 times, about 0.04 times, about 0.05 times, about 0.06 times, about 0.07 times, about 0.08 times, about 0.09 times, about 0.1 times, about 0.15 times, about 0.2 times, about 0.25 times, about 0.3 times, about 0.35 times, about 0. 4x, approximately 0.45x, approximately 0.5x, approximately 0.55x, approximately 0.6x, approximately 0.65x, approximately 0.7x, approximately 0.75x, approximately 0.8x, approximately 0.85x, approximately 0.9x, approximately 0.95x, approximately 1x, approximately 1.1x, approximately 1.15x, approximately 1 .2 times, approximately 1.25 times, approximately 1.3 times, approximately 1.35 times, approximately 1.4 times, approximately 1.45 times, approximately 1.5 times, approximately 1.55 times, approximately 1.6 times, approximately 1.65 times, approximately 1.7 times, approximately 1.75 times, approximately 1.8 times, approximately 1.85 times, approximately 1.9 times, Approximately 1.95 times, approximately 2 times, approximately 2.1 times, approximately 2.15 times, approximately 2.2 times, approximately 2.25 times, approximately 2.3 times, approximately 2.35 times, approximately 2.4 times, approximately 2.45 times, approximately 2.5 times, approximately 2.55 times, approximately 2.6 times, approximately 2.65 times, approximately 2.7 times , approximately 2.75 times, approximately 2.8 times, approximately 2.85 times, approximately 2.9 times, approximately 2.95 times, approximately 3 times, approximately 3.1 times, approximately 3.15 times, approximately 3.2 times, approximately 3.25 times, approximately 3.3 times, approximately 3.35 times, approximately 3.4 times, approximately 3.45 times, approximately 3.5 times , about 3.55 times, about 3.6 times, about 3.65 times, about 3.7 times, about 3.75 times, about 3.8 times, about 3.85 times, about 3.9 times, about 3.95 times, about 4 times, about 4.1 times, about 4.15 times, about 4.2 times, about 4.25 times, about 4.3 times, about 4.35 times, about 4.4 times, about 4.45 times, about 4.5 times, about 4.55 times, about 4.6 times, about 4.65 times, about 4.7 times, about 4.75 times, about 4.8 times, about 4.85 times, about 4.9 times, about 4.95 times, or about 5 times lower.

[0269] In some embodiments, the ability of a modified multi-segment antisense oligonucleotide to recruit an endonuclease (e.g., RNase H) and degrade a template sequence is used as a measure of biological function. In some embodiments, the degradation of the template sequence by the modified multi-segment oligonucleotide is at least 0.01-fold greater than that of a reference oligonucleotide (e.g., a modified antisense oligonucleotide having the same template nucleobase sequence but without a separator segment). In some embodiments, the degradation of the template sequence by the modified multi-segment oligonucleotide is about 0.01-fold, about 0.02-fold, about 0.03-fold, about 0.04-fold, about 0.05-fold, about 0.06-fold, about 0.07-fold, about 0.08-fold, about 0.09-fold, about 0.1-fold, about 0.15-fold, about 0.2-fold, about 0.2-fold, or about 0.2-fold greater than that of a reference oligonucleotide (e.g., a modified antisense oligonucleotide having the same template nucleobase sequence but without a separator segment). 5 times, approximately 0.3 times, approximately 0.35 times, approximately 0.4 times, approximately 0.45 times, approximately 0.5 times, approximately 0.55 times, approximately 0.6 times, approximately 0.65 times, approximately 0.7 times, approximately 0.75 times, approximately 0.8 times, approximately 0.85 times, approximately 0.9 times, approximately 0.95 times, approximately 1 times, Approximately 1.1 times, approximately 1.15 times, approximately 1.2 times, approximately 1.25 times, approximately 1.3 times, approximately 1.35 times, approximately 1.4 times, approximately 1.45 times, approximately 1.5 times, approximately 1.55 times, approximately 1.6 times, approximately 1.65 times, approximately 1.7 times, approximately 1.75 times, approximately 1.8 times, approximately 1.85 times, approximately 1.9 times, approximately 1.95 times, approximately 2 times, approximately 2.1 times, approximately 2.15 times, approximately 2.2 times, approximately 2.25 times, approximately 2.3 times, approximately 2.35 times, approximately 2.4 times, approximately 2.45 times, approximately 2.5 times, approximately 2.55 times, approximately 2.6 times, approximately 2. 65 times, approximately 2.7 times, approximately 2.75 times, approximately 2.8 times, approximately 2.85 times, approximately 2.9 times, approximately 2.95 times, approximately 3 times, approximately 3.1 times, approximately 3.15 times, approximately 3.2 times, approximately 3.25 times, approximately 3.3 times, approximately 3.35 times, approximately 3.4 times, approximately 3.45 times , about 3.5 times, about 3.55 times, about 3.6 times, about 3.65 times, about 3.7 times, about 3.75 times, about 3.8 times, about 3.85 times, about 3.9 times, about 3.95 times, about 4 times, about 4.1 times, about 4.15 times, about 4.2 times, about 4.25 times, about 4.3 times, about 4.35 times, about 4.4 times, about 4.45 times, about 4.5 times, about 4.55 times, about 4.6 times, about 4.65 times, about 4.7 times, about 4.75 times, about 4.8 times, about 4.85 times, about 4.9 times, about 4.95 times, or about 5 times larger.

[0270] Kits and manufactured products The present disclosure provides a kit comprising the modified multi-segment antisense oligonucleotide described herein and a pharmaceutical composition comprising the same. In some embodiments, the modified multi-segment antisense oligonucleotide comprises any one of the structures of the oligonucleotide sequences described herein.

[0271] Target Nucleic Acids, Target Regions, and Nucleotide Sequences The modified multi-segment antisense oligonucleotides can be directed to any target of interest.

[0272] The target can be any target whose activity is desired to be reduced. In some embodiments, the target is a mammalian target, e.g., a human or other animal target, and the target is directed to an oncogenic target, an inflammatory target, a metabolic target, a cardiovascular target, a neurological target, a neuropsychiatric target, etc. In some embodiments, the target is a gene transcript (e.g., mRNA) or an RNA that regulates the expression of a gene. Exemplary genes include, but are not limited to, DMPK, AAT, TTR, PCSK9, ApoB, ApoCIII, TNF-alpha, AGT, CFB, DGAT2, PNPLA3, ALAS1, or hydroxyacid oxidase 1. In some embodiments, the gene is DMPK, AAT, TTR, or PCSK9.

[0273] In some embodiments, the gene is selected from the group consisting of Mat1α, (pro)renin receptor / (P)RR, COVID-19 5′UTR, Acc1, Acc2, ACE2, androgen receptor, ApoB, ASGR1, ASO, ATXN2, B1AR, B2AR, C9ORF72, caspase 2, CD19, CD4, chikungunya virus, CLPro, complement component 5, COVID19, COX2, CTGF, DGAT2, DMD, DMPK, DNM2, DUX4, E2 gene, EGFR, envelope, EphA2, epithelial sodium channel alpha subunit, exon 100 dystrophin, exon 101 dystrophin, exon 102 dystrophin. dystrophin, exon 103 dystrophin, exon 104 dystrophin, exon 105 dystrophin, exon 106 dystrophin, exon 107 dystrophin, exon 108 dystrophin, exon 109 dystrophin, exon 110 dystrophin, exon 111 dystrophin, exon 112 dystrophin, exon 113 dystrophin, exon 114 dystrophin, exon 115 dystrophin, exon 116 dystrophin, exon 117 dystrophin, exon 118 Dystrophin, exon 119 dystrophin, exon 120 dystrophin, exon 121 dystrophin, exon 122 dystrophin, exon 123 dystrophin, exon 124 dystrophin, exon 125 dystrophin, exon 126 dystrophin, exon 127 dystrophin, exon 128 dystrophin, exon 129 dystrophin, exon 130 dystrophin, exon 131 dystrophin, exon 132 dystrophin, exon 133 dystrophin, exon Exon 134 dystrophin, exon 135 dystrophin, exon 136 dystrophin, exon 137 dystrophin, exon 138 dystrophin, exon 139 dystrophin, exon 140 dystrophin, exon 141 dystrophin, exon 142 dystrophin, exon 143 dystrophin, exon 144 dystrophin, exon 145 dystrophin, exon 146 dystrophin, exon 147 dystrophin, exon 148 dystrophin, exon 149 dystrophin,Exon 150 dystrophin, exon 151 dystrophin, exon 152 dystrophin, exon 44 dystrophin, exon 53 dystrophin, exon 54 dystrophin, exon 55 dystrophin, exon 56 dystrophin, exon 57 dystrophin, exon 58 dystrophin, exon 59 dystrophin, exon 60 dystrophin, exon 61 dystrophin, exon 62 dystrophin, exon 63 dystrophin, exon 64 dystrophin, exon 65 dystrophin, exon 66 dystrophin dystrophin, exon 67 dystrophin, exon 68 dystrophin, exon 69 dystrophin, exon 70 dystrophin, exon 71 dystrophin, exon 72 dystrophin, exon 73 dystrophin, exon 74 dystrophin, exon 75 dystrophin, exon 76 dystrophin, exon 77 dystrophin, exon 78 dystrophin, exon 79 dystrophin, exon 80 dystrophin, exon 81 dystrophin, exon 82 dystrophin, exon 83 dystrophin, exon 84 Dystrophin, exon 85 dystrophin, exon 86 dystrophin, exon 87 dystrophin, exon 88 dystrophin, exon 89 dystrophin, exon 90 dystrophin, exon 91 dystrophin, exon 92 dystrophin, exon 93 dystrophin, exon 94 dystrophin, exon 95 dystrophin, exon 96 dystrophin, exon 97 dystrophin, exon 98 dystrophin, exon 99 dystrophin, Fabp3, Factor VII, Factor XI, FAK, FGFR4, FO XP3, FUS, FXII, GFAP, GFP, glycogen synthase, H1N1, HBV, heat shock protein 47, heat shock protein 48, heat shock protein 49, heat shock protein 50, heat shock protein 51, heat shock protein 52, heat shock protein 53, heat shock protein 54, heat shock protein 55, heat shock protein 56, heat shock protein 57, heat shock protein 58, heat shock protein 59, helicase, HIV, HOXB13, HPRT, HPV, Hsd11β1, HTT,Hydroxyacid oxidase 1, hydroxysteroid 17β-dehydrogenase 13, hypoxia-inducible factor 2α, kinesin spindle protein and vascular endothelial growth factor, KRAS, leader, leucine-rich repeat kinase 2 (LRRK2), MERS-CoV, MGMT, miR-16, miR-21, MMP-2, MMP-9, MTL-CEBPA, MuRF1, Mycobacterium tuberculosis, COVID-19 N, rabies N 123, rabies N 749, rabies N 903, rabies N1082, rabies N53, rabies N8, chikungunya virus ns1 gene, chikungunya virus ns2 gene, chikungunya virus ns3 gene, chikungunya virus ns4 gene, ORF1 b, ORF1 a, HIV P24, rabies P330, P53, rabies P721, rabies P91, PCSK9, PDGF, PDL1, COVID-19 PLP, PNPLA3, polo-like kinase 1, protein kinase N3, RAF1, RAF-1, RDRP, RSV, SMN2, SNCA, Covid-19 spike, STAT3, TAU, TGFB1, TGFB1 and Cox2, TMPRSS2, TMPRSS6, TNFα, transthyretin, VEGF, VEGFR2, VER2, xanthine dehydrogenase, or YAP1.

[0274] Exemplary nucleic acid target sequences include, but are not limited to, those shown in Tables 8.1, 8.2, and 8.3. In some embodiments, the nucleic acid target comprises any one of the sequences set forth in Tables 8.1, 8.2, and 8.3. Each of the residues in Tables 8.1, 8.2, and 8.3 reflects only the sequence information without modified nucleosides.

[0275] [Table 4]

[0276] [Table 5-1]

[0277] [Table 5-2]

[0278] [Table 5-3]

[0279] [Table 5-4]

[0280] [Table 5-5]

[0281] HBV Certain embodiments provide methods, modified multi-segment antisense oligonucleotides, and compositions for inhibiting HBV mRNA expression.

[0282] Certain embodiments provide modified multi-segment antisense oligonucleotides that target HBV nucleic acid sequences. Exemplary HBV nucleic acid sequences include, but are not limited to, those shown in Table 8.3.

[0283] [Table 6-1]

[0284] [Table 6-2]

[0285] [Table 6-3]

[0286] [Table 6-4]

[0287] Table 6-5

[0288] Table 6-6

[0289] Table 6-7

[0290] Table 6-8

[0291] Table 6-9

[0292] Table 6-10

[0293] Table 6-11

[0294] Table 6-12

[0295] Table 6-13

[0296] Table 6-14

[0297] In certain embodiments, the HBV nucleic acid is the sequence set forth in GENBANK Accession No. U95551.1 (incorporated herein as SEQ ID NO:3). In some embodiments, the modified multi-segment antisense oligonucleotide targets the sequence set forth in SEQ ID NO:3 or a portion thereof. In some embodiments, the modified multi-segment antisense oligonucleotide targets the sequence from positions 1583 to 1602 of SEQ ID NO:3.

[0298] PCSK9 Proprotein convertase subtilisin / kexin type 9 (PCSK9) may also be referred to as subtilisin / kexin-like protease PC9. Human PCSK9 has a cytogenetic location of 1p32.3, with genomic coordinates on chromosome 1 at positions 55,039,548-55,064,852 on the normal strand. Human PCSK9 has an NCBI gene ID of 255738, a reference sequence accession number of NM_174936.4, a reference sequence accession number of NP_777596.2, and an Ensembl gene ID of ENSG00000169174.

[0299] PCSK9 activity is primarily found in the liver, and PCSK9 is associated with dyslipidemia, PCSK9-associated familial hypercholesterolemia, hypercholesterolemia (familial), gastric papillary adenocarcinoma, homozygous familial hypercholesterolemia, and nasopharyngitis. PCSK9-associated familial hypercholesterolemia is a genetic disease (autosomal dominant) in which the body dangerously increases blood cholesterol levels due to a lack of receptors for low-density lipoprotein cholesterol. PCSK9-associated familial hypercholesterolemia affects 1 in 500 heterozygous people to 1 in 1,000,000 homozygous people worldwide, and is more common in Afrikaner, French-Canadian, Lebanese Christian, and Finnish populations. Common symptoms of PCSK9-associated familial hypercholesterolemia include elevated circulating cholesterol contained in either low-density lipoproteins alone or very-low-density lipoproteins in combination. Current treatments for PCSK9-associated familial hypercholesterolemia include the administration of statins to inhibit hydroxymethylglutaryl CoA reductase (HMG-CoA reductase) in the liver. Another option for treating PCSK9-associated familial hypercholesterolemia is ezetimibe, which inhibits cholesterol absorption in the intestine.

[0300] In some embodiments, the PCSK9 nucleic acid is a sequence set forth in the NCBI gene ID of NM_174936.4, or a portion thereof.

[0301] In some embodiments, the PCSK9 target comprises a sequence set forth in SEQ ID NO: 1454, 1458, or 1472, or a portion thereof, or a variant thereof. In some embodiments, the modified multi-segment antisense oligonucleotide is at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or at least 100% complementary to SEQ ID NO: 1454, 1458, or 1472.

[0302] In some embodiments, the modified multi-segment antisense oligonucleotide comprises the nucleobase sequence of SEQ ID NO: 1432, 1436, or 1450. In some embodiments, the modified multi-segment antisense oligonucleotide is at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or at least 100% identical to SEQ ID NO: 1432, 1436, or 1450.

[0303] DMPK DM1 protein kinase (DMPK) may also be referred to as DM, DM1, DMK, MDPK, DM1PK, or MT-PK. Human DMPK has a cytogenetic location of 19q13.32, with genomic coordinates of 45,769,709-45,782,490 on opposite strands of chromosome 19. Human DMPK has an NCBI gene ID of 1760, a reference sequence accession number of NM_004409.5, a reference sequence accession number of NP_004400.4, and an Ensembl gene ID of ENSG00000104936.

[0304] DMPK activity is primarily found in muscles, and DMPK is associated with myotonic dystrophy 1 and adult-onset Steinert myotonic dystrophy. The genetic defect in DM1 results from an amplified trinucleotide repeat in the 3-prime untranslated region of the protein kinase gene. Disease severity varies with the number of repeats; normal individuals have 5–37 repeats, mildly affected individuals have 50–150 repeats, patients with classic DM have 100–1,000 repeats, and patients with onset at birth may have more than 2,000 repeats. This disorder exhibits genetic anticipation, and the expansion of repeat number depends on the sex of the transmitting parent. Alleles with 40–80 repeats are usually expanded when transmitted by males, but only alleles longer than 80 repeats tend to be expanded in maternal transmission. Repeat contraction events occur 4.2–6.4% of the time. The prevalence of DM1 exceeds 1 / 1000 in certain populations and ranges from 1 / 9 / 100,000 worldwide, particularly in Europe, Italy, Japan, Ireland, Norway, and the United States. DM1 is primarily characterized by muscle tone, muscular dystrophy, cataracts, hypogonadism, frontal baldness, and ECG changes. Currently approved medications for DM1 include methylphenidate, dopamine, lamotrigine, mexiletine, and metformin.

[0305] In some embodiments, the DMPK nucleic acid is the sequence set forth in the NCBI gene ID of NM_004409.5, or a portion thereof.

[0306] In some embodiments, the DMPK target comprises the sequence set forth in SEQ ID NO: 1455, 1459, 1460, or 1461, or a portion thereof, or a variant thereof. In some embodiments, the modified multi-segment antisense oligonucleotide is at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or at least 100% complementary to SEQ ID NO: 1455, 1459, 1460, or 1461.

[0307] In some embodiments, the modified multi-segment antisense oligonucleotide has the nucleobase sequence of SEQ ID NO: 1433, 1437, 1438, or 1439. In some embodiments, the modified multi-segment antisense oligonucleotide is at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or at least 100% identical to SEQ ID NO: 1433, 1437, 1438, or 1439.

[0308] TTR Transthyretin (TTR) may also be referred to as CTS, TTN, ATTR, CTS1, PALB, TBPA, HEL111, or HsT2651. Human TTR has a cytogenetic location of 18q12.1, with genomic coordinates of 31,591,877-31,598,821 on chromosome 18 on the normal strand. Human TTR has an NCBI gene ID of 7276, a reference sequence accession number of NM_000371.4, a reference sequence accession number of NP_000362.1, and an Ensembl gene ID of ENSG00000118271.

[0309] Transthyretin synthesized in the liver is secreted into the blood, while transthyretin derived from the choroid plexus is directed to the cerebrospinal fluid (CSF). Transthyretin is associated with both localized and systemic amyloidosis, disorders characterized by the extracellular systemic deposition of mutant or wild-type transthyretin as amyloid fibrils. Senile systemic amyloidosis is a sporadic disorder resulting from the extracellular deposition of wild-type transthyretin fibrils in the heart and other tissues. Over 80 mutations in transthyretin are associated with familial amyloid polyneuropathy and cardiomyopathy. The variants Val122Ile, Val30Met, and Gluo.1Lys are common amyloidogenic mutations worldwide. In most of these cases, inheritance is autosomal dominant. The prevalence of Val30Met amyloidosis is 6-9 / 10,000 in Portugal and 1-9 / 1,000,000 in Japan and Cyprus. Patients with transthyretin amyloidosis typically present with polyneuropathy, carpal tunnel syndrome, autonomic dysfunction, cardiomyopathy, and gastrointestinal features, sometimes accompanied by vitreous opacities and renal insufficiency. In the later stages of the disease, severe diarrhea with malabsorption, cachexia, disabling neuropathy, severe cardiac involvement, and marked orthostatic hypotension dominate the clinical picture. Death usually occurs 5-15 years after symptom onset. The FDA has approved butrisiran (Amvuttra), patisiran (Onpattro), and inotersen (Tegsedi) for the treatment of polyneuropathy caused by hATTR in adults. Tafamidis (Vyndamax) and tafamidis meglumine (Vyndaqel) are FDA-approved for transthyretin-mediated amyloidotic cardiomyopathy (ATTR-CM).

[0310] In some embodiments, the TTR nucleic acid is the sequence set forth in the NCBI gene ID of NM_000371.4, or a portion thereof.

[0311] In some embodiments, the TTR target comprises the sequence set forth in SEQ ID NO: 1456, 1462, or 1474, or a portion thereof, or a variant thereof. In some embodiments, the modified multi-segment antisense oligonucleotide is at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or at least 100% complementary to SEQ ID NO: 1456, 1462, or 1474.

[0312] In some embodiments, the modified multi-segment antisense oligonucleotide has the nucleobase sequence of SEQ ID NO: 1434, 1440, or 1452. In some embodiments, the modified multi-segment antisense oligonucleotide is at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or at least 100% identical to SEQ ID NO: 1434, 1440, or 1452.

[0313] AAT Serpin family A member 1 (SERPINA1) may also be referred to as PI, A1A, AAT, PI1, A1AT, nNIF, PRO2275, or alpha1AT. Human SERPINA1 has a cytogenetic location of 14q32.13, with genomic coordinates of 94,376,747-94,390,635 on the opposite strand of chromosome 14. Human SERPINA1 has an NCBI gene ID of 5265, a reference sequence accession number of NM_000295.5, a reference sequence accession number of NP_000286.3, and an Ensembl gene ID of ENSG00000197249.

[0314] AAT is primarily produced by hepatocytes and secreted into the blood, although low levels of AAT expression are also detected in the lungs and intestine. SERPINA1-associated disorders include alpha-1 antitrypsin deficiency and bleeding disorders caused by the alpha-1 antitrypsin Pittsburgh mutation. Alpha-1 antitrypsin deficiency is a genetic disorder that can cause lung and liver disease. Alpha-1 antitrypsin deficiency occurs worldwide, but its prevalence varies among populations. The disorder affects approximately 1 in 1,500–3,500 individuals of European descent. It is rare in people of Asian descent. Many individuals with alpha-1 antitrypsin deficiency, especially those with a lung condition called chronic obstructive pulmonary disease (COPD), are likely to be undiagnosed. COPD can be caused by alpha-1 antitrypsin deficiency. However, alpha-1 antitrypsin deficiency often goes undiagnosed. Some people with alpha-1 antitrypsin deficiency are misdiagnosed as having asthma. Treatment for lung disease may include bronchodilators, inhaled steroids, and antibiotics if infection occurs. Intravenous infusion of A1AT protein or, in severe cases, lung transplantation may also be recommended. For people with severe liver disease, liver transplantation may be an option. Avoiding smoking and receiving vaccinations against influenza, pneumococcal disease, and hepatitis are also recommended.

[0315] In some embodiments, the AAT nucleic acid is the sequence set forth in the NCBI gene ID of NM_000295.5, or a portion thereof.

[0316] In some embodiments, the AAT target comprises the sequence set forth in SEQ ID NO: 1457 or 1463, or a portion thereof, or a variant thereof. In some embodiments, the modified multi-segment antisense oligonucleotide is at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or at least 100% complementary to SEQ ID NO: 1457 or 1463.

[0317] In some embodiments, the modified multi-segment antisense oligonucleotide has the nucleobase sequence of SEQ ID NO: 1435 or 1441. In some embodiments, the modified multi-segment antisense oligonucleotide is at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or at least 100% identical to SEQ ID NO: 1435 or 1441.

[0318] Generally, the target of the modified multi-segment antisense oligonucleotide of the present disclosure is a gene transcript. In certain embodiments, the target region is a structurally defined region of the target nucleic acid. For example, the target region can include a 3'UTR, a 5'UTR, an exon, an intron, an exon / intron junction, a coding region, a translation initiation region, a translation termination region, or other defined nucleic acid region. The structurally defined region of the target can be obtained by accession number from a sequence database such as NCBI, and such information is incorporated herein by reference. In certain embodiments, the target region can include the sequence from the 5' target site of one target segment within the target region to the 3' target site of another target segment within the same target region.

[0319] Targeting involves determining at least one target segment to which the antisense compound hybridizes, resulting in a desired effect. In certain embodiments, the desired effect is a reduction in the level of mRNA target nucleic acid. In certain embodiments, the desired effect is a reduction in the level of the protein encoded by the target nucleic acid or a change in the phenotype associated with the target nucleic acid.

[0320] A target region may contain one or more target segments. Multiple target segments within a target region may overlap. Alternatively, they may not overlap. In certain embodiments, target segments within a target region are separated by no more than about 300 nucleotides. In certain embodiments, target segments within a target region are separated by about, no more than, or no more than about 250, 200, 150, 100, 90, 80, 70, 60, 50, 40, 30, 20, or 10 nucleotides on the target nucleic acid, or a range defined by any two of the foregoing values. In certain embodiments, target segments within a target region are separated by no more than, or no more than about, 5 nucleotides on the target nucleic acid. In certain embodiments, target segments are contiguous. Target regions defined by a range having a starting nucleic acid that is any of the 5' or 3' target sites listed herein are contemplated.

[0321] Suitable target segments can be found within the 5'UTR, coding region, 3'UTR, intron, exon, or exon / intron junction. Target segments containing a start codon or a stop codon are also suitable target segments. Suitable target segments can specifically exclude certain structurally defined regions, such as the start codon or the stop codon.

[0322] The determination of suitable target segment can include comparing the sequence of target nucleic acid with other sequences throughout the genome.For example, can use BLAST algorithm to identify the similarity region between different nucleic acids.This comparison can prevent the selection of antisense oligonucleotide sequence that can hybridize in a non-specific manner with sequences other than the selected target nucleic acid (i.e., non-target sequence or off-target sequence).

[0323] Hybridization In some embodiments, hybridization occurs between the antisense oligonucleotides disclosed herein and the target nucleic acid. The most common mechanism of hybridization involves hydrogen bonding (e.g., Watson-Crick, Hoogsteen, or reversed Hoogsteen hydrogen bonding) between complementary nucleobases of nucleic acid molecules.

[0324] Hybridization can occur under a variety of conditions. Stringent conditions are sequence-dependent and are determined by the nature and composition of the nucleic acid molecules to be hybridized.

[0325] Methods for determining whether a sequence is specifically hybridizable to a target nucleic acid are well known in the art. In certain embodiments, the antisense oligonucleotides provided herein are specifically hybridizable to the target nucleic acid.

[0326] Complementarity An antisense oligonucleotide and a target nucleic acid are complementary to one another if a sufficient number of nucleobases of the antisense oligonucleotide are capable of hydrogen bonding with corresponding nucleobases of the target nucleic acid, thereby producing a desired effect (e.g., antisense inhibition of the target nucleic acid).

[0327] Non-complementary nucleobases between the antisense oligonucleotide and the target nucleic acid can be tolerated, provided that the antisense oligonucleotide remains capable of specifically hybridizing to the target nucleic acid. Furthermore, the antisense oligonucleotide can hybridize across one or more segments of the target nucleic acid such that intervening or adjacent segments are not involved in the hybridization event (e.g., a loop structure, mismatch, or hairpin structure).

[0328] In certain embodiments, the antisense oligonucleotides provided herein, or designated portions thereof, are 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% complementary to a target nucleic acid, target region, target segment, or designated portion thereof. The percent complementarity of an antisense oligonucleotide with a target nucleic acid can be determined using conventional methods.

[0329] For example, antisense oligonucleotide, in which 18 of the 20 nucleobases of antisense oligonucleotide are complementary to target region, and therefore specifically hybridize, shows 90% complementarity.In this example, the remaining non-complementary nucleobases can be clustered with complementary nucleobases or can be scattered, and do not need to be adjacent to each other or complementary nucleobases.Therefore, the antisense oligonucleotide that is 18 nucleobases long, has 4 non-complementary nucleobases adjacent to two regions of complete complementarity with target nucleic acid, has 77.8% overall complementarity with target nucleic acid, and therefore falls within the scope of the present invention. The percent complementarity of antisense oligonucleotides with a region of target nucleic acid can be routinely determined using BLAST (basic local alignment search tool) and PowerBLAST programs known in the art (Altschul et al., J. Mol. Biol., 1990, 215, 403-410; Zhang and Madden, Genome Res., 1997, 7, 649-656).Percent homology, sequence identity, or complementarity can be determined, for example, by the Gap program (Wisconsin Sequence Analysis Package, Version 8 for Unix, Genetics Computer Group, University Research Park, Madison Wis.) using the default settings of the Smith and Waterman algorithm (Adv. Appl. Math., 1981, 2, 482-489).

[0330] In certain embodiments, the antisense oligonucleotides provided herein or their designated portions are fully complementary (i.e., 100% complementary) to the target nucleic acid or its designated portion. As used herein, "fully complementary" means that each nucleobase of the antisense oligonucleotide can precisely base pair with the corresponding nucleobase of the target nucleic acid. For example, a 20-nucleobase antisense oligonucleotide is fully complementary to a target sequence that is 400 nucleobases long, as long as there is a corresponding 20-nucleobase portion of the target nucleic acid that is fully complementary to the antisense oligonucleotide. "Fully complementary" can also be used in reference to a designated portion of a first and / or second nucleic acid. For example, the 20-nucleobase portion of a 30-nucleobase antisense oligonucleotide can be "fully complementary" to a target sequence that is 400 nucleobases long. The 20-nucleobase portion of a 30-nucleobase oligonucleotide is fully complementary to a target sequence if the target sequence has a corresponding 20-nucleobase portion, and each nucleobase is complementary to the 20-nucleobase portion of the antisense oligonucleotide. At the same time, the entire 30 nucleobase antisense oligonucleotide may or may not be perfectly complementary to the target sequence, depending on whether the remaining 10 nucleobases of the antisense oligonucleotide are also complementary to the target sequence.

[0331] The non-complementary nucleobase may be located at the 5'-end or 3'-end of the antisense oligonucleotide. Alternatively, the non-complementary nucleobase may be located at an internal position of the antisense oligonucleotide. When two or more non-complementary nucleobases are present, they may be contiguous (i.e., linked) or non-contiguous. In one embodiment, the non-complementary nucleobase is located in the wing segment of a gapmer antisense oligonucleotide. In one embodiment, the non-complementary nucleoba...

Claims

1. 1. A modified multi-segment antisense oligonucleotide that is complementary to a nucleotide sequence in a gene transcript, said modified multi-segment antisense oligonucleotide comprising, 5' to 3': 5'W1-G1-S1-G2-W2 3' (Formula I), During the ceremony, W1 is the 5' wing segment; W2 is the 3' wing segment; G1 is the first gap segment; S1 is the first separator segment; G2 is the second gap segment; each segment is joined by an internucleoside linkage; A modified multi-segment antisense oligonucleotide, wherein at least one nucleoside of each of W1, W2, and S1 is a modified nucleoside.

2. The modified multisegment antisense oligonucleotide comprises, 5' to 3': 5'W1-G1-S1-G2-S2-G3-W2 3' (Formula II), During the ceremony, S2 is a second separator segment; 2. The modified multi-segment antisense oligonucleotide of claim 1, wherein G3 is a third gap segment and at least one nucleoside of S2 is a modified nucleoside.

3. The modified multi-segment antisense oligonucleotide of claim 1 or 2, wherein W1 comprises 2 to 25 linked nucleosides.

4. 4. The modified multi-segment antisense oligonucleotide of claim 3, wherein the first 5' nucleoside of W1 is a modified nucleoside and / or the last 3' nucleoside of W1 is a modified nucleoside.

5. The modified multi-segment antisense oligonucleotide of any one of claims 1 to 4, wherein W2 comprises 2 to 35 linked nucleosides.

6. 6. The modified multi-segment antisense oligonucleotide of claim 5, wherein the first 5' nucleoside of W2 is a modified nucleoside and / or the last 3' nucleoside of W2 is a modified nucleoside.

7. The modified multi-segment antisense oligonucleotide of any one of claims 1 to 6, wherein G1 comprises 1 to 10 linked deoxynucleosides.

8. The modified multi-segment antisense oligonucleotide of any one of claims 1 to 7, wherein G2 comprises 1 to 10 linked deoxynucleosides.

9. The modified multi-segment antisense oligonucleotide of any one of claims 1 to 8, wherein S1 comprises 1 to 3 linked nucleosides.

10. The modified multi-segment antisense oligonucleotide of any one of claims 2 to 9, wherein G3 comprises 1 to 10 linked deoxynucleosides.

11. The modified multi-segment antisense oligonucleotide of any one of claims 2 to 10, wherein S2 comprises 1 to 3 linked nucleosides.

12. The modified multi-segment antisense oligonucleotide of any one of claims 1 to 11, wherein the modified multi-segment antisense oligonucleotide is 11 to 50 nucleobases in length.

13. The modified multi-segment antisense oligonucleotide of any one of claims 1 to 11, wherein the modified multi-segment antisense oligonucleotide is 11 to 30 nucleobases in length.

14. The modified multi-segment antisense oligonucleotide of any one of claims 1 to 11, wherein the modified multi-segment antisense oligonucleotide is at least 20 nucleobases in length.

15. The modified multi-segment antisense oligonucleotide of any one of claims 1 to 11, wherein the modified multi-segment antisense oligonucleotide is 20 nucleobases in length.

16. 16. The modified multi-segment antisense oligonucleotide of claim 15, wherein W1 comprises 2 to 8 linked nucleosides.

17. 16. The modified multi-segment antisense oligonucleotide of claim 15, wherein W1 comprises 2 to 6 linked nucleosides.

18. 16. The modified multi-segment antisense oligonucleotide of claim 15, wherein W1 comprises 4, 5, or 6 linked nucleosides.

19. 16. The modified multi-segment antisense oligonucleotide of claim 15, wherein W1 comprises five linked nucleosides.

20. 20. The modified multi-segment antisense oligonucleotide of any one of claims 15 to 19, wherein G1 comprises 2 to 7 linked deoxynucleosides.

21. 20. The modified multi-segment antisense oligonucleotide of any one of claims 15 to 19, wherein G1 comprises four linked deoxynucleosides.

22. 20. The modified multi-segment antisense oligonucleotide of any one of claims 15 to 19, wherein G1 comprises five linked deoxynucleosides.

23. 20. The modified multi-segment antisense oligonucleotide of any one of claims 15 to 19, wherein G1 comprises 6 linked deoxynucleosides.

24. 24. The modified multi-segment antisense oligonucleotide of any one of claims 15 to 23, wherein G2 comprises 1 to 7 linked deoxynucleosides.

25. 24. The modified multi-segment antisense oligonucleotide of any one of claims 15 to 23, wherein G2 comprises 2 to 7 linked deoxynucleosides.

26. 24. The modified multi-segment antisense oligonucleotide of any one of claims 15 to 23, wherein G2 comprises 1 to 2 linked deoxynucleosides.

27. 24. The modified multi-segment antisense oligonucleotide of any one of claims 15 to 23, wherein G2 comprises four linked deoxynucleosides.

28. 24. The modified multi-segment antisense oligonucleotide of any one of claims 15 to 23, wherein G2 comprises five linked deoxynucleosides.

29. 24. The modified multi-segment antisense oligonucleotide of any one of claims 15 to 23, wherein G2 comprises 6 linked deoxynucleosides.

30. 24. The modified multi-segment antisense oligonucleotide of any one of claims 15 to 23, wherein G2 comprises 7 linked deoxynucleosides.

31. 31. The modified multi-segment antisense oligonucleotide of any one of claims 15 to 30, wherein G3 comprises 2 to 7 linked deoxynucleosides.

32. The modified multi-segment antisense oligonucleotide of any one of claims 15 to 30, wherein G3 comprises four linked deoxynucleosides.

33. 31. The modified multi-segment antisense oligonucleotide of any one of claims 15 to 30, wherein G3 comprises five linked deoxynucleosides.

34. 31. The modified multi-segment antisense oligonucleotide of any one of claims 15 to 30, wherein G3 comprises 6 linked deoxynucleosides.

35. 31. The modified multi-segment antisense oligonucleotide of any one of claims 15 to 30, wherein G3 comprises 7 linked deoxynucleosides.

36. The modified multi-segment antisense oligonucleotide of any one of claims 15 to 35, wherein S1 comprises at least one linked nucleoside.

37. The modified multi-segment antisense oligonucleotide of any one of claims 15 to 35, wherein S1 comprises one linked nucleoside.

38. The modified multi-segment antisense oligonucleotide of any one of claims 15 to 35, wherein S1 comprises two linked nucleosides.

39. 39. The modified multi-segment antisense oligonucleotide of any one of claims 15 to 38, wherein S2 comprises at least one linked nucleoside.

40. 39. The modified multi-segment antisense oligonucleotide of any one of claims 15 to 38, wherein S2 comprises one linked nucleoside.

41. 39. The modified multi-segment antisense oligonucleotide of any one of claims 15 to 38, wherein S2 comprises two linked nucleosides.

42. 42. The modified multi-segment antisense oligonucleotide of any one of claims 15 to 41, wherein W2 comprises from 2 to 8 linked nucleosides.

43. 42. The modified multi-segment antisense oligonucleotide of any one of claims 15 to 41, wherein W2 comprises 2 to 6 linked nucleosides.

44. 42. The modified multi-segment antisense oligonucleotide of any one of claims 15 to 41, wherein W2 comprises three linked nucleosides.

45. 42. The modified multi-segment antisense oligonucleotide of any one of claims 15 to 41, wherein W2 comprises four linked nucleosides.

46. 42. The modified multi-segment antisense oligonucleotide of any one of claims 15 to 41, wherein W2 comprises 5 linked nucleosides.

47. 42. The modified multi-segment antisense oligonucleotide of any one of claims 15 to 41, wherein W2 comprises 6 linked nucleosides.

48. The modified multi-segment antisense oligonucleotide of any one of claims 15 to 47, wherein S1 is at position 10 of the modified multi-segment antisense oligonucleotide.

49. The modified multi-segment antisense oligonucleotide of any one of claims 15 to 47, wherein S1 is at position 11 of the modified multi-segment antisense oligonucleotide.

50. W1 comprises four linked nucleosides; G1 comprises five linked deoxynucleosides; S1 comprises one linked nucleoside; G2 comprises five linked deoxynucleosides; 49. The modified multi-segment antisense oligonucleotide of claim 48, wherein W2 comprises 5 linked nucleosides.

51. W1 comprises 5 linked nucleosides; G1 comprises four linked deoxynucleosides; S1 comprises one linked nucleoside; G2 comprises five linked deoxynucleosides; 49. The modified multi-segment antisense oligonucleotide of claim 48, wherein W2 comprises 5 linked nucleosides.

52. W1 comprises 2 to 8 linked nucleosides; G1 comprises 2 to 7 linked deoxynucleosides; S1 comprises 1 to 2 linked nucleosides; G2 comprises 2 to 7 linked deoxynucleosides; The modified multi-segment antisense oligonucleotide of claim 1, wherein W2 comprises 2 to 8 linked nucleosides.

53. W1 comprises 2 to 6 linked nucleosides; G1 comprises 2 to 7 linked deoxynucleosides; S1 comprises one linked nucleoside; G2 comprises 2 to 7 linked deoxynucleosides; 53. The modified multi-segment antisense oligonucleotide of claim 52, wherein W2 comprises 2 to 6 linked nucleosides.

54. W1 comprises 2 to 8 linked nucleosides; G1 comprises 2 to 7 linked deoxynucleosides; S1 comprises one linked nucleoside; G2 comprises 1 to 2 linked deoxynucleosides; S2 comprises one linked nucleoside; G3 comprises 2 to 7 linked deoxynucleosides; The modified multi-segment antisense oligonucleotide of claim 2, wherein W2 comprises 2 to 8 linked nucleosides.

55. W1 comprises 2 to 6 linked nucleosides; G1 comprises 2 to 7 linked deoxynucleosides; S1 comprises one linked nucleoside; G2 comprises 1 to 2 linked deoxynucleosides; S2 comprises one linked nucleoside; G3 comprises 2 to 7 linked deoxynucleosides; 55. The modified multi-segment antisense oligonucleotide of claim 54, wherein W2 comprises 2 to 6 linked nucleosides.

56. 54. The modified multi-segment antisense oligonucleotide of any one of claims 52 to 53, wherein the length of the G1-S1-G2 segment is 8 to 12 nucleobases in length.

57. 55. The modified multi-segment antisense oligonucleotide of any one of claims 54 to 54, wherein the length of G1-S1-G2-S2-G3 is 8 to 12 nucleobases in length.

58. 58. The modified multi-segment antisense oligonucleotide of any one of claims 1 to 57, wherein G1, G2, and / or G3 comprise nucleosides containing modifications to 2'-deoxynucleosides.

59. 59. The modified multi-segment antisense oligonucleotide of claim 58, wherein G1, G2, and / or G3 comprise nucleosides containing a 2'-deoxy 5-methylcytidine sugar modification.

60. 60. The modified multi-segment antisense oligonucleotide of any one of claims 1 to 59, wherein S1 and / or S2 comprise a nucleoside comprising a 2'-O-methoxyethyl sugar modification.

61. 60. The modified multi-segment antisense oligonucleotide of any one of claims 1 to 59, wherein S1 and / or S2 comprise a nucleoside containing 5-methylcytidine.

62. 60. The modified multi-segment antisense oligonucleotide of any one of claims 1 to 59, wherein S1 and / or S2 comprise a nucleoside comprising a 2'-O-methyl sugar modification.

63. 60. The modified multi-segment antisense oligonucleotide of any one of claims 1 to 59, wherein S1 and / or S2 comprise a nucleoside comprising a 2'-O-ethyl sugar modification.

64. 64. The modified multi-segment antisense oligonucleotide of any one of claims 1 to 63, wherein S1 and / or S2 comprise a nucleoside comprising a 2'-OH sugar modification.

65. 65. The modified multi-segment antisense oligonucleotide of any one of claims 1 to 64, wherein S1 and / or S2 comprise a nucleoside comprising a 2'-fluoro sugar modification.

66. 66. The modified multi-segment antisense oligonucleotide of any one of claims 1 to 65, wherein S1 and / or S2 comprise a nucleoside containing a 2'-fluoro-arabinonucleic acid (2'-fluoro-ANA) sugar modification.

67. 67. The modified multi-segment antisense oligonucleotide of any one of claims 1 to 66, wherein S1 and / or S2 comprise glycol nucleic acid (GNA).

68. 68. The modified multi-segment antisense oligonucleotide of any one of claims 1 to 67, wherein S1 and / or S2 comprise a locked nucleic acid (LNA).

69. 69. The modified multi-segment antisense oligonucleotide of any one of claims 1 to 68, wherein W1 comprises a nucleoside comprising a 2'-deoxy sugar modification.

70. 70. The modified multi-segment antisense oligonucleotide of any one of claims 1 to 69, wherein W1 comprises a nucleoside comprising a 2'-O-methoxyethyl sugar modification.

71. 71. The modified multi-segment antisense oligonucleotide of any one of claims 1 to 70, wherein W1 comprises a nucleoside comprising a 2'-O-methyl sugar modification.

72. 72. The modified multi-segment antisense oligonucleotide of claim 71, wherein W1 comprises 2'-O-methyl 5-methylcytidine.

73. 72. The modified multi-segment antisense oligonucleotide of any one of claims 1 to 71, wherein W1 comprises a nucleoside comprising a 2'-fluoro sugar modification.

74. 74. The modified multi-segment antisense oligonucleotide of any one of claims 1 to 73, wherein W1 comprises a nucleoside that includes a 2'-fluoro-arabinonucleic acid (2'-fluoro-ANA) modification.

75. 75. The modified multi-segment antisense oligonucleotide of any one of claims 1 to 74, wherein W1 comprises glycol nucleic acid (GNA).

76. 76. The modified multi-segment antisense oligonucleotide of any one of claims 1 to 75, wherein W1 comprises a locked nucleic acid (LNA).

77. 77. The modified multi-segment antisense oligonucleotide of any one of claims 1 to 76, wherein W1 comprises a nucleoside comprising a 2'-O-ethyl sugar modification.

78. 78. The modified multi-segment antisense oligonucleotide of any one of claims 1 to 77, wherein W2 comprises a nucleoside comprising a 2'-deoxy sugar modification.

79. 79. The modified multi-segment antisense oligonucleotide of any one of claims 1 to 78, wherein W2 comprises a nucleoside comprising a 2'-O-methoxyethyl sugar modification.

80. 80. The modified multi-segment antisense oligonucleotide of claim 79, wherein W2 comprises 2'-O-methoxyethyl 5-methylcytidine.

81. 79. The modified multi-segment antisense oligonucleotide of any one of claims 1 to 78, wherein W2 comprises a nucleoside comprising a 2'-fluoro sugar modification.

82. 82. The modified multi-segment antisense oligonucleotide of claim 81, wherein W2 comprises a nucleoside comprising a 2'-fluoro-arabinonucleic acid (2'-fluoro-ANA) modification.

83. 83. The modified multi-segment antisense oligonucleotide of any one of claims 1 to 82, wherein W2 comprises glycol nucleic acid (GNA).

84. 84. The modified multi-segment antisense oligonucleotide of any one of claims 1 to 83, wherein W2 comprises a locked nucleic acid (LNA).

85. 85. The modified multi-segment antisense oligonucleotide of any one of claims 1 to 84, wherein W2 comprises a nucleoside comprising a 2'-O-ethyl sugar modification.

86. 86. The modified multi-segment antisense oligonucleotide of any one of claims 1 to 85, wherein at least one internucleoside linkage is a phosphodiester linkage.

87. 87. The modified multi-segment antisense oligonucleotide of any one of claims 1 to 86, wherein at least one internucleoside linkage is a modified internucleoside linkage.

88. 88. The modified multi-segment antisense oligonucleotide of claim 87, wherein said modified internucleoside linkage is a phosphorothioate internucleoside linkage.

89. 88. The modified multi-segment antisense oligonucleotide of claim 87, wherein said modified internucleoside linkages comprise phosphorodiamidate morpholino modifications.

90. a. EC of cells treated with modified multisegment antisense oligonucleotides ５０ / EC of cells treated with modified antisense oligonucleotides having the same template nucleobase sequence without a separator segment ５０ is from about 0.05 to about 250; b. the MTT CC25 value of cells treated with the modified multisegment antisense oligonucleotide is between about 10 nM and about 250 nM; c. the CCK8 CC30 value of cells treated with the modified multisegment antisense oligonucleotide is between about 10 nM and about 250 nM; d. the ratio of CC30 fold / EC50 fold of cells treated with the modified multisegment antisense oligonucleotide compared to a modified antisense oligonucleotide having the same template nucleobase sequence without a separator segment is from about 0.01 to about 50; e. exhibits at least a 1.1-fold decrease in melting temperature compared to a modified antisense oligonucleotide having the same template nucleobase sequence without a separator segment; and / or f. The modified multi-segment antisense oligonucleotide of any one of claims 1 to 89, which exhibits at least a 1.1-fold increase in endonuclease recruitment compared to a modified antisense oligonucleotide having the same template nucleobase sequence without a separator segment.

91. 91. The modified multi-segment antisense oligonucleotide of any one of claims 1 to 90, wherein the modified multi-segment antisense oligonucleotide is complementary to a nucleotide sequence in a viral gene transcript, and optionally the viral gene is selected from the group consisting of an HPV, HBV, EBV, HSV, HIV, and RSV gene.

92. 91. The modified multi-segment antisense oligonucleotide of any one of claims 1 to 90, wherein the modified multi-segment antisense oligonucleotide is complementary to a human gene transcript.

93. 93. The modified multi-segment antisense oligonucleotide of claim 92, wherein the modified multi-segment antisense oligonucleotide is complementary to a human gene transcript selected from the group consisting of DM1 protein kinase (DMPK), alpha-1 antitrypsin (AAT), transthyretin (TTR), proprotein convertase subtilisin / kexin type 9 (PCSK9), apolipoprotein B (ApoB), apolipoprotein C-III (ApoCIII), TNF-alpha, SERPINA8 (AGT), complement factor B (CFB), diacylglycerol O-acyltransferase 2 (DGAT2), patatin-like phospholipase domain-containing 3 (PNPLA3), 5'-aminolevulinic acid synthase 1 (ALAS1), and hydroxyacid oxidase 1 gene transcripts.

94. The modified multisegment antisense oligonucleotide is selected from the group consisting of DM1 protein kinase (DMPK), alpha-1 antitrypsin (AAT), transthyretin (TTR), proprotein convertase subtilisin / kexin type 9 (PCSK9), apolipoprotein B (ApoB), apolipoprotein C-III (ApoCIII), TNF-alpha, SERPINA8 (AGT), complement factor B (CFB), diacylglycerol O-acyltransferase 2 (DGAT2), patatin-like phospholipase domain-containing 3 (PNPLA3), 5'-aminolevulinic acid synthase 1 (ALAS1), and hydroxyacid oxidase 1, Mat1α, (pro)renin receptor / (P)RR, and COVID-19. 5'UTR, Acc1, Acc2, ACE2, androgen receptor, ApoB, ASGR1, ASO, ATXN2, B1AR, B2AR, C9ORF72, caspase 2, CD19, CD4, chikungunya virus, CLPro, complement component 5, COVID19, COX2, CTGF, DGAT2, DMD, DMPK, DNM2, DUX4, E2 gene, EGFR, envelope, EphA2, epithelial sodium channel α subunit, exon 100 dystrophin, exon 101 dystrophin, exon 102 dystrophin, exon 103 dystrophin, exon 104 dystrophin, exon 105 dystrophin, exon 106 dystrophin, exon 107 dystrophin, exon 108 dystrophin, exon 109 Dystrophin, exon 110 dystrophin, exon 111 dystrophin, exon 112 dystrophin, exon 113 dystrophin, exon 114 dystrophin, exon 115 dystrophin, exon 116 dystrophin, exon 117 dystrophin, exon 118 dystrophin, exon 119 dystrophin, exon 120 dystrophin, exon 121 dystrophin, exon 122 dystrophin, exon 123 dystrophin, exon 124 dystrophin, exon 125 dystrophin, exon 126 dystrophin, exon 127 dystrophin, exon 128 dystrophin, exon 129 dystrophin, exon 130 dystrophin, exon 131 dystrophin,Exon 132 dystrophin, Exon 133 dystrophin, Exon 134 dystrophin, Exon 135 dystrophin, Exon 136 dystrophin, Exon 137 dystrophin, Exon 138 dystrophin, Exon 139 dystrophin, Exon 140 dystrophin, Exon 141 dystrophin, Exon 142 dystrophin, Exon 143 dystrophin, Exon 144 dystrophin, Exon 145 dystrophin, Exon 146 dystrophin, Exon 147 dystrophin, Exon 148 dystrophin dystrophin, exon 149 dystrophin, exon 150 dystrophin, exon 151 dystrophin, exon 152 dystrophin, exon 44 dystrophin, exon 53 dystrophin, exon 54 dystrophin, exon 55 dystrophin, exon 56 dystrophin, exon 57 dystrophin, exon 58 dystrophin, exon 59 dystrophin, exon 60 dystrophin, exon 61 dystrophin, exon 62 dystrophin, exon 63 dystrophin, exon 64 dystrophin, exon exon 65 dystrophin, exon 66 dystrophin, exon 67 dystrophin, exon 68 dystrophin, exon 69 dystrophin, exon 70 dystrophin, exon 71 dystrophin, exon 72 dystrophin, exon 73 dystrophin, exon 74 dystrophin, exon 75 dystrophin, exon 76 dystrophin, exon 77 dystrophin, exon 78 dystrophin, exon 79 dystrophin, exon 80 dystrophin, exon 81 dystrophin, exon 82 dystrophin, exon 83 dystrophin, exon 84 dystrophin, exon 85 dystrophin, exon 86 dystrophin, exon 87 dystrophin, exon 88 dystrophin, exon 89 dystrophin, exon 90 dystrophin, exon 91 dystrophin, exon 92 dystrophin, exon 93 dystrophin, exon 94 dystrophin, exon 95 dystrophin, exon 96 dystrophin, exon 97 dystrophin, exon 98 dystrophin, exon 99 dystrophin, Fabp3, factor VII,Factor XI, FAK, FGFR4, FOXP3, FUS, FXII, GFAP, GFP, glycogen synthase, H1N1, HBV, heat shock protein 47, heat shock protein 48, heat shock protein 49, heat shock protein 50, heat shock protein 51, heat shock protein 52, heat shock protein 53, heat shock protein 54, heat shock protein 55, heat shock protein 56, heat shock protein 57, heat shock protein 58, heat shock protein Protein 59, helicase, HIV, HOXB13, HPRT, HPV, Hsd11β1, HTT, hydroxyacid oxidase 1, hydroxysteroid 17β-dehydrogenase 13, hypoxia-inducible factor 2α, kinesin spindle protein and vascular endothelial growth factor, KRAS, leader, leucine-rich repeat kinase 2 (LRRK2), MERS-CoV, MGMT, miR-16, miR-21, MMP-2, MMP-9, MTL-CEBPA, MuRF1, Mycobacterium tuberculosis tuberculosis), COVID-19 N, rabies N 123, rabies N 749, rabies N 903, rabies N1082, rabies N53, rabies N8, chikungunya virus ns1 gene, chikungunya virus ns2 gene, chikungunya virus ns3 gene, chikungunya virus ns4 gene, ORF1 b, ORF1 a, HIV P24, rabies P330, P53, rabies P721, rabies P91, PCSK9, PDGF, PDL1, COVID-19 The modified multi-segment antisense oligonucleotide of any one of claims 1 to 90, which is complementary to a gene transcript selected from the group consisting of PLP, PNPLA3, polo-like kinase 1, protein kinase N3, RAF1, RAF-1, RDRP, RSV, SMN2, SNCA, Covid-19 spike, STAT3, TAU, TGFB1, TGFB1 and Cox2, TMPRSS2, TMPRSS6, TNFα, transthyretin, VEGF, VEGFR2, VER2, xanthine dehydrogenase, and YAP1.

95. 91. The modified multi-segment antisense oligonucleotide of any one of claims 1 to 90, wherein the modified multi-segment antisense oligonucleotide is complementary to a gene transcript selected from the group consisting of an oncogenic gene transcript, an inflammatory gene transcript, a metabolic disease gene transcript, a cardiovascular gene transcript, a liver disease gene transcript, an infectious disease gene transcript, a neurological disease gene transcript, a neuromuscular disease gene transcript, an ocular disease gene transcript, a renal disease gene transcript, a respiratory disease gene transcript, a hematological disease gene transcript, a wound healing gene transcript, a transplantation gene transcript, an autoimmune disease gene transcript, and a neuropsychiatric gene transcript.

96. A modified multi-segment antisense oligonucleotide comprising the structure of any one of SEQ ID NOs: 11-1453 or 5000-30983.

97. A modified multi-segment antisense oligonucleotide comprising the structure of a modified multi-segment antisense oligonucleotide of any one of SEQ ID NOs: 11-1453 or 5000-30983, or an oligonucleotide having at least 70%, at least 80%, at least 90%, at least 95%, or at least 99% sequence identity thereto.

98. A modified multi-segment antisense oligonucleotide comprising the structure of a modified multi-segment antisense oligonucleotide of any one of SEQ ID NOs: 11-1453 or 5000-30983, or an oligonucleotide comprising at least 1, 2, 3, 4, or 5 additional modifications thereto.

99. A pharmaceutical composition comprising the modified multi-segment antisense oligonucleotide of any one of claims 1 to 98, or a salt thereof, and at least one pharmaceutically acceptable carrier or diluent.

100. 100. A method for treating a disease, disorder, or condition, comprising administering to a subject a modified multi-segment antisense oligonucleotide according to any one of claims 1 to 98, or a pharmaceutical composition according to claim 99.

101. 101. The method of claim 100, wherein administration to the subject is parenteral.

102. 101. The method of claim 100, wherein the administration to the subject is subcutaneous.

103. 101. The method of claim 100, wherein administration to the subject is transdermal.

104. 101. The method of claim 100, wherein the administration to the subject is intraocular.

105. 101. The method of claim 100, wherein the administration to the subject is intramuscular.

106. 101. The method of claim 100, wherein the administration to the subject is intravenous.

107. The method of any one of claims 100 to 106, wherein the subject is a human.

108. 107. The method of any one of claims 100 to 106, wherein the subject is a non-human primate.

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