Modified oligomeric compounds containing tricyclic DNA nucleosides and uses thereof

Abstract

The present invention relates to compositions comprising an oligomeric compound comprising one or more tricyclo-deoxyribonucleic acid (tc-DNA) nucleosides and one or more lipid moieties, wherein the one or more lipid moieties are covalently linked to the oligomeric compound, either directly or via a spacer, and preferably the oligomeric compound comprises 5 to 40 monomeric subunits, as well as pharmaceutical compositions thereof and their use in the prevention or treatment of neuromuscular or musculoskeletal diseases, such as Duchenne muscular dystrophy or Steinert's disease.

Description

[Technical Field]

[0001] STATEMENT OF RELATED APPLICATIONS This application claims priority to European Application No. 17167427.8, filed April 20, 2017, and U.S. Provisional Application No. 62 / 562,124, filed September 22, 2017, both of which are incorporated by reference in their entireties. [Background technology]

[0002] Antisense technology is an effective means for reducing the expression of specific gene products, and therefore can be useful in therapeutic, diagnostic and research applications.Generally, the principle behind antisense technology is that antisense compounds (nucleotide or its analogue sequence) hybridize with target nucleic acid and modulate the activity or function of gene expression, such as transcription and / or translation.Despite its specific mechanism, its sequence specificity makes antisense compounds attractive as a tool for target validation and gene functionalization, and as a therapeutic agent for selectively modulating the expression of genes involved in the pathogenesis of disease.

[0003] Antisense compounds typically incorporate chemically modified nucleosides to enhance nucleoside properties such as nuclease resistance, pharmacokinetics or affinity for the target RNA. These chemically modified antisense oligonucleotides (AONs) include structural modifications of natural RNA such as 2'-OH modifications, locked nucleic acids (LNAs), peptide nucleic acids (PNAs), hexitol nucleic acids (HNAs), and tricyclic DNA (tcDNA), as well as many others (reviewed in: Bennett CF and Swayze EE, Annu. Rev. Pharmacol. Toxicol., 2010, 50, 259-293; Deleaviey GF and Damha M, Chemistry & Biology, 2012, 19, 937-954; Sharma VK et al., Med. Chem. Commun., 2014, 5, 1454-1471; Siva et al., Nucleic Acid Therapeutics, 2014, 24, 69-86; Goyenvalle A. et al., Journal of Neuromuscular Diseases, 2016, vol. 3, pp. 157-167, the disclosures of which are incorporated herein by reference. Additionally, backbone modifications such as phosphorothioate (PS) linkages, in which one of the non-bridging oxygen atoms of a phosphodiester linkage is replaced with a sulfur atom, are one of the most widely investigated nucleic acid chemical modifications in oligonucleotide therapeutics.

[0004] Duchenne muscular dystrophy (DMD) is an X-linked recessive disorder affecting 1 in 3,500 live male births (Emery. Neuromuscul. Disord., 1991). DMD is caused by mutations in the gene encoding dystrophin, a large protein (427 kDa) found in various tissues, particularly striated muscle fibers and neurons in specific regions of the central nervous system (Kunkel et al., PNAS., 1985; Muntoni F et al., Lancet Neurol., 2003). Dystrophin is located close to the inner surface of the cell membrane and connects the actin cytoskeleton to the extracellular matrix through a membrane dystrophin-associated glycoprotein complex (Culligan et al., 1988). The absence of dystrophin renders muscle fibers particularly vulnerable to mechanical stress and subject to recurrent cycles of necrosis. As a result, patients exhibit progressive wasting of skeletal muscle, which is replaced over time by fatty fibrous tissue, leading to loss of ambulation by age 12 and premature death from either respiratory failure or cardiomyopathy. In addition, approximately one-third of DMD patients also exhibit cognitive impairment suggestive of significant disruption of neuronal and brain function (Bresolin et al., Neuromuscul. Disord., 1994).

[0005] Full-length dystrophin, translated from a primary 14-kb mRNA transcript composed of 79 exons, is a regulatory protein that can fortunately support multiple exon deletions as long as the open reading frame is preserved (Koenig et al., Cell, 1987). This phenomenon occurs in the clinically mild disease Becker muscular dystrophy (BMD), in which deletions that maintain the open reading frame result in the synthesis of a truncated, semifunctional form of dystrophin (Monaco et al., Genomics., 1988). Human DMD is caused by a heterogeneous group of mutations across the 79 exons, with the two highest incidence regions observed in exons 3-7 and exons 45-55. Muntoni, Neuromuscul. Disord., 2009, 20, 355-362. DMD is caused by out-of-frame deletions or inappropriate stop codons in the dystrophin mRNA. This results in out-of-frame truncated mRNA, unstable and incomplete dystrophin, and ultimately the clinical manifestations of DMD. It is known that in-frame truncated mRNA results from different mutations that result in semi-functional dystrophin and a much milder form of myopathy known as Becker muscular dystrophy (BMD). Therefore, 15 years ago, it was proposed that disrupting the splicing process of selected exons by using antisense oligonucleotides (AONs) could be an appropriate therapeutic approach for DMD (Matsuo M., Brain Dev., 1996). The goal of oligonucleotide therapy for DMD is generally to induce exon skipping (e.g., exon 23 or exon 51, or any other exon), restore dystrophin production, and convert DMD to BMD, thereby reducing mortality and potentially improving cognition. In addition to DMD, many other diseases, including those described below, can also be treated by oligonucleotide-mediated exon skipping techniques.

[0006] Two types of compounds have been extensively tested for antisense-induced exon skipping: 2'-O-methyl-modified ribose oligomers (2'OMe-PS) with full-length phosphorothioate backbones and phosphorodiamidate morpholino oligomers (PMOs). Both types of antisense molecules have been shown to rescue dystrophin in skeletal muscle after systemic delivery in animal models of DMD and in clinical trials (van Deutekom et al., New. Engl. J. Med., 2007; Kinali et al., Lancet Neurol., 2009; Goemans et al., New. Engl. J. Med., 2011; Cirak et al., Lancet., 2011). However, recent studies using 2'OMe-PS and PMO AONs targeting exon 51 in DMD patients failed to demonstrate significant clinical benefit, likely due to insufficient levels of dystrophin rescue (Lu et al., Mol. Ther. Nucleic Acids, 2014). Furthermore, preclinical studies in mice have shown that these two chemicals were not suitable for addressing cardiac and cognitive abnormalities, which represents a significant challenge for successful implementation in many neuromuscular disorders. More recent studies in the mdx mouse model of DMD demonstrated that tcDNA AONs with a complete PS backbone effectively skipped exon 23 at levels 5–6-fold higher than those achieved with 2'OMe-PS and PMO AONs (Goyenvalle et al., Nat. Med., 2015).

[0007] Importantly, this translated into higher rescue of dystrophin protein levels, particularly in the diaphragm and heart, which reached 50% and 40%, respectively, compared to wild-type mice after 12 weeks of treatment. However, while significantly improving biodistribution, oxygen-to-sulfur substitutions in the phosphate backbone have also been shown to contribute to nonspecific protein binding and activation of the innate immune system, particularly complement activation, which can result in acute toxicity at worst and long-term toxicity at best (Dirin and Winkler, Expert Opin. Biol. Ther., 2013). This significant hurdle, which undermines or at least limits the therapeutic potential of this class of molecules, has prompted us to search for new agents that are more efficient and potentially less toxic. [Prior art documents] [Non-patent literature]

[0008] [Non-Patent Document 1] Bennett CF and Swayze EE, Annu. Rev. Pharmacol. Toxicol., 2010, 50, 259-293 Summary of the Invention [Means for solving the problem]

[0009] The present invention relates to compositions comprising an oligomeric compound containing one or more tricyclo-deoxyribonucleic acid (tc-DNA) nucleosides and one or more lipid moieties covalently linked to the oligomeric compound, either directly or via a spacer. Methods for treating several diseases, including Duchenne muscular dystrophy (DMD), spinal muscular atrophy (SMA), CNS-related symptoms, Pompe disease, and myotonic dystrophy type I (DM1), by using the compositions of the present invention are also disclosed.

[0010] Surprisingly, we have found that the compositions of the present invention, comprising an oligomeric compound and one or more lipid moieties covalently linked to the oligomeric compound, are much more efficient and active in penetrating skeletal muscle, cardiac tissue, and the CNS after systemic delivery than their tcDNA counterparts that do not contain the one or more lipid moieties. The latter is regardless of the nature of the internucleoside linkages present in the corresponding oligomeric counterparts, and therefore regardless of whether the counterparts have a complete PS backbone. Furthermore, we have found that the compositions of the present invention exhibit the unexpected property of efficiently penetrating cardiac tissue and thus crossing the blood-brain barrier, bypassing the need for PS internucleoside linkages, potentially significantly improving safety profiles and reducing toxicity while maintaining high efficacy. This allows the dosage of the pharmaceutical compositions of the present invention to be increased to meet and satisfy medical needs.

[0011] Furthermore, it has been shown that the compositions of the present invention are transported in the bloodstream after intravenous systemic application to all skeletal muscles, cardiac muscles, and the CNS, and are taken up by these tissues.Thus, the compositions of the present invention are particularly useful as antisense oligonucleotides (AONs), particularly for achieving antisense effects in muscle and cardiac cells or in the CNS after systemic delivery.Therefore, the present invention also provides compositions for some diseases caused by abnormal gene expression in target tissues or cells.Without being bound by this theory, it is believed that the compositions of the present invention form aggregates with blood albumin, thereby not only ensuring the transport of the compositions of the present invention in the bloodstream, but also increasing the circulatory half-life, and therefore the exposure time of tissues to the compositions of the present invention, and also enhancing the resistance to degradation in plasma.

[0012] Furthermore, preferred compositions of the present invention comprising one or more lipid moieties covalently linked to the oligomeric compound remain highly effective even when the number of phosphorothioate internucleoside linkage groups within the oligomeric compound is reduced in favor of phosphorodiester internucleoside linkage groups. Thus, preferred compositions of the present invention, particularly those comprising a fatty acid or fatty diacid moiety, such as a palmitoyl moiety, and multiple phosphorodiester internucleoside linkage groups, have been found to induce significantly higher levels of exon skipping in multiple tissues compared to their corresponding tc-DNA analogs that do not have the lipid moiety linked thereto.

[0013] Thus, in a first aspect, the present invention provides a composition comprising an oligomeric compound comprising one or more tricyclo-deoxyribonucleic acid (tc-DNA) nucleosides and one or more lipid moieties, preferably exactly one lipid moiety, wherein said one or more lipid moieties are covalently linked to said oligomeric compound, either directly or via a spacer, and preferably wherein said oligomeric compound comprises 5 to 40 monomeric subunits.

[0014] In a further aspect, the present invention provides a pharmaceutical composition comprising the composition of the present invention and further comprising a pharmaceutically acceptable carrier, preferably wherein said pharmaceutical composition is for use in the prevention, treatment or diagnosis of a neuromuscular disease or a musculoskeletal disease, more preferably wherein said neuromuscular disease or said musculoskeletal disease is Duchenne muscular dystrophy, familial dysautonomia, spinal muscular atrophy, ataxia-telangiectasia, congenital disorders of glycosylation, frontotemporal dementia, Parkinsonism linked to chromosome 17, Niemann-Pick disease type C, neurofibromatosis type 1, neurofibromatosis type 2. , macrocephalic leukoencephalopathy type 1 with subcortical cysts, Pelizaeus-Merzbach disease, Pompe disease, myotonic dystrophy type 2 (DM2, or proximal myotonic myopathy) and myotonic dystrophy type 1 (DM1, or Steinert disease), and again more preferably, the neuromuscular disease or musculoskeletal disease is selected from Duchenne muscular dystrophy, spinal muscular atrophy, Pompe disease, myotonic dystrophy type 2 (DM2, or proximal myotonic myopathy) and myotonic dystrophy type 1 (DM1, or Steinert disease).

[0015] In another aspect, the present invention provides a pharmaceutical composition of the present invention comprising the composition of the present invention, further comprising a pharmaceutically acceptable carrier, preferably wherein said pharmaceutical composition is for use in the treatment of a neuromuscular disease or a musculoskeletal disease, more preferably wherein said neuromuscular disease or said musculoskeletal disease is selected from the group consisting of Duchenne muscular dystrophy, familial dysautonomia, spinal muscular atrophy, ataxia-telangiectasia, congenital disorders of glycosylation, frontotemporal dementia, Parkinsonism linked to chromosome 17, Niemann-Pick disease type C, neurofibromatosis type 1, neurofibromatosis type 2, cortical fibrosis, fibroblastic ... and preferably, the neuromuscular disease or musculoskeletal disease is selected from Duchenne muscular dystrophy, spinal muscular atrophy, Pompe disease, myotonic dystrophy type 2 (DM2, or proximal myotonic myopathy) and myotonic dystrophy type 1 (DM1, or Steinert disease).

[0016] In yet a further aspect, the present invention relates to a composition of the present invention for use as a medicament in the prevention, treatment or diagnosis of a disease, preferably wherein said disease is a neuromuscular disease or a musculoskeletal disease, more preferably wherein said neuromuscular disease or said musculoskeletal disease is selected from the group consisting of Duchenne muscular dystrophy, familial dysautonomia, spinal muscular atrophy, ataxia-telangiectasia, congenital disorders of glycosylation, frontotemporal dementia, Parkinsonism linked to chromosome 17, Niemann-Pick disease type C, neurofibromatosis type 1, neurofibromatosis type 2, large intestinal fibrosis with subcortical cysts, and rheumatoid arthritis. and myotonic dystrophy type 1 (DM1, or Steinert disease), and again more preferably, said neuromuscular disease or said musculoskeletal disease is selected from Duchenne muscular dystrophy, spinal muscular atrophy, Pompe disease, myotonic dystrophy type 2 (DM2, or proximal myotonic myopathy) and myotonic dystrophy type 1 (DM1, or Steinert disease).

[0017] In a further aspect, the present invention relates to a composition of the present invention for use as a medicament in the treatment of a disease, preferably wherein said disease is a neuromuscular disease or a musculoskeletal disease, more preferably wherein said neuromuscular disease or said musculoskeletal disease is Duchenne muscular dystrophy, familial dysautonomia, spinal muscular atrophy, ataxia-telangiectasia, congenital disorders of glycosylation, frontotemporal dementia, Parkinsonism linked to chromosome 17, Niemann-Pick disease type C, neurofibromatosis type 1, neurofibromatosis type 2, macrocephaly white matter encephalopathy with subcortical cysts, and again more preferably, said neuromuscular disease or said musculoskeletal disease is selected from Duchenne muscular dystrophy, spinal muscular atrophy, Pompe disease, myotonic dystrophy type 2 (DM2, or proximal myotonic myopathy) and myotonic dystrophy type 1 (DM1, or Steinert disease).

[0018] In a further aspect, the present invention relates to a composition of the present invention for use in a method of treating a disease, preferably wherein said disease is a neuromuscular disease or a musculoskeletal disease, more preferably wherein said neuromuscular disease or said musculoskeletal disease is Duchenne muscular dystrophy, familial dysautonomia, spinal muscular atrophy, ataxia-telangiectasia, congenital disorders of glycosylation, frontotemporal dementia, Parkinsonism linked to chromosome 17, Niemann-Pick disease type C, neurofibromatosis type 1, neurofibromatosis type 2, macrocephalic leukoencephalopathy with subcortical cysts. and again more preferably, said neuromuscular disease or said musculoskeletal disease is selected from Duchenne muscular dystrophy, spinal muscular atrophy, Pompe disease, myotonic dystrophy type 2 (DM2, or proximal myotonic myopathy) and myotonic dystrophy type 1 (DM1, or Steinert disease).

[0019] In a further aspect, the present invention relates to a composition of the present invention for use in a method for preventing, treating or diagnosing a disease, preferably wherein said disease is a neuromuscular disease or a musculoskeletal disease, more preferably wherein said neuromuscular disease or said musculoskeletal disease is Duchenne muscular dystrophy, familial dysautonomia, spinal muscular atrophy, ataxia-telangiectasia, congenital disorders of glycosylation, frontotemporal dementia, Parkinsonism linked to chromosome 17, Niemann-Pick disease type C, neurofibromatosis type 1, neurofibromatosis type 2, macrocephaly with subcortical cysts Preferably, the neuromuscular disease or musculoskeletal disease is selected from Leukoencephalopathy Type 1, Pelizaeus-Merzbach disease, Pompe disease, Myotonic Dystrophy Type 2 (DM2, or Proximal Myotonic Myopathy) and Myotonic Dystrophy Type 1 (DM1, or Steinert Disease), and again more preferably, the neuromuscular disease or musculoskeletal disease is selected from Duchenne muscular dystrophy, spinal muscular atrophy, Pompe disease, Myotonic Dystrophy Type 2 (DM2, or Proximal Myotonic Myopathy) and Myotonic Dystrophy Type 1 (DM1, or Steinert Disease).

[0020] In yet a further aspect, the present invention provides a method for treating a neuromuscular or musculoskeletal disease, comprising administering to a patient a therapeutically effective dose of a composition of the present invention or a pharmaceutical composition of the present invention, in a preferred embodiment of which the neuromuscular or musculoskeletal disease is selected from the group consisting of Duchenne muscular dystrophy, familial dysautonomia, spinal muscular atrophy, ataxia-telangiectasia, congenital disorders of glycosylation, frontotemporal dementia, Parkinsonism linked to chromosome 17, Niemann-Pick disease type C, neurofibromatosis type 1, neurofibromatosis type 2, macrocephalic leukoencephalopathy with subcortical cysts type 1, Pelizaeus-Merzbach disease, Pompe disease, myotonia, and the like. Preferably, the neuromuscular or musculoskeletal disease is selected from Duchenne muscular dystrophy, spinal muscular atrophy, Pompe disease, myotonic dystrophy type 2 (DM2, or proximal myotonic myopathy) and myotonic dystrophy type 1 (DM1, or Steinert disease), and preferably, the neuromuscular or musculoskeletal disease is selected from Duchenne muscular dystrophy, spinal muscular atrophy, Pompe disease, myotonic dystrophy type 2 (DM2, or proximal myotonic myopathy) and myotonic dystrophy type 1 (DM1, or Steinert disease).

[0021] Further aspects and embodiments of the present invention will become apparent as this description continues. In an embodiment of the present invention, for example, the following items are provided: (Item 1) a. an oligomeric compound comprising one or more tricyclo-deoxyribonucleic acid (tc-DNA) nucleosides, preferably comprising 5 to 40 monomer subunits; and b. one or more lipid moieties A composition comprising: A composition wherein said one or more lipid moieties are covalently linked to said oligomeric compound, either directly or via a spacer. (Item 2) Item 10. The composition of claim 1, wherein the one or more lipid moieties are independently selected from a fatty acid moiety, a fatty diacid moiety, an alkyl phosphate moiety, and an alkyl phosphonate moiety. (Item 3) The one or more lipid moieties, independently of one another, have the formula (I) AB- * (I) (Wherein A is C 3~32 Alkyl, C 3~32 Alkenyl, C 3~32 Alkynyl, HOOC-C 3~32 Alkylene, HOOC-C3~32 Alkenylene or HOOC-C 3~32 alkynylene, B is C(O), OP(OH), OP(O)(OH), OP(O)(SH), NH-P(O)(OH), NH-P(O)(SH), NH-C(O), or a pharmaceutically acceptable salt thereof, * ) represents the point of covalent attachment to the oligomeric compound or the spacer. Item 3. The composition of any one of items 1 or 2, wherein (Item 4) The one or more lipid moieties may, independently of one another, be represented by the formula (a) to (u) aC 3~32 Alkyl-C(O)- * 、 bC 3~32 Alkenyl-C(O)- * 、 cC 3~32 Alkynyl-C(O)- * 、 dC 3~32 Alkyl-OP(OH)- * 、 eC 3~32 Alkenyl-OP(OH)- * 、 fC 3~32 Alkynyl-OP(OH)- * 、 gC 3~32 Alkyl-OP(O)(OH)- * 、 hC 3~32 Alkenyl-OP(O)(OH)- * 、 I C 3~32 Alkynyl-OP(O)(OH)- * 、 jC 3~32 Alkyl-OP(O)(SH)- * 、 kC 3~32 Alkenyl-OP(O)(SH)- *、 l.C 3~32 Alkynyl-OP(O)(SH)- * 、 mC 3~32 Alkyl-NH-C(O)- * 、 nC 3~32 Alkenyl-NH-C(O)- * 、 oC 3~32 Alkynyl-NH-C(O)- * 、 PC 3~32 Alkyl-NH-P(O)(OH)- * 、 qC 3~32 Alkenyl-NH-P(O)(OH)- * 、 r.C 3~32 Alkynyl-NH-P(O)(OH)- * 、 s.HOOC-C 3~32 Alkylene-C(O)- * 、 t.HOOC-C 3~32 Alkenylene-C(O)- * , and u.HOOC-C 3~32 Alkynylene-C(O)- * 、 (wherein the asterisk ( * ) represents the point of covalent attachment to the oligomeric compound or the spacer. The composition according to any one of items 1 to 3, selected from any one of the following: (Item 5) The one or more lipid moieties may be independently of each other represented by formula (a) and (b) aC 3~32 Alkyl-C(O)- * 、 b.HOOC-C 3~32 Alkylene-C(O)- * (wherein the asterisk ( * ) represents the point of covalent attachment to the oligomeric compound or the spacer, and preferably, 3~32 The alkyl is an unbranched C 3~32 alkyl, and more preferably, 3~32 The alkyl is an unbranched C 3~32 alkyl, preferably 3~32 The alkylene is an unbranched C 3~32 alkylene, and more preferably, 3~32 Alkylene is an unbranched C 3~32 alkylene) 5. The composition according to any one of items 1 to 4, which is a part from any one of (Item 6) The spacer is represented by the formula a.#-NH-C 2~12 Alkylene-§, b.#-NH-C 2~12 alkylene-OP(OH)-§, c.#-NH-C 2~12 alkylene-OP(O)(SH)-§, d.#-NH-C 2~12 alkylene-OP(O)(OH)-§, e.#-NH-C 2~12 alkylene-NH-C(O)-§, f.#-NH-C 2~12 alkylene-NH-P(O)(OH)-§, and g.#-NH-C 2~12 alkylene-NH-P(O)(SH)-§, (Wherein, C 2~12 One or more -CH in alkylene 2 The - moiety is independently -O-, -S-, -NH-, -C(O)-, -C(O)O-, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, -OP(OH)O-, -OP(O)(SH)O-, -OP(O)(OH)O-, -NHP(O)(OH)O-, -NHP(O)(SH)O-, or -(O-CH 2 -CH 2 ) k - (k is an integer from 1 to 8), 2~12 One or more -CH in alkylene 2 The - moieties may, independently of each other, be one or more of -COOH, -NH 2 , -OP(O)(OH) 2 or -OH, wherein the (#) represents a point of covalent attachment to the lipid moiety, and the (§) represents a point of covalent attachment to the oligomeric compound. and preferably independently selected from any one of the formulas: (Item 7) The spacer is represented by the formula a.#-Z-NH-(CH 2 ) m -X-§ b.#-Z-NH-(CH 2 ) n -(O-CH 2 -CH 2 ) k -O-(CH 2 ) p -X-§ c.#-Z[-NH-(CH 2 ) n -(O-CH 2 -CH 2 ) k -O-(CH 2 ) p -C(O)-]-NH-(CH 2 ) q -X-§ d.#-Z[-NH-(CH 2 ) n -(O-CH 2 -CH 2 ) k -O-(CH 2 ) p -C(O)-] r -NH-(CH 2 ) q -(O-CH 2 -CH 2 ) k -X-§ (wherein, -Z- are, independently of each other, a bond or -NH-CH(COOH)-(CH 2 ) 2 -C(O)- or -NH-CH[(CH 2 ) 2 X's each independently represent OP(OH), OP(O)(SH), OP(O)(OH), NHP(O)(OH), NHP(O)(SH) or NH-C(O), k's each independently represent an integer of 1 to 8, m's each independently represent an integer of 2 to 12, n's each independently represent an integer of 2 to 4, p's each independently represent an integer of 1 to 5, q's each independently represent an integer of 1 to 3, preferably 1 or 2, and r's each independently represent an integer of 1 to 3, preferably 1 or 2, the (#) represents a point of covalent bond to the lipid moiety, and the (§) represents a point of covalent bond to the oligomeric compound. 7. The composition according to any one of items 1 to 6, comprising any one of the following, preferably independently selected from any one of the formulas: (Item 8) 8. The composition of any one of items 1 to 7, wherein the one or more lipid moieties are linked to the oligomeric compound, independently of one another, at: (i) a terminal residue of the oligomeric compound; (ii) the 5' end of the oligomeric compound; (iii) the 3' end of the oligomeric compound; or (iv) an internal residue of the oligomeric compound. (Item 9) 9. The composition of any one of items 1 to 8, wherein the oligomeric compound further comprises one or more nucleosides other than tc-DNA nucleosides. (Item 10) 10. The composition of any one of items 1 to 9, wherein the oligomeric compound comprises 5 to 40 monomeric subunits, the monomeric subunits being nucleosides, one or more of the nucleosides being the one or more tricyclo-deoxyribonucleic acid (tc-DNA) nucleosides, and the monomeric subunits being linked by a plurality of internucleoside linking groups. (Item 11) 11. The composition of claim 10, wherein the internucleoside linking groups are independently selected from phosphorothioate linking groups and phosphorodiester linking groups, and wherein no more than six of the internucleoside linking groups are phosphorothioate linking groups, and preferably no more than three of the internucleoside linking groups are phosphorothioate linking groups. (Item 12) 12. The composition of claim 10 or 11, wherein all of the internucleoside linkage groups are phosphorodiester linkage groups. (Item 13) 13. The composition according to any one of items 1 to 12, wherein the oligomeric compound comprises a sequence selected from any one of the sequences of SEQ ID NOs: 1 to 37. (Item 14) 14. A pharmaceutical composition comprising the composition according to any one of items 1 to 13 and further comprising a pharmaceutically acceptable carrier, preferably for use in the prevention, treatment or diagnosis of a neuromuscular or musculoskeletal disorder. (Item 15) 15. The composition according to any one of items 1 to 13 or the pharmaceutical composition according to item 14, for use as a medicament in the prevention, treatment or diagnosis of a disease, preferably wherein the disease is a neuromuscular disease or a musculoskeletal disease, more preferably wherein the neuromuscular disease or the musculoskeletal disease is selected from Duchenne muscular dystrophy, familial dysautonomia, spinal muscular atrophy, ataxia-telangiectasia, congenital disorders of glycosylation, frontotemporal dementia, Parkinsonism linked to chromosome 17, Niemann-Pick disease type C, neurofibromatosis type 1, neurofibromatosis type 2, macrocephalic leukoencephalopathy with subcortical cysts type 1, Pelizaeus-Merzbach disease, Pompe disease, myotonic dystrophy type 2 (DM2, or proximal myotonic myopathy), and myotonic dystrophy type 1 (DM1, or Steinert disease). [Brief explanation of the drawings]

[0022] [Figure 1] Detection of exon-23 skipped dystrophin mRNA in mdx muscles and CNS, including tibialis anterior (TA), gastrocnemius (GAS), quadriceps (QD), triceps (TRI), biceps (BI), diaphragm (DIA), heart, cortex, and cerebellum (CB1) after 4 weeks of treatment with tcDNA-PO M23D (SY-0308), tcDNA-PS M23D (SY-0210), SY-0299, SY-0442, and SY-0455 at doses of 200 mg / kg / week, and SY-0343 at doses of 178 mg / kg / week. Exon 23 skipping was quantified by Taqman qPCR and expressed as a percentage of total dystrophin, measured by the expression levels of exons 4 and 5, after normalization to endogenous controls. N = 4 mice per group; error bars are mean ± SEM.

[0023] [Figure 2-1]Prothrombin time and partial thromboplastin time are two blood tests that measure how long it takes blood to clot in order to predict bleeding problems. Two mg / ml of the following sequences: tcDNA-PO M23D (SY-0308, SEQ ID NO: 1), or tcDNA-PS M23D (SY-0210, SEQ ID NO: 23), or tcDNA-PS SYN51 (SEQ ID NO: 19, in which all nucleotides are tcDNA and all internucleoside linkages are PS linkages), or tcDNA-PO SYN51 (SEQ ID NO: 19, in which all nucleotides are tcDNA and all internucleoside linkages are PO linkages), or SY-0343, or SY-0442, or SY-0455, was incubated with 50 μl of citrated plasma C at 37°C for 30 minutes, and then prothrombin time (PT) and activated partial thromboplastin time (PTT) assays were performed in a semi-automated START max coagulometer (Stago) according to the manufacturer's instructions (A: Mouse plasma, B: Human plasma). PBS (phosphate buffered saline) was used as a negative control, and as a positive control, (tcDNA-PS SYN51) with an intact PS backbone, which is highly toxic to mice, was used. [Figure 2-2] Same as above.

[0024] [Figure 3] One hour after AON injection, blood samples were collected from all mice to measure complement C3. Complement activation in serum samples was determined using the Microvue PanSpecific-C3 Converter and SC5b-9 Plus kit (Quidel Co., San Diego, CA, USA). Mouse C3 protein was converted to human SC5b9 using a C3 converter reagent (Panspecific C3 Reagent Kit, Microvue, Quidel) and then detected by SC5b9 Elisa (Quidel). Complement activation was expressed as a percentage of the remaining C3 level in the sample, considering the C3 level in the PBS condition as 100% (no activation).

[0025] [Figure 4-1] Serum biochemistry analysis at the end of 4 weeks of treatment. There were no significant changes in serum albumin, creatinine, and urea, suggesting the absence of nephrotoxicity. Only a slight increase in ALP was observed for most palmitoyl-conjugated PO M23D. [Figure 4-2] Same as above. [Figure 4-3] Same as above. [Figure 4-4] Same as above.

[0026] [Figure 5] Restored dystrophin levels quantified by Western blot using the licor Odyssey system in gastrocnemius (Gas), diaphragm (Dia), and triceps (Tri) muscles after 4 weeks of treatment with SY-0299, SY-0455, and SY-0442 at a dose of 200 mg / kg / week.

[0027] [Figure 6-1] Biotinylated tcDNA oligonucleotides (M23D) used for coprecipitation of potentially interacting proteins from mouse and human serum. A: SY-0446; A': SY-0448, an analog of SY-0446 except that the third nucleotide of the tcDNA sequence is changed to 2'OMe-U; B: SY-0445; C: SY-0443, an analog of SY-0445 except that two PS residues are present at both ends of the tcDNA oligonucleotide; D: SY-0451. For AONs lacking palmitoyl residues (SY-0440 and SY-0427), the biotin moiety was attached to the 5' end of the oligonucleotide via a C3 linker (not shown). The characterization and definition of the compositions of the present invention are described in Table 3. [Figure 6-2] Same as above.

[0028] [Figure 7]SDS-PAGE analysis of proteins recovered from mouse (Panel A) and human (Panel B) serum using the oligonucleotides listed and depicted in Figure 6. 20 μg of immobilized AO was incubated in 50 μL of 10-fold diluted serum.

[0029] [Figure 8] The prothrombin times and partial thromboplastin times of the following sequences with variable sulfur content in the internucleoside linkages: tcDNA-PO M23D (SY-0308, SEQ ID NO: 1), or tcDNA-PS M23D (SY-0210, SEQ ID NO: 23), or tcDNA-PS SYN51 (SEQ ID NO: 19, in which all nucleotides are tc-DNA and all internucleoside linkages are PS linkages), or tcDNA-PO SYN51 (SEQ ID NO: 19, in which all nucleotides are tc-DNA and all internucleoside linkages are PO linkages), M23D-PS 25sb3 corresponding to (p-CCTCGGCTTA*C*C*T - SEQ ID NO: 42); M23D-PS 33sb3 corresponding to (p-CCTCGGCTT*A*C*C*T - SEQ ID NO: 43); M23D-PS 50sb3, corresponding to (p-CCTCG*G*C*T*T*A*C*C*T - SEQ ID NO: 44); M23D-PS 66sb3, corresponding to (p-CCT*C*G*G*C*T*T*A*C*C*T - SEQ ID NO: 45); and M23D-PS 83sb3, corresponding to (p-CCT*C*G*G*C*T*T*A*C*C*T - SEQ ID NO: 46) (wherein the "*" between the two nucleosides indicates a phosphorothioate internucleoside linkage and p represents the terminal phosphate group).

[0030] [Figure 9] Non-denaturing PAGE of oligonucleotides SY-0357, SY-0299, SY-0442, SY-0455, SY-0343, and SY-0450. Oligonucleotide SY-0221 (CTT TCA TAA TGC TGG - SEQ ID NO: 47), which has an entirely PS backbone and is toxic in mice, is used to demonstrate migration of multimeric bands on the gel (visualized by StainsAll).

[0031] [Figure 10-1] Exon 23 skipping detected by nested RT-PCR in various tissues from mice treated with SY442 (Figure 10A) or SY450 (Figure 10B) at 200 mg / kg for 72 hours, and with SY442 (Figure 10C) or SY450 (Figure 10D) for 2 weeks. The left panel represents RT-PCR between exons 20 and 26, showing the unskipped 901-bp product and the skipped 688-bp product. The right panel represents specific RT-PCR between exon 20 and junctions 22-24, showing only the skipped 401-bp product. The specific PCR shown in the right panel was only used when the skipping level was too low to be detected by conventional RT-PCR. [Figure 10-2] Same as above.

[0032] [Figure 11-1] Exon 23 skipping was detected by nested RT-PCR in various tissues from mice treated with SY442 (Figure 11A) or SY450 (Figure 11B) for 72 hours, with SY0210 (Figure 11C) as a control, and with SY442 (Figure 11D) or SY450 (Figure 11E) for 2 weeks, with SY0210 (Figure 11F) as a control. The left panel represents RT-PCR between exons 20 and 26, showing the unskipped 901-bp product and the skipped 688-bp product. The right panel represents specific RT-PCR between exon 20 and junctions 22-24, showing only the skipped 401-bp product. The specific PCR shown in the right panel was only used when the skipping level was too low to be detected by conventional RT-PCR. [Figure 11-2] Same as above.

[0033] [Figure 12]Quantification of exon 23 skipping by taqman qPCR in tissues from mice treated with 200 mg / kg tcDNA at 72 hours and 2 weeks post-injection.

[0034] [Figure 13] Quantification of exon 23 skipping by taqman qPCR in tissues from mice treated with 50 mg / kg tcDNA at 72 hours and 2 weeks post-injection.

[0035] [Figure 14] Pooled data comparing the efficacy of SY0442 and SY0450 at 72 hours and 2 weeks post-injection. A dose effect can be observed between 50 mg / kg and 200 mg / kg (approximately 4-fold).

[0036] [Figure 15] Quantification of exon 23 skipping at 200 mg / kg comparing intact PO compounds with intact PS compounds.

[0037] [Figure 16] Quantification of exon 23 skipping at 50 mg / kg comparing intact PO compounds with intact PS compounds. DETAILED DESCRIPTION OF THE INVENTION

[0038] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The headings used herein are for convenience only and should not be construed as limiting the disclosure of any of the aspects and embodiments of the invention. definition

[0039] The term "oligomeric compound," as used herein, refers to a compound comprising preferably eight or more monomeric subunits linked by internucleoside linking groups, wherein at least two of said eight or more monomeric subunits are tricyclo-deoxyribonucleic acid (tc-DNA) nucleosides.

[0040] The term "monomer subunit," as used herein, refers to α-D-ribonucleosides, β-D-ribonucleosides, α-D-2'-deoxyribonucleosides, β-D-2'-deoxyribonucleosides, naturally occurring nucleosides, modified nucleosides, herein, in particular, tricyclo-deoxyribonucleic acid (tc-DNA) nucleosides and 2'-modified ribonucleic acid (2'-modified-RNA) nucleosides, locked nucleic acid (LNA) nucleosides, peptide nucleic acid (PNA) nucleosides, 2'-deoxy2'-fluoro-arabinonucleosides, 2'-deoxy- ... The term "nucleotides" is meant to include all manner of monomer units suitable for oligomer synthesis, including, typically and preferably, monomer subunits such as nucleotides, hexitol nucleic acid (HNA) nucleosides; and phosphorodiamidate morpholino (PMO) nucleosides, nucleoside mimetics, naturally occurring nucleotides, modified nucleotides, and in this context, particularly, tricyclo-deoxyribonucleic acid (tc-DNA) nucleotides and 2'-modified ribonucleic acid (2'-modified-RNA) nucleotides, and nucleotide mimetics. Typically and preferably, the term "monomer subunit", as used herein, refers to naturally occurring nucleosides and modified nucleosides, and herein particularly refers to ribonucleosides, deoxyribonucleosides, tricyclo-deoxyribonucleic acid (tc-DNA) nucleosides, 2'-modified ribonucleic acid (2'-modified-RNA) nucleosides, locked nucleic acid (LNA) nucleosides, peptide nucleic acid (PNA) nucleosides, 2'-deoxy 2'-fluoro-arabinonucleosides, hexitol nucleic acid (HNA) nucleosides and phosphorodiamidates. It refers to morpholino (PMO) nucleosides, as well as naturally occurring and modified nucleotides, and herein refers in particular to ribonucleotides, deoxyribonucleotides, tricyclo-deoxyribonucleic acid (tc-DNA) nucleotides, 2'-modified ribonucleic acid (2'-modified-RNA) nucleotides, locked nucleic acid (LNA) nucleotides, peptide nucleic acid (PNA) nucleotides, 2'-deoxy 2'-fluoro-arabinonucleotides, hexitol nucleic acid (HNA) nucleotides, and phosphorodiamidate morpholino (PMO) nucleotides.More preferably, the term "monomer subunit" as used herein refers to modified nucleotides, and here in particular to tricyclo-deoxyribonucleic acid (tc-DNA) nucleotides and 2'-modified ribonucleic acid (2'-modified-RNA) nucleotides.

[0041] The term "lipid moiety," as used herein, typically and preferably refers to moieties derived from hydrocarbons, oils, fats (such as fatty acids, glycerides), sterols, steroids, and derivatives of these compounds. Suitable lipid moieties include fatty acids and their derivatives, hydrocarbons and their derivatives, and moieties derived from sterols, such as cholesterol. As used herein, the term lipid moiety also includes amphiphilic compound moieties, which contain both lipid moieties and hydrophilic moieties.

[0042] The term "hydrocarbon," as used herein, includes compounds consisting solely of hydrogen and carbon connected by covalent bonds. The term includes open-chain (aliphatic) hydrocarbons, including straight (unbranched) chain and branched hydrocarbons, as well as saturated, mono-, and polyunsaturated hydrocarbons. The term also includes hydrocarbons containing one or more aromatic rings; preferably, the term excludes hydrocarbons containing one or more aromatic rings. The terms "straight" and "unbranched" are used interchangeably herein.

[0043] The term "fatty acid," as used herein, refers to a hydrocarbon chain terminated in a carboxylic acid group, said hydrocarbon chain being typically and preferably either alkyl or alkenyl, typically 3 to 32 carbons in length, and therefore saturated or unsaturated, and containing one or more, preferably one, carboxylic group (-COOH), one or more, preferably one, C 1~32alkyl, optionally substituted by one or more, preferably one, phosphate groups (HOP(O)(OH)O-), one or more, preferably one, phosphonate groups (HOP(O)O-), one or more, preferably one, thiophosphate groups (HOP(O)(SH)O-), one or more, preferably one, dithiophosphate groups (HOP(S)(SH)O-), one or more, preferably one, diphosphate groups (HO-P(O)(OH)-OP(O)(OH)-O-), one or more, preferably one, triphosphate groups (HO-P(O)(OH)-OP(O)(OH)-OP(O)(OH)-O-), one or more phenyl groups (-CH), halogen, preferably iodine, or one or more phenyl groups substituted with a carboxy group. When a fatty acid contains one or more double bonds and is therefore unsaturated, it may be either cis or trans geometric isomerism. The term "fatty acid moiety" as used herein refers to a moiety derived from a fatty acid as defined herein, wherein one carboxyl group (-COOH) of the fatty acid becomes the -C(O)- group of the fatty acid moiety, and this -C(O)- group is linked to the oligonucleotide according to the present invention, either directly or via a spacer. Preferably, the term "fatty acid" as used herein refers to a hydrocarbon chain terminated by a carboxylic acid group, wherein the hydrocarbon chain is typically and preferably either alkyl or alkenyl, typically 3 to 32 carbon atoms in length, and therefore saturated or unsaturated, and comprises one or more, preferably one, carboxyl group (-COOH), one or more, preferably one, C 1~32alkyl, optionally substituted by one or more, preferably one, phosphate groups (HOP(O)(OH)O-), one or more, preferably one, phosphonate groups (HOP(O)O-), one or more, preferably one, thiophosphate groups (HOP(O)(SH)O-), one or more, preferably one, dithiophosphate groups (HOP(S)(SH)O-), one or more, preferably one, diphosphate groups (HO-P(O)(OH)-OP(O)(OH)-O-), one or more, preferably one, triphosphate groups (HO-P(O)(OH)-OP(O)(OH)-OP(O)(OH)-O-), one or more phenyl groups (-CH), halogen, preferably iodine, or one or more phenyl groups substituted with a carboxy group. Preferably, the fatty acid has an even number of carbon atoms, and the carbon atom of the carboxy group (-COOH) of the fatty acid or the -C(O)- group of the fatty acid moiety is included in counting the number of carbon atoms.

[0044] Thus, fatty acids preferably contain an even or odd number of carbon atoms, preferably an even number, in a straight chain (usually 3 to 32 carbons), can be saturated or unsaturated, and can contain a variety of substituents, preferably one or more, preferably one, carboxy group (-COOH), one or more, preferably one, C 1~32The alkyl may contain or be modified to contain one or more, preferably one, phosphate groups (HOP(O)(OH)O-), one or more, preferably one, phosphonate groups (HOP(O)O-), one or more, preferably one, thiophosphate groups (HOP(O)(SH)O-), one or more, preferably one, dithiophosphate groups (HOP(S)(SH)O-), one or more, preferably one, diphosphate groups (HO-P(O)(OH)-OP(O)(OH)-O-), one or more, preferably one, triphosphate groups (HO-P(O)(OH)-OP(O)(OH)-OP(O)(OH)-O-), one or more phenyl groups (-CH), one or more phenyl groups substituted with a halogen, preferably iodine, or a carboxy group.

[0045] The term "fatty diacid" refers to a fatty acid as defined herein, but with an additional carboxylic acid group at the omega position. Thus, a fatty diacid is a dicarboxylic acid. The term "fatty diacid moiety", as used herein, refers to a moiety derived from a fatty diacid as defined herein, wherein one carboxy group (-COOH) of the fatty diacid becomes the -C(O)- group of the fatty diacid moiety, and the -C(O)- group of the fatty diacid moiety is linked to the oligonucleotide according to the present invention, either directly or via a spacer. A preferred embodiment of a fatty diacid is one or more, preferably one, C 1~32alkyl, saturated fatty diacids optionally substituted by one or more, preferably one, phosphate groups (HOP(O)(OH)O-), one or more, preferably one, phosphonate groups (HOP(O)O-), one or more, preferably one, thiophosphate groups (HOP(O)(SH)O-), one or more, preferably one, dithiophosphate groups (HOP(S)(SH)O-), one or more, preferably one, diphosphate groups (HO-P(O)(OH)-OP(O)(OH)-O-), one or more, preferably one, triphosphate groups (HO-P(O)(OH)-OP(O)(OH)-OP(O)(OH)-O-), one or more, preferably one, phenyl groups (-CH), one or more, preferably one, phenyl groups substituted with halogen, preferably iodine, or carboxy groups. A preferred example is a hydroxyl group having one C such as 3-pentadecyl glutaric acid (PDG). 6~24 Glutaric acid optionally substituted with alkyl is included.

[0046] The term "alkyl phosphate moiety" as used herein means a C 3~32 alkyl-OP(O)(OH)-O- group, 3~32 Alkyl is independently a C as defined herein. 3~32 alkyl.

[0047] The term "alkylphosphonate moiety," as used herein, refers to a C 3~32 alkyl-OP(O)-O- group, 3~32 Alkyl is independently a C as defined herein. 3~32 alkyl.

[0048] The term "alkyl," as used herein, refers to a straight- or branched-chain hydrocarbon radical containing no unsaturation, consisting solely of carbon and hydrogen atoms, having from 1 to 32 carbon atoms (e.g., (C 1~32 ) alkyl or C 1~32alkyl), which may be, or typically is, attached to the remainder of the molecule by a single bond. Whenever it appears herein, a numerical range such as "1 to 32" refers to each integer within the given range. For example, "1 to 32 carbon atoms" means that the alkyl group can consist of 1 carbon atom, 2 carbon atoms, 3 carbon atoms, etc., up to a maximum of 32 carbon atoms, although this definition is also intended to cover occurrences of the term "alkyl" where no numerical range is specifically specified. Typical alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, 1-methylethyl (used interchangeably with isopropyl; abbreviated interchangeably herein as iPr or Pri), n-butyl, isobutyl, sec-butyl, isobutyl, tert-butyl (used interchangeably with 1,1-dimethylethyl or tert-butyl), n-pentyl, isopentyl, neopentyl, hexyl, septyl, octyl, nonyl, and decyl. Unless otherwise specified specifically in the specification, alkyl groups are optionally substituted by one or more substituents which are independently alkenyl, alkoxy, carboxy (-COOH), heteroalkyl, heteroalkenyl, hydroxyl, phosphate (-OP(O)(OH)O-), phosphonate (-OP(O)O-), halogen, preferably iodine, or a phenyl group (-CH) optionally substituted with a carboxy group. Preferably, the term "alkyl," as used herein, refers to an unsubstituted alkyl as defined herein.

[0049] The term "alkylene," as used herein, refers to a straight or branched chain hydrocarbon biradical derived from an alkyl, as defined herein, where one hydrogen of said alkyl is cleaved to produce a second radical of said alkylene. Examples of alkylene are, by way of illustration, -CH-, -CH-CH-, -CH(CH)-, -CH-CH-CH-, -CH(CH)-CH-, or -CH(CHCH)-.

[0050] The term "alkenyl," as used herein, refers to a straight- or branched-chain hydrocarbon radical group consisting solely of carbon and hydrogen atoms (i.e., (C 3~32 ) alkenyl or C 3~32 Alkenyl), which may be, or typically is, attached to the remainder of the molecule by a single bond. Whenever it appears herein, a numerical range such as "3 to 32" refers to each integer within the given range; for example, "3 to 32 carbon atoms" means that the alkenyl group can consist of 3 carbon atoms, 4 carbon atoms, etc., up to a maximum of 32 carbon atoms. Typical alkenyl groups include, but are not limited to, ethenyl (i.e., vinyl), prop-1-enyl (i.e., allyl), but-1-enyl, pent-1-enyl, and penta-1,4-dienyl. Each double bond can be in either the (E)- or (Z)-configuration. Thus, alkenyl, where applicable, each of said double bonds can include either its (E)-configuration, its (Z)-configuration, and mixtures thereof in any ratio. Unless otherwise specified specifically in the specification, an alkenyl group is optionally substituted by one or more substituents which are independently alkenyl, alkoxy, carboxy group (-COOH), heteroalkyl, heteroalkenyl, hydroxyl, phosphate group (-OP(O)(OH)O-), phosphonate group (-OP(O)O-), halogen, preferably iodine, or a phenyl group (-CH) optionally substituted with a carboxy group. Preferably, the term "alkenyl," as used herein, refers to an unsubstituted alkenyl as defined herein.

[0051] The term "alkenylene," as used herein, refers to a straight or branched chain hydrocarbon biradical derived from an alkenyl, as defined herein, where one hydrogen of said alkenyl has been cleaved to generate a second radical of said alkenylene.

[0052] The term "alkynyl" refers to a straight- or branched-chain hydrocarbon radical group, consisting solely of carbon and hydrogen atoms, containing at least one triple bond and having from 2 to 10 carbon atoms (i.e., (C 2~32 ) alkynyl or C 2~32 Alkynyl). Whenever it appears herein, a numerical range such as "2 to 32" refers to each integer within the given range; for example, "2 to 32 carbon atoms" means that the alkynyl group can consist of 2 carbon atoms, 3 carbon atoms, etc., up to a maximum of 32 carbon atoms. Typical alkynyl groups include, but are not limited to, ethynyl, propynyl, butynyl, pentynyl, and hexynyl. Unless otherwise specified specifically in the specification, an alkynyl group is optionally substituted with one or more substituents, independently, an alkenyl, a carboxy group (-COOH), a heteroalkyl, a heteroalkenyl, a phosphate group (-OP(O)(OH)O-), a phosphonate group (-OP(O)O-), a halogen, preferably iodine, or a phenyl group (-CH) optionally substituted with a carboxy group. Preferably, the term "alkynyl," as used herein, refers to an unsubstituted alkynyl as defined herein.

[0053] The term "alkynylene," as used herein, refers to a straight or branched chain hydrocarbon biradical derived from an alkynyl, as defined herein, where one hydrogen of said alkynyl has been cleaved to generate a second radical of said alkynylene.

[0054] The term "alkoxy" refers to groups of -O-alkyl, including straight, branched, and combinations thereof, of 1 to 32 carbon atoms attached to the parent structure through an oxygen. Examples include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, cyclopropyloxy, and cyclohexyloxy. "Lower alkoxy" refers to an alkoxy group containing 1 to 6 carbons, and also includes (C 1~6 ) alkoxy or OC 1~6Also called alkyl.

[0055] The term "substituted alkoxy" refers to an alkoxy in which the alkyl component is substituted (i.e., -O-(substituted alkyl)). Unless otherwise stated specifically in the specification, the alkyl portion of the alkoxy group is optionally substituted with one or more substituents which are independently alkenyl, carboxyl (-COOH), heteroalkyl, heteroalkenyl, phosphate (-OP(O)(OH)O-), phosphonate (-OP(O)O-), halogen, preferably iodine, or a phenyl group optionally substituted with a carboxyl (-CH).

[0056] The term "acyl" refers to the groups (alkyl)-C(O)-, (aryl)-C(O)-, (heteroaryl)-C(O)-, and (heteroalkyl)-C(O)-, which are attached to the parent structure through a carbonyl functionality. Unless stated otherwise specifically in the specification, the alkyl, aryl, or heteroaryl portion of the acyl group is optionally substituted with one or more substituents, independently, an alkenyl, a carboxy group (-COOH), a heteroalkyl, a heteroalkenyl, a phosphate group (-OP(O)(OH)O-), a phosphonate group (-OP(O)O-), a halogen, preferably iodine, or a phenyl group (-CH) optionally substituted with a carboxy group.

[0057] Unless otherwise specified herein, the term “amino” or “amine” refers to —N(R a )2 radical groups, and each R a are independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl. a ) Two groups have two R aWhen substituted, these may be combined with the nitrogen atom to form a 4-, 5-, 6-, or 7-membered ring. For example, —N(R a )2 is intended to include, but is not limited to, 1-pyrrolidinyl and 4-morpholinyl. Unless otherwise specified specifically in the specification, amino or amine groups are optionally substituted by one or more substituents which are independently alkenyl, carboxy (-COOH), heteroalkyl, heteroalkenyl, phosphate (-OP(O)(OH)O-), phosphonate (-OP(O)O-), halogen, preferably iodine, or a phenyl group (-CH) optionally substituted with a carboxy group.

[0058] The term "aromatic" or "aryl" or "Ar" refers to a carbocyclic (e.g., phenyl, fluorenyl, and naphthyl) aromatic radical having 6 to 10 ring atoms (e.g., C6-C8) with at least one ring having a conjugated pi-electron system. 10 Aromatic or C6-C 10 (aryl). Divalent radicals formed from substituted benzene derivatives and having a free valence on a ring atom are named substituted phenylene radicals. Divalent radicals derived from monovalent polycyclic hydrocarbon radicals, named "-yl" by removing one hydrogen atom from the free valence carbon atom, are named by adding "-idene" to the name of the corresponding monovalent radical; for example, a naphthyl group with two points of attachment is called naphthylidene. Whenever appearing herein, numerical ranges such as "6 to 10" refer to individual integers within the given range; for example, "6 to 10 ring atoms" means that the aryl group may consist of 6 ring atoms, 7 ring atoms, etc., up to a maximum of 10 ring atoms. The term includes monocyclic or fused-ring polycyclic (i.e., rings sharing adjacent pairs of ring atoms) groups.

[0059] The term "aralkyl" or "arylalkyl" refers to an (aryl)alkyl-radical, wherein the aryl and alkyl are optionally substituted with one or more substituents disclosed herein and described as suitable substituents for aryl and alkyl, respectively.

[0060] The terms "carboxyl" or "carboxylic" are used interchangeably herein and refer to the -(C=O)OH radical.

[0061] The term "cycloalkyl" refers to a monocyclic or polycyclic radical that contains only carbon and hydrogen and can be saturated or partially unsaturated. A cycloalkyl group is a group having 3 to 10 ring atoms (i.e., (C 3~10 ) cycloalkyl or C 3~10 Whenever it appears herein, a numerical range such as "3 to 10" refers to each integer within the given range; for example, "3 to 10 carbon atoms" means that the cycloalkyl group consists of 3 carbon atoms, etc., and can contain up to 10 carbon atoms. Illustrative examples of cycloalkyl groups include, but are not limited to, the following moieties: cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, norbornyl, etc.

[0062] The term "fluoroalkyl" refers to an alkyl radical, as defined above, substituted by one or more fluoro radicals, as defined above, e.g., trifluoromethyl, difluoromethyl, 2,2,2-trifluoroethyl, 1-fluoromethyl-2-fluoroethyl, etc. The alkyl portion of the fluoroalkyl radical can be optionally substituted as defined above for an alkyl group.

[0063] The term "halogen" as used herein refers to fluorine, chlorine, bromine, or iodine, preferably iodine. In a preferred embodiment, the halogen substituent is iodine.

[0064] The terms "heteroalkyl" and "heteroalkenyl," as used herein, refer to optionally substituted alkyl and alkenyl radicals having one or more skeletal chain atoms selected from atoms other than carbon, e.g., oxygen, nitrogen, sulfur, phosphorus, or combinations thereof. Numerical ranges may be given, referring to total chain length; e.g., C1-C4 heteroalkyl, in this example, is 4 atoms long.

[0065] The term "heteroaryl" or "heteroaromatic" or "HetAr" refers to a 5- to 18-membered aromatic radical (e.g., C5-C6) containing one or more ring heteroatoms selected from nitrogen, oxygen, and sulfur. 13 Heteroaryl refers to a heteroaryl group (heteroaryl), which may be a monocyclic, bicyclic, tricyclic, or tetracyclic ring system. Whenever it appears herein, a numerical range such as "5 to 18" refers to each integer within the given range; for example, "5 to 18 ring atoms" means that the heteroaryl group can contain 5 ring atoms, 6 ring atoms, etc., up to a maximum of 18 ring atoms. Divalent radicals derived from monovalent heteroaryl radicals, designated by the suffix "-yl" by the removal of one hydrogen atom from the free valence atom, are designated by the addition of "-idene" to the name of the corresponding monovalent radical; for example, a pyridyl group with two points of attachment is a pyridylidene.

[0066] The term "stereoisomers" refers to compounds which have identical chemical constitution, but differ with regard to the arrangement of the atoms or groups in space.

[0067] "Diastereomer" refers to a stereoisomer having two or more centers of chirality, where the compounds are not mirror images of one another. Diastereomers have different physical properties, such as melting points, boiling points, spectral properties, and chemical and biological reactivity. Mixtures of diastereomers can separate under high-resolution analytical procedures such as electrophoresis and chromatography.

[0068] "Enantiomers" refer to two stereoisomers of a compound which are non-superimposable mirror images of one another.

[0069] Stereochemical definitions and conventions used herein generally follow those in S.P. Parker, ed., McRaw-Hiff Dictionary of Chemical Terms (1984), McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S., "Stereochemistry of Organic Compounds," John Wiley & Sons, Inc., New York, 1994.

[0070] (in the chemical formula * The symbols (), (#), and (§) designate i) points of attachment, ii) radicals, and / or iii) unshared electrons.

[0071] The term "antisense oligonucleotide (AON)" as used herein refers to an oligonucleotide or oligomeric compound capable of altering gene expression by interacting with and / or hybridizing to pre-mRNA or mRNA having a complementary nucleotide sequence.

[0072] The term "protecting group," as used herein, is intended to mean a group that selectively blocks one or more reactive sites of a polyfunctional compound so that a chemical reaction can be selectively carried out at another, unprotected reactive site, and which can then be easily removed or deprotected after the selective reaction is complete. Various protecting groups are disclosed, for example, in Protecting Groups in Organic Synthesis, T.W. Greene and P.G.M. Wuts, 3rd Edition, John Wiley & Sons, New York, 1999.

[0073] The terms "protecting group for amino", "protecting group for amino group" or "amino-protecting group", which are used interchangeably herein, are well known in the art and include those described in detail in Protecting Groups in Organic Synthesis, T.W. Greene and P.G.M. Wuts, 3rd Edition, John Wiley & Sons, New York (1999), Greene's Protective Groups in Organic Synthesis, P.G.M. Wuts, 5th Edition, John Wiley & Sons (2014), and Current Protocols in Nucleic Acid Chemistry, edited by S.L. Beaucage et al., June 2012, herein in particular Chapter 2. Suitable "amino protecting groups" for the present invention include methyl carbamate, ethyl carbamate, 9-fluorenylmethylcarbamate (Fmoc), 9-(2-sulfo)fluorenylmethylcarbamate, 2,7-di-t-butyl-[9-(10,10-dioxo-10,10,10,10-tetrahydrothioxanthyl)]methylcarbamate (DBD-Tmoc), 4-methoxyphenacylcarbamate (Phenoc), 2,2,2-trichloroethylcarbamate (Troc), 2-trimethylsilylethylcarbamate (Teoc), 2-methyl- ... -phenylethyl carbamate (hZ), 1,1-dimethyl-2,2-dibromoethyl carbamate (DB-t-BOC), 1,1-dimethyl-2,2,2-trichloroethyl carbamate (TCBOC), benzyl carbamate (Cbz), p-methoxybenzyl carbamate (Moz) and 2,4,6-trimethylbenzyl carbamate, (4-methoxyphenyl)diphenylmethyl (MMTr); as well as formamide, acetamide, benzamide, typically and preferably independently selected from these at each occurrence.

[0074] The terms "protecting group for hydroxyl," "protecting group for a hydroxyl group," or "hydroxyl protecting group," as used interchangeably herein, are well known in the art and include those described in detail in Protecting Groups in Organic Synthesis, T.W. Greene and P.G.M. Wuts, 3rd Edition, John Wiley & Sons, New York (1999); Greene's Protective Groups in Organic Synthesis, P.G.M. Wuts, 5th Edition, John Wiley & Sons (2014); and Current Protocols in Nucleic Acid Chemistry, edited by S.L. Beaucage et al., June 2012, herein in particular Chapter 2. In certain embodiments, "hydroxyl protecting groups" of the present invention include, and are typically and preferably independently selected from, acetyl, benzoyl, benzyl, β-methoxyethoxymethyl ether (MEM), dimethoxytrityl, [bis-(4-methoxyphenyl)phenylmethyl] (DMTr), methoxymethyl ether (MOM), methoxytrityl[(4-methoxyphenyl)diphenylmethyl] (MMT), p-methoxybenzyl ether (PMB), methylthiomethyl ether, pivaloyl (Piv), tetrahydropyranyl (THP), tetrahydrofuran (THF), trityl(triphenylmethyl, Tr), silyl ethers such as t-butyldiphenylsilyl ether (TBDPS), trimethylsilyl (TMS), tert-butyldimethylsilyl (TBDMS), tri-isopropylsilyloxymethyl (TOM), and triisopropylsilyl (TIPS) ether; methyl ether, ethoxyethyl ether (EE), and each occurrence is typically and preferably independently selected from among these.

[0075] Preferred examples of the "hydroxyl protecting group" of the present invention include acetyl, t-butyl, t-butoxymethyl, methoxymethyl, tetrahydropyranyl, 1-ethoxyethyl, 1-(2-chloroethoxy)ethyl, 2-trimethylsilylethyl, p-chlorophenyl, 2,4-dinitrophenyl, benzyl, benzoyl, p-phenylbenzoyl, 2,6-dichlorobenzyl, diphenylmethyl, p-nitrobenzyl, triphenylmethyl (trityl), 4,4'-dimethoxytrityl, trimethylsilyl, triethylsilyl, t-butyldimethylsilyl (TBDMS), t-butyldiphenylsilyl (TBDPS), triphenylsilyl, triisopropylsilyl, benzoylformate, chloroacetyl, trichloroacetyl, trifluoroacetyl, pivaloyl, 9-fluorenylmethyl carbonate, mesylate, tosylate, triflate, 4-monomethoxytrityl (MMTr), 4,4'-dimethoxytrityl (DMTr), and 4,4',4''-trimethoxytrityl (TMTr), 2-cyanoethyl (CE or Cne), 2-(trimethylsilyl)ethyl (TSE), 2-(2-nitrophenyl)ethyl, 2-(4-cyanophenyl)ethyl, 2-(4-nitrophenyl)ethyl (NPE), 2-(4-nitrophenylsulfonyl)ethyl, 3,5-dichlorophenyl, 2,4-dimethylphenyl, 2-nitrophenyl, 4-nitrophenyl, 2,4,6-trimethylphenyl, 2-(2- and 9-(p-methoxyphenyl)xanthen-9-yl (MOX), independently selected from among 9-(nitrophenyl)ethyl, butylthiocarbonyl, 4,4',4''-tris(benzoyloxy)trityl, diphenylcarbamoyl, levulinyl, 2-(dibromomethyl)benzoyl (Dbmb), 2-(isopropylthiomethoxymethyl)benzoyl (Ptmt), 9-phenylxanthen-9-yl (pixyl), or 9-(p-methoxyphenyl)xanthin-9-yl (MOX).

[0076] The term "nucleobase," as used herein, is abbreviated as Bx and refers to unmodified or naturally occurring nucleobases as well as modified or non-naturally occurring nucleobases and their synthetic mimetics. A nucleobase is any heterocyclic base that contains one or more atoms or groups of atoms that are capable of hydrogen bonding to the heterocyclic base of a nucleic acid.

[0077] Typical and preferred examples of nucleobases are purine bases or pyrimidine bases, and preferably, the purine bases are purines or substituted purines, and the pyrimidine bases are pyrimidines or substituted pyrimidines. More preferably, the nucleobases are (i) adenine (A), (ii) cytosine (C), (iii) 5-methylcytosine (MeC), (iv) guanine (G), (v) uracil (U), or (vi) 5-methyluracil (MeU), or derivatives of (i), (ii), (iii), (iv), (v) or (vi). The terms "derivatives of (i), (ii), (iii), (iv), (v) or (vi)" and "nucleobase derivatives" are used interchangeably herein. Derivatives of (i), (ii), (iii), (iv), (v) or (vi) and nucleobase derivatives are known to those skilled in the art, and are described, for example, in Sharma VK et al., Med. Chem. Commun., 2014, vol. 5, pp. 1454-1471, and examples thereof include, but are not limited to, 5-hydroxymethylcytosine, xanthine, hypoxanthine, alkyl adenines such as 2-aminoadenine, 6-methyladenine, and 2-propyladenine, alkyl guanines such as 6-methylguanine and 2-propylguanine, alkynyl pyrimidine bases such as 2-thiouracil, 2-thiothymine, and 2-thiocytosine, 5-halouracil, 5-halocytosine, 5-propynyl (-C=C-CH3) uracil, and 5-propynyl (-C=C-CH3) cytosine, 6-azo uracil, 6-azo cytosine, and 6-azo 8-substituted purine bases such as 8-halo-, 8-amino-, 8-thiol-, 8-thioalkyl-, 8-hydroxyl-adenine or guanine, 5-substituted pyrimidine bases such as 5-halo-, particularly 5-bromo-, 5-trifluoromethyl-uracil or -cytosine; 7-methylguanine, 7-methyladenine, 2-F-adenine, 2-amino-adenine, 8-azaguanine and 8-azaadenine, 7-deazaguanine, 7-deazaadenine, 3-deazaguanine, 3-deazaadenine, hydrophobic bases, promiscuous bases, size-expanded bases, or fluorinated bases.In certain embodiments, the nucleobase includes, but is not limited to, tricyclic pyrimidines such as 1,3-diazaphenoxazin-2-one, 1,3-diazaphenothiazin-2-one, or 9-(2-aminoethoxy)-1,3-diazaphenoxazin-2-one (G-clamp). The term "nucleobase derivative" also includes those in which the purine or pyrimidine base is replaced by other heterocycles, such as 7-deaza-adenine, 7-deazaguanosine, 2-aminopyridine, or 2-pyridone. Additional nucleobases of the present invention include, but are not limited to, those known to those skilled in the art (e.g., U.S. Pat. No. 3,687,808; Swayze et al., The Medicinal Chemistry of Oligonucleotides, in Antisense a Drug Technology, Chapter 6, pp. 143-182 (Crooke, ST, ed., 2008); The Concise Encyclopedia of Polymer Science and Engineering, Kroschwitz, JI, ed., John Wiley & Sons, 1990, pp. 858-859; Englisch et al., Angewandte Chemie, International Edition, 1991, Vol. 30(6), pp. 613-623; Sanghvi, YS, Antisense Research and Applications, Crooke, ST and Lebleu, B., eds., CRC Press, 1993, pp. 273-302). The term "nucleobase derivative" also includes those in which the purine or pyrimidine base is replaced with a moiety corresponding to a spacer of the present invention, in particular a moiety for linking the one or more lipid moieties within the oligomeric compound, preferably the oligonucleotide. The specific linkage of the moiety corresponding to a spacer is known to those skilled in the art.Preferred nucleobase derivatives include methylated adenine, guanine, uracil and cytosine, and nucleobase derivatives, preferably nucleobase derivatives of (i), (ii), (iii) or (iv), wherein each amino group, preferably the exocyclic amino group, is protected by an acyl protecting group or a dialkylformamidino, preferably dimethylformamidino (DMF), and further include nucleobase derivatives such as 2-fluorouracil, 2-fluorocytosine, 5-bromouracil, 5-iodouracil, 2,6-diaminopurine, azacytosine, and pyrimidine analogs such as pseudoisocytosine and pseudouracil. The preparation of modified nucleobases is known in the art and is described in U.S. Pat. Nos. 3,687,808; 4,845,205; 5,130,302; 5,134,066; 5,175,273; 5,367,066; 5,432,272; 5,457,187; 5,459,255; 5,484,908; 5,502,177; Nos. 5,525,711; 5,552,540; 5,587,469; 5,594,121; 5,596,091; 5,614,617; 5,645,985; 5,750,692; 5,830,653; 5,763,588; 6,005,096; and 5,681,941.

[0078] The term "internucleoside linking group," as used herein, refers to any linking group known in the art that allows for the linkage, preferably the further linkage of said tricyclo-deoxyribonucleic acid (tc-DNA) nucleoside, either to an additional tc-DNA nucleoside, a nucleoside other than a tc-DNA nucleoside, or a non-nucleoside, including a peptide or protein. Representative patents that teach such possible linking groups include, but are not limited to, U.S. Pat. No. 5,034,506; U.S. Pat. No. 5,166,315; U.S. Pat. No. 5,185,444; U.S. Pat. No. 5,214,134; U.S. Pat. No. 5,216,141; U.S. Pat. No. 5,235,033; U.S. Pat. No. 5,264,562; U.S. Pat. No. 5,264,564; U.S. Pat. No. 5,405,938; U.S. Pat. No. 5,434,257; U.S. Pat. No. 5,466,677; U.S. Pat. No. 5,470,967; U.S. Pat. No. 5,480,968; U.S. Pat. No. 5,490,969; U.S. Pat. No. 5,505,938 ... Nos. 5,677,896; 5,541,307; 5,561,225; 5,596,086; 5,602,240; 5,608,046; 5,610,289; 5,618,704; 5,623,070; 5,663,312; 5,633,360; 5,677,437; 5,677,439; 5,646,269 and 5,792,608. Thus, the term "internucleoside linking group" includes phosphorus and non-phosphorus linking groups. The non-phosphorus linking group does not contain a phosphorus atom, and examples of the non-phosphorus linking group include, typically and preferably selected from, alkyl, aryl, preferably phenyl, benzyl or benzoyl, cycloalkyl, alkylenearyl, alkylenediaryl, alkoxy, alkoxyalkylene, alkylsulfonyl, alkyne, ether; carboxyl, amide, amine, amino, imine, thiol, sulfide, sulfoxide, sulfone, sulfamate, sulfonate, sulfonamide, siloxane or mixtures thereof, each independently of the other, optionally substituted with cyano, nitro, halogen.Typically and preferably, the internucleoside linking group is a phosphorus linking group, and the phosphorus linking group is P. III or P V " refers to a moiety containing a phosphorus atom in a valence state of . More preferably, the internucleoside linking group is a phosphorus linking group. Again more preferably, the internucleoside linking group is selected from a phosphodiester linking group, a phosphotriester linking group, a phosphorothioate linking group, a phosphorodithioate linking group, a phosphonate linking group, preferably an H-phosphonate linking group or a methylphosphonate linking group; a phosphonothioate linking group, preferably an H-phosphonothioate linking group or a methylphosphonothioate linking group; a phosphinate linking group, a phosphorthioamidate linking group, a phosphoramidate linking group, or a phosphite linking group. In another highly preferred embodiment, the internucleoside linking group is selected from a phosphodiester linking group, a phosphotriester linking group, a phosphorothioate linking group, or a phosphonate linking group, and the phosphonate is preferably an H-phosphonate linking group or a methylphosphonate linking group.

[0079] As used herein, the term "nucleoside" refers to a compound comprising a nucleobase and a sugar covalently linked to the nucleobase. Furthermore, the term "nucleoside" is intended to include all types of naturally occurring or modified nucleosides, or nucleoside mimetics, that can be incorporated into oligomers using natural or chemical oligomer synthesis. Typically and preferably, the term "nucleoside" as used herein refers to naturally occurring nucleosides, modified nucleosides, or nucleoside mimetics. The term "modified nucleoside" is intended to include modifications made to the sugar and / or nucleobase of the nucleoside known to those skilled in the art and described herein. The term "nucleoside mimetics" is intended to include those structures that are used to replace sugars and nucleobases. Examples of nucleoside mimetics include nucleosides in which the nucleobase is replaced with a phenoxazine moiety (e.g., a 9-(2-aminoethoxy)-1,3-diazaphenoxazin-2-one group) and the sugar moiety is replaced with a cyclohexenyl or bicyclo[3.1.0]hexyl moiety. The term "nucleoside" also includes combinations of modifications, such as modifications of two or more nucleobases, modifications of two or more sugars, or modifications of at least one nucleobase and at least one sugar.

[0080] Nucleoside sugars include, but are not limited to, monocyclic, bicyclic, or tricyclic ring systems, preferably tricyclic or bicyclic systems, or monocyclic ribose or deoxyribose (de(s)oxyribose). Sugar modifications further include, but are not limited to, modified stereochemical configurations, substitution of at least one group, or deletion of at least one group. Modified sugars are typically and preferably modified versions of ribosyl moieties (i.e., furanosyl moieties) naturally occurring in RNA and DNA, such as bicyclic sugars, tetrahydropyrans, 2'-modified sugars, 3'-modified sugars, 4'-modified sugars, 5'-modified sugars, or 4'-substituted sugars. Examples of suitable sugar modifications are known to those of skill in the art and include, but are not limited to, 2', 3', and / or 4' substituted nucleosides (e.g., 4'-S-modified nucleosides); 2'-O-alkyl or 2'-O-(substituted) alkyl, e.g., 2'-O-methyl, 2'-O-(2-cyanoethyl), 2'-O-(2-methoxy)ethyl (2'-MOE), 2'-O-(2-thiomethyl)ethyl, and other 2'-O-modified RNA nucleotide residues; 2'-O-(haloalkoxy)methyl, e.g., 2'-O-(2-chloroethoxy)methyl (MCEM), 2'-O-(2-thiomethyl)ethyl, and other 2'-O-substituted nucleotide residues. ,2-dichloroethoxy)methyl (DCEM); 2'-O-alkoxycarbonyl, such as 2'-O-[2-(methoxycarbonyl)ethyl] (MOCE), 2'-O-[2-(N-methylcarbamoyl)ethyl] (MCE), 2'-O-[2-(N,N-dimethylcarbamoyl))ethyl] (DMCE), especially 2'-O-methyl modified or 2'-O-methoxyethyl (2'-O-MOE); or other modified sugar moieties such as morpholino (PMO), cationic morpholino (PMOPlus), or modified morpholino groups such as PMO-X.The term "PMO-X" refers to a modified morpholino group, including at least one 3' or 5' end modification such as a 3'-fluorescent tag, a 3' quencher (e.g., 3'-carboxyfluorescein, 3'-Gene Tools Blue, 3'-lissamine, 3'-dabsyl), a 3'-affinity tag and a functional group for chemical linkage (e.g., 3'-biotin, 3'-primary amine, 3'-disulfide amide, 3'-pyridyldithio), a 5' end modification (5'-primary amine, 5'-dabsyl), a 3'-azide, a 3'-alkyne, a 5'-azide, a 5'-alkyne, or those disclosed in WO 2011 / 150408 and U.S. Patent Application Publication No. 2012 / 0065169.

[0081] "Bicyclic sugar moiety" includes bicyclic nucleosides having two interconnected ring systems, for example, a sugar moiety having a 2'-O-CH(alkyl)-4' or 2'-O-CH2-4' group, locked nucleic acid (LNA), xylo-LNA, alpha-L-LNA, beta-D-LNA, cEt (2'-O,4'-C constrained ethyl) LNA, cMOEt (2'-O,4'-C constrained methoxyethyl) LNA, ethylene-bridged nucleic acid (ENA), hexitol nucleic acid (HNA), fluorinated HNA (F-HNA), pyranosyl-RNA (p-RNA) or 3'-deoxypyranosyl-DNA (p-DNA).

[0082] In a preferred embodiment, oligomeric compound is oligonucleotide.The term " oligonucleotide " as used herein refers to the compound that comprises at least two nucleosides that are respectively linked to each other by internucleoside linking group.Therefore, the term " oligonucleotide " as used herein comprises, typically and preferably refers to the oligomeric compound that comprises at least two nucleosides that are linked to each other by internucleoside linking group, and said at least two nucleosides are independently selected from naturally occurring nucleosides, modified nucleosides or nucleoside mimetics.

[0083] Oligomeric compounds can be single-stranded or double-stranded. In one embodiment, the oligomeric compound is double-stranded (i.e., duplex). In a preferred embodiment, the oligomeric compound is single-stranded.

[0084] In preferred embodiments, the one or more lipid moieties are linked to the oligomeric compound, independently of one another, at (i) a terminal residue of the oligomeric compound, (ii) the 5' end of the oligomeric compound, (iii) the 3' end of the oligomeric compound, or (iv) an internal residue of the oligomeric compound.

[0085] The term "terminal" refers to the end or terminus of an oligomeric compound, and the integer (3', 5, etc.) indicates the carbon atom of the sugar contained in the nucleoside of the oligomeric compound. The terms "5' terminal group" or "3' terminal group," as used herein, refer to the group located at the 5' or 3' end, respectively.

[0086] The terms "natural" or "naturally occurring," used interchangeably herein, refer to compounds that are of natural origin.

[0087] The term "complementary" refers to a nucleic acid molecule that can form hydrogen bond(s) with another nucleic acid molecule, either through traditional Watson-Crick base pairing or other non-traditional types of pairing (e.g., Hoogsteen or reversed Hoogsteen hydrogen bonding) between complementary nucleosides or nucleotides. "Complementary" (or "specifically hybridizable") is a term that indicates a sufficient degree of complementarity or precise pairing such that stable, specific binding occurs between an oligomeric compound and a pre-mRNA or mRNA target. It is understood in the art that a nucleic acid molecule need not be 100% complementary to a target nucleic acid sequence to be specifically hybridizable. That is, two or more nucleic acid molecules need not be perfectly complementary. Complementarity can be indicated by the percentage of contiguous residues in a nucleic acid molecule that can form hydrogen bonds with a second nucleic acid molecule. For example, if a first nucleic acid molecule has 10 nucleotides and a second nucleic acid molecule has 10 nucleotides, then base pairing of 5, 6, 7, 8, 9, or 10 nucleotides between the first and second nucleic acid molecules represents 50%, 60%, 70%, 80%, 90%, and 100%, respectively. "Perfectly" or "fully" complementary nucleic acid molecules mean that all consecutive residues in a first nucleic acid molecule will hydrogen bond with the same number of consecutive residues in a second nucleic acid molecule, and either both nucleic acid molecules have the same number of nucleotides (i.e., have the same length), or the two molecules have different lengths.

[0088] The term "exon skipping" refers to the modification of pre-mRNA splicing by targeting splice donor and / or splice acceptor sites in pre-mRNA with one or more complementary antisense oligonucleotides or oligomeric compounds.By blocking the spliceosome's access to one or more splice donor or splice acceptor sites, or any other site in the exon or intron involved in splicing definition, the oligonucleotide can prevent the splicing reaction and cause the exon to be deleted from the fully processed mRNA.Exon skipping is achieved in the nucleus during the maturation process of pre-mRNA.Exon skipping involves masking the important sequence involved in the splicing of the target exon by using an antisense oligonucleotide that is complementary to the splice donor sequence in pre-mRNA. For example, the compositions of the present invention comprising the oligomeric compounds provided herein can be suitably used for exon skipping by masking splice sites at intron / exon junctions within dystrophin pre-mRNA, thereby facilitating the deletion of mutant exons during processing of the pre-mRNA into mature mRNA.

[0089] The term "exon inclusion" refers to an oligonucleotide-mediated process, such as base pairing of an antisense oligonucleotide to a targeted pre-mRNA, to block exon or intron splicing enhancers, block corresponding splicing repressors, and / or disrupt unfavorable secondary structures, resulting in more efficient recognition of the exon by the spliceosome and restoration of exon expression.

[0090] The term "in vivo" refers to an event that takes place within the body of a subject.

[0091] The term "in vitro" refers to events that take place outside a subject's body. In vitro assays include cell-based assays, in which live or dead cells are used, and can also include cell-free assays, in which no intact cells are used.

[0092] The term "effective amount" or "therapeutically effective amount" refers to an amount of a compound or combination of compounds described herein sufficient to effect the intended application, including, but not limited to, the treatment of a disease. A therapeutically effective amount may vary depending on the intended application (in vitro or in vivo), or the human subject and disease state being treated (e.g., the subject's weight, age, and sex), the severity of the disease state, the mode of administration, etc., and can be readily determined by one skilled in the art. The term also applies to a dose that induces a specific response in target cells (e.g., reduced platelet adhesion and / or cell migration). The specific dose will vary depending on the particular compound selected, the dosing regimen to be followed, whether the compound is administered in combination with other compounds, the timing of administration, the tissue to which it is administered, and the bodily delivery system through which the compound is delivered.

[0093] As used herein, a "therapeutic effect" encompasses a therapeutic benefit and / or a prophylactic benefit in a human subject. A prophylactic effect includes delaying or eliminating the appearance of a disease or condition, delaying or eliminating the onset of symptoms of a disease or condition, slowing, halting, or reversing the progression of a disease or condition, or any combination thereof.

[0094] The term "pharmaceutically acceptable salt" refers to salts derived from various organic and inorganic counterions known in the art. Pharmaceutically acceptable acid addition salts can be formed using inorganic and organic acids. Inorganic acids from which salts can be derived include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, and phosphoric acid. Organic acids from which salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, and salicylic acid. Pharmaceutically acceptable base addition salts can be formed using inorganic and organic bases. Inorganic bases from which salts can be derived include, for example, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, and aluminum. Organic bases from which salts can be derived include, for example, primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins. Specific examples include isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, and ethanolamine. In some embodiments, the pharmaceutically acceptable base addition salt is selected from ammonium, potassium, sodium, calcium, and magnesium salts, and preferably, the pharmaceutically acceptable salt is a sodium salt.

[0095] In the case of a hydroxyl group (OH) or thiol group (SH) present in the oligomeric compound of the present invention, preferably the oligonucleotide of the present invention, typically and preferably bound to P(III) or P(V), typically and preferably present in the one or more lipid moieties as part of the B group of the one or more lipid moieties, or present in the spacer, or typically and preferably present as part of the internucleoside linking group selected from phosphorothioates or phosphorodiesters, each of the hydroxyl group (OH) or thiol group (SH) can exist, independently of the other, in its ionic state, such as the OH group or an O-anion and a pharmaceutically acceptable cation, or the SH group or an S-anion and a pharmaceutically acceptable cation. Furthermore, any combination and any equilibrium state between the above situations in the compositions of the present invention is included, particularly taking into account oxygen- or sulfur-containing groups on the P(III) or P(V), such as (=O), (=S), another OH group, or SH group, as will be known to those skilled in the art. For convenience, aspects and embodiments of the present invention will typically describe only one of the foregoing situations. By way of example, a preferred spacer of the present invention is #-NH-C 2~12 As shown, but not limited to, spacers where the hydrogen is positioned on the oxygen, thus #-NH-C 2~12 All alkylene-OP(OH)(S)-§ and pharmaceutically acceptable salts thereof are included herein.

[0096] Thus, a pharmaceutically acceptable salt in the context of a hydroxyl group (OH) and / or a thiol group (SH) present in the oligomeric compound of the invention, preferably in the oligonucleotide of the invention, typically and preferably attached to P(III) or P(V), typically and preferably present in the one or more lipid moieties as part of the B group of the one or more lipid moieties, or present in the spacer, or as part of the internucleoside linking group, typically and preferably selected from phosphorothioate or phosphorodiester, is a pharmaceutically acceptable salt of said OH. The present invention refers to a composition of matter according to the invention, wherein one or more of said groups or said SH groups are, independently of one another, present as said OH group or in its ionic state, such as an O-anion and its pharmaceutically acceptable cation, or as said SH group or in its ionic state, such as an S-anion and its pharmaceutically acceptable cation, typically and preferably, said pharmaceutically acceptable cation is selected from protonated trimethylamine, protonated diethylamine, protonated methylamine, ammonium, sodium or potassium, more preferably said pharmaceutically acceptable cation is sodium.

[0097] "Pharmaceutically acceptable carrier" or "pharmaceutically acceptable excipient" is intended to include any solvent, dispersion medium, coating agent, antibacterial and antifungal agent, isotonic and absorption delaying agent, and inactive ingredients. The use of such pharmaceutically acceptable carriers or pharmaceutically acceptable excipients for active pharmaceutical ingredients is well known in the art. Any conventional pharmaceutically acceptable carrier or pharmaceutically acceptable excipient is contemplated for use in the therapeutic compositions of the present invention, except insofar as it is incompatible with the active pharmaceutical ingredient. Additional active pharmaceutical ingredients, such as other drugs, can also be incorporated into the compositions and methods described.

[0098] When a range is used herein to describe a physical or chemical property, such as, for example, molecular weight or chemical formula, it is intended to include all combinations and subcombinations of ranges and specific embodiments therein. The use of the term "about" when referring to a numerical value or numerical range means that the referenced numerical value or numerical range is an approximation within experimental variation (or within statistical experimental error), and thus the numerical value or numerical range may vary. The variation is typically 0% to 15%, 0% to 10%, or 0% to 5% of the stated number or numerical range.

[0099] In a first aspect, the present invention provides a composition comprising an oligomeric compound comprising one or more tricyclo-deoxyribonucleic acid (tc-DNA) nucleosides and one or more lipid moieties, preferably exactly one lipid moiety, covalently linked to the oligomeric compound, either directly or via a spacer, and preferably wherein the oligomeric compound comprises 5 to 40 monomeric subunits.

[0100] tc-DNA nucleosides In embodiments, the tc-DNA nucleoside of the oligomeric compound of the composition of the present invention is a compound of formula (1): [ka] (In the formula, Bx is a nucleobase; one of T1 and T2 is an internucleoside linking group, and the other of T1 and T2 is OR1, OR2, a 5'-terminal group, a 3'-terminal group, or an internucleoside linking group (wherein R1 is H or a hydroxyl protecting group, and R2 is a phosphorus moiety); q1, q2, q3, q4, and q5 are each independently hydrogen (H), halogen, C 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, substituted C 1~6 Alkyl, substituted C 2~6Alkenyl, substituted C 2~6 Alkynyl, and -(CH2) n -C(O)-R 6’ wherein n is 0 to 6; and R 6’ are OH, NH2, OC 1~32 Alkyl and NH-C 1~32 selected from the group consisting of alkyl; z1 and z2 each independently represent H, halogen, or C 1~6 Alkyl, C 1~6 Alkoxyl, OC 2~6 Alkenyl, OC 2~6 Alkynyl, substituted C 1~6 Alkyl, substituted C 1~6 Alkoxy, substituted OC 2~6 Alkenyl and substituted O-C 2~6 alkynyl), or a pharmaceutically acceptable salt thereof.

[0101] In embodiments, the tc-DNA nucleosides of the oligomeric compounds of the compositions of the present invention include compounds of formula (1) wherein q5 is H.

[0102] In embodiments, the tc-DNA nucleosides of the oligomeric compounds of the present invention comprise a compound of formula (1) where Bx is selected from the group consisting of thymine, adenine, guanine, and cytosine. In embodiments, the tc-DNA nucleosides of the oligomeric compounds of the present invention comprise a compound of formula (1) where Bx is a modified base. In embodiments, the tc-DNA nucleosides of the oligomeric compounds of the present invention comprise a compound of formula (1) where Bx is a modified base selected from the group consisting of 5-methylcytosine, 5-bromouracil, inosine, and 2,6-diaminopurine.

[0103] In embodiments, the tc-DNA nucleoside of the oligomeric compound of the composition of the present invention is a compound of formula (2): [ka] (In the formula, Bx is a nucleobase; One of T1 and T2 is an internucleoside linking group, and the other of T1 and T2 is OR1, OR2, a 5'-terminal group, a 3'-terminal group, or an internucleoside linking group (wherein R1 is H or a hydroxyl protecting group and R2 is a phosphorus moiety). Includes.

[0104] In embodiments, the tc-DNA nucleosides of the oligomeric compounds of the present invention comprise compounds of formula (2) where Bx is selected from the group consisting of thymine, adenine, guanine, and cytosine. In embodiments, the tc-DNA nucleosides of the oligomeric compounds of the present invention comprise compounds of formula (2) where Bx is a modified base. In embodiments, the tc-DNA nucleosides of the oligomeric compounds of the present invention comprise compounds of formula (2) where Bx is a modified base selected from the group consisting of 5-methylcytosine, 5-bromouracil, inosine, and 2,6-diaminopurine.

[0105] In embodiments, the tc-DNA nucleoside of the oligomeric compound of the composition of the present invention is a compound of formula (3) (also known as C(6')-functionalized tc-DNA): [ka] (In the formula, Bx is a nucleobase; R 6’ are OH, NH2, OC 1~32 Alkyl and NH-C 1~32 selected from the group consisting of alkyl; One of T1 and T2 is an internucleoside linking group, and the other of T1 and T2 is OR1, OR2, a 5'-terminal group, a 3'-terminal group, or an internucleoside linking group (wherein R1 is H or a hydroxyl protecting group and R2 is a phosphorus moiety). Includes.

[0106] In embodiments, the tc-DNA nucleosides of the oligomeric compounds of the present invention comprise compounds of formula (3) where Bx is selected from the group consisting of thymine, adenine, guanine, and cytosine. In embodiments, the tc-DNA nucleosides of the oligomeric compounds of the present invention comprise compounds of formula (3) where Bx is a modified base. In embodiments, the tc-DNA nucleosides of the oligomeric compounds of the present invention comprise compounds of formula (3) where Bx is a modified base selected from the group consisting of 5-methylcytosine, 5-bromouracil, inosine, and 2,6-diaminopurine.

[0107] In embodiments, the tc-DNA nucleoside of the oligomeric compound of the composition of the present invention is a compound of formula (4) (also known as 6'-fluoro-tc-DNA): [ka] (In the formula, Bx is a nucleobase; One of T1 and T2 is an internucleoside linking group, and the other of T1 and T2 is OR1, OR2, a 5'-terminal group, a 3'-terminal group, or an internucleoside linking group (wherein R1 is H or a hydroxyl protecting group and R2 is a phosphorus moiety). Includes.

[0108] In embodiments, the tc-DNA nucleosides of the oligomeric compounds of the present invention comprise compounds of formula (4) where Bx is selected from the group consisting of thymine, adenine, guanine, and cytosine. In embodiments, the tc-DNA nucleosides of the oligomeric compounds of the present invention comprise compounds of formula (4) where Bx is a modified base. In embodiments, the tc-DNA nucleosides of the oligomeric compounds of the present invention comprise compounds of formula (4) where Bx is a modified base selected from the group consisting of 5-methylcytosine, 5-bromouracil, inosine, and 2,6-diaminopurine.

[0109] In embodiments, the tc-DNA nucleoside of the oligomeric compound of the composition of the present invention is a compound of formula (5) (also known as 2'-fluoro-tc-DNA): [ka] (In the formula, Bx is a nucleobase; One of T1 and T2 is an internucleoside linking group, and the other of T1 and T2 is OR1, OR2, a 5'-terminal group, a 3'-terminal group, or an internucleoside linking group (wherein R1 is H or a hydroxyl protecting group and R2 is a phosphorus moiety). Includes.

[0110] In embodiments, the tc-DNA nucleosides of the oligomeric compounds of the present invention comprise compounds of formula (5) where Bx is selected from the group consisting of thymine, adenine, guanine, and cytosine. In embodiments, the tc-DNA nucleosides of the oligomeric compounds of the present invention comprise compounds of formula (5) where Bx is a modified base. In embodiments, the tc-DNA nucleosides of the oligomeric compounds of the present invention comprise compounds of formula (5) where Bx is a modified base selected from the group consisting of 5-methylcytosine, 5-bromouracil, inosine, and 2,6-diaminopurine.

[0111] Thus, in embodiments, the tc-DNA nucleoside of said one or more nucleosides of the oligomeric compounds of the compositions of the invention is a compound of formula (5') (also known as 2'-fluoro-tc-ANA): [ka] (In the formula, Bx is a nucleobase; One of T1 and T2 is an internucleoside linking group, and the other of T1 and T2 is OR1, OR2, a 5'-terminal group, a 3'-terminal group, or an internucleoside linking group (wherein R1 is H or a hydroxyl protecting group and R2 is a phosphorus moiety). Includes.

[0112] In embodiments, the tc-DNA nucleosides of the oligomeric compounds of the present invention comprise compounds of formula (5') where Bx is selected from the group consisting of thymine, adenine, guanine, and cytosine. In embodiments, the tc-DNA nucleosides of the oligomeric compounds of the present invention comprise compounds of formula (5') where Bx is a modified base. In embodiments, the tc-DNA nucleosides of the oligomeric compounds of the present invention comprise compounds of formula (5') where Bx is a modified base selected from the group consisting of 5-methylcytosine, 5-bromouracil, inosine, and 2,6-diaminopurine.

[0113] General methods for preparing compounds of formula (1) and formula (2) for use in oligomeric compounds are known in the art, including those described in U.S. Patent Application Publication Nos. 2015 / 0141637, 2016 / 0002280, and 2014 / 0296323, the disclosures of which are incorporated herein by reference. Standard phosphoramidite building blocks for tc-DNA are described in the art, for example, in Steffens and Leumann, Helv. Chim. Acta, 1997, 80, 2426-2439. Methods for preparing compounds of formula (3) are described, for example, in Lietard and Leumann, J. Org. Chem., 2012, 77, 4566-77, the disclosure of which is incorporated herein by reference. Methods for preparing compounds of formula (4) are described, for example, in Medvecky, Istrate, and Leumann, J. Org. Chem., 2015, Vol. 80, pp. 3556-65, the disclosure of which is incorporated herein by reference. Methods for preparing compounds of formula (5) and (5') are described, for example, in Istrate, Medvecky, and Leumann, Org. Lett., 2015, Vol. 17, pp. 1950-53, the disclosure of which is incorporated herein by reference.

[0114] 2'-Modified RNA Nucleosides and Other Nucleosides In embodiments, the oligomeric compound further comprises one or more nucleosides other than tc-DNA nucleosides, wherein said one or more nucleosides other than tc-DNA nucleosides are, independently of each other, 2'-modified ribonucleic acid (2'-modified-RNA) nucleosides.

[0115] In embodiments, one or more nucleosides other than tc-DNA nucleosides of the oligomeric compounds of the invention are RNA nucleosides of formula (6): [ka] (In the formula, Bx is a nucleobase; One of T1 and T2 is an internucleoside linking group, and the other of T1 and T2 is OR1, OR2, a 5'-terminal group, a 3'-terminal group, or an internucleoside linking group (wherein R1 is H or a hydroxyl protecting group and R2 is a phosphorus moiety). is.

[0116] In embodiments, one or more nucleosides other than tc-DNA nucleosides of the oligomeric compounds of the invention are RNA nucleosides of formula (6), where Bx is selected from the group consisting of cytosine, adenine, guanine, and uracil. In embodiments, 2'-modified-RNA nucleosides of the oligomeric compounds of the invention include compounds of formula (6), where Bx is a modified base. In embodiments, 2'-modified-RNA nucleosides of the oligomeric compounds of the invention include compounds of formula (6), where Bx is a modified base selected from the group consisting of 5-methylcytosine, 5-methyluracil, 5-bromouracil, inosine, and 2,6-diaminopurine.

[0117] In an embodiment, the 2'-modified-RNA nucleoside of the preferred oligomeric compound of the composition of the present invention is a compound of formula (7) (2'-O-methyl-RNA nucleoside): [ka] (In the formula, Bx is a nucleobase; One of T1 and T2 is an internucleoside linking group, and the other of T1 and T2 is OR1, OR2, a 5'-terminal group, a 3'-terminal group, or an internucleoside linking group (wherein R1 is H or a hydroxyl protecting group and R2 is a phosphorus moiety). Includes.

[0118] In embodiments, 2'-modified-RNA nucleosides of the oligomeric compounds of the invention include compounds of formula (7) where Bx is selected from the group consisting of cytosine, adenine, guanine, and uracil. In embodiments, 2'-modified-RNA nucleosides of the oligomeric compounds of the invention include compounds of formula (7) where Bx is a modified base. In embodiments, 2'-modified-RNA nucleosides of the oligomeric compounds of the invention include compounds of formula (7) where Bx is a modified base selected from the group consisting of 5-methylcytosine, 5-methyluracil, 5-bromouracil, inosine, and 2,6-diaminopurine.

[0119] In embodiments, the 2'-modified-RNA nucleoside of the oligomeric compound of the invention is a compound of formula (8) (2'-O-propargyl-RNA nucleoside): [ka] (In the formula, Bx is a nucleobase; One of T1 and T2 is an internucleoside linking group, and the other of T1 and T2 is OR1, OR2, a 5'-terminal group, a 3'-terminal group, or an internucleoside linking group (wherein R1 is H or a hydroxyl protecting group and R2 is a phosphorus moiety). Includes.

[0120] In embodiments, 2'-modified-RNA nucleosides of the oligomeric compounds of the invention include compounds of formula (8) where Bx is selected from the group consisting of cytosine, adenine, guanine, and uracil. In embodiments, 2'-modified-RNA nucleosides of the oligomeric compounds of the invention include compounds of formula (8) where Bx is a modified base. In embodiments, 2'-modified-RNA nucleosides of the oligomeric compounds of the invention include compounds of formula (8) where Bx is a modified base selected from the group consisting of 5-methylcytosine, 5-methyluracil, 5-bromouracil, inosine, and 2,6-diaminopurine.

[0121] In embodiments, the 2'-modified-RNA nucleoside of the oligomeric compound of the invention is a compound of formula (9) (2'-O-propylamino-RNA nucleoside): [ka] (In the formula, Bx is a nucleobase; One of T1 and T2 is an internucleoside linking group, and the other of T1 and T2 is OR1, OR2, a 5'-terminal group, a 3'-terminal group, or an internucleoside linking group (wherein R1 is H or a hydroxyl protecting group and R2 is a phosphorus moiety). Includes.

[0122] In embodiments, 2'-modified-RNA nucleosides of the oligomeric compounds of the invention include compounds of formula (9) where Bx is selected from the group consisting of cytosine, adenine, guanine, and uracil. In embodiments, 2'-modified-RNA nucleosides of the oligomeric compounds of the invention include compounds of formula (9) where Bx is a modified base. In embodiments, 2'-modified-RNA nucleosides of the oligomeric compounds of the invention include compounds of formula (9) where Bx is a modified base selected from the group consisting of 5-methylcytosine, 5-methyluracil, 5-bromouracil, inosine, and 2,6-diaminopurine.

[0123] In embodiments, the 2'-modified-RNA nucleosides of the oligomeric compounds of the invention are compounds of formula (10) (2'-amino-RNA nucleosides): [ka] (In the formula, Bx is a nucleobase; One of T1 and T2 is an internucleoside linking group, and the other of T1 and T2 is OR1, OR2, a 5'-terminal group, a 3'-terminal group, or an internucleoside linking group (wherein R1 is H or a hydroxyl protecting group and R2 is a phosphorus moiety). Includes.

[0124] In embodiments, 2'-modified-RNA nucleosides of the oligomeric compounds of the invention include compounds of formula (10) where Bx is selected from the group consisting of cytosine, adenine, guanine, and uracil. In embodiments, 2'-modified-RNA nucleosides of the oligomeric compounds of the invention include compounds of formula (10) where Bx is a modified base. In embodiments, 2'-modified-RNA nucleosides of the oligomeric compounds of the invention include compounds of formula (10) where Bx is a modified base selected from the group consisting of 5-methylcytosine, 5-methyluracil, 5-bromouracil, inosine, and 2,6-diaminopurine.

[0125] In embodiments, the 2'-modified-RNA nucleoside of the oligomeric compound of the invention is a compound of formula (11) (2'-fluoro-RNA nucleoside): [ka] (In the formula, Bx is a nucleobase; One of T1 and T2 is an internucleoside linking group, and the other of T1 and T2 is OR1, OR2, a 5'-terminal group, a 3'-terminal group, or an internucleoside linking group (wherein R1 is H or a hydroxyl protecting group and R2 is a phosphorus moiety). Includes.

[0126] In embodiments, 2'-modified-RNA nucleosides of the oligomeric compounds of the invention include compounds of formula (11) where Bx is selected from the group consisting of cytosine, adenine, guanine, and uracil. In embodiments, 2'-modified-RNA nucleosides of the oligomeric compounds of the invention include compounds of formula (11) where Bx is a modified base. In embodiments, 2'-modified-RNA nucleosides of the oligomeric compounds of the invention include compounds of formula (11) where Bx is a modified base selected from the group consisting of 5-methylcytosine, 5-methyluracil, 5-bromouracil, inosine, and 2,6-diaminopurine.

[0127] In embodiments, one or more nucleosides other than tc-DNA nucleosides of the oligomeric compounds of the invention are compounds of formula (11') (2'-deoxy 2'-fluoro-arabinonucleosides (2'-FANA)): [ka] (In the formula, Bx is a nucleobase; One of T1 and T2 is an internucleoside linking group, and the other of T1 and T2 is OR1, OR2, a 5'-terminal group, a 3'-terminal group, or an internucleoside linking group (wherein R1 is H or a hydroxyl protecting group and R2 is a phosphorus moiety). Includes.

[0128] In embodiments, the one or more nucleosides other than tc-DNA nucleosides of the oligomeric compounds of the invention comprise a compound of formula (11') where Bx is selected from the group consisting of cytosine, adenine, guanine, and uracil. In embodiments, the 2'-modified-RNA nucleosides of the oligomeric compounds of the invention comprise a compound of formula (11') where Bx is a modified base. In embodiments, the 2'-modified-RNA nucleosides of the oligomeric compounds of the invention comprise a compound of formula (11') where Bx is a modified base selected from the group consisting of 5-methylcytosine, 5-methyluracil, 5-bromouracil, inosine, and 2,6-diaminopurine.

[0129] In embodiments, the 2'-modified-RNA nucleoside of the oligomeric compound of the invention is a compound of formula (12) (2'-O-methoxyethyl-RNA nucleoside, or 2'-MOE nucleoside): [ka] (In the formula, Bx is a nucleobase; One of T1 and T2 is an internucleoside linking group, and the other of T1 and T2 is OR1, OR2, a 5'-terminal group, a 3'-terminal group, or an internucleoside linking group (wherein R1 is H or a hydroxyl protecting group and R2 is a phosphorus moiety). Includes.

[0130] In embodiments, 2'-modified-RNA nucleosides of the oligomeric compounds of the invention include compounds of formula (12) where Bx is selected from the group consisting of cytosine, adenine, guanine, and uracil. In embodiments, 2'-modified-RNA nucleosides of the oligomeric compounds of the invention include compounds of formula (12) where Bx is a modified base. In embodiments, 2'-modified-RNA nucleosides of the oligomeric compounds of the invention include compounds of formula (12) where Bx is a modified base selected from the group consisting of 5-methylcytosine, 5-methyluracil, 5-bromouracil, inosine, and 2,6-diaminopurine.

[0131] In embodiments, one or more nucleosides other than tc-DNA nucleosides of an oligomeric compound of the invention is a compound of formula (13) (morpholino nucleoside): [ka] (In the formula, Bx is a nucleobase; One of T1 and T2 is an internucleoside linking group, and the other of T1 and T2 is OR1, OR2, a 5'-terminal group, a 3'-terminal group, or an internucleoside linking group (wherein R1 is H or a hydroxyl protecting group and R2 is a phosphorus moiety). Includes.

[0132] In embodiments, the one or more nucleosides other than tc-DNA nucleosides of the oligomeric compounds of the invention comprise a compound of formula (13) where Bx is selected from the group consisting of cytosine, adenine, guanine, and uracil. In embodiments, the 2'-modified-RNA nucleosides of the oligomeric compounds of the invention comprise a compound of formula (13) where Bx is a modified base. In embodiments, the 2'-modified-RNA nucleosides of the oligomeric compounds of the invention comprise a compound of formula (13) where Bx is a modified base selected from the group consisting of 5-methylcytosine, 5-methyluracil, 5-bromouracil, inosine, and 2,6-diaminopurine.

[0133] In embodiments, the 2'-modified-RNA nucleosides of the oligomeric compounds of the invention are compounds of formula (14) (locked nucleic acid or LNA nucleosides): [ka] (In the formula, Bx is a nucleobase; One of T1 and T2 is an internucleoside linking group, and the other of T1 and T2 is OR1, OR2, a 5'-terminal group, a 3'-terminal group, or an internucleoside linking group (wherein R1 is H or a hydroxyl protecting group and R2 is a phosphorus moiety). Includes.

[0134] In embodiments, 2'-modified-RNA nucleosides of the oligomeric compounds of the invention include compounds of formula (14) where Bx is selected from the group consisting of cytosine, adenine, guanine, and uracil. In embodiments, 2'-modified-RNA nucleosides of the oligomeric compounds of the invention include compounds of formula (14) where Bx is a modified base. In embodiments, 2'-modified-RNA nucleosides of the oligomeric compounds of the invention include compounds of formula (14) where Bx is a modified base selected from the group consisting of 5-methylcytosine, 5-methyluracil, 5-bromouracil, inosine, and 2,6-diaminopurine.

[0135] General methods for the preparation of compounds of formula (6) through formula (14) for use in oligomeric compounds are known in the art and include the following: U.S. Pat. No. 4,981,957; U.S. Pat. No. 5,118,800; U.S. Pat. No. 5,319,080; U.S. Pat. No. 5,359,044; U.S. Pat. No. 5,393,878; U.S. Pat. No. 5,446,137; U.S. Pat. No. 5,466,786; U.S. Pat. No. 5,514,785; U.S. Pat. No. 5,519,134; U.S. Pat. No. 5,567,811; U.S. Pat. No. 5,576,427; U.S. Pat. No. 5,599,142; Nos. 1,722; 5,597,909; 5,610,300; 5,627,053; 5,639,873; 5,646,265; 5,670,633; 5,700,920; 5,792,847; and 6,600,032; U.S. Patent Application Publication Nos. 2015 / 0141637, 2016 / 0002280, and 2014 / 0296323; and methods described in Renneberg et al., J. Am. Chem. Soc., 2002, 124, 5993-6002, the disclosures of which are incorporated herein by reference.

[0136] In embodiments, the oligomeric compound further comprises one or more nucleosides other than tc-DNA nucleosides, wherein said one or more nucleosides other than tc-DNA nucleosides are, independently of each other: Ribonucleic acid (RNA) nucleosides; Deoxyribonucleic acid (DNA) nucleosides; 2'-modified RNA nucleosides Preferably, 2',4'-BNA (2',4'-BNA) with a 2'-ONC crosslinking system NC ), bicyclic nucleic acid (2',4'-BNA) nucleosides selected from stereoisomers of LNA-α-L-LNA and ethylene nucleic acid (ENA) nucleosides; peptide nucleic acid (PNA) nucleosides; 2'-deoxy2'-fluoro-arabino (FANA) nucleosides; Hexitol nucleic acid (HNA) nucleosides; and Phosphorodiamidate morpholino (PMO) nucleosides is selected from.

[0137] Additional nucleosides useful for the present invention will be known to those skilled in the art, such as other lipophilic 2'-O-alkylRNAs described in Biochemistry, 2005, vol. 44, pp. 9045-9057.

[0138] Non-nucleoside In embodiments, oligomeric compounds of the present invention include non-nucleosides, also known in the art as non-nucleoside linkers, non-nucleotidyl linkers, which are highly flexible substitutes for the sugar carbons of, for example, ribofuranone moieties and can be used to replace tc-DNA nucleosides and nucleosides other than tc-DNA nucleosides of the present oligomeric compounds. An exemplary non-nucleotide is the 1,3-propanediol group shown in formula (15), which is shown connecting two exemplary phosphorodiester internucleoside linkages: [ka]

[0139] The wavy lines in formula (15) represent additional oligomeric repeating nucleoside and internucleoside linking units described herein.

[0140] The non-nucleotides of the invention can be used with any of the internucleoside linkages described herein, including embodiments in which the phosphorodiester internucleoside linkage shown in formula (15) is replaced with one or more phosphorothioate internucleoside linkages.

[0141] In embodiments, the non-nucleotide is a 1,3-propanediol group. The synthesis and incorporation of 1,3-propanediol groups into oligomeric compounds is known in the art and is described, for example, in Seela and Kaiser, Nuc. Acids Res., 1987, vol. 15, pp. 3113-29. In embodiments, the oligomeric compounds of the invention contain 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 1,3-propanediol groups linked by phosphorothioate internucleoside linkages, phosphorodiester internucleoside linkages, or mixtures thereof.

[0142] Alternative non-nucleosides can also be used in the oligomeric compounds of the invention, such as ethylene glycol oligomers of various lengths (i.e., one, two, three, or more ethylene glycol units linked to form a single non-nucleoside). Various suitable ethylene glycol groups are described, for example, in Pils and Micura, Nuc. Acids Res., 2000, 28, 1859-63. The synthesis and use of non-nucleosides is also described, for example, in U.S. Pat. No. 5,573,906, the disclosure of which is incorporated herein by reference.

[0143] Internucleoside linking group In embodiments, the internucleoside linking groups of the oligomeric compounds of the invention are independently selected from the group consisting of phosphorothioate, phosphorodithioate, phosphorodiester, phosphotriester, aminoalkylphosphotriester, methylphosphonate, alkylphosphonate, 5'-alkylenephosphonate, phosphonate, phosphinate, phosphoramidate, 3'-aminophosphoramidate, aminoalkylphosphoramidate, thionophosphoramidate, thionoalkylphosphonate, thionoalkylphosphotriester, selenophosphate, and boranophosphate linkages.

[0144] In preferred embodiments, the internucleoside linkages of the oligomeric compounds of the invention are independently selected from the group consisting of phosphorothioate linkages and phosphorodiester linkages, hi embodiments, the internucleoside linkages of the oligomeric compounds of the invention comprise only phosphorodiester linkages.

[0145] An exemplary phosphorothioate linkage is represented by formula (16): [ka] As shown in.

[0146] An exemplary phosphorodiester linkage is represented by formula (17): [ka] As shown in.

[0147] The wavy lines in formulas (16) and (17) represent additional oligomeric repeating nucleosides and internucleoside linkages described herein.

[0148] General methods for the preparation of internucleoside linkages for use in oligomeric compounds are known in the art and include, but are not limited to, U.S. Pat. Nos. 3,687,808; 4,469,863; 4,476,301; 5,023,243; 5,177,196; 5,188,897; 5,264,423; 5,276,019; 5,278,302; 5,286,717; 5,321,131; 5,399,676; 5,405,939; 5,453,496; 5,4 Nos. 5,555,233; 5,466,677; 5,476,925; 5,519,126; 5,536,821; 5,541,306; 5,550,111; 5,563,253; 5,571,799; 5,587,361; 5,194,599; 5,565,555; 5,527,899; 5,721,218; 5,672,697 and 5,625,050, the disclosures of which are incorporated herein by reference. Phosphorothioates can be prepared from phosphate triesters, for example, using phenylacetyl disulfide (PADS) chemistry as described, for example, in Krotz et al., Org. Proc. R&D, 2004, 8, 852-58, as part of a solid-phase synthesis using a 3'- to 5'-elongation cycle of four reactions: detritylation, coupling, sulfurization and capping using PADS, followed by deprotection, cleavage from the support, and purification steps.

[0149] The term "phosphorus moiety" as used herein means P III or P V This refers to a moiety containing a phosphorus atom in the valence state of formula (18): [ka] (In the formula, W represents O, S, or Se, or W represents an electron pair; R3 and R4 are, independently of one another, H, halogen, OH, OR5, NR6R7, SH, SR8, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, C1-C6 aminoalkyl; R5 are, independently of one another, optionally substituted with cyano, nitro, halogen, -NHC(O)C1-C3 alkyl, -NHC(O)C1-C3 haloalkyl, C1-C3 alkylsulfonyl. , C1-C9 alkyl, C1-C6 alkoxy; aryl, C1-C6 alkylenearyl, C1-C6 alkylenediaryl, each optionally substituted with cyano, nitro, halogen, C1-C4 alkoxy, C1-C4 haloalkyl, C1-C4 haloalkoxy, NHC(O)C1-C3 alkyl, NHC(O)C1-C3 haloalkyl, C1-C3 alkylsulfonyl; acetyl; a hydroxyl protecting group. R6 and R7 are, independently of one another, hydrogen, cyano, nitro, halogen, C2-C6 alkenyl, C3-C6 cycloalkyl, C1-C9 alkyl optionally substituted with C1-C3 alkoxy; aryl optionally substituted with cyano, nitro, halogen, C1-C3 alkyl, C1-C3 alkoxy; an amino protecting group; or together with the nitrogen atom to which they are attached form a heterocyclic ring, preferably the heterocyclic ring is selected from pyrrolidinyl, piperidinyl, morpholinyl, piperazinyl and homopiperazine, the heterocyclic ring being optionally substituted with C1-C3 alkyl; R8 is a thiol protecting group; a wavy line indicates the bond to the oxygen of the OR2 group in any one of formulas (1) to (14), or in a similar manner for nucleosides not expressly represented herein by a formula. When W represents O, S or Se, then the P atom in the phosphorus moiety is V When W represents an electron pair, the P atom in the phosphorus moiety is in a valence state of IIIThe moiety of formula (18) includes any possible stereoisomers. Furthermore, the moieties represented by formula (18) include salts thereof, typically and preferably the salts are formed upon treatment with an inorganic base or an amine, typically and preferably the salts are salts derived from reaction with an OH or SH group, which are (independently of each other) the R3 and R4. Preferred inorganic bases or amines which effect the salt formation with an OH or SH group are well known in the art, typically and preferably trimethylamine, diethylamine, methylamine or ammonium hydroxide. These phosphorus moieties included in the present invention also, where appropriate, include the salts of "O - HB + ” (wherein the above HB + refers to the counter cation that is formed).

[0150] The term "phosphorus moiety," as used herein, includes moieties derived from phosphonates, phosphite triesters, monophosphates, diphosphates, triphosphates, phosphate triesters, phosphate diesters, thiophosphate esters, dithiophosphate esters, or phosphoramidites, and typically and preferably each occurrence is independently selected from these. Thus, in embodiments, OR2 in any one of Formulas (1) through (14), or in an analogous manner for nucleosides not expressly represented herein by a formula, each occurrence is independently selected from phosphonates, phosphite triesters, monophosphates, diphosphates, triphosphates, phosphate triesters, phosphate diesters, thiophosphate esters, dithiophosphate esters, or phosphoramidites. Additional phosphorus moieties that can be used in the present invention are disclosed in Tetrahedron Report Number 309 (Beaucage and Iyer, Tetrahedron, 1992, Vol. 48, pp. 2223-2311), the disclosure of which is incorporated herein by reference.

[0151] The term "phosphorus moiety," as used herein, preferably refers to the R group defined in any one of formulas (1) to (14), or in an analogous manner for nucleosides not expressly represented herein by any formula, and P III or P V which includes a phosphorus atom in a valence state of formula (19), formula (20) or formula (21): [ka] (wherein Y is O, S or Se, Y is preferably O or S, more preferably Y is O; R and R 5’ are independently, at each occurrence, independently of one another, hydrogen, each independently of one another, cyano, nitro, halogen, -NHC(O)C1-C3 alkyl, -NHC(O)C1-C3 haloalkyl, C1-C9 alkyl, C1-C6 alkoxy optionally substituted with C1-C3 alkylsulfonyl; each independently of one another, aryl, C1-C6 alkylenearyl, C1-C6 alkylenediaryl optionally substituted with cyano, nitro, halogen, C1-C4 alkoxy, C1-C4 haloalkyl, C1-C4 haloalkoxy, NHC(O)C1-C3 alkyl, -NHC(O)C1-C3 haloalkyl, C1-C3 alkylsulfonyl; a hydroxyl protecting group; R6 and R7 are independently of one another, hydrogen C1-C9 alkyl optionally substituted with hydrogen, cyano, nitro, halogen, C2-C6 alkenyl, C3-C6 cycloalkyl, C1-C3 alkoxy; aryl, preferably phenyl, optionally substituted with cyano, nitro, halogen, C1-C3 alkyl, C1-C3 alkoxy; an amino protecting group; or together with the nitrogen atom to which they are attached form a heterocyclic ring, preferably said heterocyclic ring is selected from pyrrolidinyl, piperidinyl, morpholinyl, piperazinyl and homopiperazine, said heterocyclic ring being optionally substituted with C1-C3 alkyl; R8 is a thiol protecting group; and a wavy line indicates the bond to the oxygen of the OR2 group in any one of formulas (1) to (5). Each occurrence of each of the following is expressed independently: composition

[0152] Thus, in a first aspect, the present invention provides a composition comprising an oligomeric compound comprising one or more tricyclo-deoxyribonucleic acid (tc-DNA) nucleosides and one or more lipid moieties, preferably exactly one lipid moiety, wherein said one or more lipid moieties are covalently linked to said oligomeric compound, either directly or via a spacer, and preferably wherein said oligomeric compound comprises 5 to 40 monomeric subunits.

[0153] In one embodiment, the one or more lipid moieties are independently selected from a fatty acid moiety, a fatty diacid moiety, a glycerolipid moiety, a glycerophospholipid moiety, a sphingolipid moiety, a phospholipid, an alkylphosphate moiety, and an alkylphosphonate moiety.

[0154] In one embodiment, the one or more lipid moieties are independently selected from a fatty acid moiety, a fatty diacid moiety, a phospholipid, an alkyl phosphate moiety, and an alkyl phosphonate moiety.

[0155] In one preferred embodiment, the one or more lipid moieties are, independently of one another, fatty acid moieties. In another preferred embodiment, the one or more lipid moieties are, independently of one another, fatty diacid moieties. In another embodiment, the one or more lipid moieties are, independently of one another, glycerolipid moieties. In another embodiment, the one or more lipid moieties are, independently of one another, glycerophospholipid moieties. In another embodiment, the one or more lipid moieties are, independently of one another, sphingolipid moieties. In another preferred embodiment, the one or more lipid moieties are, independently of one another, alkylphosphate moieties. In another preferred embodiment, the one or more lipid moieties are, independently of one another, alkylphosphonate moieties.

[0156] In one embodiment, the lipid moiety or moieties are negatively charged at pH 7.4, which typically corresponds to physiological pH.

[0157] In a preferred embodiment, said one or more lipid moieties are independently selected from a fatty acid moiety, a fatty diacid moiety, an alkyl phosphate moiety and an alkyl phosphonate moiety.

[0158] In a preferred embodiment, said one or more lipid moieties are, independently of each other, a fatty acid moiety or a fatty diacid moiety.

[0159] In another preferred embodiment, the one or more lipid moieties are, independently of one another, fatty acid moieties, and the fatty acid moieties are saturated fatty acid moieties. In another preferred embodiment, the one or more lipid moieties are, independently of one another, fatty acid moieties, and the fatty acid moieties are unsaturated fatty acid moieties.

[0160] In a preferred embodiment, the one or more lipid moieties are, independently of one another, fatty diacid moieties, and the fatty diacid moieties are saturated fatty diacid moieties. In another preferred embodiment, the one or more lipid moieties are, independently of one another, fatty diacid moieties, and the fatty diacid moieties are unsaturated fatty diacid moieties.

[0161] In a highly preferred embodiment, said one or more lipid moieties, independently of each other, are fatty acid moieties, and said fatty acid moieties are saturated, unbranched fatty acid moieties.

[0162] In a preferred embodiment, the one or more lipid moieties are, independently of one another, fatty acid moieties, wherein the fatty acid moieties are derived from saturated, unbranched fatty acids. In a preferred embodiment, the one or more lipid moieties are, independently of one another, fatty diacid moieties, wherein the fatty diacid moieties are derived from saturated, unbranched fatty diacids.

[0163] In a highly preferred embodiment, the one or more lipid moieties are, independently of each other, a fatty acid moiety or a fatty diacid moiety, wherein the fatty acid moiety is a saturated, unbranched fatty acid moiety and the fatty diacid moiety is a saturated, unbranched fatty diacid moiety.

[0164] In a preferred embodiment, the one or more lipid moieties, independently of each other, have the formula (I): AB- * (I) (Wherein A is C 3~32 Alkyl, C 3~32 Alkenyl, C 3~32 Alkynyl, HOOC-C 3~32 Alkylene, HOOC-C 3~32 Alkenylene or HOOC-C 3~32 alkynylene, B is C(O), OP(OH), OP(O)(OH), OP(O)(SH), NH—C(O), NH—P(O)(OH), NH—P(O)(SH), or a pharmaceutically acceptable salt thereof, * ) represents the point of covalent attachment to the oligomeric compound or the spacer. This is the part.

[0165] In a further preferred embodiment, the one or more lipid moieties independently of each other have the formula (a) to (u) aC 3~32 Alkyl-C(O)- * , bC 3~32 Alkenyl-C(O)- * , cC 3~32 Alkynyl-C(O)- * , dC 3~32 Alkyl-OP(OH)- * , eC 3~32 Alkenyl-OP(OH)- * , fC 3~32 Alkynyl-OP(OH)- * , gC 3~32Alkyl-OP(O)(OH)- * , hC 3~32 Alkenyl-OP(O)(OH)- * , I C 3~32 Alkynyl-OP(O)(OH)- * , jC 3~32 Alkyl-OP(O)(SH)- * , kC 3~32 Alkenyl-OP(O)(SH)- * , lC 3~32 Alkynyl-OP(O)(SH)- * , mC 3~32 Alkyl-NH-C(O)- * , nC 3~32 Alkenyl-NH-C(O)- * , oC 3~32 Alkynyl-NH-C(O)- * , PC 3~32 Alkyl-NH-P(O)(OH)- * , qC 3~32 Alkenyl-NH-P(O)(OH)- * , rC 3~32 Alkynyl-NH-P(O)(OH)- * , s.HOOC-C 3~32 Alkylene-C(O)- * , t.HOOC-C 3~32 Alkenylene-C(O)- * , and u.HOOC-C 3~32 Alkynylene-C(O)- * (wherein the asterisk ( * ) represents the point of covalent attachment to the oligomeric compound or the spacer. is selected from one of the following.

[0166] In a preferred embodiment, the one or more lipid moieties, independently of each other, have the formula C 3~32 Alkyl-C(O)- * (wherein the asterisk ( * ) is a moiety of ) representing said point of covalent attachment to said oligomeric compound or said spacer, and preferably said composition comprises exactly one lipid moiety.

[0167] In a preferred embodiment, the one or more lipid moieties, independently of each other, have the formula C 3~32 Alkenyl-C(O)- * (wherein the asterisk ( * ) is a moiety of ) representing said point of covalent attachment to said oligomeric compound or said spacer, and preferably said composition comprises exactly one lipid moiety.

[0168] In a preferred embodiment, the one or more lipid moieties, independently of each other, have the formula C 3~32 Alkynyl-C(O)- * (wherein the asterisk ( * ) is a moiety of ) representing said point of covalent attachment to said oligomeric compound or said spacer, and preferably said composition comprises exactly one lipid moiety.

[0169] In a preferred embodiment, the one or more lipid moieties, independently of each other, have the formula C 3~32 Alkyl-OP(OH)- * (wherein the asterisk ( * ) is a moiety of ) representing said point of covalent attachment to said oligomeric compound or said spacer, and preferably said composition comprises exactly one lipid moiety.

[0170] In a preferred embodiment, the one or more lipid moieties, independently of each other, have the formula C 3~32 Alkenyl-OP(OH)- * (wherein the asterisk ( *) is a moiety of ) representing said point of covalent attachment to said oligomeric compound or said spacer, and preferably said composition comprises exactly one lipid moiety.

[0171] In a preferred embodiment, the one or more lipid moieties, independently of each other, have the formula C 3~32 Alkynyl-OP(OH)- * (wherein the asterisk ( * ) is a moiety of ) representing said point of covalent attachment to said oligomeric compound or said spacer, and preferably said composition comprises exactly one lipid moiety.

[0172] In a preferred embodiment, the one or more lipid moieties, independently of each other, have the formula C 3~32 Alkyl-OP(O)(OH)- * (wherein the asterisk ( * ) is a moiety of ) representing said point of covalent attachment to said oligomeric compound or said spacer, and preferably said composition comprises exactly one lipid moiety.

[0173] In a preferred embodiment, the one or more lipid moieties, independently of each other, have the formula C 3~32 Alkenyl-OP(O)(OH)- * (wherein the asterisk ( * ) is a moiety of ) representing said point of covalent attachment to said oligomeric compound or said spacer, and preferably said composition comprises exactly one lipid moiety.

[0174] In a preferred embodiment, the one or more lipid moieties, independently of each other, have the formula C 3~32 Alkynyl-OP(O)(OH)- * (wherein the asterisk ( * ) is a moiety of ) representing said point of covalent attachment to said oligomeric compound or said spacer, and preferably said composition comprises exactly one lipid moiety.

[0175] In a preferred embodiment, the one or more lipid moieties, independently of each other, have the formula C 3~32 Alkyl-OP(O)(SH)- * (wherein the asterisk ( * ) is a moiety of ) representing said point of covalent attachment to said oligomeric compound or said spacer, and preferably said composition comprises exactly one lipid moiety.

[0176] In a preferred embodiment, the one or more lipid moieties, independently of each other, have the formula C 3~32 Alkenyl-OP(O)(SH)- * (wherein the asterisk ( * ) is a moiety of ) representing said point of covalent attachment to said oligomeric compound or said spacer, and preferably said composition comprises exactly one lipid moiety.

[0177] In a preferred embodiment, the one or more lipid moieties, independently of each other, have the formula C 3~32 Alkynyl-OP(O)(SH)- * (wherein the asterisk ( * ) is a moiety of ) representing said point of covalent attachment to said oligomeric compound or said spacer, and preferably said composition comprises exactly one lipid moiety.

[0178] In a preferred embodiment, the one or more lipid moieties, independently of each other, have the formula C 3~32 Alkyl-NH-C(O)- * (wherein the asterisk ( * ) is a moiety of ) representing said point of covalent attachment to said oligomeric compound or said spacer, and preferably said composition comprises exactly one lipid moiety.

[0179] In a preferred embodiment, the one or more lipid moieties, independently of each other, have the formula C 3~32 Alkenyl-NH-C(O)- * (wherein the asterisk ( *) is a moiety of ) representing said point of covalent attachment to said oligomeric compound or said spacer, and preferably said composition comprises exactly one lipid moiety.

[0180] In a preferred embodiment, the one or more lipid moieties, independently of each other, have the formula C 3~32 Alkynyl-NH-C(O)- * (wherein the asterisk ( * ) is a moiety of ) representing said point of covalent attachment to said oligomeric compound or said spacer, and preferably said composition comprises exactly one lipid moiety.

[0181] In a preferred embodiment, the one or more lipid moieties, independently of each other, have the formula C 3~32 Alkyl-NH-P(O)(OH)- * (wherein the asterisk ( * ) is a moiety of ) representing said point of covalent attachment to said oligomeric compound or said spacer, and preferably said composition comprises exactly one lipid moiety.

[0182] In a preferred embodiment, the one or more lipid moieties, independently of each other, have the formula C 3~32 Alkenyl-NH-P(O)(OH)- * (wherein the asterisk ( * ) is a moiety of ) representing said point of covalent attachment to said oligomeric compound or said spacer, and preferably said composition comprises exactly one lipid moiety.

[0183] In a preferred embodiment, the one or more lipid moieties, independently of each other, have the formula C 3~32 Alkynyl-NH-P(O)(OH)- * (wherein the asterisk ( * ) is a moiety of ) representing said point of covalent attachment to said oligomeric compound or said spacer, and preferably said composition comprises exactly one lipid moiety.

[0184] In a preferred embodiment, the one or more lipid moieties, independently of each other, have the formula HOOC-C 3~32 Alkylene-C(O)- * (wherein the asterisk ( * ) is a moiety of ) representing said point of covalent attachment to said oligomeric compound or said spacer, and preferably said composition comprises exactly one lipid moiety.

[0185] In a preferred embodiment, the one or more lipid moieties, independently of each other, have the formula HOOC-C 3~32 Alkenylene-C(O)- * (wherein the asterisk ( * ) is a moiety of ) representing said point of covalent attachment to said oligomeric compound or said spacer, and preferably said composition comprises exactly one lipid moiety.

[0186] In a preferred embodiment, the one or more lipid moieties, independently of each other, have the formula HOOC-C 3~32 Alkynylene-C(O)- * (wherein the asterisk ( * ) is a moiety of ) representing said point of covalent attachment to said oligomeric compound or said spacer, and preferably said composition comprises exactly one lipid moiety.

[0187] In a preferred embodiment, the one or more lipid moieties are independently of each other represented by formula (a) to formula (d): aC 3~32 Alkyl-C(O)- * , b.HOOC-C 3~32 Alkylene-C(O)- * , cC 3~32 Alkyl-OP(O)(OH)- * dC 3~32 Alkyl-OP(O)(SH)- * (wherein the asterisk ( *) represents the point of covalent attachment to the oligomeric compound or the spacer. and preferably, 3~32 The alkyl is an unbranched C 3~32 alkyl, and more preferably, 3~32 Alkyl is an unbranched C alkyl group with an odd number of carbon atoms. 3~32 alkyl, preferably 3~32 The alkylene is an unbranched C 3~32 alkylene, and more preferably, 3~32 Alkylene is an unbranched C 3~32 It is alkylene.

[0188] In a further preferred embodiment, said one or more lipid moieties, independently of each other, have the formula C 3~32 Alkyl-C(O)- * (wherein the asterisk ( * ) is a moiety representing the point of covalent attachment to the oligomeric compound or the spacer, and preferably 3~32 The alkyl is an unbranched C 3~32 alkyl, and more preferably, 3~32 Alkyl is an unbranched C alkyl group with an odd number of carbon atoms. 3~32 It is alkyl.

[0189] In a preferred embodiment, the one or more lipid moieties, independently of each other, have the formula C 3~32 Alkyl-C(O)- * (wherein the asterisk ( * ) is a moiety representing the point of covalent attachment to the oligomeric compound or the spacer, and preferably the composition comprises exactly one lipid moiety, and the C 3~32 The alkyl is an unbranched C 3~32 It is alkyl.

[0190] In a preferred embodiment, the one or more lipid moieties, independently of each other, have the formula C 3~32 Alkyl-C(O)- *(wherein the asterisk ( * ) is a moiety representing the point of covalent attachment to the oligomeric compound or the spacer, and preferably the composition comprises exactly one lipid moiety, and the C 3~32 Alkyl is an unbranched C alkyl group with an odd number of carbon atoms. 3~32 It is alkyl.

[0191] In a preferred embodiment, the one or more lipid moieties, independently of each other, have the formula C 3~32 Alkenyl-C(O)- * (wherein the asterisk ( * ) is a moiety representing the point of covalent attachment to the oligomeric compound or the spacer, and preferably 3~32 Alkenyl is a branched C 3~32 alkenyl, and more preferably, 3~32 Alkenyl is a branched C alkyl group with an odd number of carbon atoms. 3~32 It is alkenyl.

[0192] In a preferred embodiment, the one or more lipid moieties, independently of each other, have the formula C 3~32 Alkenyl-C(O)- * (wherein the asterisk ( * ) is a moiety representing the point of covalent attachment to the oligomeric compound or the spacer, and preferably the composition comprises exactly one lipid moiety, and the C 3~32 Alkenyl is a branched C 3~32 It is alkenyl.

[0193] In a preferred embodiment, the one or more lipid moieties, independently of each other, have the formula C 3~32 Alkenyl-C(O)- * (wherein the asterisk ( * ) is a moiety representing the point of covalent attachment to the oligomeric compound or the spacer, and preferably the composition comprises exactly one lipid moiety, and the C 3~32 Alkenyl is a branched C alkyl group with an odd number of carbon atoms. 3~32It is alkenyl.

[0194] In a further highly preferred embodiment, said one or more lipid moieties, independently of each other, are saturated C 8~26 a fatty acid moiety, preferably the saturated C 8~26 The fatty acid moiety is derived from caprylic acid (C8), capric acid (C10), lauric acid (C12), myristic acid (C14), palmitic acid (C16), stearic acid (C18), arachidic acid (C20), lignoceric acid (C22) or cerotic acid (C24).

[0195] In a further highly preferred embodiment, said one or more lipid moieties, independently of one another, are saturated fatty acid moieties, preferably said saturated fatty acid moieties are derived from caprylic acid (C8), capric acid (C10), lauric acid (C12), myristic acid (C14), palmitic acid (C16), stearic acid (C18), arachidic acid (C20), lignoceric acid (C22) and cerotic acid (C24).

[0196] In a further highly preferred embodiment, the one or more lipid moieties are, independently of one another, saturated fatty acid moieties derived from palmitic acid (C16) or stearic acid (C18), preferably the one or more lipid moieties are saturated fatty acid moieties derived from palmitic acid (C16).

[0197] In a further highly preferred embodiment, said one or more lipid moieties, independently of each other, comprise an unsaturated C 14~22 a fatty acid moiety, preferably the unsaturated C 14~22 The fatty acid moieties are derived from myristoleic acid, palmitoleic acid, sapienic acid, oleic acid, elaidic acid, vaccenic acid, linoleic acid, linoelaidic acid, α-linolenic acid, arachidonic acid, eicosapentaenoic acid, erucic acid, and docosahexaenoic acid. In a further highly preferred embodiment, the one or more lipid moieties are saturated fatty acid moieties derived from palmitoleic acid.

[0198] In a further highly preferred embodiment, the one or more lipid moieties, independently of one another, are unsaturated fatty acid moieties derived from myristoleic acid, palmitoleic acid, sapienic acid, oleic acid, elaidic acid, vaccenic acid, linoleic acid, linoelaidic acid, α-linolenic acid, arachidonic acid, eicosapentaenoic acid, erucic acid, and docosahexaenoic acid.

[0199] In a further highly preferred embodiment, said one or more lipid moieties are unsaturated fatty acid moieties derived from oleic acid.

[0200] In a further highly preferred embodiment, said one or more lipid moieties, independently of each other, have the formula (HOOC)-C 3~32 Alkylene-C(O)- * (wherein the asterisk ( * ) is a moiety representing the point of covalent attachment to the oligomeric compound or the spacer, and preferably 3~32 Alkylene is an unbranched C 3~32 alkylene, and more preferably, 3~32 Alkylene is an unbranched C alkylene having an odd number of carbon atoms. 3~32 It is alkylene.

[0201] In a further highly preferred embodiment, said one or more lipid moieties, independently of each other, have the formula (HOOC)-(CH) r -(CH)(C 5~25 alkyl)-(CH2) t -C(O)- * (wherein the asterisk ( * ) represents the point of covalent bond to the oligomeric compound or the spacer, r's are each independently an integer of 1 to 3, and t's are each independently an integer of 1 to 3.

[0202] In a further highly preferred embodiment, said one or more lipid moieties, independently of each other, have the formula (HOOC)-(CH) r -(CH)[(CH2)s CH3]-(CH2) t -C(O)- * (wherein the asterisk ( * ) represents the point of covalent bond to the oligomeric compound or the spacer, r's are each independently an integer of 1 to 3, s's are each independently an integer of 4 to 24, and t's are each independently an integer of 1 to 3.

[0203] In a further highly preferred embodiment, said one or more lipid moieties, independently of each other, have the formula (HOOC)-(CH) r -(CH)[(CH2) s CH3]-(CH2) t -C(O)- * (wherein the asterisk ( * ) represents the point of covalent bond to the oligomeric compound or the spacer, each r is independently an integer of 1 or 2, each s is independently an integer of 5 to 19, and each t is independently an integer of 1 or 2.

[0204] In a further highly preferred embodiment, said one or more lipid moieties, independently of each other, have the formula (HOOC)-(CH) r -(CH)[(CH2) s CH3]-(CH2) t -C(O)- * (wherein the asterisk ( * ) represents the point of covalent bond with the oligomeric compound or the spacer, r is 1, s's are each independently an integer of 4 to 24, preferably 5 to 19, and t is 1.

[0205] In a further highly preferred embodiment, said one or more lipid moieties, independently of each other, have the formula (HOOC)-(CH) r -(CH)[(CH2) s CH3]-(CH2) t -C(O)- * (wherein the asterisk ( *) represents the point of covalent bond to the oligomeric compound or the spacer, r is 1, s's are each independently an integer of 5 to 19, preferably 11 to 17, and t is 1.

[0206] In a further highly preferred embodiment, said one or more lipid moieties, independently of each other, have the formula (HOOC)-(CH) r -(CH)[(CH2) s CH3]-(CH2) t -C(O)- * (wherein the asterisk ( * ) represents said point of covalent attachment to said oligomeric compound or said spacer, r is 1, s is 15, and t is 1.

[0207] Thus, in a further highly preferred embodiment, the one or more lipid moieties is 3-pentadecyl glutaric acid (PDG).

[0208] In a further highly preferred embodiment, the lipid moiety is directly linked to the oligomeric compound.

[0209] In a further highly preferred embodiment, said one or more lipid moieties are linked to said oligomeric compound via a spacer.

[0210] In one embodiment, the spacer has 5 to 30 carbon atoms, preferably 5 to 25 carbon atoms, more preferably 5 to 20 carbon atoms, and most preferably 5 to 17 carbon atoms. In a further embodiment, the spacer has 4 to 20 heteroatoms, preferably 4 to 18 heteroatoms, more preferably 4 to 14 heteroatoms, and most preferably 4 to 12 heteroatoms. Particularly preferred examples of heteroatoms are nitrogen and oxygen atoms. H atoms are not heteroatoms.

[0211] In a preferred embodiment, the spacer has the formula #-NH-C 2~12Alkylene-§, #-NH-C 2~12 alkylene-OP(OH)-§, #-NH-C 2~12 alkylene-OP(O)(SH)-§, #-NH-C 2~12 alkylene-OP(O)(OH)-§, #-SH-C 2~12 Alkylene-§, #-NH-C 2~12 alkylene-NH-C(O)-§, #-NH-C 2~12 alkylene-NH-P(O)(OH)-§, and #-NH-C 2~12 Alkylene-NH-P(O)(SH)-§ (Wherein, C 2~12 One or more -CH2- moieties in the alkylene are independently -O-, -S-, -NH-, -C(O)-, -C(O)O-, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, -OP(OH)O-, OP(O)(SH)O-, OP(O)(OH)O-, NHP(O)(OH)O-, NHP(O)(SH)O-, or -(O-CH2-CH2) k - (k is an integer from 1 to 8), 2~12 One or more -CH2- moieties in the alkylene are optionally substituted, independently of one another, with one or more -COOH, -NH2, -OP(O)(OH)2 or -OH (thus, 2~12 (meaning that one or both, preferably one, hydrogen atoms in one or more -CH2- moieties in the alkylene are optionally replaced independently with one or more -COOH, -NH2, -OP(O)(OH)2 or -OH), the (#) represents the point of covalent attachment to the lipid moiety, and the (§) represents the point of covalent attachment to the oligomeric compound. and preferably, independently selected from any one of the formulas:

[0212] In a highly preferred embodiment, the spacer has the formula a.#-NH-C 2~12 Alkylene-§, b.#-NH-C 2~12 alkylene-OP(OH)-§, c.#-NH-C 2~12 alkylene-OP(O)(SH)-§, d.#-NH-C 2~12 alkylene-OP(O)(OH)-§, e.#-NH-C 2~12 alkylene-NH-C(O)-§, f.#-NH-C 2~12 alkylene-NH-P(O)(OH)-§, and g.#-NH-C 2~12 Alkylene-NH-P(O)(SH)-§ (wherein the (#) represents a point of covalent bond attachment to the lipid moiety, and the (§) represents a point of covalent bond attachment to the oligomeric compound) and preferably, independently selected from any one of the formulas:

[0213] In a highly preferred embodiment, the spacer has the formula a.#-NH-C 2~12 Alkylene-§, b.#-NH-C 2~12 alkylene-OP(OH)-§, c.#-NH-C 2~12 alkylene-OP(O)(SH)-§, d.#-NH-C 2~12 alkylene-OP(O)(OH)-§, e.#-NH-C 2~12 alkylene-NH-C(O)-§, f.#-NH-C 2~12 alkylene-NH-P(O)(OH)-§, and g.#-NH-C 2~12 Alkylene-NH-P(O)(SH)-§ (Wherein, C 2~12One or more -CH2- moieties in the alkylene are independently -O-, -S-, -NH-, -C(O)-, -C(O)O-, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, -OP(OH)O-, OP(O)(SH)O-, OP(O)(OH)O-, NHP(O)(OH)O-, NHP(O)(SH)O-, or -(O-CH2-CH2) k -(k is an integer from 1 to 8), wherein the (#) represents a point of covalent attachment to the lipid moiety, and the (§) represents a point of covalent attachment to the oligomeric compound. and preferably, independently selected from any one of the formulas:

[0214] In a highly preferred embodiment, the spacer has the formula a.#-NH-C 2~12 Alkylene-§, b.#-NH-C 2~12 alkylene-OP(OH)-§, c.#-NH-C 2~12 alkylene-OP(O)(SH)-§, d.#-NH-C 2~12 alkylene-OP(O)(OH)-§, e.#-NH-C 2~12 alkylene-NH-C(O)-§, f.#-NH-C 2~12 alkylene-NH-P(O)(OH)-§, and g.#-NH-C 2~12 Alkylene-NH-P(O)(SH)-§ (Wherein, C 2~12 One or more -CH2- moieties in the alkylene are independently -O-, -S-, -NH-, -C(O)-, -C(O)O-, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, -OP(OH)O-, OP(O)(SH)O-, OP(O)(OH)O-, NHP(O)(OH)O-, NHP(O)(SH)O-, or -(O-CH2-CH2) k - (k is an integer from 1 to 8),2~12 one or more -CH2- moieties in the alkylene are optionally substituted, independently of one another, with one or more -COOH, -NH2, -OP(O)(OH)2 or -OH, and the (#) represents a point of covalent attachment to the lipid moiety, and the (§) represents a point of covalent attachment to the oligomeric compound. and preferably, independently selected from any one of the formulas:

[0215] In a highly preferred embodiment, the spacer has the formula a.#-NH-C 2~12 Alkylene-§, b.#-NH-C 2~12 alkylene-OP(OH)-§, c.#-NH-C 2~12 alkylene-OP(O)(SH)-§, d.#-NH-C 2~12 alkylene-OP(O)(OH)-§, e.#-NH-C 2~12 alkylene-NH-C(O)-§, f.#-NH-C 2~12 alkylene-NH-P(O)(OH)-§, and g.#-NH-C 2~12 Alkylene-NH-P(O)(SH)-§ (Wherein, C 2~12 One or more -CH2- moieties in the alkylene are independently -O-, -S-, -NH-, -C(O)-, -C(O)O-, phenyl, triazolyl, cyclopentyl, cyclohexyl, succinimidyl, -OP(OH)O-, OP(O)(SH)O-, OP(O)(OH)O-, NHP(O)(OH)O-, NHP(O)(SH)O-, or -(O-CH2-CH2) k - (k is an integer from 1 to 8), 2~12one or more -CH2- moieties in the alkylene are optionally substituted, independently of one another, with one or more -COOH, -NH2, -OP(O)(OH)2 or -OH, and the (#) represents a point of covalent attachment to the lipid moiety, and the (§) represents a point of covalent attachment to the oligomeric compound. and preferably, independently selected from any one of the formulas:

[0216] In a further highly preferred embodiment, the spacer has the formula a.-NH-(CH2) m -, b.-NH-(CH2) m -X-, c.-NH-(CH2) n -(O-CH2-CH2) k -O-(CH2) p -, d.-NH-(CH2) n -(O-CH2-CH2) k -O-(CH2) p -X-, e. -NH-CH(COOH)-(CH2) q -, f.-NH-CH(COOH)-(CH2) q -X-, g.-NH-CH(COOH)-(CH2) q -C(O)-NH-(CH2) m -, h.-NH-CH(COOH)-(CH2) q -C(O)-NH-(CH2) m -X-, i.-NH-CH(COOH)-(CH2) q -C(O)-NH-(CH2) n -(O-CH2-CH2) k -O-(CH2) p -X-, j.-NH-CH(COOH)-(CH2) q -C(O)-NH-(CH2) n -(O-CH2-CH2) k -O-(CH2)p -C(O)-NH-(CH2) n -(O-CH2-CH2) r X- (wherein X's are each independently OP(OH), OP(O)(SH), or OP(O)(OH); k's are each independently an integer of 1 to 8; m's are each independently an integer of 2 to 12; n's are each independently an integer of 2 to 4; p's are each independently an integer of 1 to 5; q's are each independently an integer of 1 to 3, preferably 1 or 2; and r's are each independently an integer of 1 to 3, preferably 1 or 2. and preferably, independently selected from any one of the formulas:

[0217] In a further highly preferred embodiment, the spacer has the formula a.#-NH-(CH2) m -§, b.#-NH-(CH2) m -X-§, c.#-NH-(CH2) n -(O-CH2-CH2) k -O-(CH2) p -§, d.#-NH-(CH2) n -(O-CH2-CH2) k -O-(CH2) p -X-§, e.#-NH-CH(COOH)-(CH2) q -§, f.#-NH-CH(COOH)-(CH2) q -X-§, g.#-NH-CH(COOH)-(CH2) q -C(O)-NH-(CH2) m -§, h.#-NH-CH(COOH)-(CH2) q -C(O)-NH-(CH2) m -X-§, i.#-NH-CH(COOH)-(CH2) q -C(O)-NH-(CH2) n -(O-CH2-CH2)k -O-(CH2) p -X-§, j.#-NH-CH(COOH)-(CH2) q -C(O)-NH-(CH2) n -(O-CH2-CH2) k -O-(CH2) p -C(O)-NH-(CH2) n -(O-CH2-CH2) r -X-§ (In the formula, X's are each independently OP(OH), OP(OH)(S), or OP(O)(OH), k's are each independently an integer of 1 to 8, m's are each independently an integer of 2 to 12, n's are each independently an integer of 2 to 4, p's are each independently an integer of 1 to 5, q's are each independently an integer of 1 to 3, preferably 1 or 2, and r's are each independently an integer of 1 to 3, preferably 1 or 2, the (#) represents a point of covalent bond to the lipid moiety, and the (§) represents a point of covalent bond to the oligomeric compound.) and preferably, independently selected from any one of the formulas:

[0218] In a further highly preferred embodiment, the spacer has the formula a.-Z-NH-(CH2) m -X- b.-Z-NH-(CH2) n -(O-CH2-CH2) k -O-(CH2) p -X- c.-Z[-NH-(CH2) n -(O-CH2-CH2) k -O-(CH2) p -C(O)-]-NH-(CH2) q -X- d.-Z[-NH-(CH2) n -(O-CH2-CH2) k -O-(CH2) p -C(O)-] r -NH-(CH2) q -(O-CH2-CH2)k -X- (wherein, -Z- are, independently of one another, a bond, or -NH-CH(COOH)-(CH)-C(O)- or -NH-CH[(CH)COOH]-C(O)-; X are, independently of one another, OP(OH), OP(O)(SH), OP(O)(OH), NHP(O)(OH), NHP(O)(SH), or NH-C(O); k are, independently of one another, an integer of 1 to 8; m are, independently of one another, an integer of 2 to 12; n are, independently of one another, an integer of 2 to 4; p are, independently of one another, an integer of 1 to 5; q are, independently of one another, an integer of 1 to 6, preferably 3 or 6; and r are, independently of one another, an integer of 1 to 3, preferably 1 or 2.) and preferably, independently selected from any one of the formulas:

[0219] In a further highly preferred embodiment, the spacer has the formula a.#-Z-NH-(CH2) m -X-§ b.#-Z-NH-(CH2) n -(O-CH2-CH2) k -O-(CH2) p -X-§ c.#-Z[-NH-(CH2) n -(O-CH2-CH2) k -O-(CH2) p -C(O)-]-NH-(CH2) q -X-§ d.#-Z[-NH-(CH2) n -(O-CH2-CH2) k -O-(CH2) p -C(O)-] r -NH-(CH2) q -(O-CH2-CH2) k -X-§ (wherein, -Z-, independently of one another, represent a bond, or -NH-CH(COOH)-(CH)-C(O)- or -NH-CH[(CH)COOH]-C(O)-; X, independently of one another, represent OP(OH), OP(O)(SH), OP(O)(OH), NHP(O)(OH), NHP(O)(SH), or NH-C(O); k, independently of one another, represent an integer from 1 to 8; and m, independently of one another, represent an integer from 1 to 8. each n is independently an integer of 2 to 12; each p is independently an integer of 1 to 5; each q is independently an integer of 1 to 6, preferably 3 or 6; each r is independently an integer of 1 to 3, preferably 1 or 2; the (#) represents a point of covalent bond to the lipid moiety; and the (§) represents a point of covalent bond to the oligomeric compound. and preferably, independently selected from any one of the formulas:

[0220] In another preferred embodiment, the spacer is of the formula a.#-Z-NH-(CH2) m -X-§ b.#-Z-NH-(CH2) n -(O-CH2-CH2) k -O-(CH2) p -X-§ c.#-Z[-NH-(CH2) n -(O-CH2-CH2) k -O-(CH2) p -C(O)-]-NH-(CH2) q -X-§ d.#-Z[-NH-(CH2) n -(O-CH2-CH2) k -O-(CH2) p -C(O)-] r -NH-(CH2) q -(O-CH2-CH2) k -X-§ (In the formula, -Z- independently represent a bond, -NH-CH(COOH)-(CH)-C(O)-, or -NH-CH[(CH)COOH]-C(O)-; X independently represent OP(OH), OP(O)(SH), OP(O)(OH), NHP(O)(OH), NHP(O)(SH), or NH-C(O); k independently represent an integer of 1 or 2; m independently represent an integer of 4 to 8; n independently represent an integer of 2 to 4; p independently represent an integer of 1 or 2; q independently represent an integer of 1 to 6; and r independently represent an integer of 1 to 3, preferably 1 or 2; the (#) represents a point of covalent bond to the lipid moiety; and the (§) represents a point of covalent bond to the oligomeric compound.) and preferably, independently selected from any one of the formulas:

[0221] In another preferred embodiment, the spacer is of the formula a.#-Z-NH-(CH2) m -X-§ b.#-Z-NH-(CH2) n -(O-CH2-CH2) k -O-(CH2) p -X-§ c.#-Z[-NH-(CH2) n -(O-CH2-CH2) k -O-(CH2) p -C(O)-]-NH-(CH2) q -X-§ d.#-Z[-NH-(CH2) n -(O-CH2-CH2) k -O-(CH2) p -C(O)-] r -NH-(CH2) q -(O-CH2-CH2) k -X-§ (In the formula, -Z- independently represent a bond, -NH-CH(COOH)-(CH)-C(O)-, or -NH-CH[(CH)COOH]-C(O)-; X independently represent OP(OH), OP(O)(SH), or OP(O)(OH); k independently represent an integer of 1 or 2; m independently represent an integer of 4 to 8; n is 2; p is 1; q independently represent an integer of 1 to 6; and r independently represent an integer of 1 to 3, preferably 1 or 2; the (#) represents a point of covalent bond to the lipid moiety; and the (§) represents a point of covalent bond to the oligomeric compound.) and preferably, independently selected from any one of the formulas:

[0222] In another preferred embodiment, the spacer is of the formula a.#-Z-NH-(CH2) m -X-§ b.#-Z-NH-(CH2) n -(O-CH2-CH2) k -O-(CH2) p -X-§ c.#-Z[-NH-(CH2) n -(O-CH2-CH2) k -O-(CH2) p -C(O)-]-NH-(CH2) q -X-§ d.#-Z[-NH-(CH2) n -(O-CH2-CH2) k -O-(CH2) p -C(O)-] r -NH-(CH2) q -(O-CH2-CH2) k -X-§ (In the formula, -Z- independently represent a bond, -NH-CH(COOH)-(CH)-C(O)-, or -NH-CH[(CH)COOH]-C(O)-; X independently represent OP(O)(SH) or OP(O)(OH); k independently represent an integer of 1 or 2; m independently represent an integer of 4 to 8; n is 2; p is 1; q independently represent an integer of 1 to 6; and r independently represent an integer of 1 to 3, preferably 1 or 2; the (#) represents a point of covalent bond to the lipid moiety; and the (§) represents a point of covalent bond to the oligomeric compound.) and preferably, independently selected from any one of the formulas:

[0223] In another highly preferred embodiment, the spacer is of the formula a.#-Z-NH-(CH2) m -X-§ b.#-Z-NH-(CH2) n -(O-CH2-CH2) k -O-(CH2) p -X-§ c.#-Z[-NH-(CH2) n -(O-CH2-CH2) k -O-(CH2) p -C(O)-]-NH-(CH2) q -X-§ d.#-Z[-NH-(CH2) n -(O-CH2-CH2) k -O-(CH2) p -C(O)-] r -NH-(CH2) q -(O-CH2-CH2) k -X-§ (In the formula, -Z- independently represent a bond, -NH-CH(COOH)-(CH)-C(O)-, or -NH-CH[(CH)COOH]-C(O)-; X independently represent OP(O)(SH); k independently represent an integer of 1 or 2; m independently represent an integer of 4 to 8; n is 2; p is 1; q independently represent an integer of 1 to 6; and r independently represent an integer of 1 to 3, preferably 1 or 2; the (#) represents a point of covalent bond to the lipid moiety; and the (§) represents a point of covalent bond to the oligomeric compound.) and preferably, independently selected from any one of the formulas:

[0224] In another highly preferred embodiment, the spacer is of the formula a.#-Z-NH-(CH2) m -X-§ b.#-Z-NH-(CH2) n -(O-CH2-CH2) k -O-(CH2) p -X-§ c.#-Z[-NH-(CH2) n -(O-CH2-CH2) k -O-(CH2) p -C(O)-]-NH-(CH2) q -X-§ d.#-Z[-NH-(CH2) n -(O-CH2-CH2) k -O-(CH2) p -C(O)-] r -NH-(CH2) q -(O-CH2-CH2) k -X-§ (In the formula, -Z- independently represent a bond, -NH-CH(COOH)-(CH)-C(O)-, or -NH-CH[(CH)COOH]-C(O)-; X independently represent OP(O)(OH); k independently represent an integer of 1 or 2; m independently represent an integer of 4 to 8; n is 2; p is 1; q independently represent an integer of 1 to 6; and r independently represent an integer of 1 to 3, preferably 1 or 2; the (#) represents a point of covalent bond to the lipid moiety; and the (§) represents a point of covalent bond to the oligomeric compound.) and preferably, independently selected from any one of the formulas:

[0225] In another highly preferred embodiment, the spacer is of the formula a.#-Z-NH-(CH2) m -X-§ b.#-Z-NH-(CH2) n -(O-CH2-CH2) k -O-(CH2) p -X-§ c.#-Z[-NH-(CH2) n -(O-CH2-CH2) k -O-(CH2) p -C(O)-]-NH-(CH2) q -X-§ d.#-Z[-NH-(CH2) n -(O-CH2-CH2) k -O-(CH2) p -C(O)-] r -NH-(CH2) q -(O-CH2-CH2) k -X-§ (In the formula, -Z-, independently of each other, represent a bond, or -NH-CH(COOH)-(CH)-C(O)- or -NH-C[(CH)COOH]-C(O)-; X, independently of each other, represent OP(O)(SH) or OP(O)(OH); k is 1, m is 6, n is 2, p is 1, q, independently of each other, represent an integer of 1 to 6; r, independently of each other, represent an integer of 1 to 3, preferably 1 or 2; the (#) represents a point of covalent bond to the lipid moiety; and the (§) represents a point of covalent bond to the oligomeric compound.) and preferably, independently selected from any one of the formulas:

[0226] In another highly preferred embodiment, the spacer comprises #-Z-NH-(CH2)mX-§, preferably #-Z-NH-(CH2)mX-§, where -Z- represents a bond, X is, independently of each other, OP(O)(SH) or OP(O)(OH), m is 6, the (#) represents the point of covalent attachment to the lipid moiety, and the (§) represents the point of covalent attachment to the oligomeric compound.

[0227] In another highly preferred embodiment, the spacer comprises #-Z-NH-(CH2)mX-§, preferably #-Z-NH-(CH2)mX-§, where -Z- represents a bond, X is OP(O)(OH), m is 6, the (#) represents a point of covalent attachment to the lipid moiety, and the (§) represents a point of covalent attachment to the oligomeric compound.

[0228] In another highly preferred embodiment, the spacer comprises #-Z-NH-(CH2)mX-§, preferably #-Z-NH-(CH2)mX-§, where -Z- represents a bond, X is OP(O)(SH), m is 6, the (#) represents a point of covalent attachment to the lipid moiety, and the (§) represents a point of covalent attachment to the oligomeric compound.

[0229] In another highly preferred embodiment, said one or more lipid moieties are covalently linked to said oligomeric compound either directly or via a spacer through an -OP(O)(SH)- or -OP(O)(OH)- moiety, typically and preferably constituted by said one or more lipid moieties or said spacer, wherein said -OP(O)(SH)- or said -OP(O)(OH)- moiety is linked to the 5'- or 3'-terminal OH group of said oligomeric compound.

[0230] In another highly preferred embodiment, the one or more lipid moieties are linked, independently of one another, to the oligomeric compound at: (i) a terminal residue of the oligomeric compound; (ii) the 5' end of the oligomeric compound; (iii) the 3' end of the oligomeric compound; or (iv) an internal residue of the oligomeric compound.

[0231] In another preferred embodiment, said one or more lipid moieties, preferably said exactly one lipid moiety, are independently linked to said oligomeric compound at a terminal residue of said oligomeric compound.

[0232] In another preferred embodiment, said one or more lipid moieties, preferably said exactly one lipid moiety, are independently linked to said oligomeric compound at the 5' end of said oligomeric compound.

[0233] In another preferred embodiment, said one or more lipid moieties, preferably said exactly one lipid moiety, are independently linked to said oligomeric compound at the 3' end of said oligomeric compound.

[0234] In another preferred embodiment, said one or more lipid moieties, preferably said exactly one lipid moiety, are independently linked to said oligomeric compound at an internal residue of said oligomeric compound.

[0235] In another highly preferred embodiment, said one or more lipid moieties, preferably exactly one lipid moiety, are covalently linked to said oligomeric compound, preferably said oligonucleotide, either directly or via a spacer through an -OP(O)(SH)- or -OP(O)(OH)- or -NHP(O)(OH)- or -NHP(O)(SH)- or -NH-C(O)- moiety, typically and preferably constituted by said one or more lipid moieties or said spacer, wherein said -OP(O)(SH)- or said -OP(O)(OH)- or said -NHP(O)(OH)- or said -NHP(O)(SH)- or said -NH-C(O)- moiety is linked to the 5'- or 3'-terminal OH group of said oligomeric compound.

[0236] In another highly preferred embodiment, said one or more lipid moieties, preferably exactly one lipid moiety, are covalently linked to said oligomeric compound, preferably said oligonucleotide, either directly or via a spacer, through an -OP(O)(SH)- or -OP(O)(OH)- moiety, said -OP(O)(SH)- or said -OP(O)(OH)- moiety being linked to the 5'- or 3'-terminal OH group of said oligomeric compound, typically and preferably said -OP(O)(SH)- or said -OP(O)(OH)- moiety being constituted by said one or more lipid moieties or said spacer.

[0237] In another highly preferred embodiment, the one or more lipid moieties, preferably exactly one lipid moiety, are covalently linked to the oligomeric compound, preferably to the oligonucleotide, either directly or via a spacer, through an -OP(O)(SH)- moiety, which is linked to the 5'- or 3'-terminal OH group of the oligomeric compound, typically and preferably wherein the -OP(O)(SH)- moiety is constituted by the one or more lipid moieties or the spacer.

[0238] In another highly preferred embodiment, said one or more lipid moieties, preferably exactly one lipid moiety, are covalently linked to said oligomeric compound, preferably said oligonucleotide, either directly or via a spacer, through an -OP(O)(SH)- moiety, said -OP(O)(SH)- moiety being linked to the 5'-terminal OH group of said oligomeric compound, typically and preferably said -OP(O)(SH)- moiety being constituted by said one or more lipid moieties or said spacer.

[0239] In another highly preferred embodiment, said one or more lipid moieties, preferably exactly one lipid moiety, are covalently linked to said oligomeric compound, preferably said oligonucleotide, either directly or via a spacer, through an -OP(O)(SH)- moiety, said -OP(O)(SH)- moiety being linked to the 3'-terminal OH group of said oligomeric compound, typically and preferably said -OP(O)(SH)- moiety being constituted by said one or more lipid moieties or said spacer.

[0240] In another highly preferred embodiment, said one or more lipid moieties, preferably exactly one lipid moiety, are covalently linked to said oligomeric compound, preferably said oligonucleotide, either directly or via a spacer, through a -P(O)(OH)- moiety, said -P(O)(OH)- moiety being linked to the 5'- or 3'-terminal OH group of said oligomeric compound, typically and preferably said -P(O)(OH)- moiety being constituted by said one or more lipid moieties or said spacer.

[0241] In another highly preferred embodiment, said one or more lipid moieties, preferably exactly one lipid moiety, are covalently linked to said oligomeric compound, preferably said oligonucleotide, either directly or via a spacer, through a -P(O)(OH)- moiety, said -P(O)(OH)- moiety being linked to the 5'-terminal OH group of said oligomeric compound, typically and preferably said -P(O)(OH)- moiety being constituted by said one or more lipid moieties or said spacer.

[0242] In another highly preferred embodiment, said one or more lipid moieties, preferably exactly one lipid moiety, are covalently linked to said oligomeric compound, preferably said oligonucleotide, either directly or via a spacer, through a -P(O)(OH)- moiety, said -P(O)(OH)- moiety being linked to the 3'-terminal OH group of said oligomeric compound, typically and preferably said -P(O)(OH)- moiety being constituted by said one or more lipid moieties or said spacer.

[0243] In another preferred embodiment, the composition does not contain any nucleosides other than tc-DNA nucleosides.

[0244] In another preferred embodiment, said oligomeric compound does not contain any nucleosides other than tc-DNA nucleosides.

[0245] In another preferred embodiment, the composition further comprises one or more nucleosides other than tc-DNA nucleosides.

[0246] In another preferred embodiment, said oligomeric compound further comprises one or more nucleosides other than tc-DNA nucleosides.

[0247] In another preferred embodiment, the oligomeric compound comprises one or more tc-DNA nucleosides and one or more nucleosides other than tc-DNA nucleosides, wherein 50% or more of all nucleosides are tc-DNA nucleosides.

[0248] In another preferred embodiment, the oligomeric compound comprises one or more tc-DNA nucleosides and one or more nucleosides other than tc-DNA nucleosides, wherein 60% or more of all nucleosides are tc-DNA nucleosides.

[0249] In another preferred embodiment, the oligomeric compound comprises one or more tc-DNA nucleosides and one or more nucleosides other than tc-DNA nucleosides, wherein 70% or more of all nucleosides are tc-DNA nucleosides.

[0250] In another preferred embodiment, the oligomeric compound comprises one or more tc-DNA nucleosides and one or more nucleosides other than tc-DNA nucleosides, wherein 75% or more of all nucleosides are tc-DNA nucleosides.

[0251] In another preferred embodiment, the oligomeric compound comprises one or more tc-DNA nucleosides and one or more nucleosides other than tc-DNA nucleosides, wherein 80% or more of all nucleosides are tc-DNA nucleosides.

[0252] In another preferred embodiment, the oligomeric compound comprises one or more tc-DNA nucleosides and one or more nucleosides other than tc-DNA nucleosides, wherein 85% or more of all nucleosides are tc-DNA nucleosides.

[0253] In another preferred embodiment, the oligomeric compound comprises one or more tc-DNA nucleosides and one or more nucleosides other than tc-DNA nucleosides, wherein 90% or more of all nucleosides are tc-DNA nucleosides.

[0254] In another preferred embodiment, the oligomeric compound comprises one or more tc-DNA nucleosides and one or more nucleosides other than tc-DNA nucleosides, wherein 95% or more of all nucleosides are tc-DNA nucleosides.

[0255] In another preferred embodiment, the oligomeric compound further comprises one or more nucleosides other than tc-DNA nucleosides, wherein said one or more nucleosides other than tc-DNA nucleosides are, independently of each other: i. 2'-modified ribonucleic acid (2'-modified-RNA) nucleosides; ii. Ribonucleic acid (RNA) nucleosides; iii. Deoxyribonucleic acid (DNA) nucleosides; iv. Locked nucleic acid (LNA) nucleosides; v. Peptide nucleic acid (PNA) nucleosides; vi. 2'-deoxy 2'-fluoro-arabinonucleosides; vii. hexitol nucleic acid (HNA) nucleosides; and viii. Phosphorodiamidate morpholino (PMO) nucleosides is selected from.

[0256] In another preferred embodiment, the oligomeric compound further comprises one or more nucleosides other than tc-DNA nucleosides, wherein said one or more nucleosides other than tc-DNA nucleosides are, independently of each other, 2'-modified ribonucleic acid (2'-modified-RNA) nucleosides.

[0257] In another preferred embodiment, the 2'-modified-RNA nucleosides are incorporated at at least two adjacent positions, forming self-complementary Watson-Crick base pairs.

[0258] In another preferred embodiment, the 2'-modified-RNA nucleosides are incorporated at three or more adjacent positions, forming self-complementary Watson-Crick base pairs.

[0259] In another preferred embodiment, the oligomeric compound further comprises one or more nucleosides other than tc-DNA nucleosides, wherein said one or more nucleosides other than tc-DNA nucleosides are, independently of each other, ribonucleic acid (RNA) nucleosides.

[0260] In another preferred embodiment, the oligomeric compound further comprises one or more nucleosides other than tc-DNA nucleosides, wherein said one or more nucleosides other than tc-DNA nucleosides are, independently of each other, deoxyribonucleic acid (DNA) nucleosides.

[0261] In another preferred embodiment, the oligomeric compound further comprises one or more nucleosides other than tc-DNA nucleosides, wherein said one or more nucleosides other than tc-DNA nucleosides are, independently of each other, locked nucleic acid (LNA) nucleosides.

[0262] In another preferred embodiment, the oligomeric compound further comprises one or more nucleosides other than tc-DNA nucleosides, wherein said one or more nucleosides other than tc-DNA nucleosides are, independently of each other, peptide nucleic acid (PNA) nucleosides.

[0263] In another preferred embodiment, the oligomeric compound further comprises one or more nucleosides other than tc-DNA nucleosides, wherein said one or more nucleosides other than tc-DNA nucleosides are, independently of each other, 2'-deoxy-2'-fluoro-arabinonucleosides.

[0264] In another preferred embodiment, the oligomeric compound further comprises one or more nucleosides other than tc-DNA nucleosides, wherein said one or more nucleosides other than tc-DNA nucleosides are, independently of each other, hexitol nucleic acid (HNA) nucleosides.

[0265] In another preferred embodiment, the oligomeric compound further comprises one or more nucleosides other than tc-DNA nucleosides, wherein said one or more nucleosides other than tc-DNA nucleosides are, independently of each other, phosphorodiamidate morpholino (PMO) nucleosides.

[0266] In another preferred embodiment, the oligomeric compound further comprises one or more nucleosides other than tc-DNA nucleosides, wherein said one or more nucleosides other than tc-DNA nucleosides are, independently of each other: i.RNA nucleosides; ii. 2'-O-methyl-RNA nucleosides; iii.2'-O-propargyl-RNA nucleoside; iv. 2'-O-propylamino-RNA nucleoside; v. 2'-O-amino-RNA nucleosides; vi. 2'-fluoro-RNA nucleosides; vii. 2'-O-methoxyethyl-RNA nucleoside; viii. morpholino nucleosides; and ix. Locked nucleic acid nucleosides is selected from.

[0267] In another preferred embodiment, the oligomeric compound further comprises one or more nucleosides other than tc-DNA nucleosides, wherein said one or more nucleosides other than tc-DNA nucleosides are, independently of each other, RNA nucleosides.

[0268] In another preferred embodiment, the oligomeric compound further comprises one or more nucleosides other than tc-DNA nucleosides, wherein said one or more nucleosides other than tc-DNA nucleosides are, independently of each other, 2'-O-methyl-RNA nucleosides.

[0269] In another preferred embodiment, the oligomeric compound further comprises one or more nucleosides other than tc-DNA nucleosides, wherein said one or more nucleosides other than tc-DNA nucleosides are, independently of each other, 2'-O-propargyl-RNA nucleosides.

[0270] In another preferred embodiment, the oligomeric compound further comprises one or more nucleosides other than tc-DNA nucleosides, wherein said one or more nucleosides other than tc-DNA nucleosides are, independently of each other, 2'-O-propylamino-RNA nucleosides.

[0271] In another preferred embodiment, the oligomeric compound further comprises one or more nucleosides other than tc-DNA nucleosides, wherein said one or more nucleosides other than tc-DNA nucleosides are, independently of each other, 2'-O-amino-RNA nucleosides.

[0272] In another preferred embodiment, the oligomeric compound further comprises one or more nucleosides other than tc-DNA nucleosides, wherein said one or more nucleosides other than tc-DNA nucleosides are, independently of each other, 2'-fluoro-RNA nucleosides.

[0273] In another preferred embodiment, the oligomeric compound further comprises one or more nucleosides other than tc-DNA nucleosides, wherein said one or more nucleosides other than tc-DNA nucleosides are, independently of each other, 2'-O-methoxyethyl-RNA nucleosides.

[0274] In another preferred embodiment, the oligomeric compound further comprises one or more nucleosides other than tc-DNA nucleosides, wherein said one or more nucleosides other than tc-DNA nucleosides are, independently of each other, morpholino nucleosides.

[0275] In another preferred embodiment, the oligomeric compound further comprises one or more nucleosides other than tc-DNA nucleosides, wherein said one or more nucleosides other than tc-DNA nucleosides are, independently of each other, locked nucleic acid RNA nucleosides.

[0276] In another preferred embodiment, said oligomeric compounds are complementary to a target sequence.

[0277] In another preferred embodiment, the oligomeric compound has a length of up to 40 monomeric subunits, preferably up to 30 monomeric subunits, more preferably up to 30 monomeric subunits, again more preferably up to 20 monomeric subunits or up to 15 monomeric subunits. In a further embodiment, the oligomer comprises 5 to 40 monomeric subunits, preferably 8 to 30 monomeric subunits, more preferably 8 to 25 monomeric subunits, again more preferably 8 to 20 monomeric subunits.

[0278] In another preferred embodiment, the oligomeric compound comprises 5 to 40 nucleotides, preferably, the oligomeric compound comprises 10 to 30 nucleotides, and more preferably, the oligomeric compound comprises 10 to 25 nucleotides.

[0279] In another preferred embodiment, the oligomeric compound is an oligonucleotide, the oligomeric compound comprising one or more tricyclo-deoxyribonucleic acid (tc-DNA) nucleosides, preferably the oligomeric compound comprising 5 to 40 monomeric subunits. In another preferred embodiment, the oligomeric compound is an oligonucleotide, the oligomeric compound comprising one or more tricyclo-deoxyribonucleic acid (tc-DNA) nucleosides, the oligomeric compound comprising 5 to 40 monomeric subunits, the monomeric subunits being linked by internucleoside linking groups.

[0280] In another preferred embodiment, the monomer subunits are independently selected from naturally occurring nucleosides, modified nucleosides, and nucleoside mimetics, and preferably, the naturally occurring nucleosides, modified nucleosides, and nucleoside mimetics are independently selected from tricyclic nucleosides, ribonucleic acid (RNA) nucleosides, deoxyribonucleic acid (DNA) nucleosides, 2'-modified ribonucleic acid (2'-modified-RNA) nucleosides, locked nucleic acid (LNA) nucleosides, peptide nucleic acid (PNA) nucleosides, 2'-deoxy-2'-fluoro-arabinonucleosides, hexitol nucleic acid (HNA) nucleosides, and phosphorodiamidate morpholino (PMO) nucleosides.

[0281] In another preferred embodiment, the monomer subunits are independently selected from naturally occurring nucleosides, modified nucleosides, and nucleoside mimetics, preferably, the naturally occurring nucleosides, modified nucleosides, and nucleoside mimetics are independently selected from tricyclic nucleosides, ribonucleic acid (RNA) nucleosides, deoxyribonucleic acid (DNA) nucleosides, 2'-modified ribonucleic acid (2'-modified-RNA) nucleosides, locked nucleic acid (LNA) nucleosides, peptide nucleic acid (PNA) nucleosides, 2'-deoxy-2'-fluoro-arabinonucleosides, hexitol nucleic acid (HNA) nucleosides, and phosphorodiamidate morpholino (PMO) nucleosides, and the monomer subunits are linked by multiple internucleoside linking groups.

[0282] In another preferred embodiment, the oligomeric compound comprises 5 to 40 monomeric subunits, wherein the monomeric subunits are nucleosides, one or more of the nucleosides are the one or more tricyclo-deoxyribonucleic acid (tc-DNA) nucleosides, and the monomeric subunits are linked by a plurality of internucleoside linking groups, and preferably the oligomeric compound is an oligonucleotide.

[0283] In another preferred embodiment, the oligomeric compound comprises 5 to 40 monomeric subunits, wherein the monomeric subunits are nucleosides, one or more of the nucleosides are the one or more tricyclo-deoxyribonucleic acid (tc-DNA) nucleosides, and the monomeric subunits are linked by a plurality of internucleoside linking groups, and preferably the oligomeric compound is an oligonucleotide.

[0284] In another preferred embodiment, the oligomeric compound comprises 5 to 40 monomeric subunits, wherein the monomeric subunits are nucleosides, wherein one or more of the nucleosides are the one or more tricyclo-deoxyribonucleic acid (tc-DNA) nucleosides, and wherein the nucleosides are linked by a plurality of internucleoside linking groups, and preferably the oligomeric compound is an oligonucleotide.

[0285] In another preferred embodiment, the composition is free of nucleosides other than tc-DNA nucleosides. In another preferred embodiment, the oligomeric compound is free of nucleosides other than tc-DNA nucleosides. In another preferred embodiment, the composition further comprises one or more nucleosides other than tc-DNA nucleosides. In another preferred embodiment, the oligomeric compound further comprises one or more nucleosides other than tc-DNA nucleosides.

[0286] In another preferred embodiment, the oligomeric compound comprises one or more tc-DNA nucleosides. In another preferred embodiment, the oligomeric compound comprises one or more tc-DNA nucleosides and one or more nucleosides other than tc-DNA nucleosides.

[0287] In another preferred embodiment, the one or more tc-DNA nucleosides, or the one or more tc-DNA nucleosides and the one or more nucleosides other than tc-DNA nucleosides, are linked by multiple internucleoside linking groups.

[0288] In another preferred embodiment, the oligomeric compound comprises 5 to 40 monomeric subunits, wherein the monomeric subunits are nucleosides, all of the nucleosides are the one or more tricyclo-deoxyribonucleic acid (tc-DNA) nucleosides, and the nucleosides are linked by a plurality of internucleoside linking groups, and preferably the oligomeric compound is an oligonucleotide.

[0289] In another preferred embodiment, the oligomeric compound comprises 5 to 40 monomeric subunits, wherein the monomeric subunits are nucleosides, one of the nucleosides is a nucleoside other than a tc-DNA nucleoside, and all other of the nucleosides are the one or more tricyclo-deoxyribonucleic acid (tc-DNA) nucleosides, and the nucleosides are linked by a plurality of internucleoside linking groups, and preferably the oligomeric compound is an oligonucleotide.

[0290] In another preferred embodiment, the oligomeric compound comprises 5 to 40 monomeric subunits, wherein the monomeric subunits are nucleosides, wherein at most 5, preferably at most 4, and more preferably at most 3, of the nucleosides are nucleosides other than tc-DNA nucleosides, and all other nucleosides are the one or more tricyclo-deoxyribonucleic acid (tc-DNA) nucleosides, and the nucleosides are linked by a plurality of internucleoside linking groups ... and wherein at most 5, preferably at most 4, Preferably, at least one, more preferably at most three, nucleosides other than tc-DNA nucleosides are selected from ribonucleic acid (RNA) nucleosides, deoxyribonucleic acid (DNA) nucleosides, 2'-modified ribonucleic acid (2'-modified-RNA) nucleosides, locked nucleic acid (LNA) nucleosides, peptide nucleic acid (PNA) nucleosides, 2'-deoxy 2'-fluoro-arabinonucleosides, hexitol nucleic acid (HNA) nucleosides, and phosphorodiamidate morpholino (PMO) nucleosides, and preferably, said oligomeric compound is an oligonucleotide.

[0291] In another highly preferred embodiment, the oligomeric compound comprises 5 to 40 monomeric subunits, wherein the monomeric subunits are nucleosides, wherein at most three of the nucleosides are nucleosides other than tc-DNA nucleosides, and all other nucleosides are the one or more tricyclo-deoxyribonucleic acid (tc-DNA) nucleosides, wherein the nucleosides are linked by a plurality of internucleoside linking groups, and wherein the at most three tc-DNA nucleosides are the one or more tricyclo-deoxyribonucleic acid (tc-DNA) nucleosides. The nucleoside other than a nucleoside is selected from ribonucleic acid (RNA) nucleosides, deoxyribonucleic acid (DNA) nucleosides, 2'-modified ribonucleic acid (2'-modified-RNA) nucleosides, locked nucleic acid (LNA) nucleosides, peptide nucleic acid (PNA) nucleosides, 2'-deoxy 2'-fluoro-arabinonucleosides, hexitol nucleic acid (HNA) nucleosides and phosphorodiamidate morpholino (PMO) nucleosides, and preferably, the oligomeric compound is an oligonucleotide.

[0292] In another highly preferred embodiment, the oligomeric compound comprises 5 to 40 monomeric subunits, wherein the monomeric subunits are nucleosides, wherein at most two of the nucleosides are nucleosides other than tc-DNA nucleosides, and all other nucleosides are the one or more tricyclo-deoxyribonucleic acid (tc-DNA) nucleosides, wherein the nucleosides are linked by a plurality of internucleoside linking groups, and wherein the at most two tc-DNA nucleosides are the one or more tricyclo-deoxyribonucleic acid (tc-DNA) nucleosides. The nucleoside other than a nucleoside is selected from ribonucleic acid (RNA) nucleosides, deoxyribonucleic acid (DNA) nucleosides, 2'-modified ribonucleic acid (2'-modified-RNA) nucleosides, locked nucleic acid (LNA) nucleosides, peptide nucleic acid (PNA) nucleosides, 2'-deoxy 2'-fluoro-arabinonucleosides, hexitol nucleic acid (HNA) nucleosides and phosphorodiamidate morpholino (PMO) nucleosides, and preferably, the oligomeric compound is an oligonucleotide.

[0293] In another highly preferred embodiment, the oligomeric compound comprises 5 to 40 monomeric subunits, wherein the monomeric subunits are nucleosides, wherein at most one of the nucleosides is a nucleoside other than a tc-DNA nucleoside, and all other nucleosides are the one or more tricyclo-deoxyribonucleic acid (tc-DNA) nucleosides, wherein the nucleosides are linked by a plurality of internucleoside linking groups, and wherein the at most one tc-DNA nucleoside is a nucleoside other than a tc-DNA nucleoside. The nucleoside other than a nucleoside is selected from ribonucleic acid (RNA) nucleosides, deoxyribonucleic acid (DNA) nucleosides, 2'-modified ribonucleic acid (2'-modified-RNA) nucleosides, locked nucleic acid (LNA) nucleosides, peptide nucleic acid (PNA) nucleosides, 2'-deoxy 2'-fluoro-arabinonucleosides, hexitol nucleic acid (HNA) nucleosides and phosphorodiamidate morpholino (PMO) nucleosides, and preferably, the oligomeric compound is an oligonucleotide.

[0294] In another preferred embodiment, the oligomeric compound comprises 5 to 40 monomeric subunits, wherein the monomeric subunits are nucleosides, one of the nucleosides is a nucleoside other than a tc-DNA nucleoside, and all other of the nucleosides are the one or more tricyclo-deoxyribonucleic acid (tc-DNA) nucleosides, the nucleosides being linked by a plurality of internucleoside linking groups, and the nucleosides other than the tc-DNA nucleosides are the one or more tricyclo-deoxyribonucleic acid (tc-DNA) nucleosides. Preferably, the nucleoside is selected from ribonucleic acid (RNA) nucleosides, deoxyribonucleic acid (DNA) nucleosides, 2'-modified ribonucleic acid (2'-modified-RNA) nucleosides, locked nucleic acid (LNA) nucleosides, peptide nucleic acid (PNA) nucleosides, 2'-deoxy 2'-fluoro-arabinonucleosides, hexitol nucleic acid (HNA) nucleosides and phosphorodiamidate morpholino (PMO) nucleosides, and the oligomeric compound is an oligonucleotide.

[0295] In another highly preferred embodiment, the oligomeric compound comprises 5 to 40 monomeric subunits, wherein the monomeric subunits are nucleosides, wherein one of the nucleosides is a nucleoside other than a tc-DNA nucleoside, and all other of the nucleosides are the one or more tricyclo-deoxyribonucleic acid (tc-DNA) nucleosides, wherein the nucleosides are linked by a plurality of internucleoside linking groups, and wherein the nucleosides other than the tc-DNA nucleosides are selected from ribonucleic acid (RNA) nucleosides, deoxyribonucleic acid (DNA) nucleosides, 2'-modified ribonucleic acid (2'-modified-RNA) nucleosides, locked nucleic acid (LNA) nucleosides, and phosphorodiamidate morpholino (PMO) nucleosides, and preferably the oligomeric compound is an oligonucleotide.

[0296] In another preferred embodiment, the oligomeric compound comprises 5 to 40 monomeric subunits, wherein the monomeric subunits are nucleosides, one of the nucleosides is a nucleoside other than a tc-DNA nucleoside, and all other of the nucleosides are the one or more tricyclo-deoxyribonucleic acid (tc-DNA) nucleosides, the nucleosides being linked by a plurality of internucleoside linking groups, and the nucleosides other than the tc-DNA nucleosides are selected from ribonucleic acid (RNA) nucleosides, deoxyribonucleic acid (DNA) nucleosides, and 2'-modified ribonucleic acid (2'-modified-RNA) nucleosides, and preferably the oligomeric compound is an oligonucleotide.

[0297] In another preferred embodiment, the oligomeric compound comprises 5 to 40 monomeric subunits, wherein the monomeric subunits are nucleosides, two of the nucleosides are nucleosides other than tc-DNA nucleosides, and all other nucleosides are the one or more tricyclo-deoxyribonucleic acid (tc-DNA) nucleosides, the nucleosides are linked by a plurality of internucleoside linking groups, and the two nucleosides other than tc-DNA nucleosides are the one or more tricyclo-deoxyribonucleic acid (tc-DNA) nucleosides. The nucleosides are independently selected from ribonucleic acid (RNA) nucleosides, deoxyribonucleic acid (DNA) nucleosides, 2'-modified ribonucleic acid (2'-modified-RNA) nucleosides, locked nucleic acid (LNA) nucleosides, peptide nucleic acid (PNA) nucleosides, 2'-deoxy 2'-fluoro-arabinonucleosides, hexitol nucleic acid (HNA) nucleosides, and phosphorodiamidate morpholino (PMO) nucleosides, and preferably, said oligomeric compound is an oligonucleotide.

[0298] In another preferred embodiment, the plurality of internucleoside linking groups are independently selected from phosphorothioate linking groups, phosphorodithioate linking groups, and phosphorodiester linking groups, phosphotriester linking groups, aminoalkylphosphotriester linking groups, methylphosphonate linking groups, alkylphosphonate linking groups, 5'-alkylenephosphonate linking groups, phosphonate linking groups, phosphinate linking groups, phosphoramidate linking groups, 3'-aminophosphoramidate linking groups, aminoalkylphosphoramidate linking groups, thionophosphoramidate linking groups, thionoalkylphosphonate linking groups, thionoalkylphosphotriester linking groups, selenophosphate linking groups, or boranophosphate linking groups.

[0299] In another preferred embodiment, said plurality of internucleoside linkage groups are independently selected from phosphorothioate linkage groups and phosphorodiester linkage groups.

[0300] In another preferred embodiment, no more than 50% of the internucleoside linking groups are phosphorothioate linking groups. In another preferred embodiment, no more than 45% of the internucleoside linking groups are phosphorothioate linking groups. In another preferred embodiment, no more than 40% of the internucleoside linking groups are phosphorothioate linking groups. In another preferred embodiment, no more than 35% of the internucleoside linking groups are phosphorothioate linking groups. In another preferred embodiment, no more than 33% of the internucleoside linking groups are phosphorothioate linking groups. In another preferred embodiment, no more than 30% of the internucleoside linking groups are phosphorothioate linking groups. In another preferred embodiment, no more than 25% of the internucleoside linking groups are phosphorothioate linking groups. In another preferred embodiment, no more than 20% of the internucleoside linking groups are phosphorothioate linking groups. In another preferred embodiment, no more than 20% of the plurality of internucleoside linking groups are phosphorothioate linking groups. In another preferred embodiment, no more than 15% of the plurality of internucleoside linking groups are phosphorothioate linking groups. In another preferred embodiment, no more than 10% of the plurality of internucleoside linking groups are phosphorothioate linking groups. In another preferred embodiment, no more than 5% of the plurality of internucleoside linking groups are phosphorothioate linking groups. In another preferred embodiment, none (0%) of the plurality of internucleoside linking groups are phosphorothioate linking groups.

[0301] In another preferred embodiment, no more than eight of the plurality of internucleoside linking groups are phosphorothioate linking groups. In another preferred embodiment, no more than seven of the plurality of internucleoside linking groups are phosphorothioate linking groups. In another preferred embodiment, no more than six of the plurality of internucleoside linking groups are phosphorothioate linking groups. In another preferred embodiment, no more than five of the plurality of internucleoside linking groups are phosphorothioate linking groups. In another preferred embodiment, no more than four of the plurality of internucleoside linking groups are phosphorothioate linking groups. In another preferred embodiment, no more than three of the plurality of internucleoside linking groups are phosphorothioate linking groups. In another preferred embodiment, no more than two of the plurality of internucleoside linking groups are phosphorothioate linking groups. In another preferred embodiment, no more than one of the plurality of internucleoside linking groups is a phosphorothioate linking group. In another preferred embodiment, none of said plurality of internucleoside linkage groups (zero / 0) is a phosphorothioate linkage group.

[0302] In another preferred embodiment, said plurality of internucleoside linkage groups are independently selected from phosphorothioate linkage groups and phosphorodiester linkage groups, and no more than 30% of said plurality of internucleoside linkage groups are phosphorothioate linkage groups.

[0303] In another preferred embodiment, at least 50% of the internucleoside linking groups are phosphorodiester linkages. In another preferred embodiment, at least 55% of the internucleoside linking groups are phosphorodiester linkages. In another preferred embodiment, at least 60% of the internucleoside linking groups are phosphorodiester linkages. In another preferred embodiment, at least 65% of the internucleoside linking groups are phosphorodiester linkages. In another preferred embodiment, at least 66% of the internucleoside linking groups are phosphorodiester linkages. In another preferred embodiment, at least 70% of the internucleoside linking groups are phosphorodiester linkages. In another preferred embodiment, at least 75% of the internucleoside linking groups are phosphorodiester linkages. In another preferred embodiment, at least 80% of the internucleoside linking groups are phosphorodiester linkages. In another preferred embodiment, at least 85% of the internucleoside linkage groups are phosphorodiester linkage groups. In another preferred embodiment, at least 90% of the internucleoside linkage groups are phosphorodiester linkage groups. In another preferred embodiment, at least 95% of the internucleoside linkage groups are phosphorodiester linkage groups. In another preferred embodiment, all (100%) of the internucleoside linkage groups are phosphorodiester linkage groups.

[0304] In another preferred embodiment, at least 80% of said plurality of internucleoside linkage groups are phosphorodiester linkage groups, and preferably said plurality of internucleoside linkage groups are independently selected from phosphorothioate linkage groups and phosphorodiester linkage groups.

[0305] In another highly preferred embodiment, the internucleoside linking groups are independently selected from phosphorothioate and phosphorodiester linking groups, and no more than six of the internucleoside linking groups are phosphorothioate linking groups. In another highly preferred embodiment, the internucleoside linking groups are independently selected from phosphorothioate and phosphorodiester linking groups, and no more than five of the internucleoside linking groups are phosphorothioate linking groups. In another highly preferred embodiment, the internucleoside linking groups are independently selected from phosphorothioate and phosphorodiester linking groups, and no more than four of the internucleoside linking groups are phosphorothioate linking groups. In another highly preferred embodiment, the internucleoside linking groups are independently selected from phosphorothioate and phosphorodiester linking groups, and no more than three of the internucleoside linking groups are phosphorothioate linking groups. In another highly preferred embodiment, the internucleoside linking groups are independently selected from phosphorothioate and phosphorodiester linking groups, and no more than two of the internucleoside linking groups are phosphorothioate linking groups. In another highly preferred embodiment, the internucleoside linking groups are independently selected from phosphorothioate and phosphorodiester linking groups, and no more than one of the internucleoside linking groups is a phosphorothioate linking group.

[0306] In another highly preferred embodiment, the plurality of internucleoside linking groups are independently selected from phosphorodiester linking groups. Thus, in another highly preferred embodiment, all of the plurality of internucleoside linking groups are phosphorodiester linking groups.

[0307] In another highly preferred embodiment, the oligomeric compound comprises 5 to 40 monomeric subunits, wherein the monomeric subunits are nucleosides, wherein at most three of the nucleosides are nucleosides other than tc-DNA nucleosides, and all other nucleosides are the one or more tricyclo-deoxyribonucleic acid (tc-DNA) nucleosides, wherein the nucleosides are linked by a plurality of internucleoside linking groups, and wherein the at most three nucleosides other than tc-DNA nucleosides are selected from the group consisting of ribonucleic acid (RNA) nucleosides, deoxyribonucleic acid (DNA) nucleosides, 2'-modified ribonucleic acid (2'-modified-RNA) nucleosides, locked nucleic acid nucleosides, and .... Preferably, the oligomeric compound is an oligonucleotide, and the internucleoside linking groups are independently selected from phosphorothioate linking groups and phosphorodiester linking groups, and no more than 6, preferably no more than 5, more preferably no more than 4, again more preferably no more than 3, more preferably no more than 2, and even more preferably no more than 1 of the internucleoside linking groups is a phosphorothioate linking group. In a highly preferred embodiment, all of the internucleoside linking groups are independently selected from phosphorodiester linking groups.

[0308] In another highly preferred embodiment, the oligomeric compound comprises 5 to 40 monomeric subunits, wherein the monomeric subunits are nucleosides, wherein at most two of the nucleosides are nucleosides other than tc-DNA nucleosides, and all other nucleosides are the one or more tricyclo-deoxyribonucleic acid (tc-DNA) nucleosides, wherein the nucleosides are linked by a plurality of internucleoside linking groups, and wherein the at most two nucleosides other than tc-DNA nucleosides are selected from the group consisting of ribonucleic acid (RNA) nucleosides, deoxyribonucleic acid (DNA) nucleosides, 2'-modified ribonucleic acid (2'-modified-RNA) nucleosides, locked nucleic acids, and the like. Preferably, the oligomeric compound is an oligonucleotide, and the internucleoside linking groups are independently selected from phosphorothioate linking groups and phosphorodiester linking groups, and no more than 6, preferably no more than 5, more preferably no more than 4, again more preferably no more than 3, more preferably no more than 2, and even more preferably no more than 1 of the internucleoside linking groups is a phosphorothioate linking group. In a highly preferred embodiment, all of the internucleoside linking groups are independently selected from phosphorodiester linking groups.

[0309] In another highly preferred embodiment, the oligomeric compound comprises 5 to 40 monomeric subunits, wherein the monomeric subunits are nucleosides, wherein at most one of the nucleosides is a nucleoside other than a tc-DNA nucleoside, and all other of the nucleosides are the one or more tricyclo-deoxyribonucleic acid (tc-DNA) nucleosides, wherein the nucleosides are linked by a plurality of internucleoside linking groups, and wherein the at most one nucleoside other than the tc-DNA nucleoside is a ribonucleic acid (RNA) nucleoside, a deoxyribonucleic acid (DNA) nucleoside, a 2'-modified ribonucleic acid (2'-modified-RNA) nucleoside, a locked ... Preferably, the oligomeric compound is an oligonucleotide, and the internucleoside linking groups are independently selected from phosphorothioate linking groups and phosphorodiester linking groups, and no more than 6, preferably no more than 5, more preferably no more than 4, again more preferably no more than 3, more preferably no more than 2, and even more preferably no more than 1 of the internucleoside linking groups is a phosphorothioate linking group. In a highly preferred embodiment, all of the internucleoside linking groups are independently selected from phosphorodiester linking groups.

[0310] In another preferred embodiment, said oligomeric compounds do not contain direct tc-DNA to tc-DNA phosphorothioate internucleoside linkages.

[0311] The following table provides a highly preferred embodiment of the present invention.

[0312] [Table 1-1] [Table 1-2]

[0313] In a preferred embodiment, the oligomeric compound is selected from the oligomeric compounds listed in Table 1, and typically and preferably, the 2'-modified-RNA is a 2'-OMe-RNA.

[0314] In a preferred embodiment, the oligomeric compound, preferably the oligonucleotide, is selected from any one of the sequences SEQ ID NOs: 1 to 22, and the 2'-modified-RNA is 2'-OMe-RNA. In a preferred embodiment, the oligomeric compound, preferably the oligonucleotide, is selected from any one of the sequences SEQ ID NOs: 1 to 22, and the 2'-modified-RNA is 2'-OMe-RNA, and each of the internucleoside linkage groups of the sequences SEQ ID NOs: 1 to 22 is a phosphorodiester linkage group.

[0315] In a further preferred embodiment, said oligomeric compound comprises, preferably is, the sequence SEQ ID NO:1.

[0316] In a further preferred embodiment, said oligomeric compound comprises, preferably is, the sequence SEQ ID NO:2.

[0317] In a further preferred embodiment, the oligomeric compound comprises, preferably is, the sequence of SEQ ID NO: 3. In a further preferred embodiment, the oligomeric compound comprises, preferably is, the sequence of SEQ ID NO: 3, and the 2'-modified-RNA is 2'-OMe-RNA.

[0318] In a further preferred embodiment, the oligomeric compound comprises the sequence of SEQ ID NO: 4, preferably is the sequence of SEQ ID NO: 4. In a further preferred embodiment, the oligomeric compound comprises the sequence of SEQ ID NO: 4, preferably is the sequence of SEQ ID NO: 4, and the 2'-modified-RNA is 2'-OMe-RNA.

[0319] In a further preferred embodiment, said oligomeric compound comprises the sequence of SEQ ID NO: 5, preferably is the sequence of SEQ ID NO: 5. In a further preferred embodiment, said oligomeric compound comprises the sequence of SEQ ID NO: 5, preferably is the sequence of SEQ ID NO: 5, and said 2'-modified-RNA is 2'-OMe-RNA.

[0320] In a further preferred embodiment, the oligomeric compound comprises the sequence of SEQ ID NO: 6, preferably is the sequence of SEQ ID NO: 6. In a further preferred embodiment, the oligomeric compound comprises the sequence of SEQ ID NO: 6, preferably is the sequence of SEQ ID NO: 6, and the 2'-modified-RNA is 2'-OMe-RNA.

[0321] In a further preferred embodiment, said oligomeric compound comprises, preferably is, the sequence SEQ ID NO:7.

[0322] In a further preferred embodiment, said oligomeric compound comprises the sequence of SEQ ID NO: 8, preferably is the sequence of SEQ ID NO: 8. In a further preferred embodiment, said oligomeric compound comprises the sequence of SEQ ID NO: 8, preferably is the sequence of SEQ ID NO: 8, and said 2'-modified-RNA is 2'-OMe-RNA.

[0323] In a further preferred embodiment, said oligomeric compound comprises, preferably is, the sequence SEQ ID NO:9.

[0324] In a further preferred embodiment, said oligomeric compound comprises, preferably is, the sequence SEQ ID NO:10.

[0325] In a further preferred embodiment, said oligomeric compound comprises the sequence of SEQ ID NO: 11, preferably is the sequence of SEQ ID NO: 11. In a further preferred embodiment, said oligomeric compound comprises the sequence of SEQ ID NO: 11, preferably is the sequence of SEQ ID NO: 11, and said 2'-modified-RNA is 2'-OMe-RNA.

[0326] In a further preferred embodiment, said oligomeric compound comprises the sequence of SEQ ID NO: 12, preferably is the sequence of SEQ ID NO: 12. In a further preferred embodiment, said oligomeric compound comprises the sequence of SEQ ID NO: 12, preferably is the sequence of SEQ ID NO: 12, and said 2'-modified-RNA is 2'-OMe-RNA.

[0327] In a further preferred embodiment, said oligomeric compound comprises the sequence of SEQ ID NO: 13, preferably is the sequence of SEQ ID NO: 13. In a further preferred embodiment, said oligomeric compound comprises the sequence of SEQ ID NO: 13, preferably is the sequence of SEQ ID NO: 13, and said 2'-modified-RNA is 2'-OMe-RNA.

[0328] In a further preferred embodiment, said oligomeric compound comprises the sequence of SEQ ID NO: 14, preferably is the sequence of SEQ ID NO: 14. In a further preferred embodiment, said oligomeric compound comprises the sequence of SEQ ID NO: 14, preferably is the sequence of SEQ ID NO: 14, and said 2'-modified-RNA is 2'-OMe-RNA.

[0329] In a further preferred embodiment, said oligomeric compound comprises the sequence of SEQ ID NO: 15, preferably is the sequence of SEQ ID NO: 15. In a further preferred embodiment, said oligomeric compound comprises the sequence of SEQ ID NO: 15, preferably is the sequence of SEQ ID NO: 15, and said 2'-modified-RNA is 2'-OMe-RNA.

[0330] In a further preferred embodiment, said oligomeric compound comprises the sequence of SEQ ID NO: 16, preferably is the sequence of SEQ ID NO: 16. In a further preferred embodiment, said oligomeric compound comprises the sequence of SEQ ID NO: 16, preferably is the sequence of SEQ ID NO: 16, and said 2'-modified-RNA is 2'-OMe-RNA.

[0331] In a further preferred embodiment, said oligomeric compound comprises the sequence of SEQ ID NO: 17, preferably is the sequence of SEQ ID NO: 17. In a further preferred embodiment, said oligomeric compound comprises the sequence of SEQ ID NO: 17, preferably is the sequence of SEQ ID NO: 17, and said 2'-modified-RNA is 2'-OMe-RNA.

[0332] In a further preferred embodiment, said oligomeric compound comprises the sequence of SEQ ID NO: 18, preferably is the sequence of SEQ ID NO: 18. In a further preferred embodiment, said oligomeric compound comprises the sequence of SEQ ID NO: 18, preferably is the sequence of SEQ ID NO: 18, and said 2'-modified-RNA is 2'-OMe-RNA.

[0333] In a further preferred embodiment, said oligomeric compound comprises, preferably is, the sequence SEQ ID NO:19.

[0334] In a further preferred embodiment, said oligomeric compound comprises the sequence SEQ ID NO: 20, preferably is the sequence SEQ ID NO: 20. In a further preferred embodiment, said oligomeric compound comprises the sequence SEQ ID NO: 20, preferably is the sequence SEQ ID NO: 20, and said 2'-modified-RNA is 2'-OMe-RNA.

[0335] In a further preferred embodiment, said oligomeric compound comprises, preferably is, the sequence SEQ ID NO:21.

[0336] In a further preferred embodiment, said oligomeric compound comprises, preferably is, the sequence SEQ ID NO:22.

[0337] In a further preferred embodiment, said oligomeric compound is selected from the oligomeric compounds listed in Table 2, and typically and preferably said 2'-modified-RNA is 2'-OMe-RNA.

[0338] In a highly preferred embodiment, said oligomeric compound, preferably said oligonucleotide, is selected from any one of the sequences SEQ ID NOs: 1-37.

[0339] In a further highly preferred embodiment, said oligomeric compound, preferably said oligonucleotide, is selected from any one of the sequences SEQ ID NOs: 1 to 37, and said 2'-modified-RNA is 2'-OMe-RNA.

[0340] [Table 2-1] [Table 2-2] [Table 2-3]

[0341] In a further highly preferred embodiment, the oligomeric compound has a sequence selected from any one of SEQ ID NOs: 1 to 37, wherein each of the internucleoside linkage groups of the sequences SEQ ID NOs: 1 to 22 is a phosphorodiester linkage group, and preferably the 2'-modified-RNA is 2'-OMe-RNA. In a further highly preferred embodiment, the oligomeric compound has a sequence selected from any one of SEQ ID NOs: 1 to 37, wherein each of the internucleoside linkage groups of the sequences SEQ ID NOs: 1 to 22 is a phosphorodiester linkage group, and preferably the 2'-modified-RNA is 2'-OMe-RNA.

[0342] In a further preferred embodiment, said oligomeric compound comprises, preferably is, the sequence SEQ ID NO:23.

[0343] In a further preferred embodiment, said oligomeric compound comprises, preferably is, the sequence of SEQ ID NO: 24. In a further preferred embodiment, said oligomeric compound comprises, preferably is, the sequence of SEQ ID NO: 24, and said 2'-modified-RNA is 2'-OMe-RNA.

[0344] In a further preferred embodiment, said oligomeric compound comprises the sequence of SEQ ID NO: 25, preferably is the sequence of SEQ ID NO: 25. In a further preferred embodiment, said oligomeric compound comprises the sequence of SEQ ID NO: 25, preferably is the sequence of SEQ ID NO: 25, and said 2'-modified-RNA is 2'-OMe-RNA.

[0345] In a further preferred embodiment, said oligomeric compound comprises the sequence SEQ ID NO: 26, preferably is the sequence SEQ ID NO: 26. In a further preferred embodiment, said oligomeric compound comprises the sequence SEQ ID NO: 26, preferably is the sequence SEQ ID NO: 26, and said 2'-modified-RNA is 2'-OMe-RNA.

[0346] In a further preferred embodiment, said oligomeric compound comprises the sequence of SEQ ID NO: 27, preferably is the sequence of SEQ ID NO: 27. In a further preferred embodiment, said oligomeric compound comprises the sequence of SEQ ID NO: 27, preferably is the sequence of SEQ ID NO: 27, and said 2'-modified-RNA is 2'-OMe-RNA.

[0347] In a further preferred embodiment, said oligomeric compound comprises the sequence of SEQ ID NO: 28, preferably is the sequence of SEQ ID NO: 28. In a further preferred embodiment, said oligomeric compound comprises the sequence of SEQ ID NO: 28, preferably is the sequence of SEQ ID NO: 28, and said 2'-modified-RNA is 2'-OMe-RNA.

[0348] In a further preferred embodiment, said oligomeric compound comprises, preferably is, the sequence of SEQ ID NO: 29. In a further preferred embodiment, said oligomeric compound comprises, preferably is, the sequence of SEQ ID NO: 29, and said 2'-modified-RNA is 2'-OMe-RNA.

[0349] In a further preferred embodiment, said oligomeric compound comprises the sequence of SEQ ID NO: 30, preferably is the sequence of SEQ ID NO: 30. In a further preferred embodiment, said oligomeric compound comprises the sequence of SEQ ID NO: 30, preferably is the sequence of SEQ ID NO: 30, and said 2'-modified-RNA is 2'-OMe-RNA.

[0350] In a further preferred embodiment, said oligomeric compound comprises, preferably is, the sequence SEQ ID NO:31.

[0351] In a further preferred embodiment, said oligomeric compound comprises, preferably is, the sequence SEQ ID NO:32.

[0352] In a further preferred embodiment, said oligomeric compound comprises, preferably is, the sequence SEQ ID NO:33.

[0353] In a further preferred embodiment, said oligomeric compound comprises, preferably is, the sequence SEQ ID NO:34.

[0354] In a further preferred embodiment, said oligomeric compound comprises, preferably is, the sequence SEQ ID NO:35.

[0355] In a further preferred embodiment, said oligomeric compound comprises, preferably is, the sequence SEQ ID NO:36.

[0356] In a further preferred embodiment, said oligomeric compound comprises, preferably is, the sequence SEQ ID NO:37.

[0357] In a further highly preferred embodiment, the composition of the present invention is selected from any one of the compositions listed in Table 3.

[0358] [Table 3-1] [Table 3-2] [Table 3-3] [Table 3-4] [ka]

[0359] In a further highly preferred embodiment, the compositions of the invention comprise, and preferably are, SY-0299.

[0360] In a further highly preferred embodiment, the compositions of the invention comprise, and preferably are, SY-0343.

[0361] In a further highly preferred embodiment, the compositions of the present invention comprise, and preferably are, SY-0357.

[0362] In a further highly preferred embodiment, the compositions of the present invention comprise, preferably are, SY-0427.

[0363] In a further highly preferred embodiment, the compositions of the present invention comprise, and preferably are, SY-0440.

[0364] In a further highly preferred embodiment, the compositions of the present invention comprise, and preferably are, SY-0442.

[0365] In a further highly preferred embodiment, the compositions of the present invention comprise, and preferably are, SY-0443.

[0366] In a further highly preferred embodiment, the compositions of the present invention comprise, and preferably are, SY-0444.

[0367] In a further highly preferred embodiment, the compositions of the present invention comprise, preferably are, SY-0445.

[0368] In a further highly preferred embodiment, the compositions of the present invention comprise, preferably are, SY-0446.

[0369] In a further highly preferred embodiment, the compositions of the present invention comprise, and preferably are, SY-0448.

[0370] In a further highly preferred embodiment, the compositions of the present invention comprise, and preferably are, SY-0450.

[0371] In a further highly preferred embodiment, the compositions of the invention comprise, and preferably are, SY-0451.

[0372] In a further highly preferred embodiment, the compositions of the present invention comprise, preferably are, SY-0455.

[0373] In a further highly preferred embodiment, the compositions of the present invention comprise, preferably are, SY-0457.

[0374] In a further highly preferred embodiment, the compositions of the present invention comprise, and preferably are, SY-0458.

[0375] In a further highly preferred embodiment, the compositions of the present invention comprise, and preferably are, SY-0459.

[0376] In a further highly preferred embodiment, the compositions of the present invention comprise, and preferably are, SY-0460.

[0377] In a further highly preferred embodiment, the compositions of the present invention comprise, and preferably are, SY-0487.

[0378] In a further highly preferred embodiment, the compositions of the present invention comprise, and preferably are, SY-0488.

[0379] In a further highly preferred embodiment, the compositions of the present invention comprise, and preferably are, SY-0489.

[0380] In a further highly preferred embodiment, the compositions of the present invention comprise, and preferably are, SY-0490.

[0381] In a further highly preferred embodiment, the compositions of the invention comprise, preferably are, SY-0491.

[0382] In a further highly preferred embodiment, the compositions of the present invention comprise, and preferably are, SY-0492.

[0383] In a further highly preferred embodiment, the compositions of the present invention comprise, preferably are, SY-0493.

[0384] In a further highly preferred embodiment, the compositions of the present invention comprise, and preferably are, SY-0494.

[0385] In a further highly preferred embodiment, the compositions of the present invention comprise, preferably are, SY-0495.

[0386] In a further highly preferred embodiment, the compositions of the present invention comprise, preferably are, SY-0496.

[0387] In a further highly preferred embodiment, the compositions of the present invention comprise, preferably are, SY-0497.

[0388] In a further highly preferred embodiment, the compositions of the present invention comprise, and preferably are, SY-0498.

[0389] In a further highly preferred embodiment, the compositions of the invention comprise, and preferably are, SY-0499.

[0390] In a further highly preferred embodiment, the compositions of the present invention comprise, and preferably are, SY-0487.

[0391] In a further highly preferred embodiment, the compositions of the present invention comprise, and preferably are, SY-0500.

[0392] In a further highly preferred embodiment, the composition of the invention comprises, preferably is, SY-0501.

[0393] In a further highly preferred embodiment, the compositions of the present invention comprise, and preferably are, SY-0502.

[0394] In a further highly preferred embodiment, the compositions of the present invention comprise, and preferably are, SY-0503.

[0395] In a further highly preferred embodiment, the compositions of the present invention comprise, and preferably are, SY-0504.

[0396] In a further highly preferred embodiment, the composition of the present invention comprises, preferably is, SY-0505.

[0397] In a further highly preferred embodiment, the compositions of the present invention comprise, preferably are, SY-0506.

[0398] In a further highly preferred embodiment, the compositions of the present invention comprise, preferably are, SY-0507.

[0399] In a further highly preferred embodiment, the compositions of the present invention comprise, preferably are, SY-0508.

[0400] In a further highly preferred embodiment, the compositions of the present invention comprise, preferably are, SY-0509.

[0401] In a further highly preferred embodiment, the compositions of the present invention comprise, and preferably are, SY-0510.

[0402] In a further highly preferred embodiment, the compositions of the invention comprise, and preferably are, SY-0511.

[0403] In a further highly preferred embodiment, the compositions of the present invention comprise, and preferably are, SY-0512.

[0404] In a further highly preferred embodiment, the compositions of the present invention comprise, and preferably are, SY-0513.

[0405] In a further highly preferred embodiment, the compositions of the present invention comprise, and preferably are, SY-0514.

[0406] In a further highly preferred embodiment, the compositions of the present invention comprise, preferably are, SY-0515.

[0407] In a further highly preferred embodiment, the compositions of the present invention comprise, preferably are, SY-0516.

[0408] In a further highly preferred embodiment, the compositions of the present invention comprise, preferably are, SY-0517.

[0409] In a further highly preferred embodiment, the compositions of the present invention comprise, and preferably are, SY-0518.

[0410] In a further highly preferred embodiment, the compositions of the present invention comprise, and preferably are, SY-0519.

[0411] In a further highly preferred embodiment, the compositions of the present invention comprise, and preferably are, SY-0520.

[0412] In a further highly preferred embodiment, the compositions of the invention comprise, and preferably are, SY-0521.

[0413] In a further highly preferred embodiment, the compositions of the present invention comprise, and preferably are, SY-0522.

[0414] In a further highly preferred embodiment, the compositions of the present invention comprise, and preferably are, SY-0523.

[0415] In a further highly preferred embodiment, the compositions of the invention comprise, and preferably are, SY-0524.

[0416] In a further highly preferred embodiment, the compositions of the present invention comprise, preferably are, SY-0525.

[0417] In a further highly preferred embodiment, the compositions of the present invention comprise, preferably are, SY-0526.

[0418] In a further highly preferred embodiment, the compositions of the present invention comprise, preferably are, SY-0527.

[0419] In a further highly preferred embodiment, the compositions of the invention comprise, and preferably are, SY-0543.

[0420] Methods of Treating Disease The compositions described herein can be used in methods for treating diseases. In some embodiments, the disease is a disease that can be treated using exon-skipping oligomeric compounds. In some embodiments, the disease is a disease that can be treated using antisense-mediated exon inclusion oligomeric compounds. In embodiments of the present invention, the compositions described herein cross the blood-brain barrier and are therefore useful in treating central nervous system diseases, behavioral disorders, psychiatric disorders, and / or behavioral symptoms of diseases. In some embodiments, the disease is a central nervous system (CNS) disease. In some embodiments, the disease is amyotrophic lateral sclerosis (ALS), Alzheimer's disease (AD), Parkinson's disease (PD), multiple sclerosis (MS), epilepsy, Creutzfeldt-Jakob disease (CJ), Menkes disease, or Huntington's disease (HD). In some embodiments, the disease is a disease affecting cerebellar function, including, but not limited to, ataxia. In some embodiments, the disease is a disease affecting the function of the amygdala, including but not limited to Urbach-Wiete disease. In some embodiments, the disease is a disease affecting the function of the hippocampus, including but not limited to memory loss. In some embodiments, the disease to be treated is a mental or behavioral disorder, including but not limited to mood disorders, dementia, anxiety, bipolar disorder, schizophrenia, sleep disorders, post-traumatic stress disorder (PTSD), attention deficit hyperactivity disorder (ADHD), and depressive disorders.

[0421] In some embodiments of the present invention, the oligonucleotides of the present invention are used to treat sleep disorders and / or cognitive disorders and their associated symptoms. In some embodiments, the sleep disorder is insomnia or slow-wave sleep disturbance. In some embodiments, the cognitive disorder is schizophrenia. When the disease being treated is schizophrenia, both the positive and negative symptoms of schizophrenia can be treated. In some embodiments, the positive symptoms of schizophrenia are hallucinations, delusions, or disturbances in logical thinking. In some embodiments, the negative symptoms of schizophrenia include lack of motivation, lack of spontaneity, inability to think abstractly, lack of mood expression, lack of cognition, lack of ability to feel pleasure, flattened emotions, allopathic behavior, loss of motivation, dysphoric moods including anger, anxiety, and depression, disrupted sleep patterns, poor impulse control, poor judgment, abnormal psychomotor activity such as pacing or locking, and movement disorders such as tardive dyskinesia. In some embodiments, cognitive domains such as verbal memory, verbal fluency, memory consolidation, and executive function are improved by administering one or more oligonucleotide compounds of the present invention. In some embodiments, administration of one or more oligonucleotide compounds of the present invention increases slow wave sleep, thereby improving cognition. In some embodiments, the disease is selected from the group consisting of Duchenne muscular dystrophy (DMD), familial dysautonomia, spinal muscular atrophy (SMA), ataxia-telangiectasia, congenital disorders of glycosylation, frontotemporal dementia (FTD), Parkinsonism linked to chromosome 17, Niemann-Pick disease type C, neurofibromatosis type 1, neurofibromatosis type 2, macrocephalic leukoencephalopathy with subcortical cysts type 1, Pelizaeus-Merzbach disease, Pompe disease, and myotonic dystrophy type 1.Methods of using tc-DNA oligonucleotides for the treatment of Duchenne muscular dystrophy (DMD), spinal muscular atrophy (SMA), spinocerebellar ataxia type 3 (SCA3), and other diseases are known in the art and are described in, for example, U.S. Pat. Nos. 4,981,957; 5,118,800; 5,319,080; 5,359,044; 5,393,878; 5,446,137; 5,466,786; 5,514,785; 5,519,134; 5,567,811; 5,576,427; 5 ,591,722; U.S. Patent No. 5,597,909; U.S. Patent No. 5,610,300; U.S. Patent No. 5,627,053; U.S. Patent No. 5,639,873; U.S. Patent No. 5,646,265; U.S. Patent No. 5,670,633; U.S. Patent No. 5,700,920; U.S. Patent No. 5,792,847; and U.S. Patent No. 6,600,032; and U.S. Patent Application Publication Nos. 2015 / 0141637, 2016 / 0002280, 2014 / 0296323, and 2012 / 0149756, the disclosures of which are incorporated herein by reference.

[0422] Table A provides a list of certain neurodegenerative diseases and their targets for which the compositions of the present invention are useful. [Table A]

[0423] The effectiveness of the compositions described herein in treating, preventing, and / or managing the indicated diseases or disorders can be tested using various models known in the art, which provide guidance for the treatment of human diseases. Models for diseases that can be treated using exon-skipping oligomeric compounds are described, for example, in Siva et al., Nucleic Acid Therapeutics, 2014, vol. 24, pp. 69-86. Models for diseases that can be treated using antisense-mediated exon inclusion oligomeric compounds are described, for example, in Hua and Krainer, Methods Mol. Biol., 2012, vol. 867, pp. 307-323.

[0424] Genetic animal models for DMD are known in the art. The mdx mouse carries a nonsense mutation in exon 23 of the dystrophin gene, which prevents the synthesis of full-length wild-type dystrophin protein. Grounds et al., Neurobiol. Dis., 2008, 31, 1-19. The GRMD (Golden Retriever Muscular Dystrophy) canine model lacks functional dystrophin due to a splice site mutation in intron 6 that disrupts the reading frame. In the GRMD model, similar to human DMD, progressive fiber degradation leads to the collapse of the skeletal musculature, with significant endomysial and perimysial fibrosis. Other models for DMD include dystrophin / utrophin double knockout mice, humanized DMD mice, mdx52 mice (which carry a deletion of exon 52 in mouse DMD), and 4CV mice (which carry a nonsense mutation in exon 53). Goyenvalle et al., Mol. Ther., 2010, 18, 198-05; Bremmer-Bout et al., Mol. Ther., 2004, 10, 232-240; Aoki et al., Mol. Ther., 2010, 18, 1995-2005; Mitrpant et al., J. Gene. Med., 2009, 11, 46-56.

[0425] Spinal muscular atrophy (SMA) is a class of genetic disorders resulting from deletion of the survival motor neuron gene (SMN1), which maps to chromosome 5q11.2-13.3. Overall, SMA is characterized by the loss of motor neurons in the spinal cord and brainstem, resulting in muscle atrophy due to loss of nerve contact. Each type of SMA has an incidence of approximately 1 in 6,000. Type 1 SMA, also known as Werdnig-Hoffmann disease or severe childhood SMA, affects infants in the first year of life and is generally fatal. Type 2 SMA, also known as moderate SMA, affects children, causing muscle weakness that prevents patients from standing or walking, although they may be able to sit. SMA Type 3 patients are able to walk at some point in their development.

[0426] SMA is caused by the loss of a functional SMN1 gene and a mutation in exon 7 of the SMN2 paralogue, which causes substantial skipping of exon 7 and the production of only low levels of functional protein, such that the SMN2 protein cannot compensate for the loss of SMN1. Cartegni et al., Am. J. Hum. Genet., 2006, 78:63-77. Oligonucleotide-mediated exon inclusion methods for the treatment of SMA have been explored, including methods to compensate for deleterious mutations in SMN2 by masking intronic silencing sequences and / or terminal stem-loop sequences within the SMN2 gene to obtain a modified, functional SMN2 protein containing the amino acid sequence encoded by exon 7, which is capable of at least partially compensating for the non-functional SMN1 protein. See, for example, International Publication No. 2010 / 115993A1, the disclosure of which is incorporated herein by reference. In addition to SMA, numerous other diseases, including those described herein, can potentially be treated by the exon inclusion approach provided by the compositions of the present invention.

[0427] Accordingly, the present invention provides a pharmaceutical composition comprising the composition of the present invention and further comprising a pharmaceutically acceptable carrier, preferably the pharmaceutical composition being for use in the prevention, treatment or diagnosis of a neuromuscular disease or a musculoskeletal disease, more preferably the neuromuscular disease or the musculoskeletal disease being selected from the group consisting of Duchenne muscular dystrophy, familial dysautonomia, spinal muscular atrophy, ataxia-telangiectasia, congenital disorders of glycosylation, frontotemporal dementia, parkinsonism linked to chromosome 17, Niemann-Pick disease type C, neurofibromatosis type 1, neurofibromatosis type 2, and dermatomyositis. and myotonic dystrophy type 1 (DM1, or Steinert disease), and even more preferably, the neuromuscular disease or musculoskeletal disease is selected from Duchenne muscular dystrophy, spinal muscular atrophy, Pompe disease, myotonic dystrophy type 2 (DM2, or proximal myotonic myopathy), and myotonic dystrophy type 1 (DM1, or Steinert disease).

[0428] The present invention further relates to a pharmaceutical composition of the present invention, which comprises the composition of the present invention and further comprises a pharmaceutically acceptable carrier, preferably wherein said pharmaceutical composition is for use in treating a neuromuscular disease or a musculoskeletal disease, more preferably wherein said neuromuscular disease or said musculoskeletal disease is selected from the group consisting of Duchenne muscular dystrophy, familial dysautonomia, spinal muscular atrophy, ataxia-telangiectasia, congenital disorders of glycosylation, frontotemporal dementia, parkinsonism linked to chromosome 17, Niemann-Pick disease type C, neurofibromatosis type 1, neurofibromatosis type 2, subcortical capsule and myotonic dystrophy type 1 (DM1, or Steinert disease), and again more preferably, said neuromuscular disease or said musculoskeletal disease is selected from Duchenne muscular dystrophy, spinal muscular atrophy, Pompe disease, myotonic dystrophy type 2 (DM2, or proximal myotonic myopathy) and myotonic dystrophy type 1 (DM1, or Steinert disease).

[0429] The present invention further relates to the composition of the present invention for use as a medicament in the prevention, treatment or diagnosis of a disease, preferably wherein said disease is a neuromuscular disease or a musculoskeletal disease, more preferably wherein said neuromuscular disease or said musculoskeletal disease is Duchenne muscular dystrophy, familial dysautonomia, spinal muscular atrophy, ataxia-telangiectasia, congenital disorders of glycosylation, frontotemporal dementia, parkinsonism linked to chromosome 17, Niemann-Pick disease type C, neurofibromatosis type 1, neurofibromatosis type 2, macrocephalic white matter with subcortical cysts, and myotonic dystrophy type 1 (DM1, or Steinert disease), and again more preferably, said neuromuscular disease or said musculoskeletal disease is selected from Duchenne muscular dystrophy, spinal muscular atrophy, Pompe disease, myotonic dystrophy type 2 (DM2, or proximal myotonic myopathy) and myotonic dystrophy type 1 (DM1, or Steinert disease). [Example]

[0430] The embodiments encompassed herein will now be described with reference to the following examples, which are provided for illustrative purposes only, and the disclosure encompassed herein should in no way be construed as being limited to these examples, but rather as encompassing any and all variations that become evident as a result of the teachings provided herein. material and method

[0431] Animals: The mdx mouse (Bulfield et al., Proc. Natl. Acad. Sci. USA, 1984, Vol. 81, pp. 1189-1192; Ryder-Cook et al., EMBO J., 1988, Vol. 7, pp. 3017-3021) contains a single base substitution in exon 23 of the dystrophin gene (Sicinski et al., Science, 1989, Vol. 244, pp. 1578-1580) that causes premature termination of the polypeptide chain, resulting in the failure to produce the full-length 427 kDa muscle isoform of dystrophin. However, other isoforms produced from a different promoter (located 3' to the point mutation) are unaffected. Mutants are viable and fertile. There are no obvious symptoms, and the muscle remains mechanically intact throughout life unless damage is caused by mechanical or chemical insults (Reimann et al., Neuromusc. Disord., 2000, 10, 276-282; Connolly et al., Neuromusc. Disord., 2001, 11, 703-712).

[0432] At the histological level, mdx exhibits classic features of dystrophic muscle, characterized by numerous necrotic fibers with subsequent infiltration of scavenger cells (Coulton et al., Neuropathol. Appl. Neurobiol., 1988, vol. 14, pp. 299–314). However, unlike DMD, efficient, unknown compensatory mechanisms counteract degeneration, thus maintaining the regenerative process and repairing the constant mechanical damage. The number of revertant fibers is low, typically present at approximately 1% of all fibers, but their number increases with age in mice (Lu et al., J. Cell. Biol., 2000, vol. 148, pp. 985–995). Exon phasing allows the translation of a truncated dystrophin by skipping exon 23 during mRNA splicing.

[0433] Oligonucleotide synthesis: Oligonucleotides for in vivo experiments were synthesized on a 260 μmol scale using an Aekta Oligo Pilot 10 synthesizer and a NittoPhase® UnyLinker™ 200 universal solid support. For compositions of the present invention containing one or more lipid moieties and / or the spacer at the 3' end, the corresponding modified, typically and preferably Fmoc-protected, phosphoramidite was introduced in the first cycle of the synthesis. A modified synthesis cycle was typically used, using 2.2 equivalents of phosphoramidite and a 4-minute coupling time. For compositions of the present invention containing one or more lipid moieties and / or spacers at the 5' end, the corresponding modified phosphoramidite was used in the last cycle. To remove the Fmoc and cyanoethyl protecting groups, the solid support was treated with 20% diethylamine in MeCN.

[0434] Oligonucleotides for in vitro experiments were synthesized on a 1 μmol scale using the Expedite Nucleic Acids Synthesis System and the universal solid support High Load Glen UnySupport™. For compositions of the present invention containing the lipid moiety and / or spacer at the 3' end, the corresponding modified phosphoramidite was introduced in the first cycle of synthesis. For compositions of the present invention containing the biotin moiety and / or spacer at the 3' end, the corresponding biotin phosphoramidite was introduced in the first cycle of synthesis, followed by a C4 spacer phosphoramidite in the second cycle of synthesis, and the corresponding, typically and preferably Fmoc-protected, spacer phosphoramidite in the third cycle of synthesis. A modified synthesis cycle was typically used, using 9 equivalents of phosphoramidite and a 3.5-minute coupling time. For compositions of the present invention containing one or more lipid moieties and / or spacers at the 5' end, the corresponding modified phosphoramidite was used in the final cycle. To remove the Fmoc and cyanoethyl protecting groups, the solid support was treated with 20% diethylamine in MeCN.

[0435] Deprotection was carried out by heating the solid-supported oligonucleotide with saturated NH4OH at 65°C for 3 hours. The mixture was then cooled to ambient temperature, and 4M NaOH and MeOH were added to a final mixture composition of 0.4M NaOH and 60% MeOH. Treatment was continued for an additional 2.5 hours at ambient temperature. The solid was filtered off, washed with 1:1 EtOH / H2O, and the filtrate was neutralized by adding 2M NaH2PO4. The resulting crude oligonucleotide solution was desalted against water using TFF and lyophilized. For compositions of the invention containing one or more -COOH functional groups, previously described conditions were used (Surzhikov et al., Nucleic Acids Res., 2000, vol. 28, p. e29).

[0436] Oligonucleotides for in vivo experiments were purified by isocratic RP HPLC (Waters x Bridge prep C18, 5 μm, 10 × 150 mm) using 42% MeCN in ammonium acetate buffer (pH 7) at a flow rate of 4 mL / min at 75 °C. Fractions containing sufficiently pure product (>70% FLP) were combined and partially lyophilized. Approximately 20 equivalents of NaCl were added to the solution, which was subsequently desalted against water using TFF.

[0437] Oligonucleotides for in vitro experiments were purified by isocratic RP HPLC (Waters x Bridge prep C18, 5 μm, 4.6 × 150 mm) using approximately 30% iPrOH in ammonium bicarbonate buffer (pH 7) at a flow rate of 1 mL / min at 60 °C. Fractions containing sufficiently pure product were combined and subsequently desalted using SEC (GE Healthcare, HiPrep 26 / 10) with water as eluent.

[0438] Linking lipid moiety to oligomeric compound: According to the present invention, there are several approaches to link one or more lipid moieties to oligonucleotide, either directly on solid support or in solution.These approaches are based on the classical conjugation chemistry that is well known to those skilled in the art and widely described in the literature (Bioconjugate Techniques, Greg T. Hermanson, Pierce Biotechnology; Singh, Chem. Soc. Rev., 2010, 39, 2054-2070; H. Rosemeyer, Chem. Biodiversity, 2005, 2, 977-1062).As shown, for the composition of the present invention that comprises one or more lipid moieties and / or spacers at the 3'-end or 5'-end, corresponding modified phosphoramidites are introduced, typically and preferably Fmoc-protected or MMTr-protected, in the first or last cycle of synthesis.

[0439] Formula (I)AB- * (wherein the asterisk ( * Typical and preferred linking procedures for linking said one or more lipid moieties of (wherein (a) represents said point of covalent attachment to said oligomeric compound or said spacer according to the present invention) include, but are not limited to: i. When B is preferably OP(O)(OH), OP(O)(SH), and more preferably OP(O)(SH), the 5' end of the oligomeric compound is linked to the oligonucleotide, respectively, according to the following reaction scheme: 5'-HO-OLIGO-3'+AOP(O-PG)-X → 5'-AOP(O-PG)-O-OLIGO-3'+HX P(III) is subsequently oxidized to P(V), where X is a suitable leaving group and PG is a suitable protecting group; ii. When B is preferably NH-P(O)(OH), the 5'-end of the oligomeric compound is linked to the oligonucleotide, respectively, via phosphoramidate chemistry according to the following reaction scheme: 5'-HO-OLIGO-3' → 5'-H-(O-PG)(O)PO-OLIGO-3' 5'-H-(O-PG)(O)PO-OLIGO-3' + H2N-A → 5'-A-NH-(O-PG)(O)PO-OLIGO-3' (where PG is a suitable protecting group); iii. When B is preferably NH-P(S)(OH), the 5' ends of the oligomeric compounds and the oligonucleotides are linked via phosphoramidate chemistry according to the following reaction scheme: 5'-HO-OLIGO-3' → 5'-XP(O-PG)-O-OLIGO-3' 5'-XP(O-PG)-O-OLIGO-3' + H2N-A → 5'-A-NH-P(O)(O-PG)-O-OLIGO-3' Subsequently, P(III) is oxidized to P(V) using a suitable sulfur transfer reagent, where PG is a suitable protecting group; iv. When B is preferably NH—C(O), the 5′ end of the oligomeric compound is linked to the oligonucleotide, respectively, via carbamate chemistry according to the following reaction scheme: 5'-HO-OLIGO-3' → 5'-XC(O)-O-OLIGO-3' 5'-XC(O)-O-OLIGO-3' + H2N-A → 5'-A-NH-C(O)-O-OLIGO-3' + HX (where X is a suitable leaving group); or 5'-HO-OLIGO-3' + OCN-A → 5'-A-NH-C(O)-O-OLIGO-3'.

[0440] The ligation procedures described above are also applicable to ligation to the 3' ends of oligomeric compounds and oligonucleotides, typically during reverse 5' to 3' oligomeric compound and oligonucleotide synthesis, respectively.

[0441] As an example: for the synthesis of preferred compositions of the invention, such as SY-0457, SY-0458, SY-0459, and SY-0460, lipid moieties were conjugated by reaction of the amino-modified oligonucleotide with the corresponding NHS-ester on a solid support using 10 equivalents of the NHS-ester in DMSO in the presence of DIPEA (15 equivalents), or in solution using 3 x 10 equivalents of the NHS-ester in 1:1 DMSO / buffer (0.1 M NaHCO3 pH 8.5) at 37°C.

[0442] As a further example: for the synthesis of preferred compositions of the invention such as SY-0299, SY-0343, SY-0442, and SY-0455, which comprise a palmitoyl residue as one lipid moiety linked to the oligomeric compound of SEQ ID NO: 1 (having a complete PO internucleoside linkage group) via either an -NH-C6 alkylene-OP(O)(SH)- or -NH-C6 alkylene-OP(O)(OH)- spacer to the 5'-terminal OH group of the oligomeric compound, 5'-Palmitate-CE-Phosphoramidite (Link, P / N 2199) was used as the phosphoramidite in the final cycle. The moiety attached to the 3' terminus of SY-0343 and SY-0299, i.e., -P(O)(SH)O-CHCH-(CHOH)-(CH)NH, was introduced in the first cycle of synthesis by coupling the corresponding Fmoc-protected phosphoramidite to the 3'-OH group of the first nucleotide.

[0443] Certain preferred compositions of the present invention and compositions used in the experimental section are specifically characterized in Table 3 by further reference to their respective names and abbreviations typically used throughout the specification.

[0444] Analysis of Oligonucleotides by RP-HPLC-DAD-MS: The following parameters were used for analytical HPLC: A C18 column with a 1.7 μm particle size was used. The column temperature was set to 75 °C. Mobile phase A was 400 mM hexafluoroisopropanol (HFIP) and 15 mM triethylamine + 10% methanol. Mobile phase B was methanol. A gradient of 32-52% mobile phase B was applied. The flow rate was set to 0.25 mL / min. Oligonucleotides were detected using a UV spectrometer at 260 nm and a time-of-flight mass spectrometer.

[0445] Detection of multimers: Polyacrylamide gel electrophoresis (PAGE) experiments were performed to detect self-multimerization. The following chemicals were used: tris(hydroxymethyl)aminomethane (Tris) (TCI A0321); acetic acid (Merck 1.00063); acrylamide / Bis solution, 29:1 (40%, Serva 10680.01); tetramethylethylenediamine (TEMED, Sigma-Aldrich T9281); ammonium persulfate (Sigma-Aldrich 248614); and glycerol (Sigma-Aldrich G9012). Buffer A was prepared by dissolving 60 g of Tris in 200 mL of water. The pH was adjusted to 7.4 with glacial acetic acid (approximately 25-30 mL). The solution was diluted to 500 mL with water and stored at +4 °C. Buffer B was prepared by diluting 16 mL of buffer A to 800 mL with water. Ammonium persulfate 10% (w / v) was prepared by dissolving 100 mg of ammonium persulfate in 0.9 mL of water. Gels were prepared by mixing the following solutions in a glass beaker: 9.4 mL of acrylamide / bis solution; 15 mL of water; 500 μL of buffer A; 125 μL of 10% (w / v) ammonium persulfate; and 38 μL of TEMED. The test solution was 1 mg / mL in 10% glycerol, and 10 μL was applied (equivalent to 10 μg of oligonucleotide). Pre-migration settings were 40 min / 90 V in buffer B. Migration settings were 45–60 min / 120 V in buffer B, in addition to 90 min / 90 V or 15 min / 90 V. 5–7 μL of 6× DNA loading dye was also transferred. Detection was performed by placing the gel on a TLC plate and examining it under UV light at 254 nm. Staining with Stains-All (Sigma-Aldrich, 1-ethyl-2-[3-(1-ethylnaphtho[1,2- d ]thiazolin-2-ylidene)-2-methylpropenyl]naphtho[1,2- d ]thiazolium bromide, 3,3′-diethyl-9-methyl-4,5,4′,5′-dibenzothiacarbocyanine) was then performed according to the manufacturer's protocol.

[0446] Isolation of oligonucleotide-interacting proteins: Blood protein isolation was performed using biotinylated oligonucleotides immobilized on streptavidin beads (High Capacity Streptavidin Agarose, Pierce). Biotinylated oligonucleotides were immobilized on the beads (10 μl of beads for 20 μg of oligonucleotide) for 30 min, and excess oligonucleotide was removed by washing three times with phosphate-buffered saline (PBS). The beads thus prepared were incubated with 5–50 μl of mouse or human serum for 1 h, followed by four successive washes in PBS to remove unbound proteins. After the final wash, precipitated proteins were either dissolved in Laemmli sample buffer for further analysis by SDS-PAGE or directly digested with trypsin for analysis by mass spectrometry on an ORBI-TRAP instrument.

[0447] Commercially available human serum (SIGMA, St. Louis, MO) or serum samples from healthy human adults obtained in accordance with regulatory guidelines were used in the experiments. Blood samples from C57BL / 6 mice were collected from the tail vein or retro-orbital plexus. All procedures involving animals were performed in accordance with the guidelines of our institution's animal ethics committee.

[0448] Identification of captured proteins by mass spectrometry using OrbiTrap technology: For mass spectrometry analysis, precipitated proteins were directly digested with trypsin in a buffer containing 100 mM ammonium carbonate pH 8.0 and 500 ng of trypsin (Sequence Grade Trypsin, Promega) at 37°C for 16 h and stored at -20°C until use. The peptide mixture was desalted using ZipTip μ-C18 pipette tips (Millipore) and separated on an Easy nano-LC Proxeon system (Thermo Fisher Scientific) equipped with a reversed-phase C18 column (Easy-Column Proxeon C18, L 15 cm, ID 75 μm). The eluate was monitored on an LTQ Velos Orbitrap mass spectrometer (Thermo Fisher Scientific), and tandem MS (MS / MS) data were processed using Proteome Discoverer 1.4 software (Thermo Fisher Scientific) coupled with an in-house Mascot search server (Matrix Science, version 2.3.2) using the SwissProt database, as previously described (Rouillon, 2015). The relative abundance of each identified protein was estimated by label-free quantification using Progenesis LC-MS software (Nonlinear Dynamics, version 4.0). The average normalized abundance (ANA), which reflects the relative abundance of a protein in the Progenesis analysis, was used to compare the amount of protein bound to the vector.

[0449] Protein identification after SDS-PAGE separation: Protein identification was performed as previously described (Denard, Proteomics, 2009, vol. 9, pp. 3666-3676). After separating isolated proteins by SDS-PAGE (4-12% gradient, NuPAGE Novex Bis-Tris Gel 1.0 mm, Life Technologies), the gel was visualized by staining with Coomassie Blue (InstantBlue Protein Stain, Expedion), and bands of interest were sliced ​​for further analysis. After washing the gel slices in 1 ml of 96% CH2CHOH for 10 min, the alcohol was carefully removed and replaced with 20 μl of a solution containing 100 mM ammonium carbonate pH 8.0 and 100 ng of trypsin (Promega). Samples were digested at 37°C for 16 h and stored at -20°C until use. The peptide mixture was desalted using ZipTip μ-C18 pipette tips (Millipore) and deposited onto a MALDI plaque with α-cyano-4-hydroxycinnamic acid (HCCA) (5 μg) matrix in 80% acetonitrile, 0.1% formic acid.

[0450] MALDI-TOF MS analysis was performed on a MALDI TOF / TOF ABI 4800+ (AB Sciex). All spectra were acquired in positive reflector mode. The resulting mass list was searched using an in-house Mascot search server (Matrix Science, version 2.3.2 213) and the SwissProt database for human / mouse, one residual cleavage site considering partial oxidation of methionine, and a mass tolerance setting of 50 ppm.

[0451] Systemic administration of AONs: Animal procedures were performed in accordance with national and European legislation and approved by the French government (Ministère de l'Enseignement Superieur et de la Recherche, Autorisation APAFiS #6518). Mdx (C57BL / 10ScSc-Dmdmdx / J) and C57BL / 10 mice were bred in our animal facility at Platform 2Care, UFR des Sciences de la Santé, Universite de Versailles Saint Quentin, and maintained on a standard 12-h light / dark cycle with free access to food and water. Mice were weaned at 4–5 weeks postnatal age and housed 2–5 individuals per cage.

[0452] Six- to eight-week-old mdx mice were intravenously injected with different AONs (15-mer or 13-mer, PO or PS, conjugated or unconjugated) into the retroorbital sinus under general anesthesia using 1.5-2% isoflurane once a week for periods ranging from 4 to 12 weeks. Age-matched C57 / BL10 (WT) and mdx mice were included as controls, receiving an equivalent volume of sterile saline. One hour after the first injection, blood samples were collected from all mice to measure complement C and cytokine / chemokine levels. Additional blood samples were collected one week after the sixth injection (mid-treatment) and one week after the end of treatment. Mice were then placed in metabolic cages for urine collection over a 24-hour period. Two weeks after the last injection, animals were sacrificed. Muscles and tissues were harvested, flash-frozen in liquid nitrogen-cooled isopentane, and stored at -80°C until further analysis. Sample size and n values ​​are indicated in each figure legend. Researchers were blinded to RNA and protein analysis.

[0453] Serum and urine analysis: Blood samples were collected by tail bleeding under general anesthesia. Analysis of serum creatine kinase (CK), alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), bilirubin, creatinine, urea, and albumin levels was performed by the pathology laboratory at the Mary Lyon Centre, Medical Research Council, Harwell, Oxfordshire, UK. Serum cytokine and chemokine levels were analyzed by multiplex assay using Luminex® technology. The Bio-Plex Pro Mouse Cytokine 10-Plex Immunoassay Panel (Bio-Rad, Hercules, CA) was used to detect IL-1β, IL-6, IL-10, IL-12p70, IL-13, IL-17, IFN-γ, MCP-1, RANTES, and TNF-α levels according to the manufacturer's instructions. Immunoassays were read using a Bio-Plex MAGPIX Multiplex reader, and results were analyzed with Bio-Plex manager 6.1 software (Bio-Rad, France).

[0454] Urine was collected directly into refrigerated tubes (4°C) using metabolic cages over a 24-hour period. After collection, urine was centrifuged at 10,000 × g for 10 minutes, and the supernatant was aliquoted and frozen at -80°C for further analysis. Urinary creatinine was measured using a creatinine assay kit (R&D Systems, Inc., Minneapolis, MN) according to the manufacturer's instructions. Total protein in urine samples was measured as previously described (Swayze et al., Nucleic Acids Res., 2007, 35, 687-700). Briefly, protein was precipitated from urine samples by adding 40 μL of dH2O and 200 μL of pre-chilled acetone to 10 μL of urine. Samples were then incubated at -20°C for 30 minutes before being centrifuged at 14,000 × g for 15 minutes at 4°C. The pellet was resuspended in 40 μl of dH2O, and protein concentration was measured using a Pierce BCA assay (Thermo Scientific, Rockford, IL). Albumin from urine samples was measured using an albumin ELISA kit (Bethy Laboratories, Montgomery, TX) according to the manufacturer's instructions. Acute kidney injury (AKI) biomarker levels were analyzed by multiplex assay using Luminex® technology. The Multiplex Kidney Injury Panel (MKI1MAG-94K, MKI2MAG-94K, Merck-Millipore) was used according to the manufacturer's instructions to measure the levels of beta-2-microglobulin (B2M), renin, kidney injury molecule 1 (KIM-1), interferon gamma-inducible protein 10 (IP-10), vascular endothelial growth factor (VEGF), cystatin C, epidermal growth factor (EGF), lipocalin-2-NGAL, clusterin, and osteopontin (OPN). Results were read using a Bio-Plex MAGPIX Multiplex reader and analyzed with Bio-Plex manager 6.1 software (Bio-Rad, France).

[0455] Complement activation assay: Complement activation in mouse serum samples was measured by Microvue PanSpecific-C3 Converter and SC5b-9 Plus Kit (Quidel Co., San Diego, CA, USA). Briefly, mouse C3 protein was converted to human SC5b9 using the C3 converter reagent (Pan specific C3 reagent kit, Microvue, Quidel), and then detected by SC5b9 Elisa (Quidel).

[0456] For in vitro complement activation studies, AON molecules were incubated with pooled normal human serum (1:10) (Seralab, UK) at 37°C for 45 minutes. Complement activation was assessed using a human SC5b-9 Plus kit (Quidel Co., San Diego, CA, USA). 5 mg / ml zymosan (Complement Technology, Inc., Texas, USA) was used as a positive control. Complement activation was expressed as a percentage of the remaining C3 level in the sample, considering the C3 level in the PBS condition as 100% (no activation).

[0457] Clotting assay: Mouse blood samples were collected in tubes containing 3.2% sodium citrate. Whole blood samples were centrifuged at 2,500 × g for 15 minutes, after which plasma was immediately separated and stored at -80°C until use. After incubating 2 mg / mL tcDNA-AON with 50 μL of citrated plasma C at 37°C for 30 minutes, prothrombin time (PT) and activated partial thromboplastin time (aPTT) assays were performed using a semi-automated STARTmax coagulometer (Stago) according to the manufacturer's instructions.

[0458] RNA analysis: Total RNA was isolated from interstitial muscle sections collected during cryosectioning using TRIzol reagent according to the manufacturer's instructions (ThermoFisher Scientific, USA). An aliquot of 500 ng of total RNA was used for RT-PCR analysis using the Access RT-PCR System (Promega, USA) in a 50 μL reaction using the following external primers: Ex 20Fo (5'-CAGAATTCTGCCAATTGCTGAG-3' - SEQ ID NO: 38) and Ex 26Ro (5'-TTCTTCAGCTTGTGTCATCC-3' - SEQ ID NO: 39).

[0459] cDNA synthesis was carried out at 45°C for 45 minutes, followed directly by 30 cycles of primary PCR at 95°C (30 seconds), 55°C (1 minute), and 72°C (2 minutes). 2 μL of these reactions were then reamplified in a nested PCR with 22 cycles of 95°C (30 seconds), 55°C (1 minute), and 72°C (2 minutes) using the following internal primers: Ex 20Fi (5'-CCCAGTCTACCACCCTATCAGAGC-3' - SEQ ID NO: 40) and Ex 26Ri (5'-CCTGCCTTTAAGGCTTCCTT-3' - SEQ ID NO: 41).

[0460] PCR products were analyzed on 2% agarose gels. Exon 23 skipping was also measured by Taqman quantitative RT-PCR as previously described (Goyenvalle et al., Hum. Mol. Genet., 2012, 21:2559-2571; Straub et al., Lancet Neurol., 2016, 15:882-890).

[0461] Total RNA was also isolated from renal cortex samples as previously described (Frazier et al., Toxicol. Pathol., 2014, 42, 923–935) and quantitative RT-PCR was performed for the following kidney injury biomarker (KIB) genes: interferon gamma (IFNg), interleukin 6 (IL6), granzyme B (Gzmb), interferon gamma-inducible protein 10 (IP-10), tumor necrosis factor (TNF), chemokine ligand 2 (Ccl2), chemokine ligand 3 (Ccl3), beta-2-microglobulin (B2M), kidney injury molecule 1 (KIM-1), renin 1 (Ren1), and epidermal growth factor (EGF).

[0462] Statistical analysis: Data were analyzed with GraphPad Prism5 software (San Diego, California, USA) and are presented as mean ± SEM. "n" refers to the number of mice per group. Comparisons of statistical significance were assessed by the non-parametric Mann-Whitney U test. The significance level was * P<0.05, ** P<0.01, *** P was set at <0.001.

[0463] As described herein, in a preferred embodiment, the oligomeric compounds contained in the compositions of the present invention are antisense oligonucleotides (AONs) designed to be complementary to specific mRNAs or pre-mRNAs. This preferred class of compounds of the present invention can be used to treat many diseases. The exemplary diseases provided below do not limit the present invention, and the compositions of the present invention provided can be used to treat any disease that can be treated by administering AONs.

[0464] Example 1 Compositions of the Invention for the Treatment of Duchenne Muscular Dystrophy Efficacy assessment Adult mdx mice were treated weekly for 4 weeks by intravenous injection of various 13-mer AONs targeting the donor splice site (M23D:+2-11) of exon 23 of the dystrophin pre-mRNA: SY-0308, SY-0210, and SY-0299, SY-0343, SY-0442, and SY-0455 of the present invention. SY-0308 (also referred to interchangeably herein as "tcDNA-PO M23D") corresponds to p-CCTCGGCTTACCT-OH of SEQ ID NO: 1, in which all nucleotides are tcDNA, all internucleoside linkages are phosphorodiester linkages, and p is the 5'-terminal phosphate moiety. SY-0210 (also referred to interchangeably herein as "tcDNA-PS M23D") corresponds to p-CCTCGGCTTACCT-OH of SEQ ID NO: 1, in which all nucleotides are tc-DNA, all internucleoside linkages are phosphorothioate linkages, and p is the 5'-terminal phosphate moiety. Composition SY-0343 of the present invention is referred to interchangeably herein as "Palm-2PS-tcDNA-PO M23D" and is shown below: [ka]

[0465] Composition SY-0442 of the present invention is referred to interchangeably herein as "Palm-1PS-tcDNA-PO M23D" and is shown below: [ka]

[0466] Composition SY-0299 of the present invention is referred to interchangeably herein as "Palm-2PO-tcDNA-PO M23D" and is shown below: [ka]

[0467] Composition SY-0455 of the present invention is referred to interchangeably herein as "Palm-1PO-tcDNA / 2OMe-PO M23D" and is shown below: [ka]

[0468] SY-0308, SY-0210, SY-0299, SY-0442, and SY-0455 were administered at a dose of 200 mg / kg body weight per week, and SY-0343 was administered at a dose of 178 mg / kg body weight per week. Two weeks after the final injection, muscles were harvested, and RNA samples were analyzed by quantitative RT-PCR to determine the level of exon 23 skipping of the dystrophin gene (Figure 1). The restored dystrophin levels were quantified by Western blot analysis using the Licor Odyssey system (Figure 5). Figure 1 shows that the level of skipping was consistently higher with palmitoyl-conjugated AONs than with tcDNA-PO M23D. The level of exon 23 skipping was higher in most tissues from mice treated with palmitoyl-conjugated AONs compared with those treated with tcDNA-PS M23D AONs, with the most notable differences observed in the heart, cortex, and cerebellum. The percentage of dystrophin mRNA that skipped exon 23 exceeded 15% in several skeletal muscles and the heart. In the heart, the fold changes relative to SY-0308 or SY-0210 for SY-0299, SY-0343, SY-0442, and SY-0455 were 4.4–9.2 and 4.0–8.5, respectively. Importantly, skipping was clearly detected in the cortex and cerebellum immediately after 4 weeks of treatment with the palmitoyl-conjugated AON, but not yet detected with the other two compounds.

[0469] Thus, palmitoyl-conjugated tcDNA AONs, in which the internucleoside linkage group is PO, are more efficient than naked tcDNA equivalent AONs with or without PS linkages.

[0470] This example demonstrates that systemic delivery of a composition of the invention comprising an antisense oligomer linked to the 5'-palmitoyl-C6-amino lipid moiety and a spacer and having an intact PO backbone (palmitoyl-conjugated PO M23D) enabled greater rescue of dystrophin compared to an equivalent sequence made of tcDNA having either a PO or PS backbone.

[0471] Toxicity assessment No significant clinical signs were observed during the course of experiments with three AONs targeting the (M23D:+2-11) region of dystrophin in mice. However, acute adverse effects have been observed with other sequences of interest, such as the one targeting exon 51 of human dystrophin (H51:+67+81 - AAGATGGCATTTCTA - SEQ ID NO: 19), herein referred to as SYN51. Therefore, SY-0206, also referred to interchangeably herein as "tcDNA-PS SYN51" and corresponding to p-AAGATGGCATTTCT-OH of SEQ ID NO: 19, in which all nucleotides are tcDNA, all internucleoside linkages are phosphorothioate linkages, and p is the phosphate moiety at the 5' end, threatens further clinical development. Historically, PO internucleoside linkages were replaced with PS internucleoside linkages to stabilize oligomers and prevent their degradation. Because tcDNA oligonucleotides are highly stable, modification and replacement of PO with PS modifications is not a priori necessary. Nevertheless, as shown in Figure 1, such modifications significantly increased the biodistribution of tcDNA-PS after systemic delivery and enabled a superior and more extensive skipping effect compared to tcDNA-PO. This phenomenon was likely due to the increased interaction of PS residues with serum proteins.

[0472] One of the major acute effects of toxic tcDNA-PS AONs was that they caused blood coagulation, thrombosis, and complement activation.

[0473] Here, we used two complementary blood coagulation tests, prothrombin time (PT) and partial thromboplastin time (aPTT), to check for potential bleeding problems caused by various tcDNA AONs, namely, SY-0308 (tcDNA-PO M23D), SY-0210 (tcDNA-PS M23D), SY-0343 (Palm-2PS-tcDNA-PO M23D), SY-0442 (Palm-1PS-tcDNA-PO M23D), SY-0455 (Palm-1PO-tcDNA / 2OMe-PO M23D), SY-0206 (tcDNA-PS SYN51), and SY-0252. SY-0252, also referred to interchangeably herein as "tcDNA-PO SYN51," corresponds to p-AGATGGCATTTCT-OH of SEQ ID NO: 19, in which all nucleotides are tcDNA, all internucleoside linkages are phosphorodiester linkages, and p is the 5'-terminal phosphate moiety. SYN51 is known to be highly toxic to mice in the presence of PS binding.

[0474] Figure 2 shows that the PT and aPTT profiles were very similar in mouse and human plasma. In both cases, tcDNAs containing PO linkages (tcDNA-PO M23D, tcDNA-PO SYN51, and palmitoyl-conjugated AONs) did not significantly alter the prothrombin time (PT). However, tcDNAs containing PS linkages, especially tcDNA-PS SYN51, increased the PT and consistently caused acute toxicity in mice after intravenous injection. Furthermore, tcDNA-PS AONs also strongly increased the aPTT, particularly in human plasma, similar to tcDNA-POs containing the SYN51 sequence (Figure 2D). Considering the clinical signs induced by various AONs, it became clear that the PT test reflected the magnitude of acute adverse effects, at least in mice. All tcDNA-PO AONs were safe in mice and did not increase the PT. On the other hand, tcDNA-PS M23D caused only a slight increase in PT and toxicity was never noted, whereas tcDNA-PS SYN51 caused a strong increase in PT and was highly toxic.

[0475] Figure 8 shows that PT and aPTT decrease proportionally with the reduction of the PS linkages in the backbone of the oligonucleotide. Here, we used tc-DNA M23D sequences with varying amounts of internucleoside PS linkages, and thus M23D-PS 25sb3 is (p-CCTCGGCTTA * C * C * T - SEQ ID NO: 42), and M23D-PS 33sb3 corresponds to (p-CCTCGGCTT * A * C * C * T - SEQ ID NO: 43), and M23D-PS 50sb3 corresponds to (p-CCTCGGC * T * T * A * C * C * T - SEQ ID NO: 44), and M23D-PS 66sb3 corresponds to (p-CCTCG * G * C * T * T * A * C * C * T - SEQ ID NO: 45), and M23D-PS 83sb3 corresponds to (p-CCT * C * G * G * C * T * T * A * C * C * T - SEQ ID NO: 46), where " * " indicates a phosphorothioate internucleoside linkage, and p represents the terminal phosphate group. As previously shown, the aPTT test is more sensitive to PS content than to PT, reaching its maximum measurement time with six internucleoside PS (compound M23D-PS 50sb3). A lower number of PS linkages induces only a very limited increase in PT and aPTT.

[0476] Complement activation was also examined in mice after systemic injection of AONs at a high dose of 200 mg / kg.

[0477] Figure 3 shows that safe AONs containing phosphorodiester linkages as internucleoside linkers, specifically tcDNA-PO M23D, tc-DNA-PS M23D, tcDNA-PO SYN51, and the palmitoyl-conjugated AONs of the present invention (SY-0299, SY-0343, SY-0442, and SY-0455), did not significantly activate the complement cascade. Only the toxic tcDNA-PS SYN51 clearly activated C3 cleavage (50%). Note that tcDNA-PS M23D, which was considered safe, still only slightly activated C3 consumption.

[0478] Serum was collected at the end of 4 weeks of treatment with compounds SY-0299, SY-0343, SY-0442, and SY-0455, and serum biochemistry was assessed (Figure 4). There were no significant changes in serum albumin, creatinine, and urea, suggesting a lack of nephrotoxicity. Only a slight increase in ALP was observed for most palmitoyl-conjugated PO M23D (Figure 4E).

[0479] Thus, as shown, systemic delivery of the inventive compositions (palmitoyl-conjugated PO M23D, SY-0343) containing antisense oligomers linked to the 5'-palmitoyl-C6-amino lipid moiety and spacer and having a complete PO backbone enabled greater rescue of dystrophin than comparable sequences made with tcDNA having either a PO or PS backbone. Furthermore, the inventive compositions (SY-0299, SY-0343, SY-0442, and SY-0455) and AONs, respectively, did not cause adverse effects in the blood compartment after systemic delivery. Furthermore, the lack of sulfur in the phosphate backbone of AONs significantly reduced nonspecific binding to serum proteins, thereby significantly reducing the risk of adverse effects of complement activation and / or coagulation. The present compositions, which contain at least one lipid moiety, such as palmitoyl-conjugated tcDNA AONs with an internucleoside linking group that is PO, confer novel properties to PO-tcDNA containing molecules that are currently believed to preferentially bind to serum proteins such as albumin and lipoproteins, natural carriers of poorly water-soluble molecules containing fatty acids. Furthermore, and in summary, the above results indicate that tcDNA AONs that alter coagulation properties also systemically activate the complement cascade in vivo. A significant increase in PTT (>180 seconds) is a sign of potential risk, which becomes apparent when PT (>100 seconds) also increases dramatically. Finally, tcDNA-POs and the present composition Palm-tcDNA-PO are significantly safer than their related tcDNA-PSs. Example 2 Co-precipitation experiments and proteomics

[0480] The oligonucleotides of interest were synthesized as biotinylated conjugates for subsequent coprecipitation of potentially interacting serum proteins using streptavidin beads. The biotinylated derivatives used in this study were SY-0440, SY-0427, SY-0446, SY-0448, SY-0445, SY-0443, and SY-0451, which are defined and characterized in Table 3 and shown in Figure 6.

[0481] Biotinylated oligonucleotides immobilized on streptavidin beads were incubated in the presence of serum for 1 hour. The beads were collected by low-speed centrifugation, washed, and then solubilized in appropriate buffers for further protein analysis by SDS-PAGE, Orbitrap LC-MS / MS, and SDS-PAGE / MALDI-TOF.

[0482] Figure 7 illustrates SDS-PAGE analysis of protein recovery from mouse and human serum for the oligonucleotides used in this study. SY-0440, the tcDNA M23D with an intact PS, appeared to retain significantly more serum proteins than its tcDNA-PO counterpart, SY-0427. This result is consistent with previous studies (not shown) using various sequences (M23D, SYN51, and Poly T) and backbone chemistries (DNA, tcDNA, 2'OMe-PS, and PMO), which showed that oligomers lacking PS linkages did not significantly bind to serum proteins, suggesting that protein adsorption occurs preferentially at the level of the thiophosphate backbone and does not involve consensus sequence motifs.

[0483] In general, we found that tcDNA-PS oligonucleotides bind to many proteins involved in the complement pathway and coagulation. Orbitrap analysis revealed that tcDNA-PS oligonucleotides retain mannose-binding protein (MBL), which is beneficial for innate immunity via the lectin pathway. MBL binds to MASPs (MBL-associated serine proteases - MASP-1; -2; -3), which also form complexes with ficolins. When MBL binds to its target (e.g., surface mannose), the MASP proteins function to cleave the blood protein C4 into C4a and C4b. The C4b fragment then binds to surfaces and can initiate the formation of C3 convertase. The MBL / MASP-1 complex also possesses thrombin-like activity (thrombin clots fibrin, initiating blood clotting). All of these proteins were found in tcDNA-PS samples but not in complexes with oligonucleotides lacking PS internucleotide linkages.

[0484] As shown in Figure 7, our tcDNA-palmitoyl conjugates showed comparable patterns of protein recovery, which were very different from those obtained with naked tcDNA-PS and tcDNA-PO. Given the fact that tcDNA-PO did not consistently and significantly bind to serum proteins, the patterns obtained for the A, B, C, A', and D molecules were likely due to their palmitoyl and alkylphosphate moieties. SDS-PAGE MALDI-ToF MS analysis of the major proteins recovered from mouse and human serum indicated that the predominant proteins were albumin and lipoprotein components of LDL (low-density lipoprotein) and HDL (high-density lipoprotein) complexes, all involved in lipid transport (Tables 4 and 5).

[0485] [Table 4]

[0486] [Table 5]

[0487] Although the C7 moiety at the 3' end of the tcDNA sequence does not appear to contribute significantly to serum protein binding (compare A(SY-0446) with B(SY-0445)), B retained more dimeric albumin in mice. Comparison of B with C(SY-0443) showed that the presence of thiophosphates at both ends of the tcDNA sequence did not enhance protein binding; quite the opposite, it slightly reduced some proteins (fragments of factor H and complement C3) in human serum. The A'(SY-0448) molecule was similar to A, except that the third nucleotide in the sequence was replaced with 2'OMe-U in A'. The A / A' profiles were nearly identical in mice, but showed some differences in human serum. In particular, A' retained less factor H and complement C3, possibly suggesting a better safety profile. Finally, molecule D (SY-0451), lacking both C7 and C6, retained slightly less albumin and HDL components compared to A and A', leaving room for additional proteins, as indicated by the increased intensity of previously marginal bands.

[0488] The C7 motif was initially introduced to allow for the grafting of ligands for further tissue-specific targeting. Our studies have shown that the C7 moiety does not affect serum protein retention, but it may still be advantageous to maintain it at the 3' end of the AON to increase the size of the compound and thereby improve steric hindrance at the level of its pre-mRNA target. On the other side of the compound, moving the palmitoyl moiety away from the 5' end of the tcDNA sequence by C6 appears to increase preferential binding to albumin and lipoprotein complexes involved in fatty acid transport. Example 3 Stability of the compositions of the present invention in human serum

[0489] The stability of preferred compositions of the invention in human serum was studied with oligonucleotides SY-0343, SY-0442, SY-0299, SY-0450, SY-0455, and SY-0357. The compositions of the invention are defined and characterized in Table 3.

[0490] A mixture (80 μL total volume) consisting of 40 μL human serum, 20 μL of oligonucleotide stock solution (2x in PBS), and milliQ water was prepared in a 0.2 mL PCR vial to a final oligonucleotide concentration of 2 μM and incubated at 37°C. Aliquots were removed after 4, 24, and 120 h. The mixture was digested in a PCR vial with proteinase K (80 μL mixture, 100 μL of 2x proteinase K buffer (200 mM Tris-HCl, pH 8.5, 400 mM NaCl, 10 mM EDTA, 0.4% SDS), 20 μL of proteinase K (20 mg / mL in 100 mM Tris-HCl pH 8.5, 10 mM CaCl) at 55°C for 2 hours. The mixture was then cooled to room temperature, centrifuged (13,400 rpm, 10 minutes), and the supernatant was collected. A 50 μL aliquot was then taken for SPE and diluted with 50 μL of 0.2 M TEAB buffer. After 30 minutes, 50 μL of Condi solution (15 mM EtN, 100 mM HFIP) was added. An SPE column (Oasis HLB 1 cc (10 mg) extraction cartridge, Waters) was used. The oligonucleotide (186000383) was conditioned by sequentially passing 1 mL of MeOH and 2 × 500 μL of Condi solution. The above test solution was quantitatively applied to the SPE column, and the column was successively washed by passing 2 × 250 μL of Condi solution and 300 μL of TEAB (0.2 M) solution. The oligonucleotide was washed by applying 2 × 200 μL of eluent (Condi solution / MeOH 4:6) and 150 μL of MeOH, and the eluate was collected. The eluate was lyophilized to dryness in a speed-vac and then diluted with 100 μL of milliQ water. The mixture was then analyzed by HPLC-DAD-MS (Waters Acquity OST C18, 100 × 2.1 mm, 1.7 μm, mobile phase A: 400 mM HFIP, 15 mM TEA + Analysis was performed using 10% methanol, mobile phase B: methanol, flow rate 0.2 mL / min, 70°C.

[0491] As noted in Tables 6 and 7, the major degradation products after incubation in human serum were the parent oligonucleotides for all sequences tested, as shown below. [ka]

[0492] No other degradation products were detected in significant amounts. In particular, after 120 h, no cleavage was observed between the tc-nucleoside and the 2'-OMe nucleoside (sequence SY-0455) or the C7-amino group at the 3' end (sequences SY-0343, SY-0299, and SY-0357). Sequences with a PO linkage between the oligonucleotide and the spacer / lipid moiety were significantly more unstable in human serum than their PS counterparts. After 24 h, a significant amount of the spacer / lipid moiety was cleaved, and after 120 h, almost no original oligonucleotide was present in sequences linked via a PO linker. However, in the case of fatty acids connected via a PS linker, only approximately 1 and 8% of the spacer / lipid moiety were cleaved after 24 and 120 h, respectively.

[0493] [Table 6]

[0494] [Table 7] Example 4 Identification of multimers

[0495] Polyacrylamide gel electrophoresis (PAGE) experiments were performed to detect self-multimerization. The following chemicals were used: tris(hydroxymethyl)aminomethane (Tris) (TCI A0321); acetic acid (Merck 1.00063); acrylamide / bis solution, 29:1 (40%, Serva 10680.01); tetramethylethylenediamine (TEMED, Sigma-Aldrich T9281); ammonium persulfate (Sigma-Aldrich 248614); and glycerol (Sigma-Aldrich G9012). Buffer A was prepared by dissolving 60 g of Tris in 200 mL of water. The pH was adjusted to 7.4 with glacial acetic acid (approximately 25-30 mL). The solution was diluted to 500 mL with water and stored at +4 °C. Buffer B was prepared by diluting 16 mL of buffer A to 800 mL with water. Ammonium persulfate 10% (w / v) was prepared by dissolving 100 mg of ammonium persulfate in 0.9 mL of water. Gels were prepared by mixing the following solutions in a glass beaker: 9.4 mL of acrylamide / bis solution; 15 mL of water; 500 μL of buffer A; 125 μL of 10% (w / v) ammonium persulfate; and 38 μL of TEMED. The test solution was 2 mg / mL in 50% glycerol, and 10 μL was applied (equivalent to 10 μg of oligonucleotide). Pre-migration settings were 40 min / 90 V in buffer B. Migration settings were 45–60 min / 120 V in buffer B, in addition to 90 min / 90 V or 15 min / 90 V. 5–7 μL of 6x DNA loading dye was also transferred. Detection was performed by placing the gel on a TLC plate and examining it under UV light at 254 nm. Staining with Stains-All (Sigma-Aldrich, 1-ethyl-2-[3-(1-ethylnaphtho[1,2- d ]thiazolin-2-ylidene)-2-methylpropenyl]naphtho[1,2- d ]thiazolium bromide, 3,3′-diethyl-9-methyl-4,5,4′,5′-dibenzothiacarbocyanine) was then performed according to the manufacturer's protocol.

[0496] As previously mentioned, acutely toxic tc-DNA oligonucleotides were observed to exhibit an additional band migrating at a level of 40–60 bp in polyacrylamide gel electrophoresis experiments. This fairly sharp band was proposed to be a multimer of the oligonucleotide (e.g., duplex, trimer, or larger aggregate). Figure 9 shows the results of a gel electrophoresis experiment in which there is no obvious appearance of a multimer band for any of the studied oligonucleotides. An example of a multimer band is also shown, for oligonucleotide SY-0221, which exhibited toxicity in vivo. Example 5 Exon 23 skipping studies performed on mouse M23D surrogate sequences

[0497] SY0442 and SY0450 were selected for further investigation into the efficacy of exon 23 skipping. SY0210 (intact PS) and SY0308 (intact PO) were used as controls for some experiments detailed herein. The purpose of these experiments was to test efficacy and tcDNA content in tissues and to determine potential toxicity of the test compounds.

[0498] Mdx23 mice were injected with the compound at 50 mg / kg / week or 200 mg / kg / week for 4 weeks. Group 1 was analyzed 72 hours after the last injection, and Group 2 was analyzed 2 weeks after the last injection. The in vitro and mouse model studies discussed in Examples 1 and 2 were performed and the results were consistent with those examples.

[0499] Nested RT-PCR was used to assess exon 23 skipping levels. Total RNA was extracted from specific tissues in the mouse model and reverse-transcribed using the SSIII cDNA synthesis kit. RT-PCR was performed between exons 20 and 26, and specific RT-PCR was also performed to detect only the skipped product. Figures 10A-D demonstrate the RT-PCR results for 200 mg / kg / week at 72 hours (Figures 10A and 10B) and 2 weeks (Figures 10C and 10D). Clear bands corresponding to the exon 23 skipping product can be observed in all muscles treated with the compound. Skipping levels were particularly high in the heart in all cases. Using specific nested PCR, exon 23 skipping was also detected in the CNS, gastrointestinal smooth muscle, and retina. Similar results were obtained in the 50 mg / kg / week cohort (Figure 11).

[0500] Exon 23 skipping was quantified even more accurately using the more sensitive Taqman qPCR protocol. Figures 12A and 12B demonstrate similar results across the 200 mg / kg (Figure 12A) and 50 mg / kg (Figure 12B) cohorts. The calculated ratios between the 72-hour and 2-week cohorts were in most cases close to 1 for SY0442 and SY0450, respectively, at each dose: [Table 8]

[0501] The ratio of exon skipping was also calculated between SY0442 and SY0450, demonstrating that SY0442 generally had a slightly higher overall level of skipping than SY0450: [Table 9]

[0502] The data are further expanded in Figure 14, showing pooled data comparing the efficacy of SY0442 and SY0450 at 72 hours and 2 weeks post-injection. A dose effect can be observed between 50 mg / kg and 200 mg / kg (approximately 4-fold). Quantitation of exon 23 skipping at 200 mg / kg (Figure 15) and 50 mg / kg (Figure 16) comparing the intact PO compound with the intact PS compound was also measured.

[0503] To identify the most potent compounds, the potency / amount ratio of compounds found in tissues was calculated for 200 mg / kg: [Table 10]

[0504] Also calculated for 50mg / kg: [Table 11]

Claims

1. (a) an oligomeric portion comprising 8 to 20 deoxyribonucleic acid nucleoside monomer subunits, wherein a plurality of the nucleosides are tricyclo-deoxyribonucleic acid (tc-DNA) nucleosides; and (b) one or more lipid moieties selected from saturated C8-26 fatty acid moieties A compound comprising the one or more lipid moieties are covalently linked to the oligomeric moiety at the 5' or 3' end of the oligomeric moiety, either directly or via a spacer; At least one tc-DNA nucleoside has the formula (1): 【Chemistry 1】 (In formula (1), Bx is a nucleobase; T １ and T ２ is an internucleoside linking group, and T １ and T ２ The other is OR １ , OR ２ , a 5' terminal group, a 3' terminal group or an internucleoside linking group, １ is H or a hydroxyl protecting group, and R ２ is the phosphorus moiety); q １ , q ２ , q ３ , q ４ and q ５ are each independently hydrogen (H), halogen, C １～６ Alkyl, C ２～６ Alkenyl, C ２～６ Alkynyl, substituted C １～６ Alkyl, substituted C ２～６ Alkenyl, substituted C ２～６ Alkynyl, and —(CH ２ ) ｎ -C(O)-R ６’ wherein n is 0 to 6 and R ６’ OH, NH ２ , O-C １～３２ Alkyl and NH—C １～３２ alkyl); z １ and z ２ are each independently H, halogen, or C １～６ Alkyl, C １～６ Alkoxyl, O-C ２～６ Alkenyl, O-C ２～６ Alkynyl, substituted C １～６ Alkyl, substituted C １～６ Alkoxy, substituted O—C ２～６ Alkenyl, and substituted O—C ２～６ alkynyl) or a pharmaceutically acceptable salt thereof. compound.

2. The spacer is represented by the formula (a) to (h): (a) #-NH-C ２～１２ Alkylene-§, (b) #-NH-C ２～１２ alkylene-OP(OH)-§, (c) #-NH-C ２～１２ alkylene-OP(O)(SH)-§, (d) #-NH-C ２～１２ alkylene-OP(O)(OH)-§, (e) #-NH-C ２～１２ alkylene-NH-C(O)-§, (f) #-NH-C ２～１２ alkylene-NH-P(O)(OH)-§, (g) #-NH-C ２～１２ alkylene-NH-P(O)(SH)-§, and (h) #-NH-C ２～１２ Alkylene-OP(S)(OH)-§ (In the formula, C ２～１２ One or more —CH ２ The - moiety is unsubstituted or independently selected from -O-, -S-, -NH-, -C(O)-, -C(O)O-, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, -OP(OH)O-, -OP(O)(SH)O-, -OP(S)(OH)-, -OP(O)(OH)O-, -NHP(O)(OH)O-, -NHP(O)(SH)O-, or -(O-CH ２ -CH ２ ) ｋ - (k is an integer from 1 to 8), and C ２～１２ One or more —CH ２ The - moieties are, independently of each other, unsubstituted or substituted with one or more -COOH, -NH ２ , -OP(O)(OH) ２ or -OH, wherein the (#) symbol represents the point of covalent attachment to the lipid moiety, and the (§) symbol represents the point of covalent attachment to the oligomer moiety.

2. The compound of claim 1, comprising any one of:

3. The spacer is represented by the formula (a) to (d) (a)#-Z-NH-(CH ２ ) ｍ -X-§、 (b)#-Z-NH-(CH ２ ) ｎ -(O-CH ２ -CH ２ ) ｋ -O-(CH ２ ) ｐ -X-§、 (c)#-Z[-NH-(CH ２ ) ｎ -(O-CH ２ -CH ２ ) ｋ -O-(CH ２ ) ｐ -C(O)-]-NH-(CH ２ ) ｑ -X-§、および (d)#-Z[-NH-(CH ２ ) ｎ -(O-CH ２ -CH ２ ) ｋ -O-(CH ２ ) ｐ -C(O)-] ｒ -NHH-(CH ２ ) ｑ -(O-CH ２ -CH ２ ) ｋ -X-§ (Wherein, -Z- is independently a bond or -NH-CH(COOH)-(CH ２ ) ２ -C(O)- or -NH-CH[(CH ２ ) ２ each X independently represents OP(OH), OP(O)(SH), OP(S)(OH), OP(O)(OH), NHP(O)(OH), NHP(O)(SH) or NH—C(O); each k independently represents an integer of 1 to 8; each m independently represents an integer of 2 to 12; each n independently represents an integer of 2 to 4; each p independently represents an integer of 1 to 5; each q independently represents an integer of 1 to 3; each r independently represents an integer of 1 to 3; the (#) symbol represents a point of covalent bond to the lipid moiety; and the (§) symbol represents a point of covalent bond to the oligomer moiety.

3. The compound of claim 1 or 2, comprising any one of:

4. The compound of any one of claims 1 to 3, wherein the oligomeric portion comprises one or more nucleosides other than tc-DNA nucleosides.

5. 5. The compound of any one of claims 1 to 4, wherein the monomer subunits are linked by a plurality of phosphorodiester linkage groups.

6. The compound of any one of claims 1 to 5, wherein the oligomeric portion comprises a sequence selected from any one of the sequences SEQ ID NOs: 1 to 37.

7. 10. A pharmaceutical composition comprising a compound according to any one of claims 1 to 6 and further comprising a pharmaceutically acceptable carrier, the pharmaceutical composition being for use in the prevention, treatment or diagnosis of a neuromuscular or musculoskeletal disorder.

8. 10. A composition comprising a compound according to any one of claims 1 to 6 or a pharmaceutical composition according to claim 7 for use as a medicament in the prevention, treatment or diagnosis of a disease, wherein the disease is a neuromuscular or musculoskeletal disease selected from Duchenne muscular dystrophy, familial dysautonomia, spinal muscular atrophy, ataxia-telangiectasia, congenital disorders of glycosylation, frontotemporal dementia, Parkinsonism linked to chromosome 17, Niemann-Pick disease type C, neurofibromatosis type 1, neurofibromatosis type 2, macrocephalic leukoencephalopathy with subcortical cysts type 1, Pelizaeus-Merzbach disease, Pompe disease, myotonic dystrophy type 2 (DM2, or proximal myotonic myopathy) and myotonic dystrophy type 1 (DM1, or Steinert disease).

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