Nucleic acid compound

Abstract

The present invention provides novel nucleic acid compounds suitable for therapeutic use. Further, the present invention provides methods for making these compounds and methods for using the compounds to treat various diseases and conditions.

Description

Technical Field

[0001] The present invention provides novel nucleic acid compounds suitable for therapeutic use. Further, the present invention provides methods for making these compounds and methods for using the compounds to treat various diseases and conditions.

Background Art

[0002] Nucleic acid compounds have important therapeutic applications in medicine. Nucleic acids can be used to silence genes that are the cause of a particular disease. Gene silencing prevents the formation of proteins by inhibiting translation. Importantly, gene silencing agents are promising alternatives to conventional small organic compounds that inhibit the function of disease-related proteins. siRNA, antisense RNA, and microRNA are oligonucleotides / oligonucleosides that prevent protein formation by gene silencing.

[0003] Several modified siRNA compounds, including siRNA / RNAi therapeutic agents for treating various diseases including central nervous system diseases, inflammatory diseases, metabolic disorders, oncology, infectious diseases, and eye diseases, have been developed particularly in the past 20 years for diagnostic and therapeutic purposes.

Summary of the Invention

[0004] The present invention relates to nucleic acid compounds for use in the treatment and / or prevention of diseases.

[0005] Description of the Invention A nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region including a first strand that is at least partially complementary to a portion of RNA transcribed from the target gene and a second strand that is at least partially complementary to the first strand, wherein the nucleosides of the second strand are as follows (5'-3'): Me-Me-Me-Me-Me-Me-F-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-F-Me-Me, or Me-Me-Me-Me-Me-F-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, or Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me A nucleic acid comprising a 2'-sugar modification pattern of

[0006] A nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region including a first strand that is at least partially complementary to a portion of RNA transcribed from the target gene, and a second strand that is at least partially complementary to the first strand, wherein the nucleosides of the second strand are as follows (5'-3'): Me(s)Me(s)Me-Me-Me-Me-F-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-F-Me-Me, or Me(s)Me(s)Me-Me-Me-F-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, or Me(s)Me(s)Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, or Me(s)Me(s)Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, or Me-Me-Me-Me-Me-Me-F-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-F(s)Me(s)Me, or Me-Me-Me-Me-Me-F-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me(s)Me(s)Me, or Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me(s)Me(s)Me, or Me-Me-Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me(s)Me(s)Me [wherein (s) is a phosphorothioate internucleoside linkage] A nucleic acid comprising 2'-sugars and modification patterns of linkages.

[0007] A nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region including a first strand that is at least partially complementary to a portion of RNA transcribed from the target gene and a second strand that is at least partially complementary to the first strand, wherein the nucleosides of the second strand are as follows (5'-3'): ia-ia-Me-Me-Me-Me-Me-Me-F-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-F-Me-Me, or ia-ia-Me-Me-Me-Me-Me-F-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, or ia-ia-Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, or ia-ia-Me-Me-Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, or Me-Me-Me-Me-Me-Me-F-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-F-Me-Me-ia-ia, or Me-Me-Me-Me-Me-F-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-ia-ia, or Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-ia-ia, or Me-Me-Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me-ia-ia [Wherein, ia represents an inverted abasic nucleoside, and when the inverted abasic nucleoside represented by ia-ia is present at the 3'-end of the second strand, the inverted abasic nucleoside is typically present in an overhang of two nucleosides] A nucleic acid comprising 2'-sugars and abasic modification patterns.

[0008] A nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region including a first strand that is at least partially complementary to a portion of RNA transcribed from the target gene and a second strand that is at least partially complementary to the first strand, wherein the nucleosides of the second strand are as follows (5'-3'): ia-ia-Me(s)Me(s)Me-Me-Me-Me-F-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-F-Me-Me, or ia-ia-Me(s)Me(s)Me-Me-Me-F-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, or ia-ia-Me(s)Me(s)Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, or ia-ia-Me(s)Me(s)Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, Me-Me-Me-Me-Me-Me-F-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-F(s)Me(s)Me-ia-ia, or Me-Me-Me-Me-Me-F-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me(s)Me(s)Me-ia-ia, or Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me(s)Me(s)Me-ia-ia, or Me-Me-Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me(s)Me(s)Me-ia-ia [wherein (s) is a phosphorothioate internucleoside linkage, ia represents an inverted abasic nucleoside, and when the inverted abasic nucleoside represented by ia-ia is present at the 3'-end of the second strand, the inverted abasic nucleoside is typically present in an overhang of two nucleosides] A nucleic acid comprising the 2'-sugar, abasic, and linkage modification pattern of

[0009] The nucleic acids described in this specification typically include a first strand containing a modification pattern selected from the following or any combination thereof, where the 1st position is the 5'-terminal nucleoside of the first strand and the counting direction is 5'-3': 2'-F sugar modifications at least at positions 2, 14, and 16, and / or 2'-Me sugar modifications at positions 17 - 23, or the first strand includes at least 8 2'-F sugar modifications, such as 2'-F sugar modifications at least at positions 2, 4, 6, 12, 14, 16, 18, and 20, and / or 2'-Me sugar modifications at positions 1, 3 - 5, 10 - 13, or the first strand includes at least 8 2'-F sugar modifications, such as 2'-F sugar modifications at least at positions 2, 4, 6, 12, 14, 16, 18, and 20, and / or 2'-Me sugar modification at position 7, or a heat - destabilizing modification at position 7, such as typically a modified unlocked nucleic acid or glycol nucleic acid, and / or 2'-F sugar modification or heat - destabilizing modification at position 6, such as typically a modified unlocked nucleic acid or glycol nucleic acid, and / or Positions 8 and 9 may be modifications selected from 2'-Me sugar modification and 2'-F sugar modification, and may typically be the same 2'-sugar modification, whereby typically the first strand has the following modification pattern (5'-3'): Me - F - Me - Me - Me - (M)4 - Me - Me - Me - Me - F - Me - F - Me - Me - Me - Me - Me - Me [wherein M represents a modification selected from 2'-Me sugar modification, 2'-F sugar modification, and heat - destabilizing modification, such as typically a modified unlocked nucleic acid or glycol nucleic acid, the heat - destabilizing modification may typically be present at position 6, and the 2'-Me sugar modification may typically be present at position 7] may include, or whereby typically the first strand has the following modification pattern (5'-3'): Me-F-Me-Me-Me-(M1)-(M2)3-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me [Wherein, M1 represents a thermolabile modification, for example typically a modified unlocked nucleic acid or glycol nucleic acid, and M2 represents a modification selected from 2'-Me sugar modification and 2'-F sugar modification] comprises, or consists of, typically the first strand has the following modification pattern (5'-3'): Me-F-Me-Me-Me-(M1)-Me-(M2)2-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me [Wherein, M1 represents a modification selected from 2'-Me sugar modification, 2'-F sugar modification and thermolabile modification, for example typically a modified unlocked nucleic acid or glycol nucleic acid, and M2 represents a modification selected from 2'-Me sugar modification and 2'-F sugar modification] comprises, or consists of, typically the first strand has the following modification pattern (5'-3'): Me-F-Me-Me-Me-(M1)-Me-(M2)2-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me [Wherein, M1 represents a thermolabile modification, for example typically a modified unlocked nucleic acid or glycol nucleic acid, and M2 represents a modification selected from 2'-Me sugar modification and 2'-F sugar modification, typically M2 can be the same 2'-sugar modification] comprises.

[0010] Typically, (M)4 shown above has the following 2'-sugar modification pattern (5'-3'): F-Me-Me-F Me-F-Me-F F-Me-F-Me F-F-F-F Me-F-F-Me Me-Me-F-F F-F-Me-Me Me-Me-Me-Me represents any one of.

[0011] Typically, the two phosphorothioate internucleoside linkages are each present between three consecutive positions in both the 5'-terminal region and the 3'-terminal region of the first strand described herein, whereby the terminal nucleosides in each of the 5'-terminal region and the 3'-terminal region of the first strand are each attached by a phosphorothioate internucleoside linkage to the respective 5' and 3' adjacent penultimate nucleosides, and the respective 5' and 3' penultimate nucleosides are attached by a phosphorothioate internucleoside linkage to the respective 5' and 3' adjacent antepenultimate nucleosides. Optionally, two phosphorothioate internucleoside linkages may further be present between three consecutive positions in the 3'-terminal region of the second strand, whereby the 3'-terminal nucleoside is attached by a phosphorothioate internucleoside linkage to the adjacent penultimate nucleoside, and the penultimate nucleoside is attached by a phosphorothioate internucleoside linkage to the adjacent antepenultimate nucleoside, and / or optionally, two phosphorothioate internucleoside linkages may further be present between three consecutive positions in the 5'-terminal region of the second strand, whereby the 5'-terminal nucleoside is attached by a phosphorothioate internucleoside linkage to the adjacent penultimate nucleoside, and the penultimate nucleoside is attached by a phosphorothioate internucleoside linkage to the adjacent antepenultimate nucleoside.

[0012] A nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region including a first strand that is at least partially complementary to a portion of RNA transcribed from the target gene and a second strand that is at least partially complementary to the first strand, wherein the nucleosides of the second strand are as follows: Second strand (5'-3'): ia-ia-Me-Me-Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me comprising a 2'-sugar and a non-base-modified pattern, or a sugar modification in which the 7-position on the second strand is a 2'-Me modification, the 1-position is the 5'-terminal nucleoside of the second strand, the counting direction is 5'-3', and typically there are two inverted non-base nucleosides in the 5'-terminal region of the second strand, a nucleic acid, wherein the second strand is typically used together with the first strand as defined herein.

[0013] A nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region including a first strand that is at least partially complementary to a portion of RNA transcribed from the target gene and a second strand that is at least partially complementary to the first strand, wherein the nucleosides of the second and first strands are as follows (5'-3'): Modification pattern 1: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-F-Me-Me, First strand (5'-3'): Me-F-Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Or modification pattern 2: Second strand (5'-3'): Me-Me-Me-Me-Me-F-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me-F-Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Or modification pattern 3: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me-F-Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Or modification pattern 4: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me-F-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Or modification pattern 5: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me-F-Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me A nucleic acid comprising a 2'-sugar modification pattern of.

[0014] A nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region including a first strand that is at least partially complementary to a portion of RNA transcribed from the target gene and a second strand that is at least partially complementary to the first strand, wherein the nucleosides of the second and first strands are as follows (5'-3'): Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me-F-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Or Second strand (5'-3'): Me-Me-Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me-F-Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me A nucleic acid comprising a 2'-sugar modification pattern of.

[0015] A nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region including a first strand that is at least partially complementary to a portion of RNA transcribed from the target gene and a second strand that is at least partially complementary to the first strand, wherein the nucleosides of the second and first strands are as follows: (5'-3'): Modification pattern 1: Second strand (5'-3'): Me(s)Me(s)Me-Me-Me-Me-F-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-F-Me-Me, First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modification pattern 2: Second strand (5'-3'): Me(s)Me(s)Me-Me-Me-F-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modification pattern 3: Second strand (5'-3'): Me(s)Me(s)Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modification pattern 4: Second strand (5'-3'): Me(s)Me(s)Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modification pattern 5: Second strand (5'-3'): Me(s)Me(s)Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me(s)F(s)Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modification pattern 6: Second strand (5'-3'): Me(s)Me(s)Me-Me-Me-Me-F-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me(s)F(s)Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me [wherein, (s) is a phosphorothioate internucleoside linkage] A nucleic acid comprising the 2'-sugar and the binding modification pattern of the above.

[0016] A nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region including a first strand that is at least partially complementary to a portion of RNA transcribed from the target gene and a second strand that is at least partially complementary to the first strand, wherein the nucleosides of the second and first strands are as follows (5'-3'): Second strand (5'-3'): Me(s)Me(s)Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or Second strand (5'-3'): Me(s)Me(s)Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me(s)F(s)Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me [where (s) is a phosphorothioate internucleoside linkage] A nucleic acid comprising a 2'-sugar and a modified linkage pattern as described above.

[0017] A nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region including a first strand that is at least partially complementary to a portion of RNA transcribed from the target gene and a second strand that is at least partially complementary to the first strand, wherein the nucleosides of the second and first strands are as follows: (5'-3'): Modification pattern 1: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-F(s)Me(s)Me, First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modification pattern 2: Second strand (5'-3'): Me-Me-Me-Me-Me-F-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me(s)Me(s)Me, First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modification pattern 3: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me(s)Me(s)Me, First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modification pattern 4: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me(s)Me(s)Me, First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modification pattern 5: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me(s)Me(s)Me, First strand (5'-3'): Me(s)F(s)Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modification pattern 6: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me(s)Me(s)Me, First strand (5'-3'): Me(s)F(s)Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me [wherein, (s) is a phosphorothioate nucleoside internucleoside linkage] A nucleic acid comprising the 2'-sugar and the linkage modification pattern of.

[0018] A nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region including a first strand that is at least partially complementary to a portion of RNA transcribed from the target gene and a second strand that is at least partially complementary to the first strand, wherein the nucleosides of the second and first strands are as follows (5'-3'): Modification pattern 1: Second strand (5'-3'): ia-ia-Me-Me-Me-Me-Me-Me-F-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-F-Me-Me, First strand (5'-3'): Me-F-Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Or modification pattern 2: Second strand (5'-3'): ia-ia-Me-Me-Me-Me-Me-F-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me-F-Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Or modification pattern 3: Second strand (5'-3'): ia-ia-Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me-F-Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Or modification pattern 4: Second strand (5'-3'): ia-ia-Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me-F-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Or modification pattern 5: Second strand (5'-3'): ia-ia-Me-Me-Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me-F-Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Or modification pattern 6: Second strand (5'-3'): ia-ia-Me-Me-Me-Me-Me-Me-F-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me First strand (5'-3'): Me-F-Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me [wherein, ia represents an inverted abasic nucleoside] A nucleic acid comprising the 2'-sugar and abasic modification pattern of .

[0019] A nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region including a first strand that is at least partially complementary to a portion of RNA transcribed from the target gene and a second strand that is at least partially complementary to the first strand, wherein the nucleosides of the second and first strands are as follows (5'-3'): Second strand (5'-3'): ia-ia-Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me-F-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me or Second strand (5'-3'): ia-ia-Me-Me-Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me-F-Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me [wherein, ia represents an inverted abasic nucleoside] A nucleic acid comprising the 2'-sugar and abasic modification pattern of .

[0020] A nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region including a first strand that is at least partially complementary to a portion of RNA transcribed from the target gene and a second strand that is at least partially complementary to the first strand, wherein the nucleosides of the second and first strands are as follows (5'-3'): Modification pattern 1: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-F-Me-Me-ia-ia, First strand (5'-3'): Me-F-Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Or modification pattern 2: Second strand (5'-3'): Me-Me-Me-Me-Me-F-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-ia-ia, First strand (5'-3'): Me-F-Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Or modification pattern 3: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-ia-ia, First strand (5'-3'): Me-F-Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Or modification pattern 4: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-ia-ia, First strand (5'-3'): Me-F-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Or modification pattern 5: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me-ia-ia, First strand (5'-3'): Me-F-Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Or modification pattern 6: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me-ia-ia First strand (5'-3'): Me-F-Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me [Wherein, ia represents an inverted abasic nucleoside, and when the inverted abasic nucleoside represented by ia-ia is present at the 3'-end of the second strand, the inverted abasic nucleoside is typically present in an overhang of two nucleosides] A nucleic acid comprising the 2'-sugar and abasic modification pattern of.

[0021] A nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region including a first strand that is at least partially complementary to a portion of RNA transcribed from the target gene and a second strand that is at least partially complementary to the first strand, wherein the nucleosides of the second and first strands are as follows (5'-3'): Modification pattern 1: Second strand (5'-3'): ia-ia-Me(s)Me(s)Me-Me-Me-Me-F-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-F-Me-Me, First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modification pattern 2: Second strand (5'-3'): ia-ia-Me(s)Me(s)Me-Me-Me-F-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modification pattern 3: Second strand (5'-3'): ia-ia-Me(s)Me(s)Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modification pattern 4: Second strand (5'-3'): ia-ia-Me(s)Me(s)Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modification pattern 5: Second strand (5'-3'): ia-ia-Me(s)Me(s)Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me(s)F(s)Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modification pattern 6: Second strand (5'-3'): ia-ia-Me(s)Me(s)Me-Me-Me-Me-F-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me(s)F(s)Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me [Wherein, (s) is a phosphorothioate nucleoside internucleoside linkage, and ia represents an inverted abasic nucleoside] A nucleic acid comprising 2'-sugar, abasic, and linkage modification patterns of

[0022] A nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region including a first strand that is at least partially complementary to a portion of RNA transcribed from the target gene and a second strand that is at least partially complementary to the first strand, wherein the nucleosides of the second and first strands are as follows (5'-3'): Second strand (5'-3'): ia-ia-Me(s)Me(s)Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me or Second strand (5'-3'): ia-ia-Me(s)Me(s)Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me(s)F(s)Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me [wherein, (s) represents a phosphorothioate internucleoside linkage, and ia represents an inverted abasic nucleoside] A nucleic acid comprising 2'-sugar, abasic, and linkage modification patterns as described above.

[0023] A nucleic acid for inhibiting the expression of ZPI or HCII, comprising a double-stranded region including a first strand that is at least partially complementary to a portion of RNA transcribed from ZPI or HCII and a second strand that is at least partially complementary to the first strand, wherein the nucleosides of the second and first strands are as follows (5'-3'): Second strand (5'-3'): ia-ia-Me(s)Me(s)Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me [wherein, (s) represents a phosphorothioate internucleoside linkage, and ia represents an inverted abasic nucleoside] A nucleic acid comprising a 2'-sugar, abasic, and linkage modification pattern of.

[0024] A nucleic acid for inhibiting the expression of B4GALT1, comprising a double-stranded region including a first strand that is at least partially complementary to a portion of RNA transcribed from B4GALT1 and a second strand that is at least partially complementary to the first strand, wherein the nucleosides of the second and first strands are as follows (5'-3'): Second strand (5'-3'): ia-ia-Me(s)Me(s)Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me(s)F(s)Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me [wherein, (s) represents a phosphorothioate internucleoside linkage, and ia represents an inverted abasic nucleoside] A nucleic acid comprising a 2'-sugar, abasic, and linkage modification pattern of.

[0025] A nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region including a first strand that is at least partially complementary to a portion of RNA transcribed from the target gene and a second strand that is at least partially complementary to the first strand, wherein the nucleosides of the second and first strands are as follows (5'-3'): Modification pattern 1: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-F(s)Me(s)Me-ia-ia, First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modification pattern 2: Second strand (5'-3'): Me-Me-Me-Me-Me-F-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me(s)Me(s)Me-ia-ia, First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modification pattern 3: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me(s)Me(s)Me-ia-ia, First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modification pattern 4: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me(s)Me(s)Me-ia-ia, First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modification pattern 5: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me(s)Me(s)Me-ia-ia, First strand (5'-3'): Me(s)F(s)Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modification pattern 6: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me(s)Me(s)Me-ia-ia, First strand (5'-3'): Me(s)F(s)Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me [wherein (s) is a phosphorothioate nucleoside internucleoside linkage, ia represents an inverted abasic nucleoside, and when the inverted abasic nucleoside represented by ia-ia is present at the 3'-end of the second strand, the inverted abasic nucleoside is typically present in an overhang of two nucleosides] A nucleic acid comprising the 2'-sugar, abasic, and linkage modification patterns of .

[0026] A particularly preferred nucleic acid for inhibiting the expression of a target gene according to the present invention, comprising a first strand that is at least partially complementary to a portion of RNA transcribed from the target gene and a second strand that is at least partially complementary to the first strand, and comprising a double-stranded region, wherein counting from the 1-position at the 5'-end of the second strand, which is the 5'-most nucleoside without an abasic nucleoside, the 7-position on the second strand comprises a sugar modification that is a 2'-Me modification, or the second strand has the following modification pattern: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me A nucleic acid comprising .

[0027] A particularly preferred nucleic acid according to the present invention is as follows: Second strand (5'-3'): ia-ia-Me-Me-Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me which is combined with a first strand comprising a modification pattern selected from the following (5'-3') (the 1-position is the 5'-terminal nucleoside of the first strand, and the counting direction is 5'-3'): (5'-3')Me-F-Me-Me-Me-(M)4-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me [wherein M represents a modification selected from 2'-Me sugar modification, 2'-F sugar modification, and thermolabile modification, for example, typically a modified unlocked nucleic acid or glycol nucleic acid, the thermolabile modification may typically be present at the 6-position, and the 2'-Me sugar modification may typically be present at the 7-position] or thereby, typically the first strand has the following modification pattern (5'-3'): (5'-3')Me-F-Me-Me-Me-(M1)-(M2)3-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me [wherein M1 represents a thermolabile modification, for example, typically a modified unlocked nucleic acid or glycol nucleic acid, and M2 represents a modification selected from 2'-Me sugar modification and 2'-F sugar modification] and includes or thereby, typically the first strand has the following modification pattern (5'-3'): (5'-3')Me-F-Me-Me-Me-(M1)-Me-(M2)2-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me [wherein M1 represents a modification selected from 2'-Me sugar modification, 2'-F sugar modification, and thermolabile modification, for example, typically a modified unlocked nucleic acid or glycol nucleic acid, and M2 represents a modification selected from 2'-Me sugar modification and 2'-F sugar modification] and includes or thereby, typically the first strand has the following modification pattern (5'-3'): (5'-3')Me-F-Me-Me-Me-(M1)-Me-(M2)2-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me [wherein M1 represents a thermolabile modification, for example, typically a modified unlocked nucleic acid or glycol nucleic acid, M2 represents a modification selected from 2'-Me sugar modification and 2'-F sugar modification, and typically M2 can be the same 2'-sugar modification] includes.

[0028] A nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region including a first strand that is at least partially complementary to a portion of RNA transcribed from the target gene and a second strand that is at least partially complementary to the first strand, wherein the second strand contains two consecutive abasic nucleosides in the 5' or 3' terminal region of the second strand, and counting from the 1st position of the 5' end of the second strand, which is the most 5'-side nucleoside without an abasic nucleoside, the 7th position on the second strand contains a sugar modification that is a 2'-Me modification.

[0029] Particularly preferred according to the present invention is a nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region including a first strand that is at least partially complementary to a portion of RNA transcribed from the target gene and a second strand that is at least partially complementary to the first strand, wherein the second and first strands have the following modification patterns: Second strand (5'-3'): ia-ia-Me-Me-Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me First strand (5'-3') Me-F-Me-Me-Me-(M1)-Me-(M2)2-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me [wherein, M1 represents a thermolabile modification, for example, typically a modified unlocked nucleic acid or glycol nucleic acid, and M2 represents a modification selected from 2'-Me sugar modification and 2'-F sugar modification, and typically M2 can be the same 2' sugar modification] and is a nucleic acid.

[0030] A nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region including a first strand that is at least partially complementary to a portion of RNA transcribed from the target gene and a second strand that is at least partially complementary to the first strand, wherein the second strand does not contain an abasic nucleoside and is the most 5'-terminal nucleoside, and counting from the 1st position of the 5'-end of the second strand, there are phosphorothioate internucleoside linkages between the 1st and 2nd positions and between the 2nd and 3rd positions of the second strand respectively; or the second strand does not contain an abasic nucleoside and is the most 3'-terminal nucleoside, and counting from the 1st position of the 3'-end of the second strand, there are phosphorothioate internucleoside linkages between the 1st and 2nd positions and between the 2nd and 3rd positions of the second strand respectively; and counting from the 1st position of the 5'-end of the second strand which does not contain an abasic nucleoside and is the most 5'-terminal nucleoside, the 7th position on the second strand contains a sugar modification that is a 2'-Me modification.

[0031] A nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region including a first strand that is at least partially complementary to a portion of RNA transcribed from the target gene and a second strand that is at least partially complementary to the first strand, wherein the second strand does not contain an abasic nucleoside and is the most 5'-terminal nucleoside, the 7th position on the second strand contains a sugar modification that is a 2'-Me modification, and at the 3'-end of the second strand, the nucleic acid is directly or indirectly conjugated to one or more ligand moieties, and the ligand moiety preferably includes one or more N-acetylgalactosamine (GalNAc) ligands, and / or one or more N-acetylgalactosamine (GalNAc) ligand derivatives, and / or one or more N-acetylgalactosamine (GalNAc) ligands and / or their derivatives conjugated to the nucleic acid through a linker.

[0032] Each of the sequences of the second strand and the constructs can be used together with any of the first strands described herein.

[0033] A nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region including a first strand that is at least partially complementary to a portion of RNA transcribed from the target gene and a second strand that is at least partially complementary to the first strand, wherein the second strand has the following modification pattern: Modification pattern 1: Optionally combined with a first strand (5'-3'): Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me, Second strand (5'-3'): ia-ia-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F, Or modification pattern 2: Optionally combined with a first strand (5'-3'): Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me, Second strand (5'-3'): F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-ia-ia [Wherein, ia represents an inverted abasic nucleoside, and when the inverted abasic nucleoside represented by ia-ia is present at the 3'-end of the second strand, the inverted abasic nucleoside is typically present in an overhang of two nucleosides] A nucleic acid having the above.

[0034] A nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region including a first strand that is at least partially complementary to a portion of RNA transcribed from the target gene and a second strand that is at least partially complementary to the first strand, wherein the second strand has the following modification pattern: Modification pattern 1: Optionally combined with a first strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me(s)F(s)Me, Second strand (5'-3'): ia-ia-F(s)Me(s)F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F, or modification pattern 2: Optionally, the first strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me(s)F(s)Me combined with, Second strand (5'-3'): F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F(s)Me(s)F-ia-ia [wherein, (s) is a phosphorothioate internucleoside linkage, ia represents an inverted abasic nucleoside, and when the inverted abasic nucleoside represented by ia-ia is present at the 3'-end of the second strand, the inverted abasic nucleoside is typically present in an overhang of two nucleosides] A nucleic acid having.

[0035] A further nucleic acid according to the present invention is A first strand that is at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand that is at least partially complementary to the first strand A nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region, wherein the nucleosides of the second and first strands comprise the following 2'-sugar and linkage modification patterns (5'-3'): Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, or The 2'-Me or 2'-F modified nucleosides of the first strand comprise any one of the following modification patterns (5'-3'): First strand (5'-3'): Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me, or First strand (5'-3'): Me-F-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me, or First strand (5'-3'): Me-F-Me-Me-Me-F-F-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me, or First strand (5'-3'): Me-F-Me-Me-Me-Me-F-F-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me, or First strand (5'-3'): Me-F-Me-Me-Me-Me-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me, or First strand (5'-3'): Me-F-Me-Me-Me-F-F-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me, or First strand (5'-3'): Me-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me(s)F(s)Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me, or First strand (5'-3'): Me(s)F(s)Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me, or First strand (5'-3'): Me(s)F(s)Me-Me-Me-F-F-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me, or First strand (5'-3'): Me(s)F(s)Me-Me-Me-Me-F-F-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me, or First strand (5'-3'): Me(s)F(s)Me-Me-Me-Me-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me, or First strand (5'-3'): Me(s)F(s)Me-Me-Me-F-F-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me, or First strand (5'-3'): Me(s)F(s)Me-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me [wherein, (s) is a phosphorothioate internucleoside linkage].

[0036] A further nucleic acid according to the present invention comprises a first strand that is at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand that is at least partially complementary to the first strand, and contains a double-stranded region, and is a nucleic acid for inhibiting the expression of a target gene, wherein the nucleosides of the second and first strands contain the following 2'-sugar, abasic, and linkage modification patterns (5'-3'): Second strand (5'-3'): Me(s)Me(s)Me-Me-Me-Me-F-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, or Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me(s)Me(s)Me, or Second strand (5'-3'): ia-ia-Me-Me-Me-Me-Me-Me-F-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, or Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me-ia-ia, or Second strand (5'-3'): ia-ia-Me(s)Me(s)Me-Me-Me-Me-F-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, or Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me(s)Me(s)Me-ia-ia The 2'-Me or 2'-F modified nucleoside of the first strand includes any one of the following modification patterns (5'-3'): First strand (5'-3'): Me-F-Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me, or First strand (5'-3'): Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me, or First strand (5'-3'): Me-F-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me, or First strand (5'-3'): Me-F-Me-Me-Me-F-F-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me, or First strand (5'-3'): Me-F-Me-Me-Me-Me-F-F-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me, or First strand (5'-3'): Me-F-Me-Me-Me-Me-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me, or First strand (5'-3'): Me-F-Me-Me-Me-F-F-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me, or First strand (5'-3'): Me-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me(s)F(s)Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me, or First strand (5'-3'): Me(s)F(s)Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me, or First strand (5'-3'): Me(s)F(s)Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me, or First strand (5'-3'): Me(s)F(s)Me-Me-Me-F-F-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me, or First strand (5'-3'): Me(s)F(s)Me-Me-Me-Me-F-F-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me, or First strand (5'-3'): Me(s)F(s)Me-Me-Me-Me-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me, or First strand (5'-3'): Me(s)F(s)Me-Me-Me-F-F-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me, or First strand (5'-3'): Me(s)F(s)Me-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me [wherein, (s) is a phosphorothioate internucleoside linkage, ia represents an inverted abasic nucleoside, and when the inverted abasic nucleoside represented by ia-ia is present at the 3'-end of the second strand, the inverted abasic nucleoside is typically present in an overhang of two nucleosides].

[0037] The nucleic acid according to the present invention may further comprise a first strand comprising at least 17 consecutive nucleosides that differ from any one of the sequences of the first strand listed in Table 2 by 0 or 1 nucleoside.

[0038] The nucleic acid according to the present invention may further include a first strand comprising at least 17 consecutive nucleosides that differ from any one of the sequences of the first strand listed in Table 3 by 0 or 1 nucleoside.

[0039] Typically, the above-mentioned first strand contains nucleosides 2 to 18 of any one of the sequences defined in Table 2 or 3.

[0040] The nucleic acid according to the present invention may further include a second strand comprising a nucleoside sequence of at least 17 consecutive nucleosides that differ from any one of the sequences of the second strand listed in Table 2 by 0 or 1 nucleoside, and the second strand has a region of at least 85% complementarity with the first strand over 17 consecutive nucleosides.

[0041] The nucleic acid according to the present invention may further include a second strand comprising a nucleoside sequence of at least 17 consecutive nucleosides that differ from any one of the sequences of the second strand listed in Table 2 by 0 or 1 nucleoside, and the double-stranded region includes at least 14, 15, 16 or 17 complementary base pairs.

[0042] The nucleic acid according to the present invention may further include a second strand comprising a nucleoside sequence of at least 17 consecutive nucleosides that differ from any one of the sequences of the second strand listed in Table 4 by 0 or 1 nucleoside, and the second strand has a region of at least 85% complementarity with the first strand over 17 consecutive nucleosides.

[0043] The nucleic acid according to the present invention may further include a second strand comprising a nucleoside sequence of at least 17 consecutive nucleosides that differ from any one of the sequences of the second strand listed in Table 4 by 0 or 1 nucleoside, and the double-stranded region includes at least 14, 15, 16 or 17 complementary base pairs.

[0044] A nucleic acid according to the present invention, wherein the first strand comprises any one of the sequences of the first strand listed in Table 2.

[0045] A nucleic acid according to the present invention, wherein the first strand comprises any one of the sequences of the first strand listed in Table 3.

[0046] A nucleic acid according to the present invention, wherein the second strand comprises any one of the sequences of the second strand listed in Table 2.

[0047] A nucleic acid according to the present invention, wherein the second strand comprises any one of the sequences of the second strand listed in Table 4.

[0048] A nucleic acid according to the present invention, wherein the first strand and the second strand form any one of the double strands listed in Table 5.

[0049] A nucleic acid according to the present invention, wherein the nucleic acid is an siRNA oligonucleoside.

[0050] A nucleic acid according to the present invention, wherein the nucleic acid is directly or indirectly conjugated to one or more ligand moieties, and optionally, the ligand moiety is present in the terminal region of the second strand, typically in its 3'-terminal region, and typically, one or more N-acetylgalactosamine (GalNAc) ligands, and / or one or more derivatives of N-acetylgalactosamine (GalNAc) ligands, and / or one or more N-acetylgalactosamine (GalNAc) ligands and / or derivatives thereof conjugated to the nucleic acid through a linker may be included. Typically, one or more GalNAc ligands and / or GalNAc ligand derivatives are directly or indirectly conjugated at the 5' or 3'-terminal region of the second strand of the nucleic acid, typically at its 3'-terminal region.

[0051] The following structure:

[0052]

Chemical formula

[0053] A nucleic acid according to the invention, comprising a ligand moiety comprising

[0054] the following structure:

[0055]

Chem.

[0056] [wherein, each occurrence of R1 is independently selected from the group consisting of hydrogen, methyl and ethyl, R2 is hydrogen, hydroxy, -OC 1~3 alkyl, -C(=O)OC 1~3 alkyl, halo and nitro, each occurrence of X1 and X2 is independently selected from the group consisting of methylene, oxygen and sulfur, m is an integer from 1 to 6, n is an integer from 1 to 10, q, r, s, t, v are independently integers from 0 to 4, provided that both q and r cannot be 0 simultaneously, and s, t and v cannot all be 0 simultaneously, Z is an oligonucleoside] A nucleic acid according to the invention, comprising a ligand moiety comprising

[0057] Structure

[0058]

Chem.

[0059] [wherein, [oligonucleotide] represents consecutive nucleosides of the second strand] A nucleic acid according to the invention, comprising

[0060] alternatively, the following structure:

[0061]

Chem.

[0062] [wherein, r and s are each independently an integer selected from 1 to 16, and Z is an oligonucleoside] A nucleic acid according to the present invention, comprising a ligand moiety containing

[0063] Structure

[0064] [Chemical formula]

[0065] [wherein, [oligonucleotide] represents consecutive nucleosides of the second strand] A nucleic acid according to the present invention, containing

[0066] The present invention further provides a pharmaceutical composition comprising the nucleic acid described herein in combination with a pharmaceutically acceptable excipient or carrier.

[0067] The present invention further provides a nucleic acid or pharmaceutical composition described herein for use in therapy.

[0068] The present invention further provides a nucleic acid or pharmaceutical composition described herein for use in the prevention or treatment of diseases related to hemostatic disorders, such as diseases related to hemostatic disorders such as hemophilia.

[0069] The present invention further provides a nucleic acid or pharmaceutical composition described herein for use in the prevention or treatment of cardiovascular diseases.

[0070] The present invention further provides a nucleic acid or pharmaceutical composition described herein for use in the prevention or treatment of diabetes.

Brief Description of the Drawings

[0071]

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5-1

Figure 5-2

Figure 5-3

Figure 5-4

Figure 5-5

Figure 5-6

Figure 5-7

Figure 5-8

Figure 6-1

Figure 6-2

Figure 6-3

Figure 6-4

Figure 6-5

Figure 7

Figure 8

Figure 9a

Figure 9b-1

Figure 9b-2

Figure 10

Figure 11

Figure 12

Figure 13

Figure 14

Figure 15

Figure 16-1

Figure 16-2

Figure 17

Figure 18

BRIEF DESCRIPTION OF THE DRAWINGS

[0072] Definitions As used herein, the "first strand," also referred to herein as the antisense strand or guide strand and used interchangeably herein, refers to a nucleic acid strand, such as a strand of siRNA, such as dsiRNA, that contains a region that is substantially complementary to a target sequence, such as mRNA. As used herein, the term "complementary region" refers to a region on the antisense strand that is substantially complementary to a sequence, such as a target sequence. When the complementary region is not completely complementary to the target sequence, mismatches can typically be in the internal or terminal regions of the molecule. In some embodiments, the double-stranded nucleic acids of the invention, such as siRNA agents, contain nucleoside mismatches in the antisense strand.

[0073] The "second strand" (also referred to herein as the sense strand or passenger strand and used interchangeably herein) refers to a strand of a nucleic acid, such as siRNA, that contains a region that is substantially complementary to a region of the antisense strand as defined herein.

[0074] In the context of a molecule comprising a nucleic acid optionally further comprising a linker moiety and a ligand moiety, the nucleic acids of the invention may be referred to as oligonucleosides or oligonucleoside moieties.

[0075] An oligonucleotide is a short nucleic acid polymer. An oligonucleotide contains phosphodiester bonds between its nucleoside components (base + sugar), but the present invention is not limited to oligonucleotides always linked by such phosphodiester bonds between adjacent nucleosides, and other oligomers of nucleosides linked by bonds that are bonds other than phosphodiester bonds are contemplated. For example, the bond between nucleosides can be a phosphorothioate bond. Thus, the term "oligonucleoside" as used herein encompasses both oligonucleotides and other oligomers of nucleosides. Oligonucleosides that are nucleic acids having at least a portion that is an oligonucleotide are preferred according to the present invention. Oligonucleosides having one or more, or a majority of, phosphodiester backbone bonds between nucleosides are also preferred according to the present invention. Oligonucleosides having one or more, or a majority of, phosphodiester backbone bonds between nucleosides and also having one or more phosphorothioate backbone bonds between nucleosides (typically in the terminal regions of the first and / or second strands) are also preferred according to the present invention.

[0076] It is preferred herein that the nucleic acid according to the present invention is a double-stranded oligonucleoside containing one or more phosphorothioate backbone bonds between nucleosides. Thus, in all cases where the present application refers to an oligonucleotide, particularly in the chemical structures disclosed herein, the oligonucleotide can likewise be an oligonucleoside as defined herein.

[0077] In some embodiments, the double-stranded nucleic acid of the present invention, such as an siRNA agent, contains a nucleoside mismatch in the sense strand. In some embodiments, the nucleoside mismatch is, for example, within 5, 4, 3, 2, or 1 nucleoside from the 3' end of the nucleic acid, such as an siRNA.

[0078] In another embodiment, the nucleoside mismatch is, for example, at the 3' terminal nucleoside of the nucleic acid, such as an siRNA.

[0079] The "target sequence" (which may also be referred to as target RNA or target mRNA) refers to a continuous portion of the nucleoside sequence of an mRNA molecule formed during gene transcription, including the mRNA that is the product of RNA processing of the primary transcript.

[0080] The target sequence may be about 10 to 35 nucleosides in length, for example, about 15 to 30 nucleosides in length. For example, the target sequence may be about 15 to 30 nucleosides, 15 to 29, 15 to 28, 15 to 27, 15 to 26, 15 to 25, 15 to 24, 15 to 23, 15 to 22, 15 to 21, 15 to 20, 15 to 19, 15 to 18, 15 to 17, 18 to 30, 18 to 29, 18 to 28, 18 to 27, 18 to 26, 18 to 25, 18 to 24, 18 to 23, 18 to 22, 18 to 21, 18 to 20, 19 to 30, 19 to 29, 19 to 28, 19 to 27, 19 to 26, 19 to 25, 19 to 24, 19 to 23, 19 to 22, 19 to 21, 19 to 20, 20 to 30, 20 to 29, 20 to 28, 20 to 27, 20 to 26, 20 to 25, 20 to 24, 20 to 23, 20 to 22, 20 to 21, 21 to 30, 21 to 29, 21 to 28, 21 to 27, 21 to 26, 21 to 25, 21 to 24, 21 to 23, or 21 to 22 nucleosides in length. Ranges and lengths intermediate to those listed above are also contemplated as part of the present invention.

[0081] The term "ribonucleoside" or "nucleoside" may also refer to modified nucleosides, which are further detailed below.

[0082] The nucleic acid may be DNA or RNA and may contain modified nucleosides. RNA is the preferred nucleic acid.

[0083] The terms "iRNA", "siRNA", "RNAi agent", and "iRNA agent", "RNA interference agent" when used interchangeably herein, refer to an agent that contains RNA and mediates the targeted cleavage of an RNA transcript via the RNA-induced silencing complex (RISC) pathway. siRNA directs the sequence-specific degradation of mRNA through RNA interference (RNAi).

[0084] Double-stranded RNA is referred to herein as "double-stranded siRNA (dsiRNA) agent", "double-stranded siRNA (dsiRNA) molecule", "double-stranded RNA (dsRNA) agent", "double-stranded RNA (dsRNA) molecule", "dsiRNA agent", "dsiRNA molecule", or "dsiRNA", which refers to a complex of ribonucleic acid molecules having a double-stranded structure comprising two anti-parallel and substantially complementary nucleic acid strands that are said to have "sense" and "antisense" orientations with respect to the target RNA.

[0085] The majority of the nucleosides of each strand of a nucleic acid, such as a dsiRNA molecule, are preferably ribonucleosides, but in that case, each or both strands can also contain one or more non-ribonucleosides, such as deoxyribonucleosides or modified nucleosides. Further, as used herein, "siRNA" can include ribonucleosides having chemical modifications.

[0086] The term "modified nucleoside" independently refers to a nucleoside having a modified sugar moiety, a modified internucleoside linkage, or a modified nucleobase, or any combination thereof. Thus, the term modified nucleoside encompasses substitutions, additions or removals to the internucleoside linkage, sugar moiety, or nucleobase, for example functional groups or atoms. Any such modification, when used in an siRNA-type molecule, is encompassed by "iRNA" or "RNAi agent" or "siRNA" or "siRNA agent" for the purposes of this specification and the claims.

[0087] The two strands forming the double-stranded structure may be different parts of one larger molecule, or they may be separate molecules, such as RNA molecules.

[0088] The term "nucleoside overhang" refers to at least one unpaired nucleoside extending from the double-stranded structure of a nucleic acid according to the present invention. The nucleic acid according to the present invention may include an overhang of at least one nucleoside; alternatively, the overhang may include at least two nucleosides, at least three nucleosides, at least four nucleosides, at least five nucleosides, or more. The nucleoside overhang may include or consist of nucleoside / nucleoside analogs including deoxynucleosides. The overhang(s) may be on the sense strand, the antisense strand, or any combination thereof. Further, the nucleoside(s) of the overhang may be present at the 5'-end, 3'-end, or both ends of either the antisense or sense strand.

[0089] In certain embodiments, the antisense strand has an overhang of 1 to 10 nucleosides, such as 0 to 3, 1 to 3, 2 to 4, 2 to 5, 4 to 10, 5 to 10, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleosides at the 3'-end or 5'-end.

[0090] "Blunt" or "blunt end" means that there are no unpaired nucleosides at that end of the double-stranded nucleic acid, i.e., there is no nucleoside overhang. The nucleic acids of the present invention include those having no nucleoside overhang at one end or no nucleoside overhang at either end.

[0091] Unless otherwise indicated, the term "complementary," when used to describe a first nucleoside sequence in relation to a second nucleoside sequence, refers to the ability of an oligonucleoside containing the first nucleoside sequence to hybridize under certain conditions with an oligonucleoside containing the second nucleoside sequence to form a double-stranded structure, as would be understood by one of ordinary skill in the art. Such conditions may be, for example, stringent conditions, which may include 400 mM NaCl, 40 mM PIPES pH 6.4, 1 mM EDTA, at 50 °C or 70 °C for 12 - 16 hours, followed by washing (see, e.g., "Molecular Cloning: A Laboratory Manual, Sambrook, et al. (1989) Cold Spring Harbor Laboratory Press").

[0092] Complementary sequences within a nucleic acid, such as a dsiRNA, as described herein, include base pairing of an oligonucleoside containing a first nucleoside sequence with an oligonucleoside containing a second nucleoside sequence over the entire length of one or both of the nucleoside sequences. Such sequences can be said to be "fully complementary" to each other herein. However, when a first sequence is said to be "substantially complementary" or "partially complementary" to a second sequence herein, the two sequences may be fully complementary, or they may form one or more mismatched base pairs, such as 2, 4, or 5, but preferably no more than 5 mismatched base pairs, while maintaining the ability to hybridize under the conditions most relevant to their ultimate application, such as inhibition of gene expression via the RISC pathway. Overhangs are not considered mismatches with respect to determination of complementarity. For example, a nucleic acid, such as a dsiRNA, containing one oligonucleoside 17 nucleosides in length and another oligonucleoside 19 nucleosides in length, where the longer oligonucleoside contains a 17-nucleoside sequence that is fully complementary to the shorter oligonucleoside, can still be said to be "fully complementary."

[0093] A "complementary" array, as used herein, may also include, or may consist entirely of, non-Watson-Crick base pairs, or base pairs formed from non-natural and modified nucleosides, so long as the above requirements regarding its ability to hybridize are met. Such non-Watson-Crick base pairs include, but are not limited to, G:U wobbles or Hoogsteen base pairing.

[0094] The terms "complementary," "fully complementary," and "substantially / partially complementary" as used herein can be used with respect to base matching between nucleic acids, such as between the sense and antisense strands of a dsiRNA, or between the antisense strand of a double-stranded nucleic acid, such as an siRNA agent, and a target sequence.

[0095] In the present invention, the second strand of the nucleic acid according to the present invention is at least partially complementary to the first strand of the nucleic acid. In certain embodiments, the first and second strands of the nucleic acid according to the present invention are partially complementary when they form a double-stranded region having a length of at least 17 base pairs and containing no more than 1, 2, 3, 4, or 5 mismatched base pairs.

[0096] In certain embodiments, the first and second strands of the nucleic acid according to the present invention are partially complementary when they form a double-stranded region having a length of 19 base pairs and containing no more than 1, 2, 3, 4, or 5 mismatched base pairs. In certain embodiments, the first and second strands of the nucleic acid according to the present invention are partially complementary when they form a double-stranded region having a length of 21 base pairs and containing no more than 1, 2, 3, 4, or 5 mismatched base pairs.

[0097] Alternatively, the first and second strands of the nucleic acid according to the present invention are partially complementary when they form a double-stranded region having a length of at least 17 base pairs, and at least 14, 15, 16, or 17 of said base pairs are complementary base pairs, particularly Watson-Crick base pairs.

[0098] In certain embodiments, the first and second strands of the nucleic acid according to the present invention form a double-stranded region having a length of 19 base pairs, and are partially complementary when at least 14, 15, 16, 17, 18 or all 19 base pairs are complementary base pairs, particularly Watson-Crick base pairs. In certain embodiments, the first and second strands of the nucleic acid according to the present invention form a double-stranded region having a length of 21 base pairs, and are partially complementary when at least 16, 17, 18, 19, 20 or all 21 base pairs are complementary base pairs, particularly Watson-Crick base pairs.

[0099] As used herein, a nucleic acid that is "substantially complementary" or "partially complementary" to at least a portion of messenger RNA (mRNA) refers to a nucleic acid that is substantially or partially complementary to a continuous portion of the mRNA of interest (e.g., the mRNA encoding a gene). In certain embodiments, the continuous portion of the mRNA is any one of the sequences listed in Table 1, i.e., any one of SEQ ID NOs: 3-42 and 443-448. For example, a nucleic acid is complementary to at least a portion of the mRNA of a gene if its sequence is substantially or partially complementary to an uninterrupted portion of the mRNA encoding the gene of interest.

[0100] Thus, in some preferred embodiments, the antisense oligonucleosides disclosed herein are completely complementary to the target gene sequence.

[0101] In other embodiments, the antisense oligonucleosides disclosed herein are substantially or partially complementary to the target RNA sequence and include a continuous nucleoside sequence that is at least about 80% complementary, e.g., at least about 85%, 86%, 87%, 88%, 89%, about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% complementary, or 100% complementary, to the equivalent region of the target RNA sequence over its entire length.

[0102] In certain embodiments, the first (antisense) strand of the nucleic acid according to the invention is partially or fully complementary to a continuous portion of the RNA transcribed from the HCII gene. In certain embodiments, the first strand of the nucleic acid according to the invention is partially or fully complementary to a continuous portion of at least 17 nucleosides of the HCII mRNA. In certain embodiments, the first strand of the nucleic acid according to the invention is partially or fully complementary to a continuous portion of 17, 18, 19, 20, 21, 22 or 23 nucleosides of the HCII mRNA. In certain embodiments, the first strand of the nucleic acid according to the invention is partially or fully complementary to a continuous portion of 17, 18, 19, 20, 21, 22 or 23 nucleosides of any one of the sequences listed in Table 1, namely any one of SEQ ID NOs: 3 to 22 and 445 to 446.

[0103] In certain embodiments, the first (antisense) strand of the nucleic acid according to the invention comprises a contiguous nucleoside sequence of at least 17 nucleosides, and at least 14, 15, 16 or 17 nucleosides of said contiguous nucleoside sequence are partially complementary to a contiguous portion of the HCII mRNA when at least 14, 15, 16 or 17 nucleosides of said contiguous nucleoside sequence are complementary to a contiguous portion of the HCII mRNA. In certain embodiments, the first strand of the nucleic acid according to the invention comprises a contiguous nucleoside sequence of at least 17 nucleosides, and at least 14, 15, 16 or 17 nucleosides of said contiguous nucleoside sequence are complementary to a contiguous portion of any one of the sequences listed in Table 1, i.e., any one of SEQ ID NOs: 3-22 and 445-446. In certain embodiments, the first strand of the nucleic acid according to the invention comprises a contiguous nucleoside sequence of 19 nucleosides, and at least 14, 15, 16, 17, 18 or all 19 nucleosides of said contiguous nucleoside sequence are complementary to a contiguous portion of any one of the sequences listed in Table 1, i.e., any one of SEQ ID NOs: 3-22 and 445-446. In certain embodiments, the first strand of the nucleic acid according to the invention comprises a contiguous nucleoside sequence of 23 nucleosides, and at least 18, 19, 20, 21, 22 or all 23 nucleosides of said contiguous nucleoside sequence are complementary to a contiguous portion of any one of the sequences listed in Table 1, i.e., any one of SEQ ID NOs: 3-22 and 445-446.

[0104] In certain embodiments, the first (antisense) strand of the nucleic acid according to the invention is partially or completely complementary to a continuous portion of the RNA transcribed from the ZPI gene. In certain embodiments, the first strand of the nucleic acid according to the invention is partially or completely complementary to a continuous portion of at least 17 nucleosides of the ZPI mRNA. In certain embodiments, the first strand of the nucleic acid according to the invention is partially or completely complementary to a continuous portion of 17, 18, 19, 20, 21, 22 or 23 nucleosides of the ZPI mRNA. In certain embodiments, the first strand of the nucleic acid according to the invention is partially or completely complementary to a continuous portion of 17, 18, 19, 20, 21, 22 or 23 nucleosides of any one of the sequences listed in Table 1, i.e., any one of SEQ ID NOs: 23 to 42 and 443 to 444.

[0105] In certain embodiments, the first (antisense) strand of the nucleic acid according to the invention comprises a continuous nucleoside sequence of at least 17 nucleosides, and at least 14, 15, 16 or 17 nucleosides of said continuous nucleoside sequence are partially complementary to a continuous portion of ZPI mRNA when at least 14, 15, 16 or 17 nucleosides of said continuous nucleoside sequence are complementary to a continuous portion of ZPI mRNA. In certain embodiments, the first strand of the nucleic acid according to the invention comprises a continuous nucleoside sequence of at least 17 nucleosides, and at least 14, 15, 16 or 17 nucleosides of said continuous nucleoside sequence are complementary to a continuous portion of any one of the sequences listed in Table 1, namely, any one of SEQ ID NOs: 23-42 and 443-444. In certain embodiments, the first strand of the nucleic acid according to the invention comprises a continuous nucleoside sequence of 19 nucleosides, and at least 14, 15, 16, 17, 18 or all 19 nucleosides of said continuous nucleoside sequence are complementary to a continuous portion of any one of the sequences listed in Table 1, namely, any one of SEQ ID NOs: 23-42 and 443-444. In certain embodiments, the first strand of the nucleic acid according to the invention comprises a continuous nucleoside sequence of 23 nucleosides, and at least 18, 19, 20, 21, 22 or all 23 nucleosides of said continuous nucleoside sequence are complementary to a continuous portion of any one of the sequences listed in Table 1, namely, any one of SEQ ID NOs: 23-42 and 443-444.

[0106] In certain embodiments, the first (antisense) strand of the nucleic acid according to the invention is partially or fully complementary to a continuous portion of the RNA transcribed from the B4GALT1 gene. In certain embodiments, the first strand of the nucleic acid according to the invention is partially or fully complementary to a continuous portion of at least 17 nucleosides of the B4GALT1 mRNA. In certain embodiments, the first strand of the nucleic acid according to the invention is partially or fully complementary to a continuous portion of 17, 18, 19, 20, 21, 22 or 23 nucleosides of the B4GALT1 mRNA. In certain embodiments, the first strand of the nucleic acid according to the invention is partially or fully complementary to a continuous portion of 17, 18, 19, 20, 21, 22 or 23 nucleosides of any one of the sequences listed in Table 1, i.e., any one of SEQ ID NOs: 447 to 448.

[0107] In certain embodiments, the first (antisense) strand of the nucleic acid according to the invention comprises a continuous nucleoside sequence of at least 17 nucleosides, and at least 14, 15, 16 or 17 nucleosides of said continuous nucleoside sequence are partially complementary to a continuous portion of B4GALT1 mRNA, if at least 17 nucleosides of said continuous nucleoside sequence are complementary to a continuous portion of B4GALT1 mRNA. In certain embodiments, the first strand of the nucleic acid according to the invention comprises a continuous nucleoside sequence of at least 17 nucleosides, and at least 14, 15, 16 or 17 nucleosides of said continuous nucleoside sequence are complementary to a continuous portion of any one of the sequences listed in Table 1, i.e., any one of SEQ ID NOs: 447 to 448. In certain embodiments, the first strand of the nucleic acid according to the invention comprises a continuous nucleoside sequence of 19 nucleosides, and at least 14, 15, 16, 17, 18 or all 19 nucleosides of said continuous nucleoside sequence are complementary to a continuous portion of any one of the sequences listed in Table 1, i.e., any one of SEQ ID NOs: 447 to 448. In certain embodiments, the first strand of the nucleic acid according to the invention comprises a continuous nucleoside sequence of 23 nucleosides, and at least 18, 19, 20, 21, 22 or all 23 nucleosides of said continuous nucleoside sequence are complementary to a continuous portion of any one of the sequences listed in Table 1, i.e., any one of SEQ ID NOs: 447 to 448.

[0108] In some embodiments, the nucleic acid of the invention, e.g., siRNA, comprises a sense strand that is substantially or partially complementary to an antisense oligonucleotide, which in turn is complementary to the target gene sequence and comprises a continuous nucleoside sequence. The nucleoside sequence of the sense strand is typically at least about 80% complementary, e.g., about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% complementary, or 100% complementary, to the equivalent region of the nucleoside sequence of the antisense strand over its entire length.

[0109] In some embodiments, the nucleic acids of the invention, such as siRNAs, comprise an antisense strand that is substantially or partially complementary to the target sequence and at least 80% complementary to the target sequence over its entire length, for example, about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% complementary, or 100% complementary, and includes a continuous nucleoside sequence.

[0110] As used herein, a "subject" is an animal, such as a mammal, such as a primate (e.g., human, non-human primate, such as monkey and chimpanzee), or non-primate or bird, that endogenously or heterologously expresses a target gene when the target gene sequence has sufficient complementarity with a nucleic acid, such as an siRNA agent, to promote target knockdown. In certain preferred embodiments, the subject is a human.

[0111] The term "treating" or "treatment" refers to beneficial or desired results, including, but not limited to, alleviation or amelioration of one or more symptoms associated with gene expression. "Treatment" can also mean prolonging survival as compared to expected survival in the absence of treatment. Treatment can include prevention of the occurrence of complications, e.g., reduction of liver damage in a subject having a liver infection.

[0112] "Therapeutically effective amount" as used herein is intended to include an amount of a nucleic acid, such as an siRNA, that is sufficient to effect treatment of a disease in a subject when administered to the patient (e.g., by reducing, ameliorating, or maintaining an existing disease or one or more symptoms of the disease or its associated complications).

[0113] The phrase "pharmaceutically acceptable" as used herein refers to compounds, materials, compositions, or dosage forms that are suitable for use in contact with the tissues of human subjects and animal subjects without excessive toxicity, irritation, allergic response, or other problems or complications commensurate with a reasonable benefit / risk ratio.

[0114] As used herein, the phrase "pharmaceutically acceptable carrier" refers to a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, manufacturing aid, or solvent encapsulation material, involved in transporting or delivering a subject compound from one organ or part of the body to another organ or another part of the body. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the subject being treated.

[0115] It is intended that when values or ranges of values of parameters are recited, values intermediate to the recited values and ranges are also intended to be part of the present invention.

[0116] The articles "a" and "an" are used herein to refer to one or more (i.e., at least one) of the grammatical objects of the article.

[0117] The term "including" is used herein to mean the phrase "including but not limited to" and is used interchangeably therewith.

[0118] The term "or" is used herein to mean the term "and / or" and is used interchangeably therewith, unless the context clearly indicates otherwise. For example, "sense strand or antisense strand" is understood to mean "sense strand or antisense strand or sense strand and antisense strand".

[0119] The term "about" is used herein to mean within the typical acceptable ranges in the art. For example, "about" can be understood to be about two standard deviations from the average. In certain embodiments, about means +10%. In certain embodiments, about means +5%. It is understood that when "about" is present before a series of numbers or ranges, "about" can modify each of the numbers in the series or range.

[0120] The term "at least" before a number or series of numbers is understood to include the number adjacent to the term "at least" and all subsequent numbers or integers that may logically be included as apparent from the context. For example, the number of nucleosides in a nucleic acid molecule must be an integer. For example, "at least 18 nucleosides out of 21 nucleoside nucleic acid molecules" means that 18, 19, 20, or 21 nucleosides have the indicated property. When "at least" is present before a series of numbers or a range, it is understood that "at least" may modify each of the numbers in the series or range.

[0121] As used herein, "not exceeding" or "less than" is understood to be from the logical lower value or integer up to zero, adjacent to the value in the phrase and as logical from the context. For example, a double-stranded having an overhang of "not exceeding 2 nucleosides" has 2, 1, or 0 nucleoside overhangs. When "not exceeding" is present before a series of numbers or a range, it is understood that "not exceeding" may modify each of the numbers in the series or range.

[0122] The terminal region of the strand is the last 5 nucleosides from the 5' or 3' end.

[0123] The various embodiments of the present invention can be combined as determined to be appropriate by one of ordinary skill in the art.

[0124] Abasic nucleoside In certain embodiments, one, such as two, such as three, such as four, or more abasic nucleosides are present in the nucleic acid according to the present invention. An abasic nucleoside is a modified nucleoside because it lacks the base normally found at the 1-position of the sugar moiety. Typically, hydrogen is present at the 1-position of the sugar moiety of the abasic nucleoside present in the nucleic acid according to the present invention.

[0125] The abasic nucleoside is in the terminal region of the second strand and is preferably located within 5 nucleosides from the end of the strand. The terminal region may be the terminal 5 nucleosides including the abasic nucleoside.

[0126] The second strand may include the following as preferred features (all of these are specifically contemplated in combination, unless mutually exclusive): Two or more abasic nucleosides in the terminal region of the second strand, and / or Two or more abasic nucleosides in either the 5' or 3' terminal region of the second strand, and / or Two or more abasic nucleosides in either the 5' or 3' terminal region of the second strand, where the abasic nucleosides are present in the overhangs described herein, and / or Two or more consecutive abasic nucleosides in the terminal region of the second strand, preferably where one of the abasic nucleosides is the terminal nucleoside, and / or Two or more consecutive abasic nucleosides in either the 5' or 3' terminal region of the second strand, preferably where one of the abasic nucleosides is the terminal nucleoside in either the 5' or 3' terminal region of the second strand, and / or An inverted nucleoside internucleoside linkage connects at least one abasic nucleoside to an adjacent base nucleoside in the terminal region of the second strand, and / or An inverted nucleoside internucleoside linkage connects at least one abasic nucleoside to an adjacent base nucleoside in either the 5' or 3' terminal region of the second strand, and / or An abasic nucleoside as the second last nucleoside that is connected to a nucleoside that is not the terminal nucleoside (referred to herein as the third last nucleoside) via an inverted linkage, and / or An abasic nucleoside as two terminal nucleosides connected via a 5'-3' linkage when the strand is read in the direction towards the end including the terminal nucleoside. When the strand is read in the direction towards the end containing the terminal nucleoside, an abasic nucleoside as two terminal nucleosides connected via a 3'-5' linkage, An abasic nucleoside at two terminal positions, wherein the second last nucleoside is connected to the third last nucleoside via a reverse linkage, and the reverse linkage is a 5-5' reverse linkage or a 3'-3' reverse linkage, An abasic nucleoside at two terminal positions, wherein the second last nucleoside is connected to the third last nucleoside via a reverse linkage, and is any of the following: (1) The reverse linkage is a 5-5' reverse linkage, and the linkage between the terminal abasic nucleoside and the second last abasic nucleoside is 3'5' when read in the direction towards the end containing the terminal abasic nucleoside and the second last abasic nucleoside, or (2) The reverse linkage is a 3-3' reverse linkage, and the linkage between the terminal abasic nucleoside and the second last abasic nucleoside is 5'3' when read in the direction towards the end containing the terminal abasic nucleoside and the second last abasic nucleoside.

[0127] Preferably, an abasic nucleoside is present at the end of the second strand.

[0128] Preferably, two or at least two abasic nucleosides are present in the terminal region of the second strand, preferably at the end and the second last position.

[0129] Preferably, two or more abasic nucleosides are consecutive, for example, all abasic nucleosides may be consecutive. For example, one or two or three or four nucleosides at the end may be abasic nucleosides.

[0130] A nucleoside without a base may also be linked to an adjacent nucleoside through a 5'-3' phosphodiester linkage or a reverse linkage, except when there is only one nucleoside without a base at the end. When there is only one nucleoside without a base at the end, it has a reverse linkage to the adjacent nucleoside.

[0131] Reverse linkage (which may also be called inverted linkage and is also found in the art) includes any of 5'-5', 3'-3', 3'-2' or 2'-3' phosphodiester linkages between adjacent sugar moieties of nucleosides.

[0132] A nucleoside without a base that is not at the end has two phosphodiester bonds with each adjacent nucleoside, and these may be reverse linkages, or 5'-3 phosphodiester linkages, or one each.

[0133] A preferred embodiment includes two nucleosides without a base at the end and the second-to-last position of the second strand, and the reverse nucleoside internucleoside linkage is located between the second-to-last (base-free) nucleoside and the third-to-last nucleoside.

[0134] Preferably, there are two nucleosides without a base at the end and the second-to-last position of the second strand, the second-to-last nucleoside is linked to the third-to-last nucleoside through a reverse nucleoside internucleoside linkage, and is linked to the terminal nucleoside through a 5'-3' or 3'-5' phosphodiester linkage (read in the direction of the end of the molecule).

[0135] Preferably, the nucleic acid according to the present invention contains one or more nucleosides without a base, and optionally, one or more nucleosides without a base are in the terminal region of the second strand, and / or at least one nucleoside without a base is linked to an adjacent base nucleoside through a reverse nucleoside internucleoside linkage.

[0136] Typically, the second strand contains two consecutive abasic nucleosides in the 5' terminal region of the second strand, one of the abasic nucleosides being the terminal nucleoside in the 5' terminal region of the second strand and the other abasic nucleoside being the penultimate nucleoside in the 5' terminal region of the second strand, (a) the penultimate abasic nucleoside is connected through a reverse nucleoside internucleoside linkage to the adjacent first base nucleoside in the adjacent 5' terminal proximal region, (b) the reverse linkage is a 5-5' reverse linkage, and (c) the linkage between the terminal abasic nucleoside and the penultimate abasic nucleoside is 3'5' when read towards the end containing the terminal abasic nucleoside and the penultimate abasic nucleoside. More typically, (i) the first strand and the second strand each have a length of 23 nucleosides, (ii) two phosphorothioate nucleoside internucleoside linkages are respectively between three consecutive positions in the 5' terminal proximal region of the second strand, the first phosphorothioate nucleoside internucleoside linkage is between the adjacent first base nucleoside in the 5' terminal proximal region of the second strand as in (a) and the adjacent second base nucleoside, and the second phosphorothioate nucleoside internucleoside linkage is between the adjacent second base nucleoside and the adjacent third base nucleoside in the 5' terminal proximal region of the second strand, (iii) two phosphorothioate nucleoside internucleoside linkages are respectively between three consecutive positions in both the 5' terminal region and the 3' terminal region of the first strand, whereby the terminal nucleosides in the 5' terminal region and the 3' terminal region of the first strand are respectively attached by phosphorothioate nucleoside internucleoside linkages to their respective 5' and 3' adjacent penultimate nucleosides, and each of the first 5' and 3' penultimate nucleosides is attached by phosphorothioate nucleoside internucleoside linkages to their respective 5' and 3' adjacent antepenultimate nucleosides, and (iv) the second strand of the nucleic acid is directly or indirectly conjugated to one or more ligand moieties in the 3' terminal region of the second strand.

[0137] Alternatively, the second strand preferably contains two consecutive abasic nucleosides in the overhang in the 3'-terminal region of the second strand, wherein one of the abasic nucleosides is the terminal nucleoside in the 3'-terminal region of the second strand, and the other abasic nucleoside is the penultimate nucleoside in the 3'-terminal region of the second strand, (a) the penultimate abasic nucleoside is connected to the adjacent first base nucleoside in the adjacent 3'-terminal proximal region through a reverse nucleoside internucleoside linkage, (b) the reverse linkage is a 3-3' reverse linkage, and (c) the linkage between the terminal abasic nucleoside and the penultimate abasic nucleoside is 5'-3' when read towards the end containing the terminal abasic nucleoside and the penultimate abasic nucleoside. More typically, (i) the first strand and the second strand each have a length of 23 nucleosides, (ii) two phosphorothioate internucleoside linkages are respectively between three consecutive positions in the 3'-terminal proximal region of the second strand, the first phosphorothioate internucleoside linkage is between the adjacent first base nucleoside and the adjacent second base nucleoside in the 3'-terminal proximal region of the second strand as in (a), and the second phosphorothioate internucleoside linkage is between the adjacent second base nucleoside and the adjacent third base nucleoside in the 3'-terminal proximal region of the second strand, (iii) two phosphorothioate internucleoside linkages are respectively between three consecutive positions in both the 5'-terminal region and the 3'-terminal region of the first strand, whereby the terminal nucleosides in the 5'-terminal region and the 3'-terminal region of the first strand are respectively attached to their 5' and 3' adjacent penultimate nucleosides by phosphorothioate internucleoside linkages, and each of the first 5' and 3' penultimate nucleosides is attached to its 5' and 3' adjacent antepenultimate nucleosides by phosphorothioate internucleoside linkages, and (iv) the second strand of the nucleic acid is directly or indirectly conjugated to one or more ligand moieties in the 5'-terminal region of the second strand.

[0138] Examples of structures are as follows (the specific RNA nucleosides shown are not limiting and can be any RNA nucleoside):

[0139] A 3'-3' inverted bond (also showing the 5'-3' direction of the last phosphodiester bond between two abasic molecules, reading towards the end of the molecule)

[0140] [Chemical formula]

[0141] B Exemplify a 5'-5' inverted bond (also showing the 3'-5' direction of the last phosphodiester bond between two abasic molecules, reading towards the end of the molecule)

[0142] [Chemical formula]

[0143] The abasic nucleoside(s) present in the nucleic acid are provided in the presence of an inverted nucleoside linkage(s), i.e., a 5'-5' or 3'-3' inverted nucleoside linkage. The inverted linkage results from a change in the orientation of adjacent nucleoside sugars such that the sugar has a 3'-5' orientation as opposed to the conventional 5'-3' orientation (referring to the numbering of the ring atoms on the nucleoside sugar). The abasic nucleoside(s) present in the nucleic acid of the present invention preferably contain such inverted nucleoside sugars.

[0144] In the case of a terminal nucleoside having an inverted orientation, this results in a stereoconfiguration of the "inverted" terminus for the overall nucleic acid. Certain structures shown and referred to herein are represented using the conventional 5'-3' direction (with reference to the numbering of the ring atoms on the nucleoside sugar), but the change in orientation and the presence of a terminal nucleoside having a proximal 3'-3' reverse linkage results in a nucleic acid having an overall 5'-5' terminus structure (i.e., the conventional 3' terminal nucleoside becomes the 5' terminal nucleoside). Alternatively, it is understood that the change in orientation and the presence of a terminal nucleoside having a proximal 5'-5' reverse linkage results in a nucleic acid having an overall 3'-3' terminus structure.

[0145] The proximal 3'-3' or 5'-5' reverse linkages described herein can include a reverse linkage that is directly adjacent / attached to a terminal nucleoside having an inverted orientation, such as a single terminal nucleoside having an inverted orientation. Alternatively, the proximal 3'-3' or 5'-5' reverse linkages described herein can include a reverse linkage that is adjacent to two or more nucleosides having an inverted orientation, such as two or more terminal region nucleosides having an inverted orientation, such as the terminal nucleoside and the second last nucleoside. In this way, the reverse linkage can be attached to the second last nucleoside having an inverted orientation. One of ordinary skill in the art will understand that the inverted orientation described above can result in a nucleic acid molecule having an overall 3'-3' or 5'-5' terminus structure as described herein, but due to the presence of one or more additional reverse linkages and / or nucleosides having an inverted orientation, it is also understood that the overall nucleic acid can have a 3'-5' terminus structure corresponding to the conventionally arranged 5' / 3' terminus.

[0146] In one aspect, the nucleic acid may have a 3'-3' reverse linkage, and the terminal sugar moiety may contain a 5'OH rather than a 5' phosphate group at the 5' position of the terminal sugar.

[0147] Thus, one of ordinary skill in the art will clearly understand that, when reverse linkage(s) is / are present, 5'-5', 3'-3', and 3'-5' (read in the direction of their termini) terminal variants of the conventional 5'-3' structure (referring to the numbering of the ring atoms on the terminal nucleoside sugar) as shown herein are within the scope of this disclosure.

[0148] For example, in the situation of one or more nucleosides having an inverted orientation that results in reverse nucleoside-nucleoside linkages and / or inverted termini, and also where the relative position of a linkage (e.g., to a linker) or the position of an internal feature (e.g., a modified nucleoside) is defined relative to the 5' or 3' terminus of the nucleic acid, the 5' or 3' terminus is the conventional 5' or 3' terminus that would have been present if the reverse linkage was not in a fixed position, and the conventional 5' or 3' terminus is determined by considering the orientation of the majority of the internal nucleoside linkages and / or nucleoside orientations within the nucleic acid. From these internal linkages and / or nucleoside orientations, it is possible to identify which terminus of the nucleic acid would constitute the conventional 5' and 3' termini of a molecule without reverse linkages (referring to the numbering of the ring atoms on the terminal nucleoside sugar).

[0149] For example, in the structure shown below, abasic residues are present at the first two positions located at the 5' terminus. When the terminal nucleoside has an inverted orientation, the 5' terminus shown in the following schematic, which is the conventional 5' terminus, may actually contain a 3'OH considering the inverted nucleoside at the terminal position. Nevertheless, the majority of the molecule contains conventional nucleoside-nucleoside linkages that run from the 3'OH of one sugar to the 5' phosphate of the next sugar when read in the standard 5'[PO4] to 3'[OH] direction of the nucleic acid molecule (referring to the numbering of the ring atoms on the nucleoside sugar), which can be used to determine the conventional 5' and 3' termini that would be seen in the absence of the stereochemistry of the inverted terminus.

[0150] 5' A-A-Me-Me-Me-Me-Me-Me-F-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me 3'

[0151] The reverse linkage is preferably located at the end of a nucleic acid, such as RNA, which is distal to the ligand portion of the molecule, such as the GalNAc-containing portion.

[0152] A GalNAc-siRNA construct having 5'-GalNAc on the sense strand may have a reverse linkage on the opposite end of the sense strand.

[0153] A GalNAc-siRNA construct having 3'-GalNAc on the sense strand may have a reverse linkage on the opposite end of the sense strand.

[0154] In certain embodiments, the invention provides a first strand that is at least partially complementary to a portion of an RNA transcribed from a target gene, and a second strand that is at least partially complementary to the first strand wherein the second strand contains two consecutive abasic nucleosides in the 5'-terminal region of the second strand, one of the abasic nucleosides being the terminal nucleoside in the 5'-terminal region of the second strand and the other abasic nucleoside being the penultimate nucleoside in the 5'-terminal region of the second strand, and wherein: (a) the penultimate abasic nucleoside is connected through a reverse nucleoside internucleoside linkage to the adjacent first base nucleoside in the adjacent 5'-terminal proximal region, (b) the reverse linkage is a 5-5' reverse linkage, (c) the linkage between the terminal abasic nucleoside and the penultimate abasic nucleoside is 3'-5' when read towards the end containing the terminal abasic nucleoside and the penultimate abasic nucleoside, relates to a nucleic acid.

[0155] In certain embodiments, the invention provides A first strand that is at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand that is at least partially complementary to the first strand A nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region, (i) Preferably, the first strand and the second strand each have a length of 23 nucleosides (this length for the second strand includes 2 abasic nucleosides), (ii) The second strand contains 2 consecutive abasic nucleosides in the 5'-terminal region of the second strand, one of the abasic nucleosides being the terminal nucleoside in the 5'-terminal region of the second strand, and the other abasic nucleoside being the second last nucleoside in the 5'-terminal region of the second strand, (a) The second last abasic nucleoside is connected to the adjacent first base nucleoside in the adjacent 5'-terminal vicinity region through a reverse nucleoside internucleoside linkage, (b) The reverse linkage is a 5-5' reverse linkage, (c) The linkage between the terminal abasic nucleoside and the second last abasic nucleoside is 3'-5' when read towards the end including the terminal abasic nucleoside and the second last abasic nucleoside, (iii) Two phosphorothioate internucleoside linkages are respectively present between three consecutive positions in the 5'-terminal vicinity region of the second strand, the first phosphorothioate internucleoside linkage is present between the first base nucleoside in (a) and the adjacent second base nucleoside in the 5'-terminal vicinity region of the second strand, and the second phosphorothioate internucleoside linkage is present between the second base nucleoside and the adjacent third base nucleoside in the 5'-terminal vicinity region of the second strand, (iv) Two phosphorothioate internucleoside linkages are present between three consecutive positions in both the 5'-terminal region and the 3'-terminal region of the first strand, whereby the terminal nucleosides in each of the 5'-terminal region and the 3'-terminal region of the first strand are attached by the phosphorothioate internucleoside linkages to the respective 5'- and 3'-adjacent penultimate nucleosides, and the respective 5'- and 3'-penultimate nucleosides are attached by the phosphorothioate internucleoside linkages to the respective 5'- and 3'-adjacent antepenultimate nucleosides. (v) The second strand of the nucleic acid is conjugated directly or indirectly to one or more ligand moieties in the 3'-terminal region of the second strand. Relating to a nucleic acid.

[0156] In certain embodiments, the present invention a first strand that is at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand that is at least partially complementary to the first strand A nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region, wherein the second strand has the following 5'-terminal motif

[0157] [Chemical formula]

[0158] [wherein B represents a nucleoside base, T represents H, OH or a 2'-ribose modification, Z represents the remaining nucleosides of the second strand] Relating to a nucleic acid comprising two consecutive abasic nucleosides in the 5'-terminal region of the second strand, present as

[0159] In certain embodiments, the present invention A first strand that is at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand that is at least partially complementary to the first strand A nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region, wherein the second strand has the following 5'-terminal motif

[0160] [Chemical formula]

[0161] [wherein, B represents a nucleoside base, T represents H, OH or a 2'-ribose modification, V represents O or S (preferably O), R represents H or C 1~4 alkyl (preferably H), and Z represents the remaining nucleosides of the second strand], more preferably the following 5'-terminal motif

[0162] [Chemical formula]

[0163] [wherein, B represents a nucleoside base, T represents H, OH or a 2'-ribose modification, and Z represents the remaining nucleosides of the second strand] relates to a nucleic acid comprising two consecutive abasic nucleosides in the 5'-terminal region of the second strand, present as such.

[0164] In certain embodiments, the present invention provides A first strand that is at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand that is at least partially complementary to the first strand A nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region, The second strand has the following 5'-end motif

[0165] [Chemical formula]

[0166] [wherein B represents a nucleoside base, T represents H, OH or a 2'-ribose modification, V represents O or S (preferably O), R represents H or C 1~4 alkyl (preferably H), Z contains 11 to 26 consecutive nucleosides, preferably 15 to 21 consecutive nucleosides, more preferably 19 consecutive nucleosides], more preferably the following 5'-end motif

[0167] [Chemical formula]

[0168] [wherein B represents a nucleoside base, T represents H, OH or a 2'-ribose modification, Z contains 11 to 26 consecutive nucleosides, preferably 15 to 21 consecutive nucleosides, more preferably 19 consecutive nucleosides] relates to a nucleic acid comprising two consecutive abasic nucleosides in the 5'-terminal region of the second strand, present as

[0169] In some embodiments, the modification pattern of the second (sense) strand of the nucleic acid according to the invention is ia-ia-Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me [wherein ia represents an inverted abasic nucleoside] comprises or consists of

[0170] In such an embodiment, the second strand preferably has the following 5'-end motif

[0171]

Chemical formula

[0172] [wherein, B represents the nucleoside base of the first base nucleoside in the 5'-end region of the second strand, T represents a 2'-Me ribose modification, Z represents the remaining consecutive base nucleosides of the second strand] and includes.

[0173] In such an embodiment, the modification pattern of the first strand of the nucleic acid preferably Me-F-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me includes or consists of.

[0174] In some embodiments, the modification pattern of the second (sense) strand of the nucleic acid according to the present invention is ia-ia-Me-Me-Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me [wherein, ia represents an inverted abasic nucleoside] includes or consists of.

[0175] In such an embodiment, the second strand preferably has the following 5'-end motif

[0176]

Chemical formula

[0177] [wherein, B represents the nucleoside base of the first base nucleoside in the 5'-end region of the second strand, T represents a 2'-Me ribose modification, Z represents the remaining consecutive base nucleosides of the second strand comprises.

[0178] In such embodiments, the modification pattern of the first strand of the nucleic acid is preferably Me-F-Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me comprises or consists of.

[0179] In some embodiments, the modification pattern of the second (sense) strand of the nucleic acid according to the invention is ia-ia-Me(s)-Me(s)-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me [where (s) is a phosphorothioate internucleoside linkage and ia represents an inverted abasic nucleoside] comprises or consists of.

[0180] In such embodiments, the second strand preferably has the following 5'-terminal motif

[0181]

Chemical formula

[0182] [wherein B represents the nucleoside bases of the first two base nucleosides in the 5'-terminal region of the second strand, T represents a 2'-Me ribose modification, V represents O or S (preferably O), R represents H or C 1~4 alkyl (preferably H), Z comprises 11 to 26 consecutive base nucleosides, preferably 15 to 21 consecutive base nucleosides, more preferably 19 consecutive base nucleosides], more preferably the following 5'-terminal motif

[0183] [Chemical formula]

[0184] [wherein, B represents the nucleoside bases of the first two base nucleosides in the 5'-terminal region of the second strand, T represents a 2'-Me ribose modification, Z represents the remaining 19 consecutive base nucleosides of the second strand] comprises.

[0185] In such embodiments, the modification pattern of the first strand of the nucleic acid is preferably Me(s)F(s)Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me [wherein, (s) is a phosphorothioate internucleoside linkage] comprises or consists of.

[0186] In some embodiments, the modification pattern of the second (sense) strand of the nucleic acid according to the present invention is ia-ia-Me(s)-Me(s)-Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me [wherein, (s) is a phosphorothioate internucleoside linkage and ia represents an inverted abasic nucleoside] comprises or consists of.

[0187] In such embodiments, the second strand preferably has the following 5'-terminal motif

[0188] [Chemical formula]

[0189] [wherein, B represents the nucleoside bases of the first two base nucleosides in the 5'-terminal region of the second strand, T represents a 2'-Me ribose modification, V represents O or S (preferably O), R represents H or C 1~4 alkyl (preferably H), Z contains 11 to 26 consecutive base nucleosides, preferably 15 to 21 consecutive base nucleosides, more preferably 19 consecutive base nucleosides, more preferably the following 5'-end motif

[0190]

Chemical formula

[0191] [wherein, B represents the nucleoside bases of the first two base nucleosides in the 5'-end region of the second strand, T represents a 2'-Me ribose modification, Z represents the remaining 19 consecutive base nucleosides of the second strand] and includes.

[0192] In such embodiments, the modification pattern of the first strand of the nucleic acid is preferably Me(s)F(s)Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me [wherein, (s) is a phosphorothioate internucleoside linkage] and includes or consists of.

[0193] The reverse linkage is preferably located at the end of a nucleic acid, such as RNA, distal to the ligand portion of the molecule, such as the GalNAc-containing portion.

[0194] A GalNAc-siRNA construct having 5'-GalNAc on the sense strand can have a reverse linkage on the opposite end of the sense strand.

[0195] A GalNAc-siRNA construct having 3'-GalNAc on the sense strand can have a reverse linkage on the opposite end of the sense strand.

[0196] In a preferred embodiment, the modification pattern of the second (sense) strand of the nucleic acid according to the present invention is ia-ia-Me(s)-Me(s)-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, or ia-ia-Me(s)-Me(s)-Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me [wherein, (s) is a phosphorothioate nucleoside internucleoside linkage, and ia represents an inverted abasic nucleoside] comprises or consists of.

[0197] In such an embodiment, the second strand preferably has the following 5'-terminal motif

[0198]

Chemical formula

[0199] [wherein, B represents the nucleoside bases of the first two base nucleosides in the 5'-terminal region of the second strand, T represents a 2'-Me ribose modification, V represents O or S (preferably O), R represents H or C 1~4 alkyl (preferably H), Z comprises 11 to 26 consecutive base nucleosides, preferably 15 to 21 consecutive base nucleosides, more preferably 19 consecutive base nucleosides], more preferably the following 5'-terminal motif

[0200]

Chemical formula

[0201] [wherein, B represents the nucleoside bases of the first two base nucleosides in the 5'-terminal region of the second strand, T represents a 2'-Me ribose modification, Z represents the remaining 19 consecutive base nucleosides of the second strand] comprises.

[0202] In such embodiments, the modification pattern of the first strand of the nucleic acid is preferably Me(s)F(s)Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me [where (s) is a phosphorothioate internucleoside linkage], or Me(s)F(s)Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me [where (s) is a phosphorothioate internucleoside linkage] comprises or consists of.

[0203] Length of the nucleic acid In one aspect, i) the first strand of the nucleic acid has a length of 17 to 30 nucleosides, preferably 19 to 25 nucleosides, more preferably 19 or 23 nucleosides, and / or ii) the second strand of the nucleic acid has a length of 17 to 30 nucleosides, preferably 19 to 25 nucleosides, more preferably 19 or 21 nucleosides.

[0204] Typically, the double-stranded region of the nucleic acid is 17 to 30 nucleosides in length, more preferably 19 or 21 nucleosides in length. Similarly, the complementary region between the first strand and a portion of the RNA transcribed from the target gene is 17 to 30 nucleosides in length.

[0205] Nucleic acid modification In certain embodiments, the nucleic acids of the invention, such as RNA, such as dsiRNA, do not include further modifications, such as chemical modifications or conjugations known in the art and described herein.

[0206] In other preferred embodiments, the nucleic acids of the invention, such as RNA, such as dsiRNA, are further chemically modified to enhance stability or other beneficial characteristics.

[0207] In certain embodiments of the invention, substantially all of the nucleosides are modified.

[0208] The nucleic acids characterized by the present invention can be synthesized or modified by methods well established in the art, such as those described in “Current protocols in nucleic acid chemistry,” Beaucage, S.L. et al. (Eds.), John Wiley & Sons, Inc., New York, NY, USA, which is incorporated herein by reference.

[0209] Modifications include, for example, terminal modifications, such as 5′-end modifications (phosphorylation, conjugation, inverted linkage) or 3′-end modifications (conjugation, DNA nucleosides within RNA, or RNA nucleosides within DNA, inverted linkage, etc.); base modifications, such as stabilizing bases, destabilizing bases, or replacement of bases that base pair with an extended repertoire of partners, conjugated bases; sugar modifications (e.g., at the 2′ or 4′ position) or replacement of sugars; or backbone modifications, including modifications or replacements of the phosphodiester linkage.

[0210] Specific examples of nucleic acids such as siRNA compounds useful in the embodiments described herein include, but are not limited to, RNAs containing a modified backbone, or RNAs lacking a native internucleoside linkage. Nucleic acids such as RNAs having a modified backbone include, inter alia, those having no phosphorus atom in the backbone. For the purposes of this specification, and as sometimes referred to in the art, modified nucleic acids, such as RNAs, having no phosphorus atom in their internucleoside backbone can also be considered oligonucleosides. In some embodiments, the modified nucleic acid, such as siRNA, has a phosphorus atom in its internucleoside backbone.

[0211] Modified nucleic acids, such as RNA backbones, include, for example, phosphorothioates, chiral phosphorothioates, phosphorodithioates, phosphotriesters, aminoalkylphosphotriesters, methyl and other alkyl phosphonates, such as 3'-alkylene phosphonates and chiral phosphonates, phosphinates, phosphoramidates, such as 3'-aminophosphoramidates and aminoalkylphosphoramidates, thionophosphoramidates, thionoalkylphosphonates, thionoalkylphosphotriesters, and boranophosphates, 2'-5' linkage analogs thereof, and those having an inverted polarity with adjacent pairs of nucleoside units linked 5'-3' or 5'-2'. Also included are various salts, mixed salts, and free acid forms.

[0212] Modified nucleic acids, such as RNA, can also contain one or more substituted sugar moieties. Nucleic acids characterized herein, such as siRNA, such as dsiRNA, may include at the 2'-position one of the following: OH; F; O-, S-, or N-alkyl; O-, S-, or N-alkenyl; O-, S-, or N-alkynyl; or O-alkyl-O-alkyl, where the alkyl, alkenyl, and alkynyl may be substituted or unsubstituted. 2'-O-methyl and 2'-F are preferred modifications.

[0213] In certain preferred embodiments, the nucleic acid comprises at least one modified nucleoside.

[0214] The nucleic acids of the invention may comprise one or more modified nucleosides on the first strand and / or the second strand.

[0215] In some embodiments, substantially all of the nucleosides of the sense strand and all of the nucleosides of the antisense strand comprise a modification.

[0216] In some embodiments, all of the nucleosides of the sense strand and substantially all of the nucleosides of the antisense strand comprise a modification.

[0217] In some embodiments, all of the nucleosides of the sense strand and all of the nucleosides of the antisense strand comprise a modification.

[0218] In one embodiment, at least one of the modified nucleosides is a deoxy-nucleoside, a 3'-terminal deoxy-thymidine (dT) nucleoside, a 2'-O-methyl modified nucleoside (also referred to herein as 2'-Me, where Me is methoxy), a 2'-fluoro modified nucleoside, a 2'-deoxy-modified nucleoside, a locked nucleoside, an unlocked nucleoside, a nucleoside with a restricted conformation, a constrained ethyl nucleoside, a nucleobase-free nucleoside, a 2'-amino-modified nucleoside, a 2'-O-allyl-modified nucleoside, a 2'-C-alkyl-modified nucleoside, a 2'-hydroxyl-modified nucleoside, a 2'-methoxyethyl modified nucleoside, a 2'-O-alkyl-modified nucleoside, a morpholino nucleoside, a phosphoramidate, a nucleoside containing an unnatural base, a tetrahydropyran modified nucleoside, a 1,5-anhydrohexitol modified nucleoside, a cyclohexenyl modified nucleoside, a nucleoside containing a phosphorothioate group, a nucleoside containing a methylphosphonate group, a nucleoside containing a 5'-phosphate, and a nucleoside containing a 5'-phosphate mimetic. In another embodiment, the modified nucleoside comprises a short sequence of 3'-terminal deoxy-thymidine nucleoside (dT).

[0219] The modification on the nucleoside may preferably be selected from the group consisting of, but not limited to, LNA, HNA, CeNA, 2'-methoxyethyl, 2'-O-alkyl, 2'-O-allyl, 2'-C-allyl, 2'-fluoro, 2'-deoxy, 2'-hydroxyl, and combinations thereof. In another embodiment, the modification on the nucleoside is a 2'-O-methyl ("2'-Me") or 2'-fluoro modification.

[0220] One preferred modification is a modification at the 2'-OH group of the ribose sugar, optionally selected from 2'-Me or 2'-F modifications.

[0221] Preferred nucleic acids contain one or more nucleosides on the first strand and / or the second strand that are modified to form modified nucleosides as follows:

[0222] A nucleic acid, wherein the modification is a modification at the 2'-OH group of the ribose sugar, optionally selected from 2'-Me or 2'-F modifications.

[0223] A nucleic acid, wherein the first strand contains a 2'-F modification at any one of positions 2, 6, 14, or any combination thereof, counted from the 1st position of the first strand.

[0224] A nucleic acid, wherein the second strand contains a 2'-F modification at any one of positions 7, 9, 11, or any combination thereof, counted from the 1st position of the second strand.

[0225] A nucleic acid, wherein each of the first and second strands contains 2'-Me and 2'-F modifications.

[0226] Preferably, a nucleic acid containing at least one thermally destabilizing modification at one or more of positions 1 to 9 of the first strand, counted from the 1st position of the first strand, and / or at one or more of the positions on the second strand aligned with positions 1 to 9 of the first strand, wherein the destabilizing modification is selected from modified unlocked nucleic acid (UNA) and glycol nucleic acid (GNA), preferably glycol nucleic acid, more preferably (S)-glycol nucleic acid.

[0227] A nucleic acid containing at least one thermally destabilizing modification at the 7th position of the first strand, counted from the 1st position of the first strand.

[0228] An siRNA oligonucleoside, wherein the second strand contains 3 or more 2'-F modifications at positions 6 to 12 of the second strand, counted from the 1st position of the second strand, for example 4, 5, 6, or 7 2'-F modifications at positions 6 to 12 of the second strand.

[0229] An siRNA oligonucleoside, wherein the second strand contains at least 3, for example 4, 5, or 6 2'-Me modifications at positions 1 to 6 of the second strand, counted from the 1st position of the second strand.

[0230] The nucleic acid is an siRNA oligonucleoside in which the first strand preferably contains at least five consecutive 2'-Me modifications in the 3'-terminal region, including the terminal nucleoside in the 3'-terminal region or at least one or two nucleosides within the terminal nucleoside in the 3'-terminal region.

[0231] The nucleic acid is an siRNA oligonucleoside in which the first strand preferably contains seven consecutive 2'-Me modifications in the 3'-terminal region, including the terminal nucleoside in the 3'-terminal region.

[0232] The nucleic acid is an siRNA oligonucleoside in which each of the first and second strands contains an alternating modification pattern, preferably a completely alternating modification pattern along the entire length of each of the first and second strands, and the nucleosides of the first strand are modified by (i) 2'-Me modification on the nucleosides numbered odd from position 1 of the first strand and (ii) 2'-F modification on the nucleosides numbered even from position 1 of the first strand, and the nucleosides of the second strand are modified by (i) 2'-F modification on the nucleosides numbered odd from position 1 of the second strand and (ii) 2'-Me modification on the nucleosides numbered even from position 1 of the second strand. Typically, such a completely alternating modification pattern is present in blunt-ended oligonucleosides, and each of the first and second strands is 19 nucleosides in length.

[0233] Position 1 of the first or second strand is closest to the end of the nucleic acid (ignoring any abasic nucleosides), and with respect to the bond between the sugar moieties of the backbone, reading in the direction away from that end of the molecule, is the nucleoside linked via an internal 3'-to-5' bond to the adjacent nucleoside (at position 2).

[0234] Thus, the "first position of the sense strand" can be regarded as the most 5'-terminal nucleoside (excluding abasic nucleosides) at the conventional 5'-end of the sense strand. Typically, the nucleoside at this first position of the sense strand is equivalent to the 5'-nucleoside of the selected target nucleic acid sequence. More generally, the sense strand has nucleosides equivalent to those of the target nucleic acid sequence starting from this first position of the sense strand, allowing for tolerated mismatches between sequences.

[0235] As used herein, the "first position of the antisense strand" is the most 5'-terminal nucleoside (excluding abasic nucleosides) at the conventional 5'-end of the antisense strand. As described above herein, there is a complementary region between the sense strand and the antisense strand, and thus the antisense strand also has a complementary region to the target nucleic acid sequence as mentioned above.

[0236] In certain embodiments, the nucleic acid, such as an siRNA agent, further comprises at least one phosphorothioate or methylphosphonate internucleoside linkage. For example, the phosphorothioate or methylphosphonate internucleoside linkage may be at the 3'-end or terminal region of one strand, i.e., the sense strand or the antisense strand, or at the termini of both strands, the sense strand and the antisense strand.

[0237] In certain embodiments, the phosphorothioate or methylphosphonate internucleoside linkage is at the 5'-end or terminal region of one strand, i.e., the sense strand or the antisense strand, or at the termini of both strands, the sense strand and the antisense strand.

[0238] In certain embodiments, the phosphorothioate or methylphosphonate internucleoside linkage is at both the 5'-end and 3'-end or terminal regions of one strand, i.e., the sense strand or the antisense strand, or at the termini of both strands, the sense strand and the antisense strand.

[0239] Any nucleic acid may contain one or more phosphorothioate (PS) modifications within the nucleic acid, for example, at least two PS internucleoside linkages at the ends of the strand.

[0240] At least one of the oligoribonucleoside strands preferably contains at least two consecutive phosphorothioate modifications at the last three nucleosides of the oligonucleoside.

[0241] Accordingly, the present invention also relates to: nucleic acids disclosed herein that contain phosphorothioate internucleoside linkages between at least two or three consecutive positions, such as in the 5' and / or 3' terminal regions and / or regions near the ends of the second strand, whereby the region near the end is preferably adjacent to the terminal region where the one or more abasic nucleosides of the second strand are located.

[0242] Nucleic acids disclosed herein that contain phosphorothioate internucleoside linkages between at least two or three consecutive positions in the 5' and / or 3' terminal regions of the first strand, whereby preferably the terminal positions in the 5' and / or 3' terminal regions of the first strand are attached to their adjacent positions by phosphorothioate internucleoside linkages.

[0243] The nucleic acid strand may be an RNA that contains a phosphorothioate internucleoside linkage between three nucleosides adjacent to the abasic nucleosides located at the two ends.

[0244] Preferred nucleic acids are double-stranded RNAs that contain two adjacent abasic nucleosides at the 5' end of the second strand and a ligand portion that contains one or more GalNAc ligand portions at the opposite 3' end of the second strand. More preferably, the same nucleic acid may also contain a phosphorothioate bond between the nucleotides at positions 3-4 and 4-5 of the second strand, read from position 1 of the second strand. Even more preferably, the same nucleic acid may also contain 2'F modifications at positions 7, 9, and 11 of the second strand.

[0245] Preferred modifications are as follows: The modified nucleoside of the second strand is as follows (5'-3'): Me-Me-Me-Me-Me-Me-F-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-F-Me-Me, or Me-Me-Me-Me-Me-F-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, or Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, or Me-Me-Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, or Me-Me-Me-Me-Me-Me-F-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me A nucleic acid having a modification pattern by any one of the above.

[0246] The modified nucleoside of the second strand is as follows (5'-3'): Me(s)Me(s)Me-Me-Me-Me-F-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-F-Me-Me, or Me(s)Me(s)Me-Me-Me-F-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, or Me(s)Me(s)Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, or Me(s)Me(s)Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, or Me(s)Me(s)Me-Me-Me-Me-F-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, or Me-Me-Me-Me-Me-Me-F-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-F-Me(s)Me(s), or Me-Me-Me-Me-Me-F-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me(s)Me(s), or Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me(s)Me(s), or Me-Me-Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me(s)Me(s), or Me-Me-Me-Me-Me-Me-F-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me(s)Me(s) [where (s) is a phosphorothioate internucleoside linkage] A nucleic acid having a modification pattern by any one of the following.

[0247] The modified nucleoside of the second strand is as follows (5'-3'): ia-ia-Me-Me-Me-Me-Me-Me-F-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-F-Me-Me, or ia-ia-Me-Me-Me-Me-Me-F-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, or ia-ia-Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, or ia-ia-Me-Me-Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, or ia-ia-Me-Me-Me-Me-Me-Me-F-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, or Me-Me-Me-Me-Me-Me-F-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-F-Me-Me-ia-ia, or Me-Me-Me-Me-Me-F-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-ia-ia, or Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-ia-ia, or Me-Me-Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me-ia-ia, or Me-Me-Me-Me-Me-Me-F-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me-ia-ia [wherein, ia represents an inverted abasic nucleoside, and when the inverted abasic nucleoside represented by ia-ia is present at the 3'-end of the second strand, the inverted abasic nucleoside is present in the overhang of two nucleosides] A nucleic acid having a modification pattern by any one of the following.

[0248] The modified nucleoside of the second strand is as follows (5'-3'): ia-ia-Me(s)Me(s)Me-Me-Me-Me-F-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-F-Me-Me, or ia-ia-Me(s)Me(s)Me-Me-Me-F-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, or ia-ia-Me(s)Me(s)Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, or ia-ia-Me(s)Me(s)Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, or ia-ia-Me(s)Me(s)Me-Me-Me-Me-F-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, or Me-Me-Me-Me-Me-Me-F-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-F-Me(s)Me(s)ia-ia, or Me-Me-Me-Me-Me-F-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me(s)Me(s)ia-ia, or Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me(s)Me(s)ia-ia, or Me-Me-Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me(s)Me(s)ia-ia, or Me-Me-Me-Me-Me-Me-F-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me(s)Me(s)ia-ia [wherein, (s) is a phosphorothioate nucleoside internucleoside linkage, ia represents an inverted abasic nucleoside, and when the inverted abasic nucleoside represented by ia-ia is present at the 3'-end of the second strand, the inverted abasic nucleoside is present in an overhang of two nucleosides] A nucleic acid having a modification pattern by any one of the following:

[0249] The modified nucleoside has the following modification patterns: Modification pattern 1: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-F-Me-Me, First strand (5'-3'): Me-F-Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Or modification pattern 2: Second strand (5'-3'): Me-Me-Me-Me-Me-F-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me-F-Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Or modified pattern 3: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me-F-Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Or modified pattern 4: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me-F-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Or modified pattern 5: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me-F-Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Or modified pattern 6: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me-F-Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me A nucleic acid having

[0250] The modified nucleoside has the following modified pattern: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me-F-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Or Second strand (5'-3'): Me-Me-Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, first strand (5'-3'): Me-F-Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me A nucleic acid having

[0251] The modified nucleosides have the following modification patterns: Modification pattern 1: Second strand (5'-3'): Me(s)Me(s)Me-Me-Me-Me-F-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-F-Me-Me, first strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modification pattern 2: Second strand (5'-3'): Me(s)Me(s)Me-Me-Me-F-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, first strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modification pattern 3: Second strand (5'-3'): Me(s)Me(s)Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, first strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modification pattern 4: Second strand (5'-3'): Me(s)Me(s)Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, first strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modified pattern 5: Second strand (5'-3'): Me(s)Me(s)Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me(s)F(s)Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modified pattern 6: Second strand (5'-3'): Me(s)Me(s)Me-Me-Me-Me-F-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me(s)F(s)Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me [where (s) is a phosphorothioate internucleoside linkage] A nucleic acid having

[0252] The modified nucleoside has the following modified pattern: Second strand (5'-3'): Me(s)Me(s)Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or Second strand (5'-3'): Me(s)Me(s)Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me(s)F(s)Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me [where (s) is a phosphorothioate internucleoside linkage] A nucleic acid having

[0253] The modified nucleoside has the following modified pattern: Modified pattern 1: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-F-Me(s)Me(s), First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modified pattern 2: Second strand (5'-3'): Me-Me-Me-Me-Me-F-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me(s)Me(s), First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modified pattern 3: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me(s)Me(s), First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modified pattern 4: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me(s)Me(s), First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modified pattern 5: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me(s)Me(s), First strand (5'-3'): Me(s)F(s)Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modified pattern 6: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me(s)Me(s), First strand (5'-3'): Me(s)F(s)Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me [wherein, (s) is a phosphorothioate internucleoside linkage] A nucleic acid having

[0254] The modified nucleoside has the following modified patterns: Modified pattern 1: Second strand (5'-3'): ia-ia-Me-Me-Me-Me-Me-Me-F-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-F-Me-Me, First strand (5'-3'): Me-F-Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Or modified pattern 2: Second strand (5'-3'): ia-ia-Me-Me-Me-Me-Me-F-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me-F-Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Or modified pattern 3: Second strand (5'-3'): ia-ia-Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me-F-Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Or modified pattern 4: Second strand (5'-3'): ia-ia-Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me-F-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Or modified pattern 5: Second strand (5'-3'): ia-ia-Me-Me-Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me-F-Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Or modified pattern 6: Second strand (5'-3'): ia-ia-Me-Me-Me-Me-Me-Me-F-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me-F-Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me [Wherein, ia represents an inverted abasic nucleoside] A nucleic acid having

[0255] The modified nucleoside has the following modified pattern: Second strand (5'-3'): ia-ia-Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me-F-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Or Second strand (5'-3'): ia-ia-Me-Me-Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me-F-Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me [Wherein, ia represents an inverted abasic nucleoside] A nucleic acid having

[0256] The modified nucleoside has the following modified pattern: Modified pattern 1: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-F-Me-Me-ia-ia, first strand (5'-3'): Me-F-Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Or modified pattern 2: Second strand (5'-3'): Me-Me-Me-Me-Me-F-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-ia-ia, first strand (5'-3'): Me-F-Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Or modified pattern 3: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-ia-ia, first strand (5'-3'): Me-F-Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Or modified pattern 4: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-ia-ia, first strand (5'-3'): Me-F-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Or modified pattern 5: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me-ia-ia, first strand (5'-3'): Me-F-Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Or modified pattern 6: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me-ia-ia, first strand (5'-3'): Me-F-Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me [Wherein, ia represents an inverted abasic nucleoside, and when the inverted abasic nucleoside represented by ia-ia is present at the 3'-end of the second strand, the inverted abasic nucleoside is present in an overhang of two nucleosides] A nucleic acid having the above structure.

[0257] The modified nucleoside has the following modified patterns: Modified pattern 1: Second strand (5'-3'): ia-ia-Me(s)Me(s)Me-Me-Me-Me-F-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-F-Me-Me, first strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modified pattern 2: Second strand (5'-3'): ia-ia-Me(s)Me(s)Me-Me-Me-F-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, first strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modified pattern 3: Second strand (5'-3'): ia-ia-Me(s)Me(s)Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, first strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modified pattern 4: Second strand (5'-3'): ia-ia-Me(s)Me(s)Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modified pattern 5: Second strand (5'-3'): ia-ia-Me(s)Me(s)Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me(s)F(s)Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modified pattern 6: Second strand (5'-3'): ia-ia-Me(s)Me(s)Me-Me-Me-Me-F-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me(s)F(s)Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me [Wherein, (s) represents a phosphorothioate nucleoside internucleoside linkage, and ia represents an inverted abasic nucleoside] A nucleic acid having

[0258] The modified nucleoside has the following modified pattern: Second strand (5'-3'): ia-ia-Me(s)Me(s)Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or Second strand (5'-3'): ia-ia-Me(s)Me(s)Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me(s)F(s)Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me [wherein, (s) represents a phosphorothioate internucleoside linkage, and ia represents an inverted abasic nucleoside] A nucleic acid having

[0259] The modified nucleosides have the following modification patterns: Modification pattern 1: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-F-Me(s)Me(s)ia-ia, First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modification pattern 2: Second strand (5'-3'): Me-Me-Me-Me-Me-F-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me(s)Me(s)ia-ia, First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modification pattern 3: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me(s)Me(s)ia-ia, First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modified pattern 4: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me(s)Me(s)ia-ia, First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modified pattern 5: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me(s)Me(s)ia-ia, First strand (5'-3'): Me(s)F(s)Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modified pattern 6: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me(s)Me(s)ia-ia, First strand (5'-3'): Me(s)F(s)Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me [Wherein, (s) is a phosphorothioate nucleoside internucleoside linkage, ia represents an inverted abasic nucleoside, and when the inverted abasic nucleoside represented by ia-ia is present at the 3'-end of the second strand, the inverted abasic nucleoside is present in an overhang of two nucleosides] A nucleic acid having the above structure.

[0260] Particularly preferred is that the modified nucleoside has the following modified pattern: Second strand (5'-3'): ia-ia-Me(s)Me(s)Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or Second strand (5'-3'): ia-ia-Me(s)Me(s)Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me(s)F(s)Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me [Wherein, (s) is a phosphorothioate internucleoside linkage, and ia represents an inverted abasic nucleoside] is a nucleic acid having.

[0261] Conjugation Another modification of the nucleic acids of the present invention, such as RNA, such as siRNA, is, for example, to enhance the activity, cellular distribution, or cellular uptake of the nucleic acid, such as siRNA, for example into cells, and includes conjugating the nucleic acid, such as siRNA, with one or more ligand moieties.

[0262] In some embodiments, the described ligand moiety may be attached to a nucleic acid, such as a siRNA oligonucleoside, via a linker that may or may not be cleavable. The term "linker" or "linking group" means an organic moiety that connects two moieties of a compound, such as covalently bonding two moieties of a compound.

[0263] The ligand may be attached to the 3' or 5' end of the sense strand.

[0264] The ligand is preferably conjugated to the 3' end of the sense strand of a nucleic acid, such as a siRNA agent.

[0265] Accordingly, in a further aspect, the present invention relates to a conjugate for inhibiting the expression of a target gene in a cell, wherein the conjugate comprises a nucleic acid moiety and one or more ligand moieties, and the nucleic acid moiety comprises a nucleic acid disclosed herein.

[0266] In one aspect, the second strand of the nucleic acid is conjugated directly or indirectly (e.g., via a linker) to one or more ligand moieties, which are typically present in the terminal region of the second strand, preferably in its 3' terminal region.

[0267] In certain embodiments, the ligand moiety comprises GalNAc or a GalNAc derivative attached to the nucleic acid, e.g., dsiRNA, through a linker.

[0268] Accordingly, the present invention relates to conjugates comprising a ligand moiety that is i) one or more GalNAc ligands, and / or ii) one or more GalNAc ligand derivatives, and / or iii) one or more GalNAc ligands conjugated to said nucleic acid through a linker .

[0269] The GalNAc ligand may be conjugated directly or indirectly, preferably at the 3' terminal region, to the 5' or 3' terminal region of the second strand of the nucleic acid.

[0270] GalNAc ligands are well known in the art and are described, inter alia, in EP3775207A1.

[0271] In some embodiments, the GalNAc ligand is included in any one of the linkers shown in FIGS. 1-4 or FIG. 5 (Formula XI), and the "oligonucleotide" may be any nucleic acid disclosed herein. Thus, the "oligonucleotide" may include linkages other than phosphodiester linkages, e.g., one or more phosphorothioate linkages. Preferably, the nucleic acid according to the present invention is a double-stranded oligonucleoside as defined herein, and the linker is conjugated to the second strand, more preferably to the 3' terminal region of the second strand, via a phosphodiester linkage.

[0272] In some embodiments, the GalNAc ligand is included in the linker shown in FIG. 3, and the "oligonucleotide" may be any nucleic acid disclosed herein. Thus, the "oligonucleotide" may include linkages other than phosphodiester linkages, such as one or more phosphorothioate linkages. Preferably, the nucleic acid according to the invention is a double-stranded oligonucleoside as defined herein, and the linker is conjugated via a phosphodiester bond to the second strand, more preferably to the 3'-terminal region of the second strand.

[0273] In some embodiments, the GalNAc ligand is included in the linker shown in FIG. 5 (Formula XI), and the "oligonucleotide" may be any nucleic acid disclosed herein. Thus, the "oligonucleotide" may include linkages other than phosphodiester linkages, such as one or more phosphorothioate linkages. Preferably, the nucleic acid according to the invention is a double-stranded oligonucleoside as defined herein, and the linker is conjugated via a phosphodiester bond to the second strand, more preferably to the 3'-terminal region of the second strand.

[0274] In some embodiments, the GalNAc ligand is included in any one of the linkers shown in FIGS. 1-4 or FIG. 5 (Formula XI), and the "oligonucleotide" represents the nucleic acid according to the invention, and the nucleic acid according to the invention has the following modification pattern (5'-3'): ia-ia-Me(s)Me(s)Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me or ia-ia-Me(s)Me(s)Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me [wherein, (s) is a phosphorothioate internucleoside linkage, and ia represents an inverted abasic nucleoside] and includes a modified second strand having the above, and preferably, the linker is conjugated to the 3'-terminal region of the second strand via a phosphodiester bond.

[0275] In some embodiments, the GalNAc ligand is included in the linker shown in FIG. 3, the "oligonucleotide" represents a nucleic acid according to the present invention, and the nucleic acid according to the present invention has the following modification pattern (5'-3'): ia-ia-Me(s)Me(s)Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me or ia-ia-Me(s)Me(s)Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me [wherein, (s) is a phosphorothioate internucleoside linkage, and ia represents an inverted abasic nucleoside] and includes a modified second strand having the above, preferably the linker is conjugated to the 3'-terminal region of the second strand via a phosphodiester bond.

[0276] In some embodiments, the GalNAc ligand is included in the linker shown in FIG. 5 (Formula XI), the "oligonucleotide" represents a nucleic acid according to the present invention, and the nucleic acid according to the present invention has the following modification pattern (5'-3'): ia-ia-Me(s)Me(s)Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me or ia-ia-Me(s)Me(s)Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me [wherein, (s) is a phosphorothioate internucleoside linkage, and ia represents an inverted abasic nucleoside] and includes a modified second strand having the above, preferably the linker is conjugated to the 3'-terminal region of the second strand via a phosphodiester bond.

[0277] In some embodiments, the GalNAc ligand is included in any one of the linkers shown in FIGS. 1-4 or FIG. 5 (Formula XI), and "oligonucleotide" represents the nucleic acid according to the present invention, and the nucleic acid according to the present invention has the following modification pattern (5'-3'): ia-ia-Me(s)Me(s)Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me or ia-ia-Me(s)Me(s)Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me [wherein, (s) is a phosphorothioate internucleoside linkage, and ia represents an inverted abasic nucleoside] and includes a modified second strand having:

[0278]

Chemical formula

[0279] [wherein, T represents a 2'-Me ribose modification, B represents the nucleoside bases of the first two base nucleosides in the 5'-terminal region of the second strand, and Z represents the remaining 19 consecutive base nucleosides of the second strand] having.

[0280] In some embodiments, the GalNAc ligand is included in the linker shown in FIG. 3, and "oligonucleotide" represents the nucleic acid according to the present invention, and the nucleic acid according to the present invention has the following modification pattern (5'-3'): ia-ia-Me(s)Me(s)Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me or ia-ia-Me(s)Me(s)Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me [wherein, (s) is a phosphorothioate nucleoside internucleoside linkage, and ia represents an inverted abasic nucleoside] comprises a modified second strand having

[0281]

Chemical formula

[0282] [wherein, T represents a 2'-Me ribose modification, B represents the nucleoside bases of the first two base nucleosides in the 5'-terminal region of the second strand, and Z represents the remaining 19 consecutive base nucleosides of the second strand] having

[0283] In some embodiments, the GalNAc ligand is included in the linker shown in Figure 5 (Formula XI), and the "oligonucleotide" represents a nucleic acid according to the present invention, and the nucleic acid according to the present invention has the following modification pattern (5'-3'): ia-ia-Me(s)Me(s)Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me or ia-ia-Me(s)Me(s)Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me [wherein, (s) is a phosphorothioate nucleoside internucleoside linkage, and ia represents an inverted abasic nucleoside] comprises a modified second strand having

[0284]

Chemical formula

[0285] [wherein, T represents a 2'-Me ribose modification, B represents the nucleoside bases of the first two base nucleosides in the 5'-terminal region of the second strand, Z represents the remaining 19 consecutive base nucleosides of the second strand] having.

[0286] Vectors and cells In one aspect, the present invention provides a cell containing a nucleic acid described herein, such as inhibitory RNA [RNAi].

[0287] In one aspect, the present invention provides a cell containing a vector described herein.

[0288] Pharmaceutically acceptable compositions In one aspect, the present invention provides a pharmaceutical composition for inhibiting the expression of a target gene, the pharmaceutical composition comprising a nucleic acid disclosed herein.

[0289] The pharmaceutically acceptable composition may contain excipients and / or carriers.

[0290] Some examples of materials that can serve as pharmaceutically acceptable carriers include: (1) sugars, such as lactose, glucose, and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) lubricants, such as magnesium stearate, sodium lauryl sulfate, and talc; (8) excipients, such as cocoa butter and suppository wax; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitol, and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffering agents, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethyl alcohol; (20) pH buffering solutions; (21) polyesters, polycarbonates, and / or polyanhydrides; (22) bulking agents, such as polypeptides and amino acids; (23) serum components, such as serum albumin, HDL, and LDL; and (22) other non-toxic, compatible substances used in pharmaceutical formulations.

[0291] Typical pharmaceutical carriers include, but are not limited to, binders (such as pregelatinized corn starch, polyvinylpyrrolidone, or hydroxypropylmethylcellulose), fillers (such as lactose and other sugars, microcrystalline cellulose, pectin, gelatin, calcium sulfate, ethyl cellulose, polyacrylate, or calcium hydrogen phosphate), lubricants (such as magnesium stearate, talc, silica, colloidal silicon dioxide, stearic acid, metal stearates, hydrogenated vegetable oil, corn starch, polyethylene glycol, sodium benzoate, sodium acetate), disintegrants (such as starch, sodium starch glycolate), and wetting agents (such as sodium lauryl sulfate).

[0292] Pharmaceutically acceptable organic or inorganic excipients that do not react detrimentally with the nucleic acid and are suitable for non - oral administration can also be used to formulate the compositions of the present invention. Suitable pharmaceutically acceptable excipients include, but are not limited to, water, salt solutions, alcohols, polyethylene glycol, gelatin, lactose, amylose, magnesium stearate, talc, silicic acid, viscous paraffin, hydroxymethylcellulose, polyvinylpyrrolidone, and the like.

[0293] Formulations for topical administration of nucleic acids can include sterile and non - sterile aqueous solutions, non - aqueous solutions in common solvents such as alcohols, or solutions of nucleic acids in liquid or solid oil - based vehicles. The solutions can also contain buffers, diluents, and other suitable additives. Pharmaceutically acceptable organic or inorganic excipients that do not react detrimentally with the nucleic acid and are suitable for non - oral administration can be used.

[0294] In one embodiment, the nucleic acid or composition is administered in a non - buffered solution. In certain embodiments, the non - buffered solution is saline or water. In other embodiments, the nucleic acid, e.g., an siRNA agent, is administered in a buffered solution. In such embodiments, the buffered solution can contain acetate, citrate, prolamine, carbonate, or phosphate, or any combination thereof. For example, the buffered solution can be phosphate - buffered saline (PBS).

[0295] Dosage The pharmaceutical compositions of the present invention can be administered in a dosage sufficient to inhibit gene expression. Generally, a suitable dosage of the nucleic acids of the present invention, e.g., siRNA, ranges from about 0.001 to about 200.0 milligrams per kilogram of the recipient's body weight per day, generally in the range of about 1 to 50 mg per kilogram of body weight per day. Typically, a suitable dosage of the nucleic acids of the present invention, e.g., siRNA, ranges from about 0.1 mg / kg to about 5.0 mg / kg, for example about 0.3 mg / kg and about 3.0 mg / kg.

[0296] The repeated dosing regimen can include the administration of a therapeutic amount of a nucleic acid, such as siRNA, at regular intervals, such as every other day or once a year. In certain embodiments, the nucleic acid, such as siRNA, is administered from once a month to once every quarter (i.e., once every 3 months).

[0297] In various embodiments, the nucleic acid, such as an siRNA agent, is administered at a dose of about 0.01 mg / kg to about 10 mg / kg or about 0.5 mg / kg to about 50 mg / kg. In some embodiments, the nucleic acid, such as an siRNA agent, is administered at a dose of about 10 mg / kg to about 30 mg / kg. In certain embodiments, the nucleic acid, such as an siRNA agent, is administered at a dose selected from about 0.5 mg / kg, 1 mg / kg, 1.5 mg / kg, 3 mg / kg, 5 mg / kg, 10 mg / kg, and 30 mg / kg. In certain embodiments, the nucleic acid, such as an agent, is administered at a dose of about 0.1 mg / kg to about 5.0 mg / kg once a week, once a month, once every two months, or once every quarter (i.e., once every 3 months). In certain embodiments, the nucleic acid, such as an siRNA agent, is administered to the subject once a week. In certain embodiments, the nucleic acid, such as an siRNA agent, is administered to the subject once a month. In certain embodiments, the nucleic acid, such as an siRNA agent, is administered once every quarter (i.e., every 3 months).

[0298] After the initial treatment regimen, the treatment can be administered at a less frequent rate. For example, after administering weekly or bi-weekly for 3 months, the administration can be repeated once a month for 6 months or 1 year, or longer.

[0299] The pharmaceutical composition can be administered once a day, or as two, three, or more partial doses at appropriate intervals throughout the day, or even by delivery through a sustained infusion or controlled release formulation. In that case, the nucleic acid, e.g., siRNA, contained in each partial dose will correspondingly have to be smaller in order to achieve the total daily dose. Dosage units can also be constituted for delivery over several days, e.g., using conventional sustained release formulations that provide for the sustained release of the nucleic acid, e.g., siRNA, over several days. Sustained release formulations are well known in the art and are particularly useful for the delivery of drugs at specific sites, such as those that can be used with the agents of the present invention. In this embodiment, the dosage unit contains the corresponding multiple daily doses.

[0300] In other embodiments, a single dose of the pharmaceutical composition can be of long duration such that subsequent doses are administered at intervals not exceeding 3, 4, or 5 days, or intervals not exceeding 1, 2, 3, or 4 weeks. In some embodiments of the present invention, a single dose of the pharmaceutical composition of the present invention is administered once a week. In other embodiments of the present invention, a single dose of the pharmaceutical composition of the present invention is administered every other month. In certain embodiments, the siRNA is administered from about once a month to about once every quarter (i.e., about once every 3 months), or even every 6 months or 12 months.

[0301] The determination of the effective dosage and in vivo half-life for the individual nucleic acids, e.g., siRNA, encompassed by the present invention can be made using conventional methods or based on in vivo tests using appropriate animal models as known in the art.

[0302] The pharmaceutical composition of the present invention can be administered in several ways depending on whether local treatment is desired or systemic treatment is desired and on the area to be treated. Administration can be local (e.g., by transdermal patch), pulmonary, e.g., by inhalation or insufflation of powder or aerosol, e.g., by nebulizer, intratracheal, intranasal, epidermal and transdermal, oral or parenteral. Parenteral administration can include intravenous, intraarterial, subcutaneous, intraperitoneal, or intramuscular injection or infusion; subdermal, e.g., via an implant device; or intracranial, e.g., by parenchymal, intrathecal or intraventricular administration. In certain preferred embodiments, the composition is administered by intravenous infusion or injection. In certain embodiments, the composition is administered by subcutaneous injection.

[0303] In one embodiment, a nucleic acid, e.g., an agent, is administered subcutaneously to a subject.

[0304] A nucleic acid, e.g., siRNA, can be delivered in a manner that targets a specific tissue (e.g., particularly in liver cells).

[0305] Method for inhibiting target gene expression The present invention also provides a method for inhibiting the expression of a target gene in a cell. The method includes contacting the cell with a nucleic acid of the present invention, e.g., an siRNA agent, e.g., a double-stranded siRNA agent, in an amount effective to inhibit the expression of the target gene in the cell, thereby inhibiting the expression of the target gene in the cell.

[0306] Contacting a cell with a nucleic acid, such as siRNA, such as a double-stranded siRNA agent, may be performed in vitro or in vivo. Contacting a cell with a nucleic acid in vivo includes, for example, contacting a cell or cell population within a subject, such as a human subject, with a nucleic acid, such as siRNA. Combinations of in vitro and in vivo methods of contacting a cell are also possible. Contacting a cell may be direct or indirect as discussed above. Further, contacting a cell may be achieved via a targeting ligand moiety that includes any ligand moiety described herein or known in the art. In a preferred embodiment, the targeting ligand moiety is a carbohydrate moiety, such as a GalNAc3 ligand, or any other ligand moiety that directs the siRNA agent to the site of interest.

[0307] The term "inhibiting" as used herein is used interchangeably with "reducing," "silencing," "downregulating," "suppressing," and other similar terms and includes any level of inhibition.

[0308] In some embodiments of the methods of the invention, the expression of the target gene is inhibited by at least 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%, or below the level of detection of the assay, when preferably determined by qPCR as described herein and / or when the siRNA is introduced into the target cell by transfection. In certain embodiments, the method includes a clinically relevant inhibition of the expression of the target gene, as evidenced by a clinically relevant result, for example, after treatment of a subject with an agent that reduces the expression of the gene.

[0309] In some embodiments, when transfected into cells, the nucleic acids of the invention preferably inhibit the expression of the target gene with an IC50 value lower than 2000 pM, 1900 pM, 1800 pM, 1700 pM, 1600 pM, 1500 pM, 1400 pM, 1300 pM, 1200 pM, 1100 pM, 1000 pM, 900 pM, 800 pM, 700 pM, 600 pM, 500 pM, 400 pM, 300 pM, 200 pM or 100 pM, as determined by qPCR, more preferably by reverse transcriptase (RT)-qPCR, as described herein.

[0310] In preferred embodiments, when transfected into cells, the nucleic acids of the invention inhibit the expression of the target gene with an IC50 value lower than 2000 pM. In more preferred embodiments, when transfected into cells, the nucleic acids of the invention inhibit the expression of the target gene with an IC50 value lower than 1000 pM. In even more preferred embodiments, when transfected into cells, the nucleic acids of the invention inhibit the expression of the target gene with an IC50 value lower than 500 pM. In the most preferred embodiments, when transfected into cells, the nucleic acids of the invention inhibit the expression of the target gene with an IC50 value lower than 100 pM.

[0311] Inhibition of the target gene can be quantified by the following methods: Huh7 cells (a human hepatocyte-derived cell line, obtained from the JCRB cell bank) can be maintained in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% FBS at 37°C in a 5% CO2 atmosphere. The cells can then be transfected with siRNA duplexes targeting the mRNA transcribed from the target gene or negative control siRNA (siRNA control; sense strand 5'-UUCUCCGAACGUGUCACGUTT-3' (SEQ ID NO: 487), antisense strand 5'-ACGUGACACGUUCGGAGAATT-3' (SEQ ID NO: 486)) using 10×3-fold serial dilutions over a final double-stranded concentration range of 20 nM to 1 pM. Transfection can be carried out by adding 9.7 μL of Opti-MEM (ThermoFisher) + 0.3 μL of Lipofectamine RNAiMAX (ThermoFisher) to 10 μL of each siRNA duplex. The mixture is incubated at room temperature for 15 minutes and then added to 100 μL of complete growth medium containing 20,000 Huh7 cells. The cells are incubated at 37°C / 5% CO2 for 24 hours, after which total RNA purification can be performed using the RNeasy 96 Kit (Qiagen). Each duplex can be tested by transfection in duplicate wells in a single experiment.

[0312] cDNA synthesis can be carried out using the FastQuant RT (with gDNase) Kit (Tiangen). Real-time quantitative PCR (qPCR) can be performed using the FastStart Universal Probe Master Kit (Roche) with primers specific for the target gene and human GAPDH (Hs02786624_g1) on an ABI Prism 7900HT or ABI QuantStudio 7.

[0313] qPCR is performed in duplicate on cDNA from each well, and the average cycle threshold (Ct) can be calculated. Relative target gene expression can be calculated relative to untreated cells, normalized to GAPDH, from the average Ct values using the comparative Ct (ΔΔCt) method. The maximum percent inhibition of target gene expression and the IC50 value can be calculated using a four-parameter (variable slope) model with GraphPad Prism 9.

[0314] Alternatively or additionally, inhibition of target gene expression may be characterized by a decrease in the average relative expression of the target gene.

[0315] In some embodiments, when the cells are transfected with 0.1 nM of the nucleic acid of the invention, the average relative expression of the target gene is preferably less than 1, 0.9, 0.8, 0.7, 0.6, 0.5, or 0.4, as determined by qPCR, more preferably by reverse transcriptase (RT)-qPCR, as described herein.

[0316] In some embodiments, when the cells are transfected with 5 nM of the nucleic acid of the invention, the average relative expression of the target gene is preferably less than 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, or 0.3, as determined by qPCR, more preferably by reverse transcriptase (RT)-qPCR, as described herein.

[0317] The average relative expression of the target gene can be quantified by the following method: Huh7 cells (a human hepatocyte-derived cell line obtained from the JCRB cell bank) can be maintained in Dulbecco's Modified Eagle Medium (DMEM) supplemented with 10% FBS at 37°C in a 5% CO2 atmosphere. The cells can be transfected with siRNA duplexes targeting mRNA or negative control siRNA (siRNA control; sense strand 5'-UUCUCCGAACGUGUCACGUTT-3' (SEQ ID NO: 487), antisense strand 5'-ACGUGACACGUUCGGAGAATT-3' (SEQ ID NO: 486)) at final duplex concentrations of 5 nM and 0.1 nM. Transfection can be carried out by adding 9.7 μL of Opti-MEM (ThermoFisher) + 0.3 μL of Lipofectamine RNAiMAX (ThermoFisher) to 10 μL of each siRNA duplex. The mixture is incubated at room temperature for 15 minutes and then added to 100 μL of complete growth medium containing 20,000 Huh7 cells. The cells are incubated at 37°C / 5% CO2 for 24 hours, after which total RNA purification can be performed using the RNeasy 96 Kit (Qiagen). Each duplex can be tested by transfection in duplicate wells in two independent experiments.

[0318] cDNA synthesis can be carried out using the FastQuant RT (with gDNase) Kit (Tiangen). Real-time quantitative PCR (qPCR) can be performed using the FastStart Universal Probe Master Kit (Roche) with primers specific for the target gene and human GAPDH (Hs02786624_g1) on an ABI Prism 7900HT or ABI QuantStudio 7.

[0319] qPCR is performed in duplicate for the cDNA from each well, and the average Ct can be calculated. Relative target gene expression can be calculated relative to untreated cells, normalized to GAPDH, from the average Ct values using the comparative Ct (ΔΔCt) method.

[0320] Inhibition of target gene expression can be manifested by a decrease in the amount of target gene mRNA compared to a suitable control.

[0321] In other embodiments, inhibition of target gene expression can be evaluated with respect to parameters functionally related to gene expression, such as a decrease in protein expression or signal transduction pathways. Examples of target genes exemplified herein are HCII, ZPI, and B4GALT1.

[0322] Method for treating or preventing diseases related to target gene expression The present invention also provides a method of using a nucleic acid of the present invention, such as siRNA, or a composition containing a nucleic acid of the present invention, such as siRNA, to reduce or inhibit target gene expression in a cell. The method includes contacting the cell with a nucleic acid of the present invention, such as dsiRNA, and maintaining the cell for a time sufficient to obtain degradation of the target mRNA transcript, thereby inhibiting the expression of the target gene in the cell. The decrease in gene expression can be evaluated by any method known in the art.

[0323] In the method of the present invention, the cell may be contacted in vitro or in vivo, that is, the cell may be within a subject.

[0324] Suitable cells for treatment using the method of the present invention can be any cell that expresses a gene of interest related to a disease related to a hemostatic disorder, such as a hemostatic disorder such as hemophilia. Alternatively, suitable cells for treatment using the method of the present invention can be any cell that expresses a gene of interest related to diabetes or cardiovascular disease.

[0325] The in vivo method of the present invention may include administering to a subject a composition containing a nucleic acid of the present invention, such as siRNA, and the nucleic acid, such as siRNA, includes a nucleoside sequence that is complementary to at least a part of the RNA transcript of the target gene of the mammal to be treated.

[0326] The present invention further provides a treatment method for a subject in need of treatment. The treatment method of the present invention comprises administering a therapeutically effective amount of a pharmaceutical composition comprising a nucleic acid such as an siRNA of the present invention, or a nucleic acid targeting a gene, to a subject who would benefit from a decrease or inhibition of the expression of a target gene, for example, a subject.

[0327] The disease to be treated may relate to a hemostatic disorder, such as a disease related to a hemostatic disorder such as hemophilia, especially when the target gene is HCII or ZPI as disclosed herein.

[0328] Haemophilia, or hemophilia, is a mostly genetic hereditary disorder that impairs the body's ability to form blood clots, a process necessary to stop bleeding. This results in longer bleeding of the subject after injury, easy bruising, and an increased risk of bleeding within joints or the brain. Subjects with mild disease may only have symptoms after an accident or during surgery. Bleeding into joints, also called hemarthrosis, can cause permanent damage, while bleeding in the brain can cause long-term headache, seizures, or a decrease in the level of consciousness.

[0329] There are two main types of hemophilia: hemophilia A, which results from low levels of clotting factor VIII, and hemophilia B, which results from low levels of clotting factor IX. They are typically inherited from parents through the X chromosome carrying a non-functional gene. Rarely, new mutations can occur during early development, or hemophilia can develop later in life due to antibodies formed against the clotting factors. Other types include hemophilia C, which results from low levels of factor XI, von Willebrand disease, which results from low levels of a substance called von Willebrand factor, and parahemophilia, which results from low levels of factor V. Hemophilia A, B, and C prevent the proper functioning of the intrinsic pathway; this clotting pathway is necessary when there is damage to the endothelium of blood vessels. Acquired hemophilia is associated with cancer, autoimmune disorders, and pregnancy. Diagnosis is by testing blood for its clotting ability and its clotting factor levels.

[0330] In certain embodiments, the nucleic acids of the invention, particularly those that inhibit the expression of ZPI or HCII, are suitable for the treatment, or the treatment of hemophilia A, B, and / or C. In certain embodiments, the nucleic acids of the invention, particularly those that inhibit the expression of ZPI or HCII, are suitable for the treatment, or the treatment of hemophilia A and / or B. In certain embodiments, the nucleic acids of the invention, particularly those that inhibit the expression of ZPI or HCII, are suitable for the treatment, or the treatment of acquired hemophilia. In certain embodiments, the nucleic acids of the invention, particularly those that inhibit the expression of ZPI or HCII, are suitable for the treatment, or the treatment of von Willebrand disease. In certain embodiments, the nucleic acids of the invention, particularly those that inhibit the expression of ZPI or HCII, are suitable for the treatment, or the treatment of parahemophilia.

[0331] Although not wishing to be bound by theory, treatment with the nucleic acids of the invention can result in an increase in coagulation factor levels such that bleeding can be reduced or prevented. Thus, in a preferred embodiment, treatment with a nucleic acid of the invention, particularly a nucleic acid that inhibits the expression of ZPI or HCII, can reduce or prevent bleeding episodes in a subject suffering from hemophilia. In another preferred embodiment, treatment with a nucleic acid of the invention, particularly a nucleic acid that inhibits the expression of ZPI or HCII, can reduce or prevent bleeding into joints in a subject suffering from hemophilia. In certain embodiments, treatment with a nucleic acid of the invention, particularly a nucleic acid that inhibits the expression of ZPI or HCII, can reduce or prevent bleeding into muscle or the brain in a subject suffering from hemophilia.

[0332] The disease to be treated can be diabetes, particularly when the target gene is B4GALT1 as disclosed herein.

[0333] According to the present invention, the term “diabetes” as used herein refers to a group of metabolic diseases in which a subject has hyperglycemia because the body does not produce sufficient insulin or the cells do not respond to the insulin produced. There are three main types of diabetes: (1) Type 1 diabetes (T1D): caused by the body's inability to produce insulin, and the person currently needs to inject insulin (also called insulin-dependent diabetes mellitus, abbreviated IDDM, and juvenile diabetes). (2) Type 2 diabetes (T2D): results from insulin resistance, a condition in which cells cannot properly use insulin, and is sometimes combined with absolute insulin deficiency (formerly called non-insulin-dependent diabetes mellitus, abbreviated NIDDM, and adult-onset diabetes). (3) Gestational diabetes (GD): occurs when a pregnant woman who has never had diabetes before has high blood glucose levels during pregnancy. This can precede the onset of T2D.

[0334] In certain embodiments, a nucleic acid according to the invention, particularly a nucleic acid that inhibits the expression of B4GALT1, or a pharmaceutical composition comprising said nucleic acid, is used for the treatment of diabetes, preferably type 2 diabetes (T2D).

[0335] The disease to be treated can be a cardiovascular disease, especially when the target gene is B4GALT1 as disclosed herein.

[0336] As used herein, the term "cardiovascular disease" refers to any condition, disorder or medical condition associated with, resulting from or causing a structural or functional abnormality that impairs the normal function of the heart or the blood vessels supplying the heart. Cardiovascular diseases can include coronary artery disease, atherosclerosis, myocardial infarction, arteriosclerosis, hypertension, angina, deep vein thrombosis, stroke, congestive heart failure or arrhythmia. In a preferred embodiment, the cardiovascular disease is coronary artery disease.

[0337] In certain embodiments, the nucleic acids according to the invention, in particular nucleic acids that inhibit the expression of B4GALT1, or pharmaceutical compositions comprising said nucleic acids are used for the treatment of cardiovascular diseases, preferably coronary artery disease.

[0338] The nucleic acids of the invention, such as siRNA, may be administered as "free" nucleic acids or "free" siRNA administered in the absence of a pharmaceutical composition. The naked nucleic acid may be in a suitable buffer solution. The buffer solution may contain acetate, citrate, prolamine, carbonate, or phosphate, or any combination thereof. In one embodiment, the buffer solution is phosphate buffered saline (PBS). The pH and osmolality of the buffer solution can be adjusted to be suitable for administration to the subject.

[0339] Alternatively, the nucleic acids of the invention, such as siRNA, may be administered as a pharmaceutical composition, such as a dsiRNA liposome formulation.

[0340] In one embodiment, the method comprises administering a composition characterized herein such that the expression of the target gene is reduced, for example, by about 1, 2, 3, 4, 5, 6, 7, 8, 12, 16, 18, 24 hours, 28, 32, or about 36 hours. In one embodiment, the expression of the target gene is reduced for a long period of time, for example, at least about 2, 3, 4 days or more, for example, about 1 week, 2 weeks, 3 weeks, or 4 weeks or longer, for example, about 1 month, 2 months, or 3 months.

[0341] A therapeutically effective amount of a nucleic acid, such as siRNA, can be administered to a subject, for example, in an amount of about 0.01 mg / kg to about 200 mg / kg, to treat a disease related to a hemostatic disorder, such as a disease related to a hemostatic disorder such as hemophilia, or to treat diabetes, or to treat a cardiovascular disease.

[0342] The nucleic acid, such as siRNA, can be administered by intravenous infusion periodically over a period of time. In certain embodiments, after an initial treatment regimen, the treatment can be administered at a less frequent rate. Administration of siRNA can reduce the gene product level of the target gene, for example, in a patient's cells or tissues, by at least about 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%, or below the detection level of the assay method used. In certain embodiments, the administration results in clinical stabilization or preferably a clinically relevant decrease in at least one sign or symptom of the target gene-related disorder.

[0343] Alternatively, a nucleic acid, such as siRNA, can be administered subcutaneously, i.e., by subcutaneous injection. One or more injections can be used to deliver the desired daily dose of the nucleic acid, such as siRNA, to a subject. The injection may be repeated over a period of time. The administration may be repeated periodically. In certain embodiments, after an initial treatment regimen, the treatment can be administered less frequently. A repeated dosing regimen can include the administration of a therapeutic amount of the nucleic acid at regular intervals, such as every other day or once a year. In certain embodiments, the nucleic acid is administered from about once a month to about once every quarter (i.e., about once every three months).

[0344] In one aspect, the nucleic acids disclosed herein can be the nucleic acids defined hereinbelow in Sentences 1 to 45 of this specification:

[0345] 1. A nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region containing a first strand that is at least partially complementary to a portion of the RNA transcribed from the target gene and a second strand that is at least partially complementary to the first strand, wherein the nucleosides of the second strand are as follows (5'-3'): Me-Me-Me-Me-Me-Me-F-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-F-Me-Me, or Me-Me-Me-Me-Me-F-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, or Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me and contain a 2'-sugar modification pattern of nucleic acid.

[0346] 2. A nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region containing a first strand that is at least partially complementary to a portion of the RNA transcribed from the target gene and a second strand that is at least partially complementary to the first strand, wherein the nucleosides of the second strand are as follows (5'-3'): wherein the nucleosides of the second strand are as follows (5'-3'): Me(s)Me(s)Me-Me-Me-Me-F-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-F-Me-Me, or Me(s)Me(s)Me-Me-Me-F-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, or Me(s)Me(s)Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, or Me(s)Me(s)Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, or Me-Me-Me-Me-Me-Me-F-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-F(s)Me(s)Me, or Me-Me-Me-Me-Me-F-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me(s)Me(s)Me, or Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me(s)Me(s)Me, or Me-Me-Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me(s)Me(s)Me [wherein, (s) is a phosphorothioate nucleoside internucleoside linkage] A nucleic acid comprising the 2'-sugar and the linkage modification pattern thereof.

[0347] 3. A first strand that is at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand that is at least partially complementary to the first strand A nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region, wherein the nucleosides of the second strand are as follows (5'-3'): ia-ia-Me-Me-Me-Me-Me-Me-F-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-F-Me-Me, or ia-ia-Me-Me-Me-Me-Me-F-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, or ia-ia-Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, or ia-ia-Me-Me-Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, or Me-Me-Me-Me-Me-Me-F-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-F-Me-Me-ia-ia, or Me-Me-Me-Me-Me-F-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-ia-ia, or Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-ia-ia, or Me-Me-Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me-ia-ia [wherein, ia represents an inverted abasic nucleoside, and when the inverted abasic nucleoside represented by ia-ia is present at the 3'-end of the second strand, the inverted abasic nucleoside is typically present in an overhang of two nucleosides] a nucleic acid comprising the 2'-sugar and abasic modification pattern of

[0348] 4. A first strand that is at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand that is at least partially complementary to the first strand a nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region wherein the nucleosides of the second strand are as follows (5'-3'): ia-ia-Me(s)Me(s)Me-Me-Me-Me-F-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-F-Me-Me, or ia-ia-Me(s)Me(s)Me-Me-Me-F-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, or ia-ia-Me(s)Me(s)Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, or ia-ia-Me(s)Me(s)Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, Me-Me-Me-Me-Me-Me-F-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-F(s)Me(s)Me-ia-ia, or Me-Me-Me-Me-Me-F-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me(s)Me(s)Me-ia-ia, or Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me(s)Me(s)Me-ia-ia, or Me-Me-Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me(s)Me(s)Me-ia-ia [wherein, (s) is a phosphorothioate nucleoside internucleoside linkage, ia represents an inverted abasic nucleoside, and when the inverted abasic nucleoside represented by ia-ia is present at the 3'-end of the second strand, the inverted abasic nucleoside is typically present in an overhang of two nucleosides] A nucleic acid comprising a 2'-sugar, abasic, and linkage modification pattern of.

[0349] 5. The nucleic acid according to any preceding sentence, wherein the first strand comprises a modification pattern selected from the following or any combination thereof, the 1-position is the 5'-terminal nucleoside of the first strand, and the counting direction is 5'-3': 2'-F sugar modifications at at least positions 2, 14, and 16, and / or 2'-Me sugar modifications at positions 17-23, or said first strand comprises at least eight 2'-F sugar modifications, such as at least 2'-F sugar modifications at positions 2, 4, 6, 12, 14, 16, 18 and 20; and / or 2'-Me sugar modifications at positions 1, 3-5, 10-13, or said first strand comprises at least eight 2'-F sugar modifications, such as at least 2'-F sugar modifications at positions 2, 4, 6, 12, 14, 16, 18 and 20; and / or a 2'-Me sugar modification at position 7, or a thermodestabilizing modification at position 7, such as typically a modified unlocked nucleic acid or glycol nucleic acid, and / or 2'-F sugar modifications or thermodestabilizing modifications at position 6, such as typically modified unlocked or glycol nucleic acids, and / or positions 8 and 9 may be modified by a 2'-Me sugar modification and a 2'-F sugar modification, typically the same 2' sugar modification; Thereby, typically the first strand has the following modification pattern (5'-3'): Me-F-Me-Me-Me-(M)4-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me wherein M represents a modification selected from a 2'-Me sugar modification, a 2'-F sugar modification and a thermolabile modification, such as typically a modified unlocked nucleic acid or a glycol nucleic acid, wherein the thermolabile modification may typically be present at the 6 position and the 2'-Me sugar modification may typically be present at the 7 position. or whereby typically the first strand comprises the following modification pattern (5'-3'): Me-F-Me-Me-Me-(M1)-(M2)3-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me wherein M1 represents a thermodestabilizing modification, such as typically a modified unlocked nucleic acid or a glycol nucleic acid, and M2 represents a modification selected from a 2'-Me sugar modification and a 2'-F sugar modification. or whereby typically the first strand comprises the following modification pattern (5'-3'): Me-F-Me-Me-Me-(M1)-Me-(M2)2-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me [Wherein, M1 represents a modification selected from 2'-Me sugar modification, 2'-F sugar modification and heat destabilizing modification, for example typically a modified unlocked nucleic acid or glycol nucleic acid, and M2 represents a modification selected from 2'-Me sugar modification and 2'-F sugar modification] comprising, or thereby, typically the first strand has the following modification pattern (5'-3'): Me-F-Me-Me-Me-(M1)-Me-(M2)2-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me [Wherein, M1 represents a heat destabilizing modification, for example typically a modified unlocked nucleic acid or glycol nucleic acid, M2 represents a modification selected from 2'-Me sugar modification and 2'-F sugar modification, and typically M2 can be the same 2' sugar modification] comprising.

[0350] 6. A first strand that is at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand that is at least partially complementary to the first strand A nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region, wherein the nucleosides of the second strand are as follows: Second strand (5'-3'): ia-ia-Me-Me-Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me comprising the 2' sugar and abasic modification pattern of, or comprising a sugar modification in which the 7-position on the second strand is a 2'-Me modification, the 1-position is the 5'-terminal nucleoside of the second strand, the counting direction is 5'-3', and typically there are 2 inverted abasic nucleosides in the 5'-terminal region of the second strand, wherein the modification pattern of the first strand comprises a modification pattern selected from the following or any combination thereof, the 1-position is the 5'-terminal nucleoside of the first strand, and the counting direction is 5'-3', nucleic acid: 2'-F sugar modifications at least at positions 2, 14, and 16, and / or 2'-Me sugar modifications at positions 17 to 23, and / or 2'-Me sugar modifications at positions 1, 3 to 5, 10 to 13, and / or 2'-Me sugar modification at position 7, or a heat destabilizing modification at position 7, such as typically a modified unlocked nucleic acid or glycol nucleic acid, and / or 2'-F sugar modification or heat destabilizing modification at position 6, such as typically a modified unlocked nucleic acid or glycol nucleic acid, and / or Positions 8 and 9 may be modifications selected from 2'-Me sugar modification and 2'-F sugar modification, and typically may be the same 2'-sugar modification, whereby typically the first strand has the following modification pattern (5'-3'): Me-F-Me-Me-Me-(M)4-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me [wherein M represents a modification selected from 2'-Me sugar modification, 2'-F sugar modification, and heat destabilizing modification, such as typically a modified unlocked nucleic acid or glycol nucleic acid, the heat destabilizing modification may typically be present at position 6, and the 2'-Me sugar modification may typically be present at position 7] and may include, or whereby typically the first strand has the following modification pattern (5'-3'): Me-F-Me-Me-Me-(M1)-(M2)3-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me [wherein M1 represents a heat destabilizing modification, such as typically a modified unlocked nucleic acid or glycol nucleic acid, and M2 represents a modification selected from 2'-Me sugar modification and 2'-F sugar modification] and may include, or whereby typically the first strand has the following modification pattern (5'-3'): Me-F-Me-Me-Me-(M1)-Me-(M2)2-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me [wherein, M1 represents a modification selected from 2'-Me sugar modification, 2'-F sugar modification, and heat destabilizing modification, for example, typically a modified unlocked nucleic acid or glycol nucleic acid, and M2 represents a modification selected from 2'-Me sugar modification and 2'-F sugar modification] comprising, or thereby, typically the first strand having the following modification pattern (5'-3'): Me-F-Me-Me-Me-(M1)-Me-(M2)2-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me [wherein, M1 represents a heat destabilizing modification, for example, typically a modified unlocked nucleic acid or glycol nucleic acid, M2 represents a modification selected from 2'-Me sugar modification and 2'-F sugar modification, and typically M2 can be the same 2' sugar modification] is included.

[0351] 7. (M)4 has the following 2' sugar modification pattern (5'-3'): F-Me-Me-F Me-F-Me-F F-Me-F-Me F-F-F-F Me-F-F-Me Me-Me-F-F F-F-Me-Me Me-Me-Me-Me The nucleic acid according to the fifth or sixth sentence, wherein any one of the above represents.

[0352] 8. Two phosphorothioate internucleoside linkages are respectively present between three consecutive positions in both the 5' terminal region and the 3' terminal region of the first strand, whereby the terminal nucleosides in each of the 5' terminal region and the 3' terminal region of the first strand are respectively attached to the 5' and 3' adjacent second last nucleosides by the phosphorothioate internucleoside linkages, and the 5' and 3' second last nucleosides are respectively attached to the 5' and 3' adjacent third last nucleosides by the phosphorothioate internucleoside linkages, Optionally, there may be further two phosphorothioate internucleoside linkages between three consecutive positions in the 3'-terminal region of the second strand, whereby the 3'-terminal nucleoside is attached to the adjacent penultimate nucleoside by a phosphorothioate internucleoside linkage, and the penultimate nucleoside is attached to the adjacent antepenultimate nucleoside by a phosphorothioate internucleoside linkage, and / or Optionally, there may be further two phosphorothioate internucleoside linkages between three consecutive positions in the 5'-terminal region of the second strand, whereby the 5'-terminal nucleoside is attached to the adjacent penultimate nucleoside by a phosphorothioate internucleoside linkage, and the penultimate nucleoside is attached to the adjacent antepenultimate nucleoside by a phosphorothioate internucleoside linkage, the nucleic acid according to any one of the fifth to seventh sentences.

[0353] 9. A first strand that is at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand that is at least partially complementary to the first strand A nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region, wherein the nucleosides of the second and first strands are as follows (5'-3'): Modification pattern 1: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-F-Me-Me, First strand (5'-3'): Me-F-Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Or modification pattern 2: Second strand (5'-3'): Me-Me-Me-Me-Me-F-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me-F-Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Or modification pattern 3: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me-F-Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Or modification pattern 4: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me-F-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Or modification pattern 5: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me-F-Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me A nucleic acid comprising a 2'-sugar modification pattern of

[0354] 10. A first strand that is at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand that is at least partially complementary to the first strand A nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region, wherein the nucleosides of the second and first strands are as follows (5'-3'): Modification pattern 1: Second strand (5'-3'): Me(s)Me(s)Me-Me-Me-Me-F-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-F-Me-Me, First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modification pattern 2: Second strand (5'-3'): Me(s)Me(s)Me-Me-Me-F-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modification pattern 3: Second strand (5'-3'): Me(s)Me(s)Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modification pattern 4: Second strand (5'-3'): Me(s)Me(s)Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modification pattern 5: Second strand (5'-3'): Me(s)Me(s)Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me(s)F(s)Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modification pattern 6: Second strand (5'-3'): Me(s)Me(s)Me-Me-Me-Me-F-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me(s)F(s)Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me [wherein, (s) is a phosphorothioate internucleoside linkage] A nucleic acid comprising the 2'-sugar and the binding modification pattern of

[0355] 11. A first strand that is at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand that is at least partially complementary to the first strand A nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region, wherein the nucleosides of the second and first strands are as follows (5'-3'): Modification pattern 1: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-F(s)Me(s)Me, First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modification pattern 2: Second strand (5'-3'): Me-Me-Me-Me-Me-F-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me(s)Me(s)Me, First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modification pattern 3: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me(s)Me(s)Me, First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modification pattern 4: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me(s)Me(s)Me, First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modification pattern 5: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me(s)Me(s)Me, First strand (5'-3'): Me(s)F(s)Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modification pattern 6: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me(s)Me(s)Me, First strand (5'-3'): Me(s)F(s)Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me [wherein, (s) is a phosphorothioate internucleoside linkage] A nucleic acid comprising the 2'-sugar and the binding modification pattern of.

[0356] 12. A first strand that is at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand that is at least partially complementary to the first strand A nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region, wherein the nucleosides of the second and first strands are as follows (5'-3'): Modification pattern 1: Second strand (5'-3'): ia-ia-Me-Me-Me-Me-Me-Me-F-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-F-Me-Me, First strand (5'-3'): Me-F-Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Or modification pattern 2: Second strand (5'-3'): ia-ia-Me-Me-Me-Me-Me-F-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me-F-Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Or modification pattern 3: Second strand (5'-3'): ia-ia-Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me-F-Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Or modification pattern 4: Second strand (5'-3'): ia-ia-Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me-F-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Or modification pattern 5: Second strand (5'-3'): ia-ia-Me-Me-Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me-F-Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Or modification pattern 6: Second strand (5'-3'): ia-ia-Me-Me-Me-Me-Me-Me-F-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me First strand (5'-3'): Me-F-Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me [wherein, ia represents an inverted abasic nucleoside] A nucleic acid comprising the 2'-sugar and abasic modification pattern of.

[0357] 13. A first strand that is at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand that is at least partially complementary to the first strand A nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region, wherein the nucleosides of the second and first strands are as follows: Modification pattern 1: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-F-Me-Me-ia-ia, First strand (5'-3'): Me-F-Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Or modification pattern 2: Second strand (5'-3'): Me-Me-Me-Me-Me-F-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-ia-ia, First strand (5'-3'): Me-F-Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Or modification pattern 3: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-ia-ia, First strand (5'-3'): Me-F-Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Or modification pattern 4: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-ia-ia, First strand (5'-3'): Me-F-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Or modification pattern 5: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me-ia-ia, First strand (5'-3'): Me-F-Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Or modification pattern 6: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me-ia-ia First strand (5'-3'): Me-F-Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me [Wherein, ia represents an inverted abasic nucleoside, and when the inverted abasic nucleoside represented by ia-ia is present at the 3'-end of the second strand, the inverted abasic nucleoside is typically present in an overhang of two nucleosides] A nucleic acid comprising the 2'-sugar and abasic modification patterns of

[0358] 14. A first strand that is at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand that is at least partially complementary to the first strand A nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region The nucleosides of the second and first strands are as follows (5'-3'): Modification pattern 1: Second strand (5'-3'): ia-ia-Me(s)Me(s)Me-Me-Me-Me-F-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-F-Me-Me, First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modification pattern 2: Second strand (5'-3'): ia-ia-Me(s)Me(s)Me-Me-Me-F-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modification pattern 3: Second strand (5'-3'): ia-ia-Me(s)Me(s)Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modification pattern 4: Second strand (5'-3'): ia-ia-Me(s)Me(s)Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modification pattern 5: Second strand (5'-3'): ia-ia-Me(s)Me(s)Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me(s)F(s)Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modification pattern 6: Second strand (5'-3'): ia-ia-Me(s)Me(s)Me-Me-Me-Me-F-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me(s)F(s)Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me [Wherein, (s) is a phosphorothioate internucleoside linkage, ia represents an inverted abasic nucleoside] A nucleic acid comprising the 2'-sugar, abasic, and linkage modification patterns of.

[0359] 15. A first strand that is at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand that is at least partially complementary to the first strand A nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region, Wherein the nucleosides of the second and first strands are as follows (5'-3'): Modification pattern 1: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-F-F-F-F-Me-Me-Me-Me-Me-Me-Me-F(s)Me(s)Me-ia-ia, First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modification pattern 2: Second strand (5'-3'): Me-Me-Me-Me-Me-F-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me(s)Me(s)Me-ia-ia, First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modification pattern 3: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me(s)Me(s)Me-ia-ia, First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modification pattern 4: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-Me-F-F-F-F-Me-Me-Me-Me-Me-Me-Me(s)Me(s)Me-ia-ia, First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modification pattern 5: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me(s)Me(s)Me-ia-ia, First strand (5'-3'): Me(s)F(s)Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me Or modification pattern 6: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me(s)Me(s)Me-ia-ia, First strand (5'-3'): Me(s)F(s)Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me [wherein, (s) is a phosphorothioate internucleoside linkage, ia represents an inverted abasic nucleoside. When the inverted abasic nucleoside represented by ia-ia is present at the 3'-end of the second strand, the inverted abasic nucleoside typically exists in an overhang of two nucleosides. A nucleic acid comprising the 2'-sugar, abasic, and linkage modification patterns of .

[0360] 16. A first strand that is at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand that is at least partially complementary to the first strand A nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region, Counting from the 1st position at the 5'-end of the second strand, which is the most 5'-side nucleoside without including an abasic nucleoside, the 7th position on the second strand contains a sugar modification that is a 2'-Me modification, or The second strand has the following modification pattern: Second strand (5'-3'): Me-Me-Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me A nucleic acid, wherein the first strand contains a modification pattern selected from the following or any combination thereof, the 1st position is the 5'-end nucleoside of the first strand, and the counting direction is 5'-3': 2'-F sugar modifications at least at positions 2, 14, and 16, and / or 2'-Me sugar modifications at positions 17 to 23, and / or 2'-Me sugar modifications at positions 1, 3 to 5, 10 to 13, and / or A 2'-Me sugar modification at the 7th position, or a heat destabilizing modification at the 7th position, for example, typically a modified unlocked nucleic acid or glycol nucleic acid, and / or A 2'-F sugar modification or a heat destabilizing modification at the 6th position, for example, typically a modified unlocked nucleic acid or glycol nucleic acid, and / or The 8th and 9th positions may be modifications selected from 2'-Me sugar modification and 2'-F sugar modification, and typically may be the same 2'-sugar modification, Thereby, typically the first strand has the following modification pattern (5'-3'): Me-F-Me-Me-Me-(M)4-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me [Wherein, M represents a modification selected from 2'-Me sugar modification, 2'-F sugar modification and thermally labile modification, for example typically a modified unlocked nucleic acid or glycol nucleic acid, and the thermally labile modification may typically be present at the 6-position, and the 2'-Me sugar modification may typically be present at the 7-position] may comprise, or thereby, typically the first strand has the following modification pattern (5'-3'): Me-F-Me-Me-Me-(M1)-(M2)3-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me [Wherein, M1 represents a thermally labile modification, for example typically a modified unlocked nucleic acid or glycol nucleic acid, and M2 represents a modification selected from 2'-Me sugar modification and 2'-F sugar modification] may comprise, or thereby, typically the first strand has the following modification pattern (5'-3'): Me-F-Me-Me-Me-(M1)-Me-(M2)2-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me [Wherein, M1 represents a modification selected from 2'-Me sugar modification, 2'-F sugar modification and thermally labile modification, for example typically a modified unlocked nucleic acid or glycol nucleic acid, and M2 represents a modification selected from 2'-Me sugar modification and 2'-F sugar modification] may comprise, or thereby, typically the first strand has the following modification pattern (5'-3'): Me-F-Me-Me-Me-(M1)-Me-(M2)2-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me [Wherein, M1 represents a thermally labile modification, for example typically a modified unlocked nucleic acid or glycol nucleic acid, and M2 represents a modification selected from 2'-Me sugar modification and 2'-F sugar modification, and typically M2 may be the same 2'-sugar modification] comprises.

[0361] 17. A first strand that is at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand that is at least partially complementary to the first strand A nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region, wherein the second strand contains two consecutive abasic nucleosides in the 5' or 3' terminal region of the second strand, Counting from the 1st position at the 5' end of the second strand, which is the most 5'-side nucleoside without containing an abasic nucleoside, the 7th position on the second strand contains a sugar modification that is a 2'-Me modification, Optionally, the first strand contains a modification pattern selected from the following or any combination thereof. The 1st position is the 5'-terminal nucleoside of the first strand, and the counting direction is 5'-3': nucleic acid 2'-F sugar modifications at least at positions 2, 14, and 16, and / or 2'-Me sugar modifications at positions 17 to 23, and / or 2'-Me sugar modifications at positions 1, 3 to 5, 10 to 13, and / or 2'-Me sugar modification at position 7, or a heat-labile modification at position 7, such as typically a modified unlocked nucleic acid or glycol nucleic acid, and / or 2'-F sugar modification or heat-labile modification at position 6, such as typically a modified unlocked nucleic acid or glycol nucleic acid, and / or Positions 8 and 9 may be modifications selected from 2'-Me sugar modification and 2'-F sugar modification, and typically may be the same 2'-sugar modification, Thereby, typically the first strand has the following modification pattern (5'-3'): Me-F-Me-Me-Me-(M)4-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me [Wherein, M represents a modification selected from 2'-Me sugar modification, 2'-F sugar modification, and heat-labile modification, such as typically a modified unlocked nucleic acid or glycol nucleic acid. The heat-labile modification may typically be present at position 6, and the 2'-Me sugar modification may typically be present at position 7.] may include or thereby, typically the first strand has the following modification pattern (5'-3'): Me-F-Me-Me-Me-(M1)-(M2)3-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me [wherein M1 represents a thermally labile modification, for example typically a modified unlocked nucleic acid or glycol nucleic acid, and M2 represents a modification selected from 2'-Me sugar modification and 2'-F sugar modification] may include or thereby, typically the first strand has the following modification pattern (5'-3'): Me-F-Me-Me-Me-(M1)-Me-(M2)2-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me [wherein M1 represents a modification selected from 2'-Me sugar modification, 2'-F sugar modification and thermally labile modification, for example typically a modified unlocked nucleic acid or glycol nucleic acid, and M2 represents a modification selected from 2'-Me sugar modification and 2'-F sugar modification] may include or thereby, typically the first strand has the following modification pattern (5'-3'): Me-F-Me-Me-Me-(M1)-Me-(M2)2-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me [wherein M1 represents a thermally labile modification, for example typically a modified unlocked nucleic acid or glycol nucleic acid, and M2 represents a modification selected from 2'-Me sugar modification and 2'-F sugar modification, typically M2 may be the same 2'-sugar modification] includes.

[0362] 18. A nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region including a first strand that is at least partially complementary to a portion of RNA transcribed from the target gene and a second strand that is at least partially complementary to the first strand wherein Two phosphorothioate internucleoside linkages are present between three consecutive positions in either the 5'-terminal region or the 3'-terminal region of the second strand, whereby each terminal nucleoside in either the 5'-terminal region or the 3'-terminal region of said second strand is attached by a phosphorothioate internucleoside linkage to its respective 5' or 3' adjacent second-last nucleoside, and each 5' or 3' second-last nucleoside is attached by a phosphorothioate internucleoside linkage to its respective 5' or 3' adjacent third-last nucleoside. Counting from the 1st position of the 5'-end of the second strand, which is the most 5'-side nucleoside without including abasic nucleosides, the 7th position on the second strand contains a sugar modification that is a 2'-Me modification. Optionally, the first strand contains a modification pattern selected from the following or any combination thereof, where the 1st position is the 5'-terminal nucleoside of the first strand and the counting direction is 5'-3': nucleic acid 2'-F sugar modifications at least at positions 2, 14, and 16, and / or 2'-Me sugar modifications at positions 17 to 23, and / or 2'-Me sugar modifications at positions 1, 3 to 5, 10 to 13, and / or 2'-Me sugar modification at position 7, or a heat destabilizing modification at position 7, such as typically a modified unlocked nucleic acid or glycol nucleic acid, and / or 2'-F sugar modification or heat destabilizing modification at position 6, such as typically a modified unlocked nucleic acid or glycol nucleic acid, and / or Positions 8 and 9 may be modifications selected from 2'-Me sugar modification and 2'-F sugar modification, and typically may be the same 2'-sugar modification. Thereby, typically the first strand has the following modification pattern (5'-3'): Me-F-Me-Me-Me-(M)4-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me [wherein M represents a modification selected from 2'-Me sugar modification, 2'-F sugar modification and thermolabile modification, for example typically a modified unlocked nucleic acid or glycol nucleic acid, the thermolabile modification may typically be present at the 6-position, and the 2'-Me sugar modification may typically be present at the 7-position] may comprise, or thereby, typically the first strand has the following modification pattern (5'-3'): Me-F-Me-Me-Me-(M1)-(M2)3-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me [wherein M1 represents a thermolabile modification, for example typically a modified unlocked nucleic acid or glycol nucleic acid, and M2 represents a modification selected from 2'-Me sugar modification and 2'-F sugar modification] may comprise, or thereby, typically the first strand has the following modification pattern (5'-3'): Me-F-Me-Me-Me-(M1)-Me-(M2)2-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me [wherein M1 represents a modification selected from 2'-Me sugar modification, 2'-F sugar modification and thermolabile modification, for example typically a modified unlocked nucleic acid or glycol nucleic acid, and M2 represents a modification selected from 2'-Me sugar modification and 2'-F sugar modification] may comprise, or thereby, typically the first strand has the following modification pattern (5'-3'): Me-F-Me-Me-Me-(M1)-Me-(M2)2-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me [wherein M1 represents a thermolabile modification, for example typically a modified unlocked nucleic acid or glycol nucleic acid, M2 represents a modification selected from 2'-Me sugar modification and 2'-F sugar modification, and typically M2 may be the same 2'-sugar modification] comprises.

[0363] 19. A first strand that is at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand that is at least partially complementary to the first strand A nucleic acid for inhibiting the expression of a target gene, which contains a double-stranded region and contains counting from the 1st position at the 5'-end of the second strand, which is the 5'-most nucleoside without a nucleobase nucleoside, the 7th position on the second strand contains a sugar modification that is a 2'-Me modification, at the 3'-end of the second strand, the nucleic acid is directly or indirectly conjugated to one or more ligand moieties, and the ligand moiety typically (i) one or more N-acetylgalactosamine (GalNAc) ligands, and / or (ii) one or more N-acetylgalactosamine (GalNAc) ligand derivatives, and / or (iii) one or more N-acetylgalactosamine (GalNAc) ligands and / or their derivatives conjugated to the nucleic acid through a linker including Optionally, the first strand contains a modification pattern selected from the following or any combination thereof, where the 1st position is the 5'-terminal nucleoside of the first strand and the counting direction is 5'-3': nucleic acid: 2'-F sugar modifications at least at positions 2, 14 and 16, and / or 2'-Me sugar modifications at positions 17 - 23, and / or 2'-Me sugar modifications at positions 1, 3 - 5, 10 - 13, and / or a 2'-Me sugar modification at the 7th position, or a heat destabilizing modification at the 7th position, such as typically a modified unlocked nucleic acid or glycol nucleic acid, and / or a 2'-F sugar modification or a heat destabilizing modification at the 6th position, such as typically a modified unlocked nucleic acid or glycol nucleic acid, and / or the 8th and 9th positions may be modifications selected from 2'-Me sugar modification and 2'-F sugar modification, and typically may be the same 2'-sugar modification, whereby, typically the first strand has the following modification pattern (5'-3'): Me-F-Me-Me-Me-(M)4-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me [wherein M represents a modification selected from 2'-Me sugar modification, 2'-F sugar modification and thermolabile modification, for example typically a modified unlocked nucleic acid or glycol nucleic acid, the thermolabile modification may typically be present at the 6-position, and the 2'-Me sugar modification may typically be present at the 7-position] may comprise, or thereby, typically the first strand has the following modification pattern (5'-3'): Me-F-Me-Me-Me-(M1)-(M2)3-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me [wherein M1 represents a thermolabile modification, for example typically a modified unlocked nucleic acid or glycol nucleic acid, and M2 represents a modification selected from 2'-Me sugar modification and 2'-F sugar modification] may comprise, or thereby, typically the first strand has the following modification pattern (5'-3'): Me-F-Me-Me-Me-(M1)-Me-(M2)2-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me [wherein M1 represents a modification selected from 2'-Me sugar modification, 2'-F sugar modification and thermolabile modification, for example typically a modified unlocked nucleic acid or glycol nucleic acid, and M2 represents a modification selected from 2'-Me sugar modification and 2'-F sugar modification] may comprise, or thereby, typically the first strand has the following modification pattern (5'-3'): Me-F-Me-Me-Me-(M1)-Me-(M2)2-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me [wherein M1 represents a thermolabile modification, for example typically a modified unlocked nucleic acid or glycol nucleic acid, M2 represents a modification selected from 2'-Me sugar modification and 2'-F sugar modification, and typically M2 may be the same 2'-sugar modification] comprises.

[0364] 20. A first strand that is at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand that is at least partially complementary to the first strand A nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region and containing the following, wherein the second strand has the following 2'-sugar and abasic modification pattern: Second strand (5'-3'): ia-ia-Me-Me-Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me The first strand contains a modification pattern selected from the following (5'-3'), with the 1-position being the 5'-terminal nucleoside of the first strand and the counting direction being 5'-3': (5'-3')Me-F-Me-Me-Me-(M)4-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me [wherein M represents a modification selected from 2'-Me sugar modification, 2'-F sugar modification, and heat destabilizing modification, for example, typically a modified unlocked nucleic acid or glycol nucleic acid, the heat destabilizing modification may typically be present at the 6-position, and the 2'-Me sugar modification may typically be present at the 7-position] Or thereby, typically the first strand has the following modification pattern (5'-3'): (5'-3')Me-F-Me-Me-Me-(M1)-(M2)3-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me [wherein M1 represents a heat destabilizing modification, for example, typically a modified unlocked nucleic acid or glycol nucleic acid, and M2 represents a modification selected from 2'-Me sugar modification and 2'-F sugar modification] Including or thereby, typically the first strand has the following modification pattern (5'-3'): (5'-3')Me-F-Me-Me-Me-(M1)-Me-(M2)2-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me [wherein M1 represents a modification selected from 2'-Me sugar modification, 2'-F sugar modification, and heat destabilizing modification, for example, typically a modified unlocked nucleic acid or glycol nucleic acid, and M2 represents a modification selected from 2'-Me sugar modification and 2'-F sugar modification] comprises or is thereby, typically the first strand has the following modification pattern (5'-3'): (5'-3')Me-F-Me-Me-Me-(M1)-Me-(M2)2-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me [wherein M1 represents a heat-labile modification, for example typically a modified unlocked nucleic acid or glycol nucleic acid, and M2 represents a modification selected from 2'-Me sugar modification and 2'-F sugar modification, typically M2 can be the same 2' sugar modification] comprises.

[0365] 21. A first strand that is at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand that is at least partially complementary to the first strand A nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region, wherein the second and first strands have the following modification patterns: Second strand (5'-3'): ia-ia-Me-Me-Me-Me-Me-Me-Me-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me First strand (5'-3') Me-F-Me-Me-Me-(M1)-Me-(M2)2-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me [wherein M1 represents a heat-labile modification, for example typically a modified unlocked nucleic acid or glycol nucleic acid, and M2 represents a modification selected from 2'-Me sugar modification and 2'-F sugar modification, typically M2 can be the same 2' sugar modification] A nucleic acid comprising.

[0366] 22. A first strand that is at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand that is at least partially complementary to the first strand A nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region, wherein the second strand has the following 2'-sugar and abasic modification patterns: Modification pattern 1: Optionally combined with a first strand (5'-3'): Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me A second strand (5'-3'): ia-ia-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F Or modification pattern 2: Optionally combined with a first strand (5'-3'): Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me A second strand (5'-3'): F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-ia-ia [Wherein, ia represents an inverted abasic nucleoside, and when the inverted abasic nucleoside represented by ia-ia is present at the 3'-end of the second strand, the inverted abasic nucleoside is typically present in an overhang of two nucleosides] A nucleic acid comprising

[0367] 23. A first strand that is at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand that is at least partially complementary to the first strand A nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region, wherein the second strand has the following 2'-sugar, abasic, and linkage modification patterns: Modification pattern 1: Optionally combined with a first strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me(s)F(s)Me A second strand (5'-3'): ia-ia-F(s)Me(s)F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F Or modification pattern 2: Optionally, the first strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me(s)F(s)Me combined with The second strand (5'-3'): F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F-Me-F(s)Me(s)F-ia-ia [wherein, (s) is a phosphorothioate internucleoside linkage, ia represents an inverted abasic nucleoside, and when the inverted abasic nucleoside represented by ia-ia is present at the 3'-end of the second strand, the inverted abasic nucleoside is typically present in an overhang of two nucleosides] A nucleic acid comprising

[0368] 24. A first strand that is at least partially complementary to a portion of RNA transcribed from a target gene, and a second strand that is at least partially complementary to the first strand A nucleic acid for inhibiting the expression of a target gene, comprising a double-stranded region, wherein the nucleosides of the second and first strands comprise the following 2'-sugar and linkage modification patterns (5'-3'): The second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, or wherein the 2'-Me or 2'-F modified nucleosides of the first strand comprise any one of the following modification patterns (5'-3'): The first strand (5'-3'): Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me, or The first strand (5'-3'): Me-F-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me, or First strand (5'-3'): Me-F-Me-Me-Me-Me-F-F-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me, or First strand (5'-3'): Me-F-Me-Me-Me-Me-F-F-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me, or First strand (5'-3'): Me-F-Me-Me-Me-Me-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me, or First strand (5'-3'): Me-F-Me-Me-Me-F-F-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me, or First strand (5'-3'): Me-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me(s)F(s)Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me, or First strand (5'-3'): Me(s)F(s)Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me, or First strand (5'-3'): Me(s)F(s)Me-Me-Me-F-F-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me, or First strand (5'-3'): Me(s)F(s)Me-Me-Me-Me-F-F-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me, or First strand (5'-3'): Me(s)F(s)Me-Me-Me-Me-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me, or First strand (5'-3'): Me(s)F(s)Me-Me-Me-F-F-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me, or First strand (5'-3'): Me(s)F(s)Me-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me [wherein (s) is a phosphorothioate internucleoside linkage].

[0369] 25. A nucleic acid for inhibiting the expression of a target gene, comprising a first strand that is at least partially complementary to a portion of RNA transcribed from the target gene, and a second strand that is at least partially complementary to the first strand, wherein the nucleosides of the second and first strands comprise the following 2'-sugar, abasic, and linkage modification patterns (5'-3'): Nucleic acid: Second strand (5'-3'): Me(s)Me(s)Me-Me-Me-Me-F-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, or Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me(s)Me(s)Me, or Second strand (5'-3'): ia-ia-Me-Me-Me-Me-Me-Me-F-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, or Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me-ia-ia, or Second strand (5'-3'): ia-ia-Me(s)Me(s)Me-Me-Me-Me-F-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, or Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-Me-F-F-F-Me-Me-Me-Me-Me-Me-Me-Me(s)Me(s)Me-ia-ia The 2'-Me or 2'-F modified nucleoside of the first strand includes any one of the following modification patterns (5'-3'): First strand (5'-3'): Me-F-Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me, or First strand (5'-3'): Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me, or First strand (5'-3'): Me-F-Me-Me-Me-F-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me, or First strand (5'-3'): Me-F-Me-Me-Me-F-F-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me, or First strand (5'-3'): Me-F-Me-Me-Me-Me-F-F-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me, or First strand (5'-3'): Me-F-Me-Me-Me-Me-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me, or First strand (5'-3'): Me-F-Me-Me-Me-F-F-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me, or First strand (5'-3'): Me-F-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me(s)F(s)Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me, or First strand (5'-3'): Me(s)F(s)Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me, or First strand (5'-3'): Me(s)F(s)Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me, or First strand (5'-3'): Me(s)F(s)Me-Me-Me-F-F-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me, or First strand (5'-3'): Me(s)F(s)Me-Me-Me-Me-F-F-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me, or First strand (5'-3'): Me(s)F(s)Me-Me-Me-Me-Me-F-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me, or First strand (5'-3'): Me(s)F(s)Me-Me-Me-F-F-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me, or First strand (5'-3'): Me(s)F(s)Me-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me [wherein, (s) is a phosphorothioate internucleoside linkage, ia represents an inverted abasic nucleoside, and when the inverted abasic nucleoside represented by ia-ia is present at the 3'-end of the second strand, the inverted abasic nucleoside is typically present in an overhang of two nucleosides. ]

[0370] 26. The nucleic acid according to any preceding sentence, wherein the first strand comprises at least 17 consecutive nucleosides that differ from any one of the sequences of the first strands listed in Table 2 by 0 or 1 nucleoside.

[0371] 27. The nucleic acid according to any preceding sentence, wherein the first strand comprises at least 17 consecutive nucleosides that differ from any one of the sequences of the first strand listed in Table 3 by 0 or 1 nucleoside.

[0372] 28. The nucleic acid according to sentence 26 or 27, wherein the first strand comprises nucleosides 2-18 of any one of the sequences defined in sentence 26 or 27.

[0373] 29. The nucleic acid according to any preceding sentence, wherein the second strand comprises a nucleoside sequence of at least 17 consecutive nucleosides that differ from any one of the sequences of the second strand listed in Table 2 by 0 or 1 nucleoside, and the second strand has a region of at least 85% complementarity with the first strand over 17 consecutive nucleosides.

[0374] 30. The nucleic acid according to any preceding sentence, wherein the second strand comprises a nucleoside sequence of at least 17 consecutive nucleosides that differ from any one of the sequences of the second strand listed in Table 4 by 0 or 1 nucleoside, and the second strand has a region of at least 85% complementarity with the first strand over 17 consecutive nucleosides.

[0375] 31. The nucleic acid according to any preceding sentence, wherein the first strand comprises any one of the sequences of the first strand listed in Table 2.

[0376] 32. The nucleic acid according to any preceding sentence, wherein the first strand comprises any one of the sequences of the first strand listed in Table 3.

[0377] 33. The nucleic acid according to any preceding sentence, wherein the second strand comprises any one of the sequences of the second strand listed in Table 2.

[0378] 34. The nucleic acid according to any preceding sentence, wherein the second strand comprises any one of the sequences of the second strand listed in Table 4.

[0379] 35. The nucleic acid according to any preceding sentence, wherein the nucleic acid is an siRNA oligonucleoside.

[0380] 36. The nucleic acid according to any preceding sentence, wherein the nucleic acid is directly or indirectly conjugated to one or more ligand moieties, and optionally, the ligand moiety is present in the terminal region of the second strand, typically in its 3'-terminal region.

[0381] 37. The ligand moiety is (i) one or more N-acetylgalactosamine (GalNAc) ligands, and / or (ii) one or more N-acetylgalactosamine (GalNAc) ligand derivatives, and / or (iii) one or more N-acetylgalactosamine (GalNAc) ligands and / or their derivatives conjugated to the nucleic acid through a linker The nucleic acid according to sentence 36.

[0382] 38. The nucleic acid according to sentence 37, wherein the one or more GalNAc ligands and / or GalNAc ligand derivatives are directly or indirectly conjugated in the 5' or 3'-terminal region of the second strand of the nucleic acid, typically in its 3'-terminal region.

[0383] 39. The ligand moiety has the following structure:

[0384]

Chemical formula

[0385] The nucleic acid according to any one of sentences 36 to 38.

[0386] 40. Structure:

[0387]

Chemical formula

[0388] [wherein, each occurrence of R1 is independently selected from the group consisting of hydrogen, methyl and ethyl, R2 is hydrogen, hydroxy, -OC 1~3 alkyl, -C(=O)OC 1~3 alkyl, halo and nitro, each occurrence of X1 and X2 is independently selected from the group consisting of methylene, oxygen and sulfur, m is an integer from 1 to 6, n is an integer from 1 to 10, q, r, s, t, v are independently integers from 0 to 4, provided that (i) both q and r cannot be 0 simultaneously, and (ii) s, t and v cannot all be 0 simultaneously, Z is an oligonucleoside] The nucleic acid according to any one of Sentences 36 to 39 having the same.

[0389] 41. Structure:

[0390]

Chemical formula

[0391] [wherein, r and s are independently integers selected from 1 to 16, Z is an oligonucleoside] The nucleic acid according to any one of Sentences 36 to 39 having the same.

[0392] 42. A pharmaceutical composition comprising the nucleic acid according to any one of the preceding sentences in combination with a pharmaceutically acceptable excipient or carrier.

[0393] 43. The nucleic acid or pharmaceutical composition according to any one of the preceding sentences for use in therapy.

[0394] 44. A nucleic acid or pharmaceutical composition as described in any of the preceding paragraphs for use in the prevention or treatment of a disease related to a hemostatic disorder, such as a disease related to a hemostatic disorder like hemophilia.

[0395] 45. A nucleic acid or pharmaceutical composition as described in any of the preceding paragraphs for use in the prevention or treatment of diabetes.

[0396] 46. A nucleic acid or pharmaceutical composition as described in any of the preceding paragraphs for use in the prevention or treatment of cardiovascular diseases.

[0397] In one aspect, the present invention can be applied to the compounds, methods, compositions or uses of the following sentences numbered from 1 to 101, and any reference to the formulas in Sentences 1 to 101 refers only to the formulas defined within Sentences 1 to 101. These formulas are reproduced in Figure 5. Specifically, the oligonucleoside moiety represented by Z in any of the following sentences may include a nucleic acid for inhibiting the expression of ZPI or HCII as defined below in this specification.

[0398] 1. The following structure:

[0399]

Chemical formula

[0400] [Wherein, R1 is independently selected from the group consisting of hydrogen, methyl and ethyl each time it appears, R2 is selected from the group consisting of hydrogen, hydroxy, -OC 1~3 alkyl, -C(=O)OC 1~3 alkyl, halo and nitro, X1 and X2 are independently selected from the group consisting of methylene, oxygen and sulfur each time they appear, m is an integer from 1 to 6, n is an integer from 1 to 10, q, r, s, t, v are independently integers from 0 to 4, provided that (i) q and r cannot both be 0 simultaneously, and (ii) s, t, and v cannot all be 0 simultaneously, Z is an oligonucleoside moiety] A compound comprising.

[0401] 2. The compound according to the first sentence, wherein each occurrence of R1 is hydrogen.

[0402] 3. The compound according to the first sentence, wherein R1 is methyl.

[0403] 4. The compound according to the first sentence, wherein R1 is ethyl.

[0404] 5. The compound according to any one of the first to fourth sentences, wherein R2 is hydroxy.

[0405] 6. The compound according to any one of the first to fourth sentences, wherein R2 is halo.

[0406] 7. The compound according to the sixth sentence, wherein R2 is fluoro.

[0407] 8. The compound according to the sixth sentence, wherein R2 is chloro.

[0408] 9. The compound according to the sixth sentence, wherein R2 is bromo.

[0409] 10. The compound according to the sixth sentence, wherein R2 is iodo.

[0410] 11. The compound according to the sixth sentence, wherein R2 is nitro.

[0411] 12. The compound according to any one of the first to eleventh sentences, wherein X1 is methylene.

[0412] 13. The compound according to any one of the first to eleventh sentences, wherein X1 is oxygen.

[0413] 14. A compound according to any one of the first to eleventh sentences, wherein X1 is sulfur.

[0414] 15. A compound according to any one of the first to fourteenth sentences, wherein X2 is methylene.

[0415] 16. A compound according to any one of the first to fifteenth sentences, wherein X2 is oxygen.

[0416] 17. A compound according to any one of the first to sixteenth sentences, wherein X2 is sulfur.

[0417] 18. A compound according to any one of the first to seventeenth sentences, wherein m = 3.

[0418] 19. A compound according to any one of the first to eighteenth sentences, wherein n = 6.

[0419] 20. A compound according to the thirteenth and fifteenth sentences, wherein X1 is oxygen, X2 is methylene, and preferably, q = 1, r = 2, s = 1, t = 1, v = 1. A compound according to the thirteenth and fifteenth sentences.

[0420] 21. A compound according to the twelfth and fifteenth sentences, wherein both X1 and X2 are methylene, and preferably, q = 1, r = 3, s = 1, t = 1, v = 1. A compound according to the twelfth and fifteenth sentences.

[0421] 22. Z is

[0422]

Chemical formula

[0423] [In the formula, Z1, Z2, Z3, and Z4 are each independently oxygen or sulfur each time they appear, One of the bond between P and Z2 and the bond between P and Z3 is a single bond and the other is a double bond] A compound according to any one of the first to twenty-first sentences.

[0424] 23. The compound according to sentence 22, wherein the oligonucleoside is an RNA compound capable of modulating, preferably inhibiting, the expression of a target gene.

[0425] 24. The compound according to sentence 23, wherein the RNA compound comprises an RNA duplex comprising a first and a second strand, the first strand is at least partially complementary to the RNA sequence of the target gene, the second strand is at least partially complementary to the first strand, and each of the first and second strands has 5' and 3' ends.

[0426] 25. The compound according to sentence 24, wherein the RNA compound has a phosphate attached adjacent to the 5' end of its second strand.

[0427] 26. The compound according to sentence 24, wherein the RNA compound has a phosphate attached adjacent to the 3' end of its second strand.

[0428] 27. A compound of formula (II):

[0429]

Chem.

[0430] 28. A compound of formula (III):

[0431]

Chem.

[0432] 29. An oligonucleoside comprising an RNA duplex comprising a first and a second strand, wherein the first strand is at least partially complementary to an RNA sequence of a target gene, the second strand is at least partially complementary to the first strand, each of the first and second strands has 5' and 3' ends, and the RNA duplex is attached to an adjacent phosphate at the 5' end of its second strand, a compound according to claim 27 or 28.

[0433] 30. A composition comprising a compound of formula (II) as defined in claim 27 and a compound of formula (III) as defined in claim 28, optionally dependent on claim 29.

[0434] 31. The composition according to claim 30, wherein the compound of formula (III) as defined in claim 28 is present in an amount in the range of 10 to 15% by weight of the composition.

[0435] 32. A compound of formula (IV):

[0436]

Chemical formula

[0437] 33. A compound of formula (V):

[0438]

Chemical formula

[0439] 34. An oligonucleoside comprising an RNA duplex comprising a first and a second strand, wherein the first strand is at least partially complementary to an RNA sequence of a target gene, the second strand is at least partially complementary to the first strand, each of the first and second strands has 5' and 3' ends, and the RNA duplex is attached to an adjacent phosphate at the 3' end of its second strand, a compound according to claim 32 or 33.

[0440] 35. A composition comprising a compound of formula (IV) as defined in claim 32 and a compound of formula (V) as defined in claim 33, optionally dependent on claim 34.

[0441] 36. The composition according to claim 35, wherein the compound of formula (V) as defined in claim 33 is present in an amount in the range of 10 to 15% by weight of the composition.

[0442] 37. A compound as defined in any one of claims 1 to 29, or claims 32 to 34, comprising an RNA duplex further comprising one or more riboses modified at the 2'-position, preferably a plurality of riboses modified at the 2'-position.

[0443] 38. The compound according to claim 37, wherein the modification is selected from 2'-O-methyl, 2'-deoxy-fluoro, and 2'-deoxy.

[0444] 39. A compound as defined in any one of claims 1 to 29, or claims 32 to 34, or claims 37 to 38, wherein the oligonucleoside further comprises one or more protecting moieties at one or more termini.

[0445] 40. The compound according to claim 39, wherein the one or more protecting moieties are not present at the termini of the oligonucleoside chain carrying the ligand moiety, and / or the one or more protecting moieties are selected from phosphorothioate internucleoside linkages, phosphorodithioate internucleoside linkages, and inverted abasic nucleosides, and the inverted abasic nucleoside is present at the distal terminus of the chain carrying the ligand moiety.

[0446] 41. The compound as defined in any one of claims 1 to 29, or claims 32 to 34, or claims 37 to 40, wherein the ligand moiety shown in formula (I) in claim 1 comprises one or more ligands.

[0447] 42. The compound according to claim 41, wherein the ligand moiety shown in formula (I) in claim 1 comprises one or more carbohydrate ligands.

[0448] 43. The compound according to sentence 42, wherein the one or more carbohydrates can be monosaccharides, disaccharides, trisaccharides, tetrasaccharides, oligosaccharides or polysaccharides.

[0449] 44. The compound according to sentence 43, wherein the one or more carbohydrates comprise one or more galactose moieties, one or more lactose moieties, one or more N-acetylgalactosamine moieties, and / or one or more mannose moieties.

[0450] 45. The compound according to sentence 44, wherein the one or more carbohydrates comprise one or more N-acetyl-galactosamine moieties.

[0451] 46. The compound according to sentence 45, comprising two or three N-acetylgalactosamine moieties.

[0452] 47. The compound according to any one of sentences 41 to 46, wherein the one or more ligands are attached in a linear configuration or a branched configuration.

[0453] 48. The compound according to sentence 47, wherein the one or more ligands are attached as a bifurcated or trifurcated branched configuration.

[0454] 49. The moiety shown in formula (I) in sentence 1:

[0455]

Chemical formula

[0456] is any one of formula (VIa), (VIb) or (VIc), preferably formula (VIa):

[0457]

Chemical formula

[0458] [wherein, A Iis hydrogen or a suitable hydroxy protecting group, a is an integer of 2 or 3, b is an integer of 2 to 5], or

[0459] [Chemical formula]

[0460] [wherein, A I is hydrogen or a suitable hydroxy protecting group, a is an integer of 2 or 3, c and d are independently integers of 1 to 6], or

[0461] [Chemical formula]

[0462] [wherein, A I is hydrogen or a suitable hydroxy protecting group, a is an integer of 2 or 3, e is an integer of 2 to 10] which is the compound described in Sentences 46 to 48.

[0463] 50. The said moiety shown in formula (I) in Sentence 1:

[0464] [Chemical formula]

[0465] is formula (VII):

[0466] [Chemical formula]

[0467] [wherein, A I is hydrogen, a is an integer of 2 or 3 The compound according to the 46th to 48th sentences.

[0468] 51. The compound according to the 49th or 50th sentence, where a = 2.

[0469] 52. The compound according to the 49th or 50th sentence, where a = 3.

[0470] 53. The compound according to the 49th sentence, where b = 3.

[0471] 54. The compound of formula (VIII):

[0472]

Chemical formula

[0473] 55. The compound of formula (IX):

[0474]

Chemical formula

[0475] 56. The compound according to the 54th or 55th sentence, wherein the oligonucleoside comprises an RNA duplex containing a first and a second strand, the first strand is at least partially complementary to the RNA sequence of the target gene, the second strand is at least partially complementary to the first strand, each of the first and second strands has 5' and 3' termini, and the RNA duplex is attached at the 5' terminus of its second strand to an adjacent phosphate.

[0476] 57. A composition comprising the compound of formula (VIII) defined in the 54th sentence and the compound of formula (IX) defined in the 55th sentence, optionally dependent on the 56th sentence.

[0477] 58. The composition according to the 57th sentence, wherein the compound of formula (IX) defined in the 55th sentence is present in an amount in the range of 10 to 15% by weight of the composition.

[0478] 59. Compound of formula (X):

[0479] [Chemical formula]

[0480] 60. Compound of formula (XI):

[0481] [Chemical formula]

[0482] 61. The compound according to paragraph 59 or 60, wherein the oligonucleoside comprises an RNA duplex comprising a first and a second strand, the first strand is at least partially complementary to the RNA sequence of the target gene, the second strand is at least partially complementary to the first strand, each of the first and second strands has 5' and 3' ends, and the RNA duplex is attached to an adjacent phosphate at the 3' end of its second strand.

[0483] 62. A composition comprising a compound of formula (X) as defined in paragraph 59 and a compound of formula (XI) as defined in paragraph 60, optionally dependent on paragraph 61.

[0484] 63. The composition according to paragraph 62, wherein the compound of formula (XI) as defined in paragraph 60 is present in an amount in the range of 10 to 15% by weight of the composition.

[0485] 64. The compound according to any one of paragraphs 54 to 63, wherein the oligonucleoside comprises an RNA duplex further comprising one or more riboses modified at the 2'-position, preferably a plurality of riboses modified at the 2'-position.

[0486] 65. The compound according to paragraph 64, wherein the modification is selected from 2'-O-methyl, 2'-deoxy-fluoro, and 2'-deoxy.

[0487] 66. A compound according to any one of paragraphs 54 to 65, wherein the oligonucleoside further comprises one or more deprotection moieties at one or more termini.

[0488] 67. The one or more deprotection moieties are not present at the termini of the oligonucleoside chain carrying the ligand moiety, and / or the one or more deprotection moieties are selected from phosphorothioate internucleoside linkages, phosphorodithioate internucleoside linkages and inverted abasic nucleosides, and the inverted abasic nucleoside is present at the distal terminus of the chain carrying the ligand moiety as shown in any one of formula (VIII), (IX), (X) or (XI) in any one of paragraphs 54, 55, 59 or 60, a compound according to paragraph 66.

[0489] 68. Compounds of formula (XII) and (XIII):

[0490] [Chemical formula]

[0491] [wherein, R1 is independently selected from the group consisting of hydrogen, methyl and ethyl each time it appears, R2 is selected from the group consisting of hydrogen, hydroxy, -OC 1~3 alkyl, -C(=O)OC 1~3 alkyl, halo and nitro, X1 and X2 are independently selected from the group consisting of methylene, oxygen and sulfur each time they appear, m is an integer from 1 to 6, n is an integer from 1 to 10, q, r, s, t, v are independently integers from 0 to 4, provided that (i) q and r cannot both be 0 simultaneously, and (ii) s, t and v cannot all be 0 simultaneously, Z is an oligonucleoside moiety] A step of reacting, and, if appropriate, a step of deprotecting the ligand and / or annealing the second strand for the oligonucleoside moiety, a compound described in any one of the first to 29th, 32nd to 34th, 37th to 56th, 59th to 61st, and 64th to 67th sentences, and / or a composition described in any one of the 30th, 31st, 35th, 36th, 57th, 58th, 62nd, and 63rd sentences.

[0492] 69. The compound of formula (XII) is the compound of formula (XIV) and (XV):

[0493]

Chemical formula

[0494] [wherein, Each occurrence of R1 is independently selected from the group consisting of hydrogen, methyl and ethyl, R2 is selected from the group consisting of hydrogen, hydroxy, -OC 1~3 alkyl, -C(=O)OC 1~3 alkyl, halo and nitro, Each occurrence of X1 and X2 is independently selected from the group consisting of methylene, oxygen and sulfur, q, r, s, t, v are independently integers from 0 to 4, provided that (i) q and r cannot both be 0 at the same time, and (ii) s, t and v cannot all be 0 at the same time, Z is an oligonucleoside moiety] The method according to sentence 68, which is prepared by reacting.

[0495] 70. The compound of formula (XII) is of formula (XIIa):

[0496]

Chemical formula

[0497] wherein the compound of formula (XIII) is of formula (XIIIa):

[0498] [Chemical formula]

[0499] wherein the oligonucleoside comprises an RNA duplex containing a first and a second strand, the first strand is at least partially complementary to the RNA sequence of the target gene, the second strand is at least partially complementary to the first strand, each of the first and second strands has 5' and 3' termini, and the RNA duplex is attached at the 5' terminus of its second strand to an adjacent phosphate, a compound according to any one of paragraphs 20, 25, 27, 29, 54, 56, and / or a composition according to any one of paragraphs 30, 31, 57, 58, by the method according to paragraph 68.

[0500] 71. The compound of formula (XII) is of formula (XIIb):

[0501] [Chemical formula]

[0502] wherein the compound of formula (XIII) is of formula (XIIIa):

[0503] [Chemical formula]

[0504] and the oligonucleoside comprises an RNA duplex comprising a first and a second strand, the first strand being at least partially complementary to the RNA sequence of the target gene, the second strand being at least partially complementary to the first strand, each of the first and second strands having 5' and 3' termini, and the RNA duplex being attached at the 5' terminus of its second strand to an adjacent phosphate, a compound according to any one of the 20th, 25th, 28th, 29th, 55th, 56th sentences, and / or a composition according to any one of the 30th, 31st, 57th, 58th sentences, the method according to the 68th sentence for preparing the same.

[0505] 72. The compound of formula (XII) is of formula (XIIc):

[0506]

Chemical formula

[0507] and the compound of formula (XIII) is of formula (XIIIa):

[0508]

Chemical formula

[0509] and the oligonucleoside comprises an RNA duplex comprising a first and a second strand, the first strand being at least partially complementary to the RNA sequence of the target gene, the second strand being at least partially complementary to the first strand, each of the first and second strands having 5' and 3' termini, and the RNA duplex being attached at the 3' terminus of its second strand to an adjacent phosphate, a compound according to any one of the 21st, 26th, 32nd, 34th, 59th, 61st sentences, and / or a composition according to any one of the 35th, 36th, 62nd, 63rd sentences, the method according to the 68th sentence for preparing the same.

[0510] 73. The compound of formula (XII) is of formula (XIId):

[0511] [Chemical]

[0512] and the compound of formula (XIII) is of formula (XIIIa):

[0513] [Chemical]

[0514] and the oligonucleoside contains an RNA duplex comprising a first and a second strand, the first strand is at least partially complementary to the RNA sequence of the target gene, the second strand is at least partially complementary to the first strand, each of the first and second strands has 5' and 3' ends, and the RNA duplex is attached to an adjacent phosphate at the 3' end of its second strand, a compound according to any one of sentences 21, 26, 33, 34, 60, 61, and / or a composition according to any one of sentences 35, 36, 62, 63, for the method according to sentence 68.

[0515] 74. The compound of formula (XIIIa) is of formula (XIIIb):

[0516] [Chemical]

[0517] for the method according to any one of sentences 70 to 73.

[0518] 75. The compound of formula (XIV) is either of formula (XIVa) or formula (XIVb):

[0519] [Chemical]

[0520] and the compound of formula (XV) is either of formula (XVa) or formula (XIVb):

[0521] [Chem.]

[0522] which is such that the oligonucleoside comprises an RNA duplex containing a first and a second strand, the first strand being at least partially complementary to the RNA sequence of the target gene, the second strand being at least partially complementary to the first strand, each of the first and second strands having 5' and 3' ends, and (i) the RNA duplex is attached at the 5' end of its second strand to an adjacent phosphate in formula (XVa), or (ii) the RNA duplex is attached at the 3' end of its second strand to an adjacent phosphate in formula (XVb), the method according to claim 69, dependent on claims 70 to 73.

[0523] 76. A compound of formula (XII):

[0524] [Chem.]

[0525] [wherein R1 is independently selected each time it occurs from the group consisting of hydrogen, methyl and ethyl, R2 is selected from the group consisting of hydrogen, hydroxy, -OC 1~3 alkyl, -C(=O)OC 1~3 alkyl, halo and nitro, X1 and X2 are independently selected each time they occur from the group consisting of methylene, oxygen and sulfur, q, r, s, t, v are independently integers from 0 to 4, provided that (i) q and r cannot both be 0 at the same time, and (ii) s, t and v cannot all be 0 at the same time, Z is an oligonucleoside moiety].

[0526] 77. A compound of formula (XIIa):

[0527] [Chemical formula]

[0528] 78. Compound of formula (XIIb):

[0529] [Chemical formula]

[0530] 79. Compound of formula (XIIc):

[0531] [Chemical formula]

[0532] 80. Compound of formula (XIId):

[0533] [Chemical formula]

[0534] 81. Compound of formula (XIII):

[0535] [Chemical formula]

[0536] [wherein, each R1, each time it appears, is independently selected from the group consisting of hydrogen, methyl and ethyl, m is an integer from 1 to 6, n is an integer from 1 to 10].

[0537] 82. Compound of formula (XIIIa):

[0538] [Chemical formula]

[0539] 83. Compound of formula (XIIIb):

[0540]

Chem.

[0541] 84. Compound of formula (XIV):

[0542]

Chem.

[0543] [wherein R1 is selected from the group consisting of hydrogen, methyl and ethyl, R2 is selected from the group consisting of hydrogen, hydroxy, -OC 1~3 alkyl, -C(=O)OC 1~3 alkyl, halo and nitro, X2 is selected from the group consisting of methylene, oxygen and sulfur, s, t, v are independently integers from 0 to 4, provided that s, t and v cannot all be 0 simultaneously].

[0544] 85. Compound of formula (XIVa):

[0545]

Chem.

[0546] 86. Compound of formula (XIVb):

[0547]

Chem.

[0548] 87. Compound of formula (XV):

[0549]

Chem.

[0550] [wherein, each occurrence of R1 is independently selected from the group consisting of hydrogen, methyl, and ethyl, X1 is selected from the group consisting of methylene, oxygen, and sulfur, q and r are independently integers from 0 to 4, provided that q and r cannot both be 0 at the same time, and Z is an oligonucleoside moiety].

[0551] 88. Compound of formula (XV a):

[0552]

Chemical formula

[0553] 89. Compound of formula (XV b):

[0554]

Chemical formula

[0555] 90. Use of a compound described in any one of Sentences 1 to 29, Sentences 32 to 34, Sentences 37 to 56, Sentences 59 to 61, and Sentences 64 to 67, and / or a composition described in any one of Sentences 30, 31, 35, 36, 57, 58, 62, and 63, for preparing a compound described in any one of Sentences 76, 81 to 84, and 87.

[0556] 91. Use of the compound described in Sentence 85 for preparing a compound described in any one of Sentences 1 to 29, Sentences 32 to 34, Sentences 37 to 56, Sentences 59 to 61, and Sentences 64 to 67, and / or a composition described in any one of Sentences 30, 31, 35, 36, 57, 58, 62, and 63, wherein R2 = F.

[0557] Use of the compound according to any one of the first to twenty-ninth, thirty-second to thirty-fourth, thirty-seventh to fifty-sixth, fifty-ninth to sixty-first, and sixty-fourth to sixty-seventh sentences, where R2 = OH, and / or the composition according to any one of the thirtieth, thirty-first, thirty-fifth, thirty-sixth, fifty-seventh, fifty-eighth, sixty-second, and sixty-third sentences, for preparing the compound according to the eighty-sixth sentence.

[0558] 93. Use of the compound according to any one of the twentieth, twenty-fifth, twenty-seventh, twenty-ninth, fifty-fourth, and fifty-sixth sentences, and / or the composition according to any one of the thirtieth, thirty-first, fifty-seventh, and fifty-eighth sentences, for preparing the compound according to the seventy-seventh sentence.

[0559] 94. Use of the compound according to any one of the twentieth, twenty-fifth, twenty-eighth, twenty-ninth, fifty-fifth, and fifty-sixth sentences, and / or the composition according to any one of the thirtieth, thirty-first, fifty-seventh, and fifty-eighth sentences, for preparing the compound according to the seventy-eighth sentence.

[0560] 95. Use of the compound according to any one of the twenty-first, twenty-sixth, thirty-second, thirty-fourth, fifty-ninth, and sixty-first sentences, and / or the composition according to any one of the thirty-fifth, thirty-sixth, sixty-second, and sixty-third sentences, for preparing the compound according to the seventy-ninth sentence.

[0561] 96. Use of the compound according to any one of the twenty-first, twenty-sixth, thirty-third, thirty-fourth, sixtieth, and sixty-first sentences, and / or the composition according to any one of the thirty-fifth, thirty-sixth, sixty-second, and sixty-third sentences, for preparing the compound according to the eightieth sentence.

[0562] 97. Use of the compound according to any one of the twentieth, twenty-fifth, twenty-seventh to twenty-ninth, fifty-fourth to fifty-sixth sentences, and / or the composition according to any one of the thirtieth, thirty-first, fifty-seventh, and fifty-eighth sentences, for preparing the compound according to the eighty-eighth sentence.

[0563] 98. Use of the compound according to any one of Paragraphs 21, 26, 32 to 34, 59 to 61, and / or the composition according to any one of Paragraphs 35, 36, 62, 63, for preparing the compound according to Paragraph 89.

[0564] 99. A compound or composition obtainable or obtained by the method according to any one of Paragraphs 68 to 75.

[0565] 100. A pharmaceutical composition comprising the compound according to any one of Paragraphs 1 to 29, 32 to 34, 37 to 56, 59 to 61, and 64 to 67, and / or the composition according to any one of Paragraphs 30, 31, 35, 36, 57, 58, 62, 63, together with a pharmaceutically acceptable carrier, diluent or excipient.

[0566] 101. The compound according to any one of Paragraphs 1 to 29, 32 to 34, 37 to 56, 59 to 61, and 64 to 67, and / or the composition according to any one of Paragraphs 30, 31, 35, 36, 57, 58, 62, 63, for use in therapy.

[0567] In another aspect, the present invention can be applied to the compounds, methods, compositions or uses of the following paragraphs numbered 1 to 56, and any reference to any formula in the paragraphs refers only to the formula defined within Paragraphs 1 to 56. These formulas are reproduced in Figure 6. Specifically, the oligonucleoside moiety represented by Z in any of the following paragraphs may comprise a nucleic acid for inhibiting the expression of ZPI or HCII as defined below in the present specification.

[0568] 1. The following structure:

[0569]

Chemical formula

[0570] [wherein, r and s are each independently an integer selected from 1 to 16, Z is an oligonucleoside moiety] A compound comprising.

[0571] 2. The compound according to claim 1, wherein s is an integer selected from 4 to 12.

[0572] 3. The compound according to claim 2, wherein s is 6.

[0573] 4. The compound according to any one of claims 1 to 3, wherein r is an integer selected from 4 to 14.

[0574] 5. The compound according to claim 4, wherein r is 6.

[0575] 6. The compound according to claim 4, wherein r is 12.

[0576] 7. The compound according to claim 5, which depends on claim 3.

[0577] 8. The compound according to claim 6, which depends on claim 3.

[0578] 9. Z is

[0579] [Chemical formula]

[0580] [wherein, Z1, Z2, Z3, and Z4 are each independently oxygen or sulfur each time they appear, One of the bond between P and Z2 and the bond between P and Z3 is a single bond, and the other bond is a double bond] The compound according to any one of claims 1 to 8.

[0581] 10. The compound according to any one of claims 1 to 9, wherein the oligonucleoside is an RNA compound capable of modulating, preferably inhibiting, the expression of a target gene.

[0582] 11. The compound according to claim 10, wherein the RNA compound comprises an RNA duplex comprising a first and a second strand, the first strand being at least partially complementary to the RNA sequence of the target gene, the second strand being at least partially complementary to the first strand, and each of the first and second strands having 5' and 3' termini.

[0583] 12. The compound according to claim 11, wherein the RNA compound is attached to an adjacent phosphate at the 5' terminus of its second strand, preferably also being dependent on claims 3 and 6.

[0584] 13. The compound according to claim 11, wherein the RNA compound is attached to an adjacent phosphate at the 3' terminus of its second strand, preferably also being dependent on claims 3 and 5.

[0585] 14. A compound of formula (II), preferably being dependent on claim 12:

[0586]

Chemical formula

[0587] 15. A compound of formula (III), preferably being dependent on claim 13:

[0588]

Chemical formula

[0589] 16. The compound as defined in any one of claims 1 to 15, wherein the oligonucleoside further comprises one or more riboses modified at the 2'-position, preferably a plurality of riboses modified at the 2'-position, in an RNA duplex.

[0590] 17. The compound according to claim 16, wherein the modification is selected from 2'-O-methyl, 2'-deoxy-fluoro, and 2'-deoxy.

[0591] 18. The compound according to any one of claims 1 to 17, wherein the oligonucleoside further comprises one or more deprotecting moieties at one or more termini.

[0592] 19. The one or more deprotecting moieties are not present at the termini of the oligonucleoside chain carrying the linker / ligand moiety, and / or the one or more deprotecting moieties are selected from phosphorothioate internucleoside linkages, phosphorodithioate internucleoside linkages, and inverted abasic nucleosides, and the inverted abasic nucleoside is present at the distal terminus of the same chain with respect to the terminus carrying the linker / ligand moiety. The compound according to claim 18.

[0593] 20. The compound according to any one of claims 1 to 19, wherein the ligand moiety shown in formula (I) in claim 1 comprises one or more ligands.

[0594] 21. The compound according to claim 20, wherein the ligand moiety shown in formula (I) in claim 1 comprises one or more carbohydrate ligands.

[0595] 22. The compound according to claim 21, wherein the one or more carbohydrates can be monosaccharides, disaccharides, trisaccharides, tetrasaccharides, oligosaccharides, or polysaccharides.

[0596] 23. The compound according to claim 22, wherein the one or more carbohydrates comprise one or more galactose moieties, one or more lactose moieties, one or more N-acetylgalactosamine moieties, and / or one or more mannose moieties.

[0597] 24. The compound according to claim 23, wherein the one or more carbohydrates comprise one or more N-acetyl-galactosamine moieties.

[0598] 25. The compound according to item 24, comprising 2 or 3 N-acetylgalactosamine moieties.

[0599] 26. The compound according to any one of the preceding items, wherein the one or more ligands are attached in a linear configuration or a branched configuration.

[0600] 27. The compound according to item 26, wherein the one or more ligands are attached as a bifurcated or trifurcated branched configuration.

[0601] 28. The moiety shown in formula (I) in item 1:

[0602]

Chemical formula

[0603] is any one of formula (IV), (V) or (VI), preferably formula (IV):

[0604]

Chemical formula

[0605] [wherein, A I is hydrogen or a suitable hydroxy protecting group, a is an integer of 2 or 3, b is an integer of 2 to 5], or

[0606]

Chemical formula

[0607] [wherein, A I is hydrogen or a suitable hydroxy protecting group, a is an integer of 2 or 3, c and d are independently integers of 1 to 6], or

[0608] [Chemical formula]

[0609] [wherein, A I is hydrogen or a suitable hydroxy protecting group, a is an integer of 2 or 3, e is an integer of 2 to 10] is a compound according to any one of claims 20 to 27.

[0610] 29. The said moiety represented by formula (I) in claim 1:

[0611] [Chemical formula]

[0612] is formula (VII):

[0613] [Chemical formula]

[0614] [wherein, A I is hydrogen, a is an integer of 2 or 3] is a compound according to any one of claims 1 to 28.

[0615] 30. The compound according to claim 28 or 29, wherein a = 2.

[0616] 31. The compound according to claim 28 or 29, wherein a = 3.

[0617] 32. The compound according to claim 28, wherein b = 3.

[0618] 33. The compound of formula (VIII):

[0619] [Chemical formula]

[0620] 34. Compound of formula (IX):

[0621] [Chemical formula]

[0622] 35. The compound according to item 33 or 34, comprising an RNA duplex in which the oligonucleoside further comprises one or more riboses modified at the 2'-position, preferably a plurality of riboses modified at the 2'-position.

[0623] 36. The compound according to item 35, wherein the modification is selected from 2'-O-methyl, 2'-deoxy-fluoro, and 2'-deoxy.

[0624] 37. The compound according to any one of items 33 to 36, wherein the oligonucleoside further comprises one or more deprotecting moieties at one or more termini.

[0625] 38. The one or more deprotecting moieties are not present at the termini of the oligonucleoside chain carrying the linker / ligand moiety, and / or the one or more deprotecting moieties are selected from phosphorothioate internucleoside linkages, phosphorodithioate internucleoside linkages, and inverted abasic nucleosides, and the inverted abasic nucleoside is present at the distal terminus of the same chain relative to the terminus carrying the linker / ligand moiety. The compound according to item 37.

[0626] 39. The oligonucleoside comprises an RNA duplex comprising a first and a second strand, the first strand is at least partially complementary to the RNA sequence of the target gene, the second strand is at least partially complementary to the first strand, each of the first and second strands has 5' and 3' termini, and the RNA duplex is attached to an adjacent phosphate at the 5' terminus of its second strand. The compound according to item 33.

[0627] 40. The compound according to item 34, wherein the oligonucleoside comprises an RNA duplex containing a first and a second strand, the first strand is at least partially complementary to the RNA sequence of the target gene, the second strand is at least partially complementary to the first strand, each of the first and second strands has 5' and 3' termini, and the RNA duplex is attached at the 3' terminus of its second strand to an adjacent phosphate.

[0628] 41. Compounds of formula (X) and (XI):

[0629]

Chemical formula

[0630] [Wherein, r and s are independently integers selected from 1 to 16, Z is an oligonucleoside moiety] A method for preparing a compound according to any one of items 1 to 40, comprising the step of reacting, and, where appropriate, performing deprotection of the ligand and / or annealing of the second strand for the oligonucleoside.

[0631] 42. The compound of formula (X) is of formula (Xa):

[0632]

Chemical formula

[0633] And the compound of formula (XI) is of formula (XIa):

[0634]

Chemical formula

[0635] and the oligonucleoside comprises an RNA duplex comprising a first and a second strand, the first strand being at least partially complementary to the RNA sequence of the target gene, the second strand being at least partially complementary to the first strand, each of the first and second strands having 5' and 3' ends, and the RNA duplex being attached at the 5' end of its second strand to an adjacent phosphate, a method according to claim 41 for preparing a compound according to any one of claims 6, 8 to 14, 16 to 33, and 35 to 40.

[0636] 43. A compound of formula (X) is of formula (Xb):

[0637]

Chemical formula

[0638] and a compound of formula (XI) is of formula (XIa):

[0639]

Chemical formula

[0640] and the oligonucleoside comprises an RNA duplex comprising a first and a second strand, the first strand being at least partially complementary to the RNA sequence of the target gene, the second strand being at least partially complementary to the first strand, each of the first and second strands having 5' and 3' ends, and the RNA duplex being attached at the 3' end of its second strand to an adjacent phosphate, a method according to claim 41 for preparing a compound according to any one of claims 5, 7, 9 to 13, 15 to 32, and 34 to 40.

[0641] 44. A compound of formula (XIa) is of formula (XIb):

[0642]

Chemical formula

[0643] The method according to claim 42 or 43.

[0644] 45. A compound of formula (X):

[0645] [Chem.]

[0646] [wherein, r is independently an integer selected from 1 to 16, Z is an oligonucleoside moiety].

[0647] 46. A compound of formula (Xa):

[0648] [Chem.]

[0649] 47. A compound of formula (Xb):

[0650] [Chem.]

[0651] 48. A compound of formula (XI):

[0652] [Chem.]

[0653] [wherein, s is independently an integer selected from 1 to 16, Z is an oligonucleoside moiety].

[0654] 49. A compound of formula (XIa):

[0655] [Chem.]

[0656] 50. Compound of formula (XIb):

[0657]

Chem.

[0658] 51. Use of a compound according to any one of claims 45 and 48 to 50 for preparing a compound according to any one of claims 1 to 40.

[0659] 52. Use of a compound according to claim 46 for preparing a compound according to any one of claims 6, 8 to 14, 16 to 33, and 35 to 40.

[0660] 53. Use of a compound according to claim 47 for preparing a compound according to any one of claims 5, 7, 9 to 13, 15 to 32, and 34 to 40.

[0661] 54. A compound or composition obtainable or obtained by a method according to any one of claims 41 to 44.

[0662] 55. A pharmaceutical composition comprising a compound according to any one of claims 1 to 40 together with a pharmaceutically acceptable carrier, diluent or excipient.

[0663] 56. A compound according to any one of claims 1 to 40 for use in therapy.

Example

[0664] The present invention will be more fully understood by reference to the following examples. However, they should not be construed as limiting the scope of the present invention. The examples and embodiments described herein are for illustrative purposes only, and various modifications or changes contemplated in light thereof are proposed to those skilled in the art and should be included within the spirit and scope of the present application and the scope of the appended claims.

[0665] Example 1: Synthesis of Tether 1 General experimental conditions: Thin layer chromatography (TLC) was performed on aluminum plates coated with silica using a fluorescent indicator 254 nm from Macherey-Nagel. Compounds were visualized under UV light (254 nm) or after spraying with 5% H2SO4 in methanol (MeOH) or ninhydrin reagent (from Sigma-Aldrich) by Stahl followed by heating. Flash chromatography was performed using Biotage Sfar Silica 10, 25, 50 or 100 g columns (Uppsala, Sweden) on a Biotage Isolera One flash chromatography instrument equipped with a dual variable UV wavelength detector (200 - 400 nm).

[0666] All moisture-sensitive reactions were carried out under anhydrous conditions using dried glassware, anhydrous solvents, and an argon atmosphere. All commercially available reagents were purchased from Sigma-Aldrich and solvents from Carl Roth GmbH + Co. KG. D-Galactosamine pentaacetate was purchased from AK scientific.

[0667] HPLC / ESI-MS was performed at 60 °C using an Acquity UPLC Protein BEH C4 column (300 Å, 1.7 μm, 2.1×100 mm) from Waters on a Dionex UltiMate 3000 RS UHPLC system and a Thermo Scientific MSQ Plus mass spectrometer. The solvent system consisted of solvent A with H2O containing 0.1% formic acid and solvent B with acetonitrile (ACN) containing 0.1% formic acid. A gradient from 5 - 100% B over 15 minutes at a flow rate of 0.4 mL / min was used. Detector and conditions: Corona charged aerosol detection (from esa). Nebulizer temperature: 25 °C. N2 pressure: 35.1 psi. Filter: Corona.

[0668] 1 H and 13 C NMR spectra were recorded at room temperature on a Varian spectrometer at 500 MHz ( 1 H NMR) and 125 MHz ( 13 C NMR). Chemical shifts are given in ppm, referenced to the solvent residual peak (CDCl3 - 1 H NMR: δ at 7.26 ppm, and 13 C NMR δ at 77.2 ppm; DMSO-d6 - 1H NMR: δ at 2.50 ppm, and 13 C NMR δ at 39.5 ppm). Coupling constants are given in Hertz. Signal splitting patterns are described as singlet (s), doublet (d), triplet (t) or multiplet (m).

[0669] Synthetic route for the conjugate building block TriGalNAc_tether1:

[0670]

Chem.

[0671] Preparation of Compound 2: D-Galactosamine pentaacetate (3.00 g, 7.71 mmol, 1.0 equiv) was dissolved in anhydrous dichloromethane (DCM) (30 mL) under argon, and trimethylsilyl trifluoromethanesulfonate (TMSOTf, 4.28 g, 19.27 mmol, 2.5 equiv) was added. The reaction was stirred at room temperature for 3 h. The reaction mixture was diluted with DCM (50 mL) and washed with cold saturated aqueous NaHCO3 (100 mL) and water (100 mL). The organic layer was separated, dried over Na2SO4, and concentrated to give the title compound as a yellow oil, which was purified by flash chromatography (gradient elution: 0 - 10% MeOH in DCM, 10 CV). The product was obtained as a colorless oil (2.5 g, 98%, rf = 0.45 (2% MeOH in DCM)).

[0672]

Chem.

[0673] Preparation of Compound 4: Compound 2 (2.30 g, 6.98 mmol, 1.0 equiv) and azido-PEG3-OH (1.83 g, 10.5 mmol, 1.5 equiv) were dissolved in anhydrous DCM (40 mL) under argon, and molecular sieves 3 Å (5 g) were added to the solution. The mixture was stirred at room temperature for 1 hour. Then, TMSOTf (0.77 g, 3.49 mmol, 0.5 equiv) was added to the mixture, and the reaction was stirred overnight. The molecular sieves were filtered off, the filtrate was diluted with DCM (100 mL), and washed with cold saturated aqueous NaHCO3 (100 mL) and water (100 mL). The organic layer was separated, dried over Na2SO4, and the solvent was removed under reduced pressure. The crude material was purified by flash chromatography (gradient elution: 0 - 3% MeOH in DCM for 10 CV) to give the title product as a pale yellow oil (3.10 g, 88%, rf = 0.25 (2% MeOH in DCM)). MS: C 20 H 32 N4O 11 calculated for, 504.21. Found 505.4. 1 H NMR (500 MHz, CDCl3) δ 6.21 - 6.14 (m, 1H), 5.30 (dd, J = 3.4, 1.1 Hz, 1H), 5.04 (dd, J = 11.2, 3.4 Hz,1H), 4.76 (d, J = 8.6 Hz, 1H), 4.23 - 4.08 (m, 3H), 3.91 - 3.80 (m, 3H), 3.74 - 3.59 (m, 9H), 3.49 - 3.41 (m, 2H), 2.14 (s, 3H), 2.02 (s, 3H), 1.97 (d, J = 4.2 Hz, 6H). 13 C NMR (125 MHz, CDCl3) δ 170.6 (C), 170.5 (C), 170.4 (C), 170.3 (C), 102.1 (CH), 71.6 (CH), 70.8 (CH), 70.6 (CH), 70.5 (CH), 70.3 (CH2), 69.7 (CH2), 68.5 (CH2), 66.6 (CH2), 61.5 (CH2), 23.1 (CH3), 20.7 (3xCH3).

[0674]

Chem.

[0675] Preparation of Compound 5: Compound 4 (1.00 g, 1.98 mmol, 1.0 eq) was dissolved in a mixture of ethyl acetate (EtOAc) and MeOH (30 mL 1:1 v / v), and Pd / C (100 mg) was added. The reaction mixture was degassed using a vacuum / argon cycle (3×) and hydrogenated overnight under balloon pressure. The reaction mixture was filtered through celite and washed with EtOAc (30 mL). The solvent was removed under reduced pressure to give the title compound as a colorless oil (0.95 g, quantitative yield, rf = 0.25 (10% MeOH in DCM)). The compound was used without further purification. MS: C 20 H 34 N2O 11 Calculated for, 478.2. Found 479.4.

[0676]

Chem.

[0677] Preparation of Compound 7: Tris{[2-(tert-butoxycarbonyl)ethoxy]methyl}-methylamine 6 (3.37 g, 6.67 mmol, 1.0 eq) was dissolved in a mixture of DCM / water (40 mL 1:1 v / v), and Na2CO3 (0.18 g, 1.7 mmol, 0.25 eq) was added with vigorous stirring. Benzyl chloroformate (2.94 mL, 20.7 mmol, 3.10 eq) was added dropwise to the previous mixture, and the reaction was stirred at room temperature for 24 h. The reaction mixture was diluted with CH2Cl2 (100 mL) and washed with water (100 mL). The organic layer was separated and dried over Na2SO4. The solvent was removed under reduced pressure, and the resulting crude material was purified by flash chromatography (gradient elution: 0 - 10% EtOAc in cyclohexane with 12 CV) to give the title compound as a pale yellowish oil (3.9 g, 91%, rf = 0.56 (10% EtOAc in cyclohexane)). MS: C 33 H53 NO 11 Calculated value: 639.3, Measured value: 640.9 1 H NMR (500 MHz, DMSO-d6) δ 7.38 - 7.26 (m, 5H), 4.97 (s, 2H), 3.54 (t, 6H), 3.50 (s, 6H), 2.38 (t, 6H), 1.39 (s, 27H). 13 C NMR (125 MHz, DMSO-d6) δ 170.3 (3xC), 154.5 (C), 137.1 (C), 128.2 (2xCH), 127.7 (CH), 127.6 (2xCH), 79.7 (3xC), 68.4 (3xCH2), 66.8 (3xCH2), 64.9 (C), 58.7 (CH2), 35.8 (3xCH2), 27.7 (9xCH3).

[0678]

Chem.

[0679] Preparation of Compound 8: Cbz-NH-tris-Boc-ester 7 (0.20 g, 0.39 mmol, 1.0 equiv) was dissolved in CH2Cl2 (1 mL) under argon, trifluoroacetic acid (TFA, 1 mL) was added, and the reaction mixture was stirred at room temperature for 1 hour. The solvent was removed under reduced pressure, and the residue was co-evaporated with toluene (5 mL) three times and dried under high vacuum to obtain the compound as its TFA salt (0.183 g, 98%). The compound was used without further purification. MS: C 21 H 29 NO 11 Calculated value: 471.6, Measured value: 472.4

[0680]

Chem.

[0681] Preparation of Compound 9: CbzNH-Tris-COOH 8 (0.72 g, 1.49 mmol, 1.0 equiv) and GalNAc-PEG3-NH2 5 (3.56 g, 7.44 mmol, 5.0 equiv) were dissolved in N,N-dimethylformamide (DMF) (25 mL). Then, N,N,N',N'-tetramethyl-O-(1H-benzotriazol-1-yl)uronium hexafluorophosphate (HBTU) (2.78 g, 7.44 mmol, 5.0 equiv), 1-hydroxybenzotriazole hydrate (HOBt) (1.05 g, 7.44 mmol, 5.0 equiv) and N,N-diisopropylethylamine (DIPEA) (2.07 mL, 11.9 mmol, 8.0 equiv) were added to the solution and the reaction was stirred for 72 h. The solvent was removed under reduced pressure and the residue was dissolved in DCM (100 mL) and washed with saturated aqueous NaHCO3 solution (100 mL). The organic layer was dried over Na2SO4 and the solvent was evaporated. The crude material was purified by flash chromatography (gradient elution: 0 - 5% MeOH in DCM over 14 CV). The product was obtained as a pale yellowish oil (1.2 g, 43%, rf = 0.20 (5% MeOH in DCM)). MS: C 81 H 125 N7O 41 calculated value, 1852.9. Found 1854.7. 1 H NMR (500 MHz, DMSO-d6) δ 7.90 - 7.80 (m, 10H), 7.65 - 7.62 (m, 4H), 7.47 - 7.43 (m, 3H), 7.38 - 7.32 (m, 8H), 5.24 - 5.22 (m, 3H), 5.02 - 4.97 (m, 4H), 4.60 - 4.57 (m, 3 H), 4.07 - 3.90 (m 10H), 3.67 - 3.36 (m, 70H), 3.23 - 3.07 (m, 25H), 2.18 (s, 10H), 2.00 (s, 13H), 1.89 (s, 11H), 1.80 - 1.78 (m, 17H). 1313C NMR (125 MHz, DMSO-d6) δ 170.1 (C), 169.8 (C), 169.7 (C), 169.4 (C), 169.2 (C), 169.1 (C), 142.7 (C), 126.3 (CH), 123.9 (CH), 118.7 (CH), 109.7 (CH), 100.8 (CH), 70.5 (CH), 69.8 (CH), 69.6 (CH), 69.5 (CH), 69.3 (CH2), 69.0 (CH2), 68.2 (CH2), 67.2 (CH2), 66.7 (CH2), 61.4 (CH2), 22.6 (CH2), 22.4 (3xCH3), 20.7 (9xCH3).

[0682]

Chem.

[0683] Preparation of Compound 10: The branched GalNAc compound 9 (0.27 g, 0.14 mmol, 1.0 equiv) was dissolved in MeOH (15 mL), and 3 drops of acetic acid (AcOH) and Pd / C (30 mg) were added. The reaction mixture was degassed using a vacuum / argon cycle (3×) and hydrogenated overnight under balloon pressure. Mass spectrometry was performed following the completion of the reaction, and the resulting mixture was filtered through a thin pad of celite. The solvent was evaporated, and the obtained residue was dried under high vacuum and used in the next step without further purification. The product was obtained as a pale yellowish oil (0.24 g, quantitative yield). MS: C 73 H 119 N7O 39 Calculated value, 1718.8. Measured value 1719.3.

[0684]

Chem.

[0685] Preparation of Compound 11: Commercially available bis(N-hydroxysuccinimide ester) suberate (3.67 g, 9.9 mmol, 1.0 equiv) was dissolved in DMF (5 mL), and triethylamine (1.2 mL) was added. To this solution, a solution of 3-azido-1-propylamine (1.0 g, 9.9 mmol, 1.0 equiv) in DMF (5 mL) was added dropwise. The reaction mixture was stirred at room temperature for 3 h. The reaction mixture was diluted with EtOAc (100 mL) and washed with water (50 mL). The organic layer was separated, dried over Na2SO4, and the solvent was removed under reduced pressure. The crude material was purified by flash chromatography (gradient elution: 0 - 5% MeOH in DCM for 16 CV). The product was obtained as a white solid (1.54 g, 43%, rf = 0.71 (5% MeOH in DCM)). MS: C 15 H 23 Calculated for C

[0686]

Chemical Structure

[0687] Preparation of TriGalNAc(12): The branched GalNAc compound 10 (0.35 g, 0.24 mmol, 1.0 equiv) and Compound 11 (0.11 g, 0.31 mmol, 1.5 equiv) were dissolved in DCM (5 mL) under argon, and triethylamine (0.1 mL, 0.61 mmol, 3.0 equiv) was added. The reaction mixture was stirred at room temperature overnight. The solvent was removed under reduced pressure, and the residue was dissolved in EtOAc (100 mL) and washed with water (100 mL). The organic layer was separated and dried over Na2SO4. The solvent was evaporated, and the resulting crude material was purified by flash chromatography (elution gradient: 0 - 10% MeOH in DCM for 20 CV) to give the title compound as a white fluffy solid (0.27 g, 67%, rf = 0.5 (10% MeOH in DCM)). MS: C 84 H 137 N 11 O 41 Calculated for

[0688] Conjugation of Tether 1 to siRNA strands: Monofluorocyclooctyne (MFCO) conjugation at the 5' or 3' end 5'-end MFCO conjugation

[0689]

Chem.

[0690] 3'-end MFCO conjugation

[0691]

Chem.

[0692] General conditions for MFCO conjugation: The amine-modified single strand was dissolved at 700 OD / mL in 50 mM carbonate / bicarbonate buffer pH 9.6 / dimethyl sulfoxide (DMSO) 4:6 (v / v). To this solution was added a 35 mM solution of 1 molar equivalent of MFCO-C6-NHS ester (Berry & Associates, catalog number LK4300) in DMF. The reaction was carried out at room temperature and after 1 hour, an additional 1 molar equivalent of MFCO solution was added. The reaction was allowed to proceed for a further 1 hour and monitored by LC / MS. At least 2 molar equivalents of excess MFCO NHS ester reagent relative to the amino-modified oligonucleotide were necessary to achieve quantitative consumption of the starting material. The reaction mixture was diluted 15-fold with water, filtered through a 1.2 μm filter from Sartorius, and then purified by reverse phase (RP HPLC) on an Akta Pure instrument (GE Healthcare).

[0693] Purification was carried out using a Waters XBridge C18 Prep 19×50 mm column. Buffer A was 100 mM TEAAc pH 7 and buffer B contained 95% acetonitrile in buffer A. A flow rate of 10 mL / min and a temperature of 60 °C were used. A UV trace at 280 nm was recorded. A gradient of 0 - 100% B was used within 60 column volumes.

[0694] Fractions containing full-length conjugated oligonucleotides were pooled and precipitated in the freezer using 3 M NaOAc, pH 5.2 and 85% ethanol. The collected pellet was dissolved in water. The sample was desalted by size exclusion chromatography and concentrated using a speed-vac concentrator to obtain the conjugated oligonucleotides in 40 - 80% isolated yield.

[0695] 5'-GalNAc-T1 conjugate

[0696]

Chem.

[0697] 3'-GalNAc-T1 conjugate

[0698]

Chem.

[0699] General procedure for TriGalNAc conjugation: The MFCO-modified single strand was dissolved in water at 2000 OD / mL, and a 1 equivalent solution of compound 12 (10 mM) in DMF was added to this solution. The reaction was carried out at room temperature, and after 3 hours, a 0.7 molar equivalent solution of compound 12 was added. The reaction was allowed to proceed overnight and completion was monitored by LCMS. The conjugate was diluted 15-fold with water, filtered through a 1.2 μm filter from Sartorius, and then purified by RP HPLC on an Akta Pure instrument (GE Healthcare).

[0700] RP-HPLC purification was performed using an XBridge C18 Prep 19×50 mm column from Waters. Buffer A was 100 mM triethylammonium acetate pH 7, and buffer B contained 95% acetonitrile in buffer A. A flow rate of 10 mL / min and a temperature of 60 °C were used. A UV trace at 280 nm was recorded. A gradient of 0–100% B was used within 60 column volumes.

[0701] Fractions containing the full-length conjugated oligonucleotide were pooled and precipitated in the freezer using 3 M NaOAc, pH 5.2 and 85% ethanol, and the collected pellet was dissolved in water to give an oligonucleotide solution of approximately 1000 OD / mL. O-Acetate was removed by adding 20% aqueous ammonia. Quantitative removal of these protecting groups was confirmed by LC-MS.

[0702] The conjugate was desalted by size-exclusion chromatography using an Akta Pure (GE Healthcare) instrument with Sephadex G25 Fine resin (GE Healthcare) to give the conjugated oligonucleotide in an isolated yield of 50–70%.

[0703] The following scheme further shows the synthetic route: Scheme 1:

[0704]

Chemical Structure

[0705] Scheme 2:

[0706]

Chemical Structure

[0707] Scheme 3:

[0708]

Chemical Structure

[0709] Scheme 4:

[0710]

Chem.

[0711] Scheme 5:

[0712]

Chem.

[0713] Example 2: Double-stranded annealing To generate the desired siRNA double-strand, two complementary strands were annealed by combining equimolar aqueous solutions of both strands. The mixture was placed in a water bath at 70 °C for 5 minutes and then cooled to ambient temperature within 2 hours. The double-strand was lyophilized for 2 days and stored at -20 °C.

[0714] The double-strand was analyzed by analytical SEC HPLC on a Dionex Ultimate 3000 (Thermo Fisher Scientific) HPLC system with a Superdex™ 75 Increase 5 / 150 GL column 5 × 153–158 mm (Cytiva). The mobile phase consisted of 1× PBS containing 10% acetonitrile. A homogeneous concentration gradient was run for 10 minutes at a flow rate of 1.5 mL / min at room temperature. UV traces at 260 and 280 nm were recorded. Water (LC-MS grade) was purchased from Sigma-Aldrich and phosphate-buffered saline (PBS, 10×, pH 7.4) was purchased from GIBCO (Thermo Fisher Scientific).

[0715] Example 3: Synthesis of Tether 2 General experimental conditions: Thin-layer chromatography (TLC) was performed on aluminum plates coated with silica using a fluorescent indicator 254 nm from Macherey-Nagel. The compounds were visualized under UV light (254 nm) or after spraying with 5% H2SO4 in methanol (MeOH) or ninhydrin reagent from Stahl (from Sigma-Aldrich) followed by heating. Flash chromatography was performed using a Biotage Sfar Silica 10, 25, 50 or 100 g column (Uppsala, Sweden) on a Biotage Isolera One flash chromatography instrument equipped with a dual variable UV wavelength detector (200 - 400 nm).

[0716] All moisture-sensitive reactions were carried out under anhydrous conditions using dried glassware, anhydrous solvents, and an argon atmosphere. All commercially available reagents were purchased from Sigma-Aldrich and the solvents were purchased from Carl Roth GmbH + Co. KG. D-Galactosamine pentaacetate was purchased from AK scientific.

[0717] HPLC / ESI-MS was performed at 60 °C using an Acquity UPLC Protein BEH C4 column (300 Å, 1.7 μm, 2.1×100 mm) from Waters on a Dionex UltiMate 3000 RS UHPLC system and a Thermo Scientific MSQ Plus mass spectrometer. The solvent system consisted of solvent A with H2O containing 0.1% formic acid and solvent B with acetonitrile (ACN) containing 0.1% formic acid. A gradient from 5 - 100% B over 15 minutes at a flow rate of 0.4 mL / min was used. Detector and conditions: Corona charged aerosol detection (from esa). Nebulizer temperature: 25 °C. N2 pressure: 35.1 psi. Filter: Corona.

[0718] 1 H and 13 C NMR spectra were recorded at room temperature on a Varian spectrometer at 500 MHz ( 1 H NMR) and 125 MHz (13 Recorded by 13C NMR). Chemical shifts are given in ppm, referenced to the solvent residual peak (CDCl3 - 1 1H...

Claims

1. A first strand that is at least partially complementary to a portion of the RNA transcribed from the target gene, and a second strand that is at least partially complementary to the first strand. A nucleic acid for inhibiting the expression of a target gene, which includes a double-stranded region, The nucleosides of the second and first chains are as follows (5'-3'): Second strand (5'-3'): Me-Me-Me-Me-Me-Me-F-Me-FFFF-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me-F-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me or Second strand (5'-3'): Me-Me-Me-Me-Me-Me-Me-Me-FFF-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me-F-Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me Nucleic acids containing the 2' sugar modification pattern.

2. The nucleosides of the second and first chains are, (a) Below (5'-3'): Second strand (5'-3'): Me(s)Me(s)Me-Me-Me-Me-F-Me-FFFF-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me or Second strand (5'-3'): Me(s)Me(s)Me-Me-Me-Me-Me-Me-FFF-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me(s)F(s)Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me [In the formula, (s) represents a phosphorothioate nucleoside linkage.] 2' sugar and bond modification patterns, (b) Below (5'-3'): Second strand (5'-3'): ia-ia-Me-Me-Me-Me-Me-Me-F-Me-FFFF-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me-F-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me or Second strand (5'-3'): ia-ia-Me-Me-Me-Me-Me-Me-Me-Me-FFF-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me-F-Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me [In the formula, ia represents an inverted baseless nucleoside.] The 2' sugar and base-free modification patterns, or (c) Below (5'-3'): Second strand (5'-3'): ia-ia-Me(s)Me(s)Me-Me-Me-Me-F-Me-FFFF-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me(s)F(s)Me-F-Me-F-Me-Me-Me-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me or Second strand (5'-3'): ia-ia-Me(s)Me(s)Me-Me-Me-Me-Me-Me-FFF-Me-Me-Me-Me-Me-Me-Me-Me-Me-Me, First strand (5'-3'): Me(s)F(s)Me-Me-Me-F-Me-Me-F-Me-Me-Me-Me-F-Me-F-Me-Me-Me-Me-Me(s)Me(s)Me [In the formula, (s) is a phosphorothioate nucleoside linkage, [ia represents an inverted baseless nucleoside] 2' sugar, base-free and bond modification patterns The nucleic acid according to claim 1, comprising:

3. The nucleic acid according to claim 1, wherein the first chain comprises at least 17 consecutive nucleosides that differ by 0 or 1 nucleosides from any one of the first chain sequences listed in Table 2 or Table 3.

4. The nucleic acid according to claim 3, wherein the first chain comprises nucleosides 2 to 18 of any one of the sequences defined in claim 3.

5. The nucleic acid according to claim 1, wherein the second strand comprises a nucleoside sequence of at least 17 consecutive nucleosides, each differing by 0 or 1 nucleosides from any one of the second strand sequences listed in Table 2 or Table 4, and the second strand has a region of at least 85% complementarity with the first strand across the 17 consecutive nucleosides.

6. The nucleic acid according to claim 1, wherein the nucleic acid is an siRNA oligonucleoside.

7. The nucleic acid according to claim 1, wherein the nucleic acid is directly or indirectly conjugated to one or more ligand moieties.

8. (a) The ligand portion is located in the terminal region of the second chain, typically in its 3' terminal region, or (b) The ligand portion is (i) one or more N-acetylgalactosamine (GalNAc) ligands, and / or (ii) One or more N-acetylgalactosamine (GalNAc) ligand derivatives, and / or (iii) One or more N-acetylgalactosamine (GalNAc) ligands and / or derivatives thereof conjugated to nucleic acids via a linker The nucleic acid according to claim 7, comprising:

9. The nucleic acid according to claim 8, wherein one or more GalNAc ligands and / or GalNAc ligand derivatives are conjugated, typically directly or indirectly, to the 5' or 3' terminal region of the second strand of the nucleic acid, typically at its 3' terminal region.

10. (a) The nucleic acid has the structure: 【Chemistry 1】 [In the formula, R 1 Each time it appears, it is independently selected from the group consisting of hydrogen, methyl, and ethyl. R 2 is hydrogen, hydroxyl, -OC 1～3 Alkyl, -C(=O)OC 1～3 Selected from the group consisting of alkyl, halo, and nitro, X 1 and X 2 Each time it appears, it is independently selected from the group consisting of methylene, oxygen, and sulfur. m is an integer between 1 and 6. n is an integer between 1 and 10. q, r, s, t, and v are independent integers between 0 and 4, however, (i) q and r cannot both be 0 at the same time, and (ii) s, t, and v cannot all be 0 at the same time. Z is an oligonucleoside. Having, or (b) nucleic acids, structure: 【Chemistry 2】 [In the formula, r and s are independent integers selected from 1 to 16. Z is an oligonucleoside. The nucleic acid according to claim 7, having the properties of the nucleic acid.

11. A pharmaceutical composition comprising a nucleic acid according to any one of claims 1 to 10 in combination with a pharmaceutically acceptable excipient or carrier.

12. A pharmaceutical composition according to claim 11 for use in treatment.

13. The pharmaceutical composition according to claim 11, for use in the prevention or treatment of diseases related to impaired hemostasis, such as hemophilia.

14. The pharmaceutical composition according to claim 11, for use in the prevention or treatment of cardiovascular disease.

15. A pharmaceutical composition according to claim 11 for use in the prevention or treatment of diabetes.

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