COMPOSITIONS AND METHODS FOR MODULATION OF TARGET NUCLEIC ACIDS

• US 20160122761A1
• Filed: 06/23/2014
• Published: 05/05/2016
• Est. Priority Date: 06/21/2013
• Status: Abandoned Application

• Current Assignee: Ionis Pharmaceuticals, Inc.
• Original Assignee: ISIS Pharmaceuticals Incorporated
• Inventors: Murray, Heather, Prakash, Thazha P., Lima, Walter F., Kinberger, Garth A., Swayze, Eric E., Crooke, Stanley T.
• Application Number:
  US14/898,873
• Publication Number:
  US 20160122761A1
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: A61K 31/7125 Nucleic acids or oligonucleotides having modified internucleoside linkage, i.e. other than 3'-5' phosphodiesters
• CPC Class Codes: A61K 47/549 Sugars, nucleosides, nucleotides or nucleic acids
• CPC Class Codes: A61P 3/06 Antihyperlipidemics
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C12N 15/113 Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing when used in plants C12N15/8218
• CPC Class Codes: C12N 2310/11 Antisense
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: C12N 2310/312 Phosphonates
• CPC Class Codes: C12N 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/322 2'-R Modification
• CPC Class Codes: C12N 2310/3341 5-Methylcytosine
• CPC Class Codes: C12N 2310/346 having a combination of backbone and sugar modifications
• CPC Class Codes: C12N 2310/351 Conjugate
• CPC Class Codes: C12N 2310/3515 Lipophilic moiety, e.g. cholesterol
• CPC Class Codes: C12N 2310/3525 MOE, methoxyethoxy
• CPC Class Codes: C12N 2310/3533 Halogen
• CPC Class Codes: C12N 2320/30 Special therapeutic applications
• CPC Class Codes: C12N 2320/32 Special delivery means, e.g. tissue-specific
• CPC Class Codes: C12N 2320/51 modulating the chemical stability, e.g. nuclease-resistance
• CPC Class Codes: C12N 2320/53 : reducing unwanted side-effects

Abstract

The present disclosure pertains generally to chemically-modified oligonucleotides for use in research, diagnostics, and/or therapeutics. In certain embodiments, the present disclosure describes compounds and methods for the modulation of a target nucleic acid. In certain embodiments, the present disclosure describes compounds and methods for the modulation of Apoliprotein C-III expression.

Claims

1. A compound comprising a single-stranded oligonucleotide consisting of 13 to 30 linked nucleosides and having a nucleobase sequence comprising at least 8 contiguous nucleobases complementary to an equal-length portion within a target region of a target nucleic acid, wherein the 5′

-terminal nucleoside of the single-stranded oligonucleotide comprises a stabilized phosphate moiety and an internucleoside linking group linking the 5′

-terminal nucleoside to the remainder of the oligonucleotide.

2. The compound of claim 1, wherein the target nucleic acid is Apolipoprotein C-III transcript.

3. The compound of claim 1 or 2, wherein the compound comprises a conjugate group.

4. The compound of claim 3, wherein the conjugate group is covalently attached to the oligonucleotide.

5. The compound of any of claims 1 to 4, wherein the conjugate group is attached to the oligonucleotide at a nucleoside at position 1, 2, 3, 4, 6, 7, 8, 9, 18, 19, 20, or 21 from the 5′

-end of the oligonucleotide or at position 1, 2, 3, 12, 13, 14, 15, 17, 18, 19, 20, or 21 from the 3′

-end of the oligonucleotide.

6. The compound of any of claims 1 to 5, wherein the conjugate group is attached to the oligonucleotide at a nucleoside at position 1 from the 5′

-end of the oligonucleotide.

7. The compound of any of claims 1 to 6, wherein the conjugate group is attached to the oligonucleotide at a nucleoside at position 8 from the 5′

-end of the oligonucleotide.

8. The compound of any of claims 1 to 7, wherein the Apolipoprotein C-III transcript comprises the nucleobase sequence as set forth in SEQ ID NO:

1.

9. The compound of any of claims 1 to 8, wherein the Apolipoprotein C-III transcript comprises the nucleobase sequence as set forth in SEQ ID NO:

2.

10. The compound of any of claims 1 to 9, wherein the complementary region comprises at least 10 contiguous nucleobases complementary to an equal-length portion within a target region of an Apolipoprotein C-III transcript.

11. The compound of any of claims 1 to 9, wherein the complementary region comprises at least 12 contiguous nucleobases complementary to an equal-length portion within a target region of an Apolipoprotein C-III transcript.

12. The compound of any of claims 1 to 9, wherein the complementary region comprises at least 14 contiguous nucleobases complementary to an equal-length portion within a target region of an Apolipoprotein C-III transcript.

13. The compound of any of claims 1 to 9, wherein the complementary region comprises at least 16 contiguous nucleobases complementary to an equal-length portion within a target region of an Apolipoprotein C-III transcript.

14. The compound of any of claims 1 to 9, wherein the complementary region comprises at least 18 contiguous nucleobases complementary to an equal-length portion within a target region of an Apolipoprotein C-III transcript.

15. The compound of any of claims 1 to 14, wherein the 5′

-terminal nucleoside of the single-stranded oligonucleotide has Formula I;

16. The compound of claim 15, wherein M₃ is selected from among:

O, CH═

CH, OCH₂, and OC(H)(Bx₂).

17. The compound of claim 15, wherein M₃ is O.

18. The compound of any of claims 15-17, wherein each of J₄, J₅, J₆ and J₇ is H.

19. The compound of any of claims 15-18, wherein J₄ forms a bridge with either J₅ or J₇.

20. The compound of any of claims 15-19, wherein A has the formula:

21. The compound of claim 20, wherein each of Q₁ and Q₂ is H.

22. The compound of claim 20, wherein Q₁ and Q₂ are each independently selected from among:

H and a halogen.

23. The compound of claim 20, wherein one of Q₁ and Q₂ is H and the other of Q₁ and Q₂ is F, CH₃ or OCH₃.

24. The compound of any of claims 15 to 23, wherein T₁ has the formula:

25. The compound of claim 24, wherein R_b is O and R_a and R_c are each, independently selected from among:

OCH₃, OCH₂CH₃, OCH(CH₃)₂.

26. The compound of any of claims 15 to 25, wherein G is selected from among:

a halogen, OCH₃, OCH₂F, OCHF₂, OCF₃, OCH₂CH₃, O(CH₂)₂F, OCH₂CHF₂, OCH₂CF₃, OCH₂—

CH═

CH₂, O(CH₂)₂—

OCH₃, O(CH₂)₂—

SCH₃, O(CH₂)₂—

OCF₃, O(CH₂)₃—

N(R₁₀)(R₁₁), O(CH₂)₂—

ON(R₁₀)(R₁₁), O(CH₂)₂—

O(CH₂)₂—

N(R₁₀)(R₁₁), OCH₂C(═

O)—

N(R₁₀)(R₁₁), OCH₂C(═

O)—

N(R₁₂)—

(CH₂)₂—

N(R₁₀)(R₁₁), and O(CH₂)₂—

N(R₁₂)—

C(═

NR₁₃)[N(R₁₀)(R₁₁)];

wherein R₁₀, R₁₁, R₁₂ and R₁₃ are each, independently, H or C₁-C₆ alkyl.

27. The compound of any of claims 15-26, wherein G is selected from among:

a halogen, OCH₃, OCF₃, OCH₂CH₃, OCH₂CF₃, OCH₂—

CH═

CH₂, O(CH₂)₂—

OCH₃, O(CH₂)₂—

O(CH₂)₂—

N(CH₃)₂, OCH₂C(═

O)—

N(H)CH₃, OCH₂C(═

O)—

N(H)—

(CH₂)₂—

N(CH₃)₂, and OCH₂—

N(H)—

C(═

NH)NH₂.

28. The compound of any of claims 15-27, wherein G is selected from among:

F, OCH₃, and O(CH₂)₂—

OCH₃.

29. The compound of claim 28, wherein G is O(CH₂)₂—

OCH₃.

30. The compound of any of claims 15-25, wherein G is a conjugate group.

31. The compound of claim 30, wherein the conjugate of the conjugate group is selected from among:

cholesterol, palmityl, stearoyl, lithocholic-oleyl, C₂₂ alkyl, C₂₀ alkyl, C₁₆ alkyl, C₁₈ alkyl, and C₁₀ alkyl.

32. The compound of claim 31, wherein the conjugate group comprises C₁₆ alkyl.

33. The compound of any of claims 30 to 32, wherein the conjugate group comprises a linker.

34. The compound of claim 33, wherein the linker is selected from among:

hexanamide, 8-amino-3,6-dioxaoctanoic acid (ADO), succinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC), 6-aminohexanoic acid (AHEX or AHA), substituted C₁-C₁₀ alkyl, substituted or unsubstituted C₂-C₁₀ alkenyl, and substituted or unsubstituted C₂-C₁₀ alkynyl.

35. The compound of any of claims 15-34, wherein the nucleobase is a modified nucleobase.

36. The compound of any of claims 15-35, wherein the nucleobase is a pyrimidine, substituted pyrimidine, purine or substituted purine.

37. The compound of any of claims 15-36, wherein the nucleobase is uracil, thymine, cytosine, 5-methylcytosine, adenine or guanine.

38. The compound of any of claims 15-37, wherein the 5′

-terminal nucleoside of the single-stranded oligonucleotide has Formula III;

39. The compound of claim 38, wherein A has the formula:

40. The compound of claim 39, wherein Q₁ and Q₂ are each independently selected from among:

H, F, CH₃, and OCH₃.

41. The compound of any of claims 15-40, wherein the 5′

-terminal nucleoside has Formula V;

42. The compound of any of claims 1-41, wherein the remainder of the oligonucleotide comprises at least one RNA-like nucleoside.

43. The compound of claim 42, wherein essentially each nucleoside of the remainder of the oligonucleotide is an RNA-like nucleoside.

44. The compound of claim 43, wherein each nucleoside of the remainder of the oligonucleotide is an RNA-like nucleoside.

45. The compound of any of claims 42-44, wherein each RNA-like nucleoside is independently selected from among:

a 2′

-endo furanosyl nucleoside and an RNA-surrogate nucleoside.

46. The compound of claim 45, wherein each RNA-like nucleoside is a 2′

-endo furanosyl nucleoside.

47. The compound of claim 46, wherein each RNA-like nucleoside is selected from among:

2′

-F, 2′

-MOE, 2′

-OMe, LNA, F-HNA, and cEt.

48. The compound of any of claims 1-47, wherein the remainder of the oligonucleotide comprises at least one region having sugar motif:

-[(A)_x-(B)_y-(A)_z]_q-

wherein

A is a modified nucleoside of a first type,

B is a modified nucleoside of a second type;

each x and each y is independently 1 or 2;

z is 0 or 1;

q is 1-15.

49. The compound of any of claims 1-47, wherein the remainder of the oligonucleotide comprises at least one region having sugar motif:

-[(A)_x-(B)_y-(A)_z]_q-

wherein

A is a modified nucleoside of a first type,

B is a modified nucleoside of a second type;

each x and each y is independently 1 or 2;

z is 0 or 1;

q is 1-15.

50. The compound of any of claims 1-49, wherein the remainder of the oligonucleotide comprises at least one region having sugar motif:

-[(A)_x-(A)_y-(A)_z]_q-

wherein

A is a modified nucleoside;

each x and each y is independently 1 or 2;

z is 0 or 1;

q is 1-15.

51. The compound of any of claims 1-49, wherein the remainder of the oligonucleotide comprises at least one region having sugar motif:

-[(B)_x-(B)_y-(B)_z]_q-

wherein

B is a modified nucleoside,

each x and each y is independently 1 or 2;

z is 0 or 1;

q is 1-15.

52. The compound of claim 50 or 51, wherein A is a modified nucleoside selected from among:

2′

-F, 2′

-OMe, and F-HNA.

53. The compound of claim 50 or 51, wherein B is a modified nucleoside selected from among:

2′

-F, 2′

-OMe, and F-HNA.

54. The compound of any of claims 48 to 53, wherein the modifications of the first type and the modifications of the second type are selected from among:

2′

-F, 2′

-OMe, and F-HNA.

55. The compound of any of claims 48 to 53, wherein the modifications of the first type are 2′

-F and the modifications of the second type are 2′

-OMe.

56. The compound of any of claims 48 to 53, wherein the modifications of the first type are 2′

-OMe and the modifications of the second type are 2′

-F.

57. The compound of any of claims 48 to 56, wherein each x and each y is 1.

58. The compound of any of claims 1-57, wherein the remainder of the oligonucleotide comprises 1-4 3′

terminal nucleosides, each comprising the same sugar modification, wherein the sugar modification of the 1-4 3′

terminal nucleosides is different from the sugar modification of the immediately adjacent nucleoside.

59. The compound of claim 58, wherein the 3′

-terminal nucleosides are each 2′

-MOE nucleosides.

60. The compound of claim 58 or 59 comprising two 3′

-terminal nucleosides.

61. The compound of any of claims 1-60, comprising at least one modified internucleoside linkage.

62. The compound of claim 61, wherein each internucleoside linkage is selected from phosphorothioate and phosphodiester.

63. The compound of claim 61 or 62, wherein each of the 6-10 3′

-most internucleoside linkages is phosphorothioate linkage.

64. The compound of any of claims 61 to 63, wherein the 5′

-most internucleoside linkage is a phosphorothioate linkage.

65. The compound of any of claims 61 to 64, comprising a region of alternating linkages.

66. The compound of any of claims 1-65, comprising a 5′

region having the motif;

-s-(A-s-B-o-A)_x(-s-B)_Y 

 

(Nucleoside of Formula I, III, or V)

wherein;

A is a nucleoside of a first type;

B is a nucleoside of a second type;

s is a phosphorothioate linkage;

o is a phosphodiester linkage;

X is 1-8; and

Y is 1 or 0.

67. The compound of any of claims 1-66, comprising a 3′

region having the motif;

-(A-s-B-s-A)_z(-s-B)_q-s-(D)-(s-D)_r

wherein;

s is a phosphorothioate linkage;

A is a nucleoside of a first type;

B is a nucleoside of a second type;

D is a nucleoside of a third type;

Z is 1-5;

q is 1 or 0; and

and r is 0-3.

68. The compound claim 66 or 67, wherein A is a 2′

-F nucleoside.

69. The compound of any of claims 66 to 68, wherein B is a 2′

-OMe nucleoside.

70. The compound of any of claims 67 to 69, wherein D is a 2′

-MOE nucleoside.

71. The compound of any of claims 67 to 70, wherein the oligonucleotide comprises a hybridizing region and a 3′

-terminal region, wherein the hybridizing region comprises nucleosides A and B and the terminal region comprising nucleosides D, wherein the hybridizing region is complementary to a target region of an Apoliprotein CIII transcript.

72. The compound of any of claims 1-65, comprising the motif:

-s-A-s-B-o-A-s-B-o-A-s-B-o-A-s-B-o-A-s-B-o-A-s-B-o-A-s-B-s-A-s-B-s-A-s-B-s-D-s-D-s 

 

(Nucleoside of Formula V)

wherein;

s is a phosphorothioate linkage;

A is a nucleoside of a first type;

B is a nucleoside of a second type; and

D is a nucleoside of a third type.

73. The compound of any of claims 1-65, consisting of the motif:

-s-A-s-B-o-A-s-B-o-A-s-B-o-A-s-B-o-A-s-B-o-A-s-B-o-A-s-B-s-A-s-B-s-A-s-B-s-D-s-D-s 

 

(Nucleoside of Formula V)

wherein;

s is a phosphorothioate linkage;

A is a nucleoside of a first type;

B is a nucleoside of a second type; and

D is a nucleoside of a third type.

74. The compound of any of claims 1-65, comprising the motif:

-s-A-s-B-o-A-s-B-o-A-s-B-o-A-s-B-o-A-s-B-o-A-s-B-o-A-s-B-s-A-s-B-s-A-s-B-s-D-s-D-s-X 

 

(Nucleoside of Formula V)

wherein;

s is a phosphorothioate linkage;

A is a nucleoside of a first type;

B is a nucleoside of a second type;

D is a nucleoside of a third type; and

X is a conjugate group.

75. The compound of any of claims 1-65, consisting of the motif:

-s-A-s-B-o-A-s-B-o-A-s-B-o-A-s-B-o-A-s-B-o-A-s-B-o-A-s-B-s-A-s-B-s-A-s-B-s-D-s-D-s-X 

 

(Nucleoside of Formula V)

wherein;

s is a phosphorothioate linkage;

A is a nucleoside of a first type;

B is a nucleoside of a second type;

D is a nucleoside of a third type; and

X is a conjugate group.

76. The compound of any of claims 72 to 75, wherein A is a 2′

-F nucleoside.

77. The compound of any of claims 72 to 76, wherein B is a 2′

-OMe nucleoside.

78. The compound of any of claims 72 to 77, wherein D is a 2′

-MOE nucleoside.

79. The compound of any of claims 1-75, wherein the oligonucleotide has two mismatches relative to a target region of the Apolipoprotein C-III transcript.

80. The compound of any of claims 1-79, wherein the oligonucleotide has three mismatches relative to a target region of the Apolipoprotein C-III transcript.

81. The compound of any of claims 1-79, wherein the oligonucleotide has four mismatches relative to a target region of the Apolipoprotein C-III transcript.

82. The compound of any of claims 1-81, wherein the oligonucleotide comprises a hybridizing region and 0-4 3′

-terminal nucleosides.

83. The compound of any of claims 1-81, wherein the oligonucleotide comprises a hybridizing region and 1-4 3′

-terminal nucleosides.

84. The compound of claim 82 or 83, wherein the hybridizing region is 100% complementary to a target region of the Apolipoprotein C-III transcript.

85. The compound of claim 82 or 83, wherein the hybridizing region has one mismatch relative to a target region of the Apolipoprotein C-III transcript.

86. The compound of claim 82 or 83, wherein the hybridizing region has two mismatches relative a target region of the Apolipoprotein C-III transcript.

87. The compound of claim 82 or 83, wherein the hybridizing region has three mismatches relative to a target region of the Apolipoprotein C-III transcript.

88. The compound of claim 82 or 83, wherein the hybridizing region has four mismatches relative to a target region of the Apolipoprotein C-III transcript.

89. The compound of any of claims 79 to 88, wherein one or more of the 3′

-terminal nucleosides is not complementary to the target RNA.

90. The compound of any of claims 79 to 89, wherein the nucleobase of each 3′

-terminal nucleoside is a purine.

91. The compound of claim 90, wherein the nucleobase of each 3′

-terminal nucleoside is an adenine.

92. The compound of any of claims 1-91, wherein the oligonucleotide comprises at least one modified nucleobase.

93. The compound of any of claims 1-92, wherein each cytosine residue comprises a 5-methylcytosine.

94. The compound of any of claims 1-92, wherein the nucleobase sequence of the oligonucleotide comprises a nucleobase sequence selected from among:

SEQ ID NO;

3, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, or 86.

95. The compound of any of claims 1-92, wherein the nucleobase sequence of the oligonucleotide comprises a nucleobase sequence selected from among:

SEQ ID NO;

87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, or 110.

96. The compound of any of claims 1-92, wherein the nucleobase sequence of the oligonucleotide comprises the nucleobase sequence of SEQ ID NO:

3.

97. The compound of any of claims 1-92, wherein the nucleobase sequence of the oligonucleotide comprises the nucleobase sequence of SEQ ID NO:

14.

98. The compound of any of claims 1-92, wherein the nucleobase sequence of the oligonucleotide comprises the nucleobase sequence of SEQ ID NO:

140.

99. The compound of any of claims 1-93, wherein the nucleobase sequence of the oligonucleotide consists of a nucleobase sequence selected from among:

SEQ ID NO;

3, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, or 86.

100. The compound of any of claims 1-93, wherein the nucleobase sequence of the oligonucleotide consists of a nucleobase sequence selected from among:

SEQ ID NO;

87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, or 110.

101. The compound of any of claims 1-93, wherein the nucleobase sequence of the oligonucleotide consists of the nucleobase sequence of SEQ ID NO:

3.

102. The compound of any of claims 1-93, wherein the nucleobase sequence of the oligonucleotide consists of the nucleobase sequence of SEQ ID NO:

14.

103. The compound of any of claims 1-93, wherein the nucleobase sequence of the oligonucleotide consists of the nucleobase sequence of SEQ ID NO:

140.

104. The compound of claim 1, wherein the compound comprises ISIS No. 594290.

105. The compound of claim 1, wherein the compound comprises ISIS No. 594231.

106. The compound of claim 1, wherein the compound comprises ISIS No. 722060.

107. The compound of claim 1, wherein the single-stranded oligonucleotide is ISIS No. 594290.

108. The compound of claim 1, wherein the single-stranded oligonucleotide is ISIS No. 594231.

109. The compound of claim 1, wherein the single-stranded oligonucleotide is ISIS No. 722060.

110. The compound of any of claims 1-109, wherein the oligonucleotide comprises at least one conjugate group.

115. The compound of any of claims 3 to 114, wherein the conjugate group comprises:

116. The compound of any of claims 3 to 114, wherein the conjugate group comprises:

117. The compound of any of claims 3 to 114, wherein the conjugate group comprises:

118. The compound of any of claims 3 to 114, wherein the conjugate group comprises a cell-targeting moiety having the following structure:

119. The compound of any of claims 3 to 114, wherein the conjugate group comprises a cell-targeting moiety having the following structure:

120. The compound of any of claims 3 to 114, wherein the conjugate group comprises a cell-targeting moiety having the following structure:

121. The compound of any of claims 3 to 114, wherein the conjugate group comprises a cell-targeting moiety having the following structure:

122. The compound of any of claims 3 to 114, wherein the conjugate group comprises a cell-targeting moiety having the following structure:

123. The compound of any of claims 3 to 114, wherein the conjugate group comprises a cell-targeting moiety having the following structure:

124. The compound of any of claims 3 to 114, wherein the conjugate group comprises a cell-targeting moiety having the following structure:

125. The compound of any of claims 3 to 114, wherein the conjugate group comprises a cell-targeting moiety having the following structure:

126. The compound of any of claims 3 to 114, wherein the conjugate group comprises a cell-targeting moiety having the following structure:

127. The compound of any of claims 3 to 114, wherein the conjugate group comprises a cell-targeting moiety having the following structure:

128. The compound of any of claims 3 to 114, wherein the conjugate group comprises a cell-targeting moiety having the following structure:

129. The compound of any of claims 3 to 114, wherein the conjugate group comprises the following structure:

130. The compound of any of claims 3 to 114, wherein the conjugate group comprises a cell-targeting moiety having the following structure:

131. The compound of any of claims 3 to 114, wherein the conjugate group comprises a cell-targeting moiety having the following structure:

132. The compound of any of claims 3 to 114, wherein the conjugate group comprises a cell-targeting moiety having the following structure:

133. The compound of any of claims 3 to 114, wherein the conjugate group comprises a cell-targeting moiety having the following structure:

134. The compound of any of claims 3 to 114, wherein the conjugate group comprises a cell-targeting moiety having the following structure:

135. The compound of any of claims 3 to 114, wherein the conjugate group comprises a cell-targeting moiety having the following structure:

136. The compound of any of claims 3 to 114, wherein the conjugate group comprises a cell-targeting moiety having the following structure:

137. The compound of any of claims 3 to 114, wherein the conjugate group comprises a cell-targeting moiety having the following structure:

138. The compound of any of claims 3 to 114, wherein the conjugate comprises the following structure:

139. The compound of any of claims 3 to 114, wherein the conjugate comprises the following structure:

140. The compound of any of claims 3 to 114, wherein the conjugate comprises the following structure:

141. The compound of any of claims 3 to 114, wherein the conjugate comprises the following structure:

142. The compound of any of claims 3 to 114, wherein the conjugate comprises the following structure:

143. The compound of any of claims 3 to 114, wherein the conjugate comprises the following structure:

144. The compound of any of claims 3 to 114, wherein the conjugate comprises the following structure:

145. The compound of any of claims 3 to 114, wherein the conjugate comprises the following structure:

146. The compound of any of claims 3 to 114, wherein the conjugate comprises the following structure:

147. The compound of any of claims 3 to 114, wherein the conjugate comprises the following structure:

148. The compound of any of claims 3 to 114, wherein the conjugate comprises the following structure:

149. The compound of any of claims 3 to 114, wherein the conjugate group has the following structure:

150. The compound of any of claims 3 to 114, wherein the conjugate group has the following structure:

151. The compound of any of claims 3 to 114, wherein the conjugate group has the following structure:

152. The compound of any of claims 3 to 114, wherein the conjugate group has the following structure:

153. The compound of any of claims 3 to 114, wherein the conjugate group has the following structure:

154. The compound of any of claims 3 to 114, wherein the conjugate group has the following structure:

155. The compound of any of claims 3 to 114, wherein the conjugate group has the following structure:

156. The compound of any of claims 3 to 114, wherein the conjugate group has the following structure:

157. The compound of any of claims 3 to 114, wherein the conjugate group has the following structure:

158. The compound of any of claims 3 to 114, wherein the conjugate group has the following structure:

159. The compound of any of claims 3 to 114, wherein the conjugate group has the following structure:

160. The compound of any of claims 3 to 114, wherein the conjugate group has the following structure:

161. The compound of any of claims 3 to 114, wherein the conjugate group has the following structure:

162. The compound of any of claims 3 to 114, wherein the conjugate group has the following structure:

163. The compound of any of claims 3 to 114, wherein the conjugate group has the following structure:

164. The compound of any of claims 3 to 114, wherein the conjugate group has the following structure:

165. The compound of any of claims 3 to 114, wherein the conjugate group has the following structure:

166. The compound of any of claims 3 to 114, wherein the conjugate group has the following structure:

167. The compound of any of claims 3 to 114, wherein the conjugate group has the following structure:

168. The compound of any of claims 3 to 114, wherein the conjugate group has the following structure:

169. The compound of any of claims 3 to 114, wherein the conjugate of the conjugate group is selected from among:

cholesterol, palmityl, stearoyl, lithocholic-oleyl, C₂₂ alkyl, C₂₀ alkyl, C₁₆ alkyl, C₁₈ alkyl, and C₁₀ alkyl.

170. The compound of claim 164, wherein the conjugate of the conjugate group is C₁₆ alkyl.

171. The compound of any of claims 3 to 114, wherein the conjugate comprises exactly one GalNAc ligand.

172. The compound of any of claims 3 to 114, wherein the conjugate comprises exactly two GalNAc ligands.

173. The compound of any of claims 3 to 114, wherein the conjugate comprises exactly three GalNAc ligands.

174. The compound of any of claims 3 to 148 or 169 to 173, wherein the conjugate group comprises a linker.

175. The compound of claim 174, wherein the linker is selected from among:

hexanamide, 8-amino-3,6-dioxaoctanoic acid (ADO), succinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC), 6-aminohexanoic acid (AHEX or AHA), substituted C₁-C₁₀ alkyl, substituted or unsubstituted C₂-C₁₀ alkenyl, and substituted or unsubstituted C₂-C₁₀ alkynyl.

176. The compound of claim 175, wherein the linker is hexanamide.

177. The compound of claim 175, wherein the linker does not comprise pyrollidine.

178. The compound of claim 175, wherein the linker comprises pyrollidine.

179. A pharmaceutical composition comprising at least one compound of any of claims 1-178 and a pharmaceutically acceptable carrier or diluent.

180. A method of reducing the activity or amount of a nucleic acid transcript in a cell, comprising contacting a cell with at least one compound of any of claims 1 to 179;

and thereby reducing the activity or amount of the nucleic acid transcript in the cell.

181. A method of reducing the activity or amount of an Apolipoprotein C-III transcript in a cell, comprising contacting a cell with at least one compound of any of claims 1 to 179;

and thereby reducing the activity or amount of the Apolipoprotein C-III transcript in the cell.

182. The method of claim 181, wherein the Apolipoprotein C-III transcript is Apolipoprotein C-III pre-mRNA.

183. The method of claim 181, wherein the Apolipoprotein C-III transcript is Apolipoprotein C-III mRNA.

184. The method of any of claims 181 to 183, wherein the cell is in vitro.

185. The method of any of claims 181 to 183, wherein the cell is in an animal.

186. The method of claim 185, wherein the animal is a human.

187. A method of reducing the activity or amount of an Apolipoprotein C-III protein in a cell, comprising contacting a cell with at least one compound of any of claims 1 to 179;

and thereby reducing the activity or amount of the Apolipoprotein C-III protein in the cell.

188. The method of claim 187, wherein the cell is in vitro.

189. The method of claim 187, wherein the cell is in an animal.

190. The method of claim 189, wherein the animal is a human.

191. A method of decreasing total cholesterol, comprising contacting a cell with at least one compound of any of claims 1 to 179;

and thereby decreasing total cholesterol.

192. The method of claim 191, wherein the cell is in vitro.

193. The method of claim 191, wherein the cell is in an animal.

194. The method of claim 193, wherein the animal is a human.

195. A method of decreasing triglycerides, comprising contacting a cell with at least one compound of any of claims 1 to 179;

and thereby decreasing triglycerides.

196. The method of claim 195, wherein the cell is in vitro.

197. The method of claim 195, wherein the cell is in an animal.

198. The method of claim 197, wherein the animal is a human.

199. A method of lowering LDL, comprising contacting a cell with at least one compound of any of claims 1 to 179;

and thereby lowering LDL.

200. The method of claim 199, wherein the cell is in vitro.

201. The method of claim 199, wherein the cell is in an animal.

202. The method of claim 201, wherein the animal is a human.

203. A method of increasing HDL, comprising contacting a cell with at least one compound of any of claims 1 to 179;

and thereby increasing HDL.

204. The method of claim 203, wherein the cell is in vitro.

205. The method of claim 203, wherein the cell is in an animal.

206. The method of claim 205, wherein the animal is a human.

207. Use of a compound of any of claims 1 to 178 or the pharmaceutical composition of claim 174 for the manufacture of a medicament for use in treatment of a disease.

208. A compound of any of claims 1 to 178, or the pharmaceutical composition of claim 174, for use in treatment of a disease.

209. A compound of any of claims 1 to 178, or the pharmaceutical composition of claim 174, for use in treatment of hypertriglyceridemia.

111. A double-stranded compound comprising a sense strand and an antisense strand, wherein the antisense strand comprises at least 12 contiguous nucleobases of a nucleobase sequence selected from SEQ ID NO:

3, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110; and

at least one conjugate group.

112. The double-stranded compound of claim 111, wherein the sense strand comprises at least 12 contiguous nucleobases of a nucleobase sequence selected from SEQ ID NO:

111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, or 133.

113. The double-stranded compound of claim 112, wherein the conjugate group is covalently attached to the 5′

-end of the sense strand.

114. The double-stranded compound of claim 113, wherein the conjugate group is covalently attached to the 3′

-end of the sense strand.