Formulations for oligonucleotide delivery

Abstract

The disclosure relates to compositions and methods for delivery of nucleic acid therapeutics, particularly iRNA agents to target locations and genes residing in the central nervous system (CNS) of a subject.

Description

HK Application no.: 62026122074.8 Our Ref: HKP / 2026 / 95367 cc * (Draft) * May 20, 2026 Invention Title: Formulation for Oligonucleotide Delivery Abstract This invention relates to compositions and methods for delivering nucleic acid therapeutic agents, particularly iRNA reagents, to target sites and genes located in the central nervous system (CNS) of a subject.

Claims

We claim:

1. A composition comprising (a) a double-stranded ribonucleic acid (dsRNA) comprising a sense strand and an antisense strand, wherein one of the sense strand or the antisense strand of the dsRNA comprises at least one lipophilic modification and the other strand of the dsRNA does not comprise a lipophilic modification; and (b) a divalent ion source, wherein: (i) substantially all of the sense strand or the antisense strand of the dsRNA comprising the at least one lipophilic modification in the composition is duplexed with the strand that does not comprise a lipophilic modification, or (ii) the sense strand or the antisense strand of the dsRNA comprising the at least one lipophilic modification is present at less than 1% molar excess relative to the strand that does not comprise a lipophilic modification, the sense strand and the antisense strand are present at molar equivalence, or the strand that does not comprise a lipophilic modification is present in a molar excess relative to the strand of the dsRNA comprising the at least one lipophilic modification.

2. A composition comprising (a) a double-stranded ribonucleic acid (dsRNA) comprising a sense strand and an antisense strand, wherein one of the sense strand or the antisense strand of the dsRNA comprises at least one lipophilic modification and the other strand of the dsRNA does not comprise a lipophilic modification; and (b) a divalent ion source, wherein: the sense strand or the antisense strand of the dsRNA that comprises at least one lipophilic modification is present at less than 1% molar excess relative to the strand of the dsRNA that does not comprise a lipophilic modification, the strand of the dsRNA that does not comprise a lipophilic modification is present at molar equivalence to the sense strand or the antisense strand of the dsRNA that comprises at least one lipophilic modification, or the strand of the dsRNA that does not comprise a lipophilicmodification is present in molar excess relative to the sense strand or the antisense strand of the dsRNA that comprises at least one lipophilic modification.

3. The composition of claim 1 or claim 2, wherein the composition is substantially free of inorganic phosphate.

4. The composition of any one of claims 1-3, wherein the sense strand of the dsRNA comprises at least one lipophilic modification and the antisense strand of the dsRNA does not comprise a lipophilic modification.

5. The composition of any one of claims 1-4, wherein there is a molar excess of the antisense strand relative to the sense strand.

6. The composition of claim 5, wherein there is at least a 0.1% molar excess of the antisense strand relative to the sense strand, optionally wherein there is at least a 0.2% molar excess of the antisense strand relative to the sense strand, optionally wherein there is at least a 0.3% molar excess of the antisense strand relative to the sense strand, optionally wherein there is at least a 0.4% molar excess of the antisense strand relative to the sense strand, optionally wherein there is at least a 0.5% molar excess of the antisense strand relative to the sense strand, optionally wherein there is at least a 1% molar excess of the antisense strand relative to the sense strand, optionally wherein there is at least a 2% molar excess of the antisense strand relative to the sense strand, optionally wherein there is at least a 3% molar excess of the antisense strand relative to the sense strand, optionally wherein there is at least a 4% molar excess of the antisense strand relative to the sense strand, optionally wherein there is about a 5% or greater molar excess of the antisense strand relative to the sense strand.

7. The composition of any one of claims 1-3, wherein the antisense strand of the dsRNA comprises the at least one lipophilic modification and the sense strand of the dsRNA does not comprise a lipophilic modification, optionally wherein the antisense strand of the dsRNA comprises the at least one lipophilic modification, optionally wherein there is a molar excess of the sense strand relative to the antisense strand.

8. The composition of claim 7, wherein there is at least a 0.1% molar excess of the sense strand relative to the antisense strand, optionally wherein there is at least a 0.2% molar excess ofthe sense strand relative to the antisense strand, optionally wherein there is at least a 0.3% molar excess of the sense strand relative to the antisense strand, optionally wherein there is at least a 0.4% molar excess of the sense strand relative to the antisense strand, optionally wherein there is at least a 0.5% molar excess of the sense strand relative to the antisense strand, optionally wherein there is at least a 1% molar excess of the sense strand relative to the antisense strand, optionally wherein there is at least a 2% molar excess of the sense strand relative to the antisense strand, optionally wherein there is at least a 3% molar excess of the sense strand relative to the antisense strand, optionally wherein there is at least a 4% molar excess of the sense strand relative to the antisense strand, optionally wherein there is about a 5% or greater molar excess of the sense strand relative to the antisense strand.

9. The composition of any one of the preceding claims, formulated for a route of administration selected from the group consisting of intravenous, subcutaneous, intramuscular, intradermal, intra-articular and intrathecal.

10. The composition of any one of the preceding claims, wherein the lipophilic modification is a saturated or unsaturated C4-C30hydrocarbon, optionally a C4-C30alkyl or alkenyl, optionally a linear C6-C18alkyl or alkenyl, optionally a C16alkyl, optionally wherein the lipophilic modification is attached at the 2’-ribo position of a nucleic acid residue of the dsRNA.

11. The composition of any one of the preceding claims, wherein the divalent ion source is selected from the group consisting of magnesium, calcium, copper, nickel, zinc, and strontium.

12. The composition of any one of the preceding claims, wherein the molar ratio of the divalent ion-to-the dsRNA is greater than about 2:

1.

13. The composition of any one of the preceding claims, wherein the composition is substantially free of inorganic phosphate and / or comprises less than 100 ppm of inorganic phosphate, optionally less than 50 ppm of inorganic phosphate, optionally less than 10 ppm of inorganic phosphate, optionally less than 5 ppm of inorganic phosphate, optionally wherein the composition does not comprise inorganic phosphate.

14. The composition of any one of the preceding claims, wherein the molar ratio of the divalent ion source-to-the-dsRNA is greater than about 2.5:1, optionally greater than about 3.0:1, optionally greater than about 3.5:1, optionally greater than about 4.0:1.

15. The composition of any one of the preceding claims, wherein the molar ratio of the divalent ion source-to-the-dsRNA is between about 2:1 and about 10:1, optionally between about 3:1 and about 10:1, optionally between about 3:1 and about 9:1, optionally between about 3:1 and about 8:1, optionally between about 3:1 and about 7:1, optionally between about 3:1 and about 6:1, optionally between about 3 : 1 and about 5:1.

16. The composition of any one of the preceding claims, wherein the dsRNA comprises a 5’- phosphate or 5 ’-phosphate mimic modification.

17. The composition of claim 16, wherein the 5 ’-phosphate mimic modification is where the preceding structurereplaces the 4’-CH2OH group within the ribose ring of a 5 ’-terminal nucleotide.

18. The composition of claim 16, wherein the phosphonate modification is a 5’- phosphonate modification.

19. The composition of claim 16, wherein the phosphonate modification is a 5’-vinyl phosphonate modification, optionally a 5’-(E)-vinyl phosphonate modification.

20. The composition of claim 16, wherein the phosphate mimic modification is a 5’-vinyl phosphate modification.

21. The composition of any one of the preceding claims, further comprising a diluent.

22. The composition of claim 21, wherein the composition is isotonic to cerebrospinal fluid (CSF).

23. The composition of claim 22, further comprising a sodium source, a potassium source, a magnesium source, and a calcium source.

24. The composition of claim 23, comprising sodium chloride, magnesium chloride, potassium chloride, and calcium chloride.

25. The composition of any one of the preceding claims, having a pH between about 4 and about 10, optionally wherein the pH is between about 6 and about 10, optionally wherein the pH is between about 6.5 and about 8.

0.

26. The composition of any one of the preceding claims, having an osmolality between about 200 and 400 mOsm / kg.

27. The composition of any one of the preceding claims, wherein the composition does not comprise hydrogen phosphate or dihydrogen phosphate.

28. The composition of any one of the preceding claims, wherein the composition does not comprise a buffer.

29. The composition of any one of the preceding claims, further comprising a stabilizing agent selected from the group consisting of sucrose, glucose, mannitol, sorbitol, polyethylene glycol (PEG), histidine, arginine, lysine, phospholipids, trehalose and a combination thereof.

30. The composition of any one of the preceding claims, wherein the composition comprises greater than 1 mg of the dsRNA per mL of the composition, optionally wherein the composition comprises greater than 5 mg of the dsRNA per mL of the composition, optionally wherein the composition comprises greater than 10 mg of the dsRNA per mL of the composition, optionally wherein the composition comprises greater than 25 mg of the dsRNA per mL of the composition, optionally wherein the composition comprises greater than 50 mg of the dsRNA per mL of the composition, optionally wherein the composition comprises about 60 mg of the dsRNA per mL of the composition.

31. The composition of any one of the preceding claims, wherein the dsRNA is AD-961583, AD-1395762, or AD-1498524.

32. A composition comprising:(a) a double-stranded ribonucleic acid (dsRNA) comprising a sense strand and an antisense strand, wherein one of the sense strand and antisense strand comprises at least one lipophilic modification, and the composition comprises about 50 mg to about 70 mg of the dsRNA per mL of the composition; (b) sodium chloride at about 80 mM to about 110 mM; and (c) calcium chloride at about 8.0 mM to about 20.0 mM.

33. The composition of claim 32, wherein the composition is substantially free of inorganic phosphate and / or comprises less than 100 ppm of inorganic phosphate, optionally less than 50 ppm of inorganic phosphate, optionally less than 10 ppm of inorganic phosphate, optionally less than 5 ppm of inorganic phosphate, optionally wherein the composition does not comprise inorganic phosphate.

34. The composition of claim 32 or claim 33, wherein the sense strand of the dsRNA comprises at least one lipophilic modification and the antisense strand of the dsRNA does not comprise a lipophilic modification.

35. The composition of any one of claims 32-34, wherein there is a molar excess of the antisense strand relative to the sense strand.

36. The composition of claim 35, wherein there is at least a 0.1% molar excess of the antisense strand relative to the sense strand, optionally wherein there is at least a 0.2% molar excess of the antisense strand relative to the sense strand, optionally wherein there is at least a 0.3% molar excess of the antisense strand relative to the sense strand, optionally wherein there is at least a 0.4% molar excess of the antisense strand relative to the sense strand, optionally wherein there is at least a 0.5% molar excess of the antisense strand relative to the sense strand, optionally wherein there is at least a 1% molar excess of the antisense strand relative to the sense strand, optionally wherein there is at least a 2% molar excess of the antisense strand relative to the sense strand, optionally wherein there is at least a 3% molar excess of the antisense strand relative to the sense strand, optionally wherein there is at least a 4% molar excess of the antisense strand relative to the sense strand, optionally wherein there is about a 5% or greater molar excess of the antisense strand relative to the sense strand.

37. The composition of any one of claims 32-36, further comprising: (d) potassium chloride at about 1.0 mM to about 2.5 mM; and (e) magnesium chloride at about 0.1 mM to about 1.0 mM.

38. The composition of any one of claims 32-37, wherein the composition comprises about 60 mg of the dsRNA per mL of the composition.

39. The composition of any one of claims 32-38, wherein the composition comprises sodium chloride at about 97.6 mM.

40. The composition of any one of claims 32-39, wherein the composition comprises calcium chloride at about 13.0 mM.

41. The composition of any one of claims 32-40, wherein the composition comprises potassium chloride at about 1.9 mM.

42. The composition of any one of claims 32-41, wherein the composition comprises magnesium chloride at about 0.5 mM.

43. The composition of any one of claims 32-42, wherein the composition is a pharmaceutical composition for intrathecal administration of the dsRNA to a subject.

44. The composition of claim 43, wherein the subject is a mammal, optionally wherein the subject is human.

45. The composition of any one of claims 32-44, further comprising a diluent.

46. The composition of claim 45, wherein the composition is isotonic to CSF.

47. The composition of any one of claims 32-46, having a pH between about 6 and about 10, optionally wherein the pH is between about 6.5 and about 8.

0.

48. The composition of any one of claims 32-47, having an osmolality between about 200 and 400 mOsm / kg.

49. The composition of any one of claims 32-48, wherein the composition does not comprise hydrogen phosphate or dihydrogen phosphate.

50. The composition of any one of claims 32-49, wherein the composition does not comprise a buffer.

51. The composition of any one of claims 32-50, further comprising a stabilizing agent selected from the group consisting of sucrose, glucose, mannitol, sorbitol, polyethylene glycol (PEG), histidine, arginine, lysine, phospholipids, trehalose and a combination thereof.

52. The composition of any one of claims 32-51, wherein the dsRNA AD-961583.

53. A composition for intrathecal administration comprising, (a) a dsRNA selected from the group consisting of AD-961583, AD-454973, AD-454843, AD-961584, AD-961585, and AD-961586; (b) a calcium ion source; and (c) a diluent, wherein the molar ratio of the calcium ion to the dsRNA is greater than 3 to 1.

54. A composition for intrathecal administration comprising, (a) a dsRNA selected from the group consisting of AD-1395718, AD-1395724, AD- 1395731, AD-1395738, AD-1395743, AD-1395756, AD-1395760, AD-1395762, AD- 1395764, and AD-1395771; (b) a calcium ion source; and (c) a diluent, wherein the molar ratio of the calcium ion to the dsRNA is greater than 3 to 1.

55. A composition for intrathecal administration comprising, (a) a dsRNA selected from the group consisting of AD-1019448, AD-1019465, AD- 1271082, AD-1271083, AD-1271084, AD-1271085, AD-1498524, AD-1498526, and AD-1498528; (b) a calcium ion source; and (c) a diluent, wherein the molar ratio of the calcium ion to the dsRNA is greater than 3 to 1.

56. The composition of any one of claims 53-55, wherein the composition is substantially free of inorganic phosphate and / or comprises less than 100 ppm of inorganic phosphate, optionally less than 50 ppm of inorganic phosphate, optionally less than 10 ppm of inorganic phosphate, optionally less than 5 ppm of inorganic phosphate, optionally wherein the composition does not comprise inorganic phosphate.

57. The composition of any one of claims 53-56, wherein the composition is isotonic to CSF.

58. The composition of any one of claims 53-57, further comprising a sodium source, a potassium source, a magnesium source, and a calcium source.

59. The composition of claim 58, comprising sodium chloride, magnesium chloride, potassium chloride, and calcium chloride.

60. The composition of any one of claims 53-59, having a pH between about 6 and about 10, optionally wherein the pH is between about 6.5 and about 8.

0.

61. The composition of any one of claims 53-60, having an osmolality between about 200 and 400 mOsm / kg.

62. The composition of any one of claims 53-61, wherein the composition does not comprise a hydrogen phosphate or dihydrogen phosphate.

63. The composition of any one of claims 53-62, wherein the composition does not comprise a buffer.

64. The composition of any one of claims 53-63, wherein the composition comprises greater than 1 mg of the dsRNA per mL of the composition, optionally wherein the composition comprises greater than 5 mg of the dsRNA per mL of the composition, optionally wherein the composition comprises greater than 10 mg of the dsRNA per mL of the composition, optionally wherein the composition comprises greater than 25 mg of the dsRNA per mL of the composition, optionally wherein the composition comprises greater than 50 mg of the dsRNA per mL of the composition, optionally wherein the composition comprises about 60 mg of the dsRNA per mL of the composition.

65. The composition of any one of claims 53-64, wherein the dsRNA is AD-961583.

66. The composition of any one of claims 53-65, wherein the antisense strand of the dsRNA is present at molar equivalence or in molar excess relative to the sense strand of the dsRNA.

67. A composition comprising a double-stranded ribonucleic acid (dsRNA) capable of annealing and reducing expression of an amyloid precursor protein (APP) mRNA, wherein the dsRNA comprises a sense strand and an antisense strand, wherein one of the sense strand or the antisense strand of the dsRNA comprises at least one lipophilic modification and the other strand of the dsRNA does not comprise a lipophilic modification; and wherein: (i) substantially all of the sense strand or the antisense strand of the dsRNA comprising the at least one lipophilic modification in the composition is duplexed with the strand that does not comprise a lipophilic modification, or (ii) the sense strand or the antisense strand of the dsRNA comprising the at least one lipophilic modification is present at less than 1% molar excess relative to the strand that does not comprise a lipophilic modification, the sense strand and the antisense strand are present at molar equivalence, or the strand that does not comprise a lipophilic modification is present in a molar excess relative to the strand of the dsRNA comprising the at least one lipophilic modification.

68. The composition of claim 67 further comprising a divalent ion source.

69. The composition of claim 67 or claim 68, wherein the dsRNA is selected from the group consisting of AD-961583, AD-454973, AD-454843, AD-961584, AD-961585, and AD-961586.

70. A composition comprising a double-stranded ribonucleic acid (dsRNA) capable of annealing and reducing expression of a superoxide dismutase 1 (SOD1) mRNA, wherein the dsRNA comprises a sense strand and an antisense strand, wherein one of the sense strand or the antisense strand of the dsRNA comprises at least one lipophilic modification and the other strand of the dsRNA does not comprise a lipophilic modification; and wherein: (i) substantially all of the sense strand or the antisense strand of the dsRNA comprising the at least one lipophilic modification in the composition is duplexed with the strand that does not comprise a lipophilic modification, or(ii) the sense strand or the antisense strand of the dsRNA comprising the at least one lipophilic modification is present at less than 1% molar excess relative to the strand that does not comprise a lipophilic modification, the sense strand and the antisense strand are present at molar equivalence, or the strand that does not comprise a lipophilic modification is present in a molar excess relative to the strand of the dsRNA comprising the at least one lipophilic modification.

71. The composition of claim 70 further comprising a divalent ion source.

72. The composition of claim 70 or claim 71, wherein the dsRNA is selected from the group consisting of AD-1395718, AD-1395724, AD-1395731, AD-1395738, AD-1395743, AD- 1395756, AD-1395760, AD-1395762, AD-1395764, and AD-1395771.

73. A composition comprising a double-stranded ribonucleic acid (dsRNA) capable of annealing and reducing expression of a huntingtin (HTT) mRNA, wherein the dsRNA comprises a sense strand and an antisense strand, wherein one of the sense strand or the antisense strand of the dsRNA comprises at least one lipophilic modification and the other strand of the dsRNA does not comprise a lipophilic modification; and wherein: (i) substantially all of the sense strand or the antisense strand of the dsRNA comprising the at least one lipophilic modification in the composition is duplexed with the strand that does not comprise a lipophilic modification, or (ii) the sense strand or the antisense strand of the dsRNA comprising the at least one lipophilic modification is present at less than 1% molar excess relative to the strand that does not comprise a lipophilic modification, the sense strand and the antisense strand are present at molar equivalence, or the strand that does not comprise a lipophilic modification is present in a molar excess relative to the strand of the dsRNA comprising the at least one lipophilic modification.

74. The composition of claim 73 further comprising a divalent ion source.

75. The composition of claim 73 or claim 74, wherein the dsRNA targets a sequence in exon 1 of the huntingtin gene.

76. The composition of any one of claims 73-75, wherein the dsRNA is selected from the group consisting of AD-1019448, AD-1019465, AD-1271082, AD-1271083, AD-1271084, AD- 1271085, AD-1498524, AD-1498526, and AD-1498528.

77. A composition comprising a double-stranded ribonucleic acid (dsRNA) comprising a sense strand and an antisense strand, wherein one of the sense strand or the antisense strand of the dsRNA comprises at least one lipophilic modification at one or more internal residue(s) of the sense or antisense strand and the other strand of the dsRNA does not comprise a lipophilic modification; and wherein: (i) substantially all of the sense strand or the antisense strand of the dsRNA comprising the at least one lipophilic modification in the composition is duplexed with the strand that does not comprise a lipophilic modification, or (ii) the sense strand or the antisense strand of the dsRNA comprising the at least one lipophilic modification is present at less than 1% molar excess relative to the strand that does not comprise a lipophilic modification, the sense strand and the antisense strand are present at molar equivalence, or the strand that does not comprise a lipophilic modification is present in a molar excess relative to the strand of the dsRNA comprising the at least one lipophilic modification.

78. The composition of claim 77 further comprising a divalent ion source.

79. The composition of claim 77 or claim 78, wherein the dsRNA comprises at least one lipophilic modification at one or more internal residue(s) of the sense strand, optionally wherein the dsRNA comprises at least one lipophilic modification at any one of positions 4-8 or 13-18 counting from the 5’-end of the strand, optionally wherein the dsRNA comprises at least one lipophilic modification at position 6 counting from the 5’-end of the strand.

80. The composition of claim 77 or claim 78, wherein the dsRNA comprises at least one lipophilic modification at one or more internal residue(s) of the antisense strand.

81. The composition of any one of claims 77-80, wherein the at least one lipophilic modification comprises a saturated or unsaturated C4-C30hydrocarbon, optionally a C4-C30alkyl or alkenyl, optionally a linear C6-C18alkyl or alkenyl, optionally a C16alkyl, optionally whereinthe at least one lipophilic modification is attached at the 2’-ribo position of a nucleic acid residue of the dsRNA.

82. A composition comprising a double-stranded ribonucleic acid (dsRNA) comprising a sense strand and an antisense strand, wherein one of the sense strand or the antisense strand of the dsRNA comprises at least one lipophilic modification at one or more terminal residue(s) of the sense or antisense strand and the other strand of the dsRNA does not comprise a lipophilic modification; and wherein: (i) substantially all of the sense strand or the antisense strand of the dsRNA comprising the at least one lipophilic modification in the composition is duplexed with the strand that does not comprise a lipophilic modification, or (ii) the sense strand or the antisense strand of the dsRNA comprising the at least one lipophilic modification is present at less than 1% molar excess relative to the strand that does not comprise a lipophilic modification, the sense strand and the antisense strand are present at molar equivalence, or the strand that does not comprise a lipophilic modification is present in a molar excess relative to the strand of the dsRNA comprising the at least one lipophilic modification.

83. The composition of claim 82 further comprising a divalent ion source.

84. The composition of claim 82 or claim 83, wherein the dsRNA comprises at least one lipophilic modification at the 5'-terminal and / or the 3'-terminal residue(s) of the sense strand, optionally wherein the dsRNA comprises at least one lipophilic modification at the 3'-terminal residue of the sense strand.

85. The composition of any one of claims 82-84, wherein the dsRNA comprises at least one lipophilic modification at the 5'-terminal residue of the sense strand.

86. The composition of claim 82 or claim 83, wherein the dsRNA comprises at least one lipophilic modification at the 5'-terminal and / or the 3'-terminal residue(s) of the antisense strand, optionally wherein the dsRNA comprises at least one lipophilic modification at the 3'-terminal residue of the antisense strand.

87. The composition of any one of claims 82, 83 or 86, wherein the dsRNA comprises at least one lipophilic modification at the 5'-terminal residue of the antisense strand.

88. The composition of any one of claims 67-87, wherein the at least one lipophilic modification comprises a saturated or unsaturated C4-C30hydrocarbon, optionally a C4-C30alkyl or alkenyl, optionally a linear C6-C18alkyl or alkenyl, optionally a C16alkyl, optionally wherein the at least one lipophilic modification is attached at the 2’-ribo position of a nucleic acid residue of the dsRNA.

89. A solid prepared by lyophilization of the composition of any one of the preceding claims.

90. A kit comprising: (a) a diluent comprising a divalent cation source e; and (b) a double-stranded ribonucleic acid (dsRNA) comprising a sense strand and an antisense strand, wherein the dsRNA comprises at least one modified nucleotide that is not a 2'- deoxynucleotide, wherein the molar ratio of the divalent cation source to the dsRNA is greater than 2 to 1.

91. The kit of claim 90, wherein the diluent is substantially free of inorganic phosphate and / or comprises less than 100 ppm of inorganic phosphate, optionally less than 50 ppm of inorganic phosphate, optionally less than 10 ppm of inorganic phosphate, optionally less than 5 ppm of inorganic phosphate, optionally wherein the diluent does not comprise inorganic phosphate.

92. A kit comprising (a) the solid of claim 89 and (b) a diluent.

93. The kit of claim 92, wherein the diluent is substantially free of inorganic phosphate and / or comprises less than 100 ppm of inorganic phosphate, optionally less than 50 ppm of inorganic phosphate, optionally less than 10 ppm of inorganic phosphate, optionally less than 5 ppm of inorganic phosphate, optionally wherein the diluent does not comprise inorganic phosphate.

94. A method of treating a subject having a disorder that would benefit from a reduction in expression of a target gene, the method comprising administering to the subject a therapeutically effective amount of a composition of any one of claims 1-88, thereby treating said subject.

95. The method of claim 94, wherein the subject is a human.

96. The method of claim 94 or claim 95, wherein the target gene is amyloid precursor protein (APP), superoxide dismutase 1 (SOD1), or the huntingtin gene, optionally exon 1 of the huntingtin gene.

97. The method of any one of claims 94-96, wherein the subject suffers from an APP- associated disease.

98. The method of claim 97, wherein the APP-associated disease is cerebral amyloid angiopathy (CAA).

99. The method of claim 97, wherein the APP-associated disease is early onset familial Alzheimer disease (EOFAD).

100. The method of claim 97, wherein the APP-associated disease is Alzheimer’s disease (AD), early onset Alzheimer’s disease (EOAD), familial Alzheimer’s disease, or late onset Alzheimer’s disease.

101. The method of any one of claims 96-100, wherein the APP expression is inhibited by at least about 30%.

102. The method of any one of claims 94-101, further comprising administering an additional therapeutic agent to the subject.

103. The method of any one of claims 94-102, wherein the dsRNA of the composition is administered at a dose of about 0.1 mg / kg to about 50 mg / kg.

104. The method of any one of claims 94-103, wherein the composition is administered to the subject intrathecally.

105. A method of inhibiting the expression of APP in a subject, the method comprising: administering to said subject a therapeutically effective amount of the composition of any one of claims 1-88, thereby inhibiting the expression of APP in said subject.

106. A method for treating or preventing an APP-associated disease or disorder in a subject, the method comprising administering to said subject a therapeutically effective amount of the composition of any one of claims 1-88, thereby treating or preventing an APP-associated disease or disorder in the subject.

107. The method of claim 106, wherein the APP-associated disease or disorder is selected from the group consisting of cerebral amyloid angiopathy (CAA), Alzheimer’s disease (AD), early onset familial Alzheimer disease (EOFAD), early onset Alzheimer’s disease (EOAD), familial Alzheimer’s disease, and late onset Alzheimer’s disease.

108. The method of claim 106 or claim 107, wherein the composition is administered by intrathecal injection, optionally wherein the intrathecal injection is performed in conjunction with intravenous administration.

109. The method of claim 108, wherein the intrathecal administration is used in the absence of intravenous administration.

110. The method of any one of claims 106-109, wherein administering the composition results in reduced intensity, severity, or frequency, or delayed onset of at least one symptom or feature of the APP-associated disease or disorder.

111. The method of any one of claims 106-110, wherein administering the composition results in no significant adverse effects in the subject, optionally wherein the subject does not have tremors or twitches upon administering the composition to the subject.

112. The method of any one of claims 106-111, wherein administering the composition takes place at an interval selected from once every two weeks, once every month, and once every two months, once every three months, once every four months, once every five months, and once every six months.

113. A method for administering a dsRNA to a subject in need thereof, the method comprising administering the composition of any one of claims 1-88 intrathecally to the subject, thereby administering the dsRNA to said subject.

114. The method of claim 113, wherein the subject is a human.

115. The method of claim 113 or claim 114, wherein the dsRNA targets an amyloid precursor protein (APP) gene, superoxide dismutase 1 (SOD1) gene, or the huntingtin gene, optionally wherein exon 1 of the huntingtin gene is targeted.

116. The method of any one of claims 113-115, wherein the dsRNA of the composition is administered at a dose of about 0.1 mg / kg to about 50 mg / kg.

117. A method of inhibiting the expression of SOD1 in a cell or tissue of a subject, the method comprising administering the composition of any one of claims 1-88 to the subject in an amount sufficient to reduce SOD1 expression in the cell or tissue of the subject, thereby inhibiting the expression of SOD1 in the cell or tissue of the subject.

118. The method of claim 117, wherein the subject is a human.

119. The method of claim 117 or claim 118, wherein the composition is administered intrathecally to the subject.

120. The method of any one of claims 117-119, wherein said administering reduces the level of SOD1 mRNA in the cell or tissue of the subject by at least 50%, optionally by at least 80%, as compared to an appropriate control.

121. The method of any one of claims 117-120, wherein the dsRNA of the composition is administered at a dose of about 0.1 mg / kg to about 50 mg / kg.

122. A method of inhibiting the expression of HTT in a cell or tissue of a subject, the method comprising administering the composition of any one of claims 1-66 to the subject in an amount sufficient to reduce HTT expression in the cell or tissue of the subject, thereby inhibiting the expression of HTT in the cell or tissue of the subject.

123. The method of claim 122, wherein the subject is a human.

124. The method of claim 122 or claim 123, wherein the composition is administered intrathecally to the subject.

125. The method of any one of claims 122-124, wherein the composition comprises a dsRNA that targets exon 1 of HTT.

126. The method of any one of claims 122-124, wherein said administering reduces the level of HTT mRNA in the cell or tissue of the subject by at least 50%, optionally by at least 80%, as compared to an appropriate control.

127. The method of any one of claims 94-126, wherein the dsRNA of the composition is administered at a dose of about 0.1 mg / kg to about 50 mg / kg.

128. A kit for performing the method of any one of claims 94-127, comprising a) the composition comprising dsRNA, and b) instructions for use, and c) optionally, a means for administering the composition to the subject.

129. A method for reducing or preventing particle formation in a solution comprising a divalent ion source and a double-stranded ribonucleic acid (dsRNA) having a sense strand and an antisense strand, wherein the dsRNA comprises at least one lipophilic modification of the sense strand or the antisense strand, and either the sense or the antisense strand does not comprise a lipophilic modification, the method comprising maintaining the sense strand or the antisense strand of the dsRNA comprising the lipophilic modification at less than 1% molar excess relative to the strand that does not comprise a lipophilic modification, thereby reducing or preventing particle formation in the solution comprising the divalent ion source and the double-stranded ribonucleic acid (dsRNA).

130. The method of claim 129, wherein the sense strand and the antisense strand are present at molar equivalence.

131. The method of claim 129, wherein the strand that does not comprise a lipophilic modification is present in a molar excess relative to the strand of the dsRNA comprising the at least one lipophilic modification.

132. The method of any one of claims 129-131, wherein the sense strand comprises the at least one lipophilic modification and the antisense strand does not comprise a lipophilic modification.

133. The method of any one of claims 129-131, wherein the antisense strand comprises the lipophilic modification and the sense strand does not comprise a lipophilic modification.

134. The method of any one of claims 129-133, wherein the divalent ion source is calcium, magnesium, copper, nickel, zinc, or strontium.

135. The method of any one of claims 129-134, wherein the at least one lipophilic modification is a C16 or longer lipophilic modification.

136. A method for preparing a formulation comprising annealing a sense strand and an antisense strand, wherein one of the sense strand and antisense strand contains a lipophilic modification, to form a duplex solution comprising a double stranded RNA (dsRNA); lyophilizing the duplex solution to provide a duplex composition; and dissolving the duplex composition in an injection solution, wherein the injection solution comprises a divalent cation source (e.g., calcium) and does not comprise a phosphate buffer; and the duplex composition comprises 0 – 5% molar excess (e.g., about 1 – 2 % molar excess) of antisense strand over sense strand.

137. The method of claim 136, wherein the divalent ion source is calcium, magnesium, copper, nickel, zinc, or strontium.

138. The method of claim 136 or 137, wherein the duplex composition comprises about a 1-2% molar excess of antisense strand over sense strand.

139. The method of any one of claims 136-138, wherein the sense strand comprises the at least one lipophilic modification and the antisense strand does not comprise a lipophilic modification.

140. The method of any one of claims 136-139, wherein the at least one lipophilic modification is a C16 or longer lipophilic modification.

141. The method of any one of claims 136-140, wherein the dsRNA is selected from the group consisting of AD-961583, AD-454973, AD-454843, AD-961584, AD-961585, and AD-961586.