Compositions and methods for inhibiting expression of the alas1 gene

Patent metadata

Field	Value
Publication number	US-2025034566-A1
Application number	US-202418630441-A
Country	US
Kind code	A1
Filing date	Apr 9, 2024
Priority date	Apr 10, 2012
Publication date	Jan 30, 2025
Grant date	—

Abstract

The invention relates to double-stranded ribonucleic acid (dsRNA) compositions targeting the ALAS1 gene, and methods of using such dsRNA compositions to alter (e.g., inhibit) expression of ALAS1.

First claim

1 . A double-stranded ribonucleic acid (dsRNA) for inhibiting expression of ALAS1, wherein said dsRNA comprises a sense strand and an antisense strand 15-30 base pairs in length and the antisense strand is complementary to at least 15 contiguous nucleotides of SEQ ID NO: 1 or 382.

2 . A double-stranded ribonucleic acid (dsRNA) for inhibiting expression of ALAS1, wherein said dsRNA comprises a sense strand and an antisense strand, the antisense strand comprising a region of complementarity to an ALAS1 RNA transcript, which antisense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from the antisense sequence of AD-58882 (SEQ ID NO: 3434) or one of the antisense sequences listed in any one of Tables 2, 3, 6, 7, 8, 9, 14, 15, 18, or 20.

3 . The dsRNA of claim 1 , wherein said dsRNA comprises at least one modified nucleotide.

4 . The dsRNA of claim 3 , wherein at least one of said modified nucleotides is chosen from the group consisting of: a 2′-O-methyl modified nucleotide, a nucleotide comprising a 5′-phosphorothioate group, a terminal nucleotide linked to a cholesteryl derivative or dodecanoic acid bisdecylamide group, a 2′-deoxy-2′-fluoro modified nucleotide, a 2′-deoxy-modified nucleotide, a locked nucleotide, an abasic nucleotide, 2′-amino-modified nucleotide, 2′-alkyl-modified nucleotide, morpholino nucleotide, a phosphoramidate, and a non-natural base comprising nucleotide.

5 . (canceled)

6 . The dsRNA of claim 1 , wherein the region of complementarity is at least 17 nucleotides in length, between 19 and 21 nucleotides in length, or 19 nucleotides in length.

7 .- 8 . (canceled)

9 . The dsRNA of claim 1 , wherein each strand is no more than 30 nucleotides in length.

10 . The dsRNA of claim 1 , wherein at least one strand comprises a 3′ overhang of at least 1 nucleotide.

11 . (canceled)

12 . The dsRNA of claim 1 , further comprising a GalNAc ligand.

13 .- 14 . (canceled)

15 . The dsRNA of claim 1 , wherein the ligand is conjugated to the 3′ end of the sense strand of the dsRNA.

16 . (canceled)

17 . The dsRNA of claim 1 , wherein the dsRNA comprises a sense strand consisting of a sense sequence selected from the sense sequences disclosed in Tables 2, 3, 6, 7, 8, 9, 14, 15, 18 and 20, and an antisense strand consisting of an antisense sequence selected from the antisense sequences disclosed in Tables 2, 3, 6, 7, 8, 9, 14, 15, 18 and 20.

18 . A cell containing the dsRNA of claim 1 .

19 . A pharmaceutical composition for inhibiting expression of an ALAS1 gene, the composition comprising the dsRNA of claim 1 .

20 .- 31 . (canceled)

32 . A method of inhibiting ALAS1 expression in a cell, the method comprising:
(a) introducing into the cell the dsRNA of claim 1 , and (b) maintaining the cell of step (a) for a time sufficient to obtain degradation of the mRNA transcript of an ALAS1 gene, thereby inhibiting expression of the ALAS1 gene in the cell.

33 .- 41 . (canceled)

42 . A method of treating a disorder related to ALAS1 expression comprising administering to a subject in need of such treatment a therapeutically effective amount of
the dsRNA of claim 1 .

43 . (canceled)

44 . The method of claim 42 , wherein the subject is at risk for developing, or is diagnosed with, a porphyria, or wherein the subject has an elevated level of δ-aminolevulinic acid (ALA) and/or porphopilinogen (PBG).

45 .- 58 . (canceled)

59 . A method for decreasing a level of a porphyrin or a porphyrin precursor in a cell, comprising contacting the cell with the dsRNA of claim 1 , in an amount effective to decrease the level of the porphyrin or the porphyrin precursor in the cell.

60 .- 61 . (canceled)

62 . A vector encoding at least one strand of a dsRNA of claim 1 .

63 .- 68 . (canceled)

69 . The dsRNA of claim 1 , wherein the dsRNA comprises a sense strand comprising a sequence selected from the group consisting of SEQ ID NO:330, SEQ ID NO:334, SEQ ID NO:342, SEQ ID NO:344, SEQ ID NO:346, SEQ ID NO:356, SEQ ID NO:358, SEQ ID NO:362, SEQ ID NO:366, SEQ ID NO:376, SEQ ID NO:380, SEQ ID NO:140, SEQ ID NO:144, SEQ ID NO:152, SEQ ID NO:154, SEQ ID NO:156, SEQ ID NO:166, SEQ ID NO:168, SEQ ID NO:172, SEQ ID NO:176, SEQ ID NO:186, and SEQ ID NO:190.

70 .- 72 . (canceled)

73 . The method of claim 42 , wherein said method
(i) ameliorates a symptom associated with an ALAS1 related disorder (e.g., a porphyria) (ii) inhibits ALAS1 expression in the subject, (iii) decreases a level of a porphyrin precursor or a porphyrin in the subject, (iv) decreases frequency of acute attacks of symptoms associated with a porphyria in the subject, or (v) decreases incidence of acute attacks of symptoms associated with a porphyria in the subject when the subject is exposed to a precipitating factor.

74 .- 77 . (canceled)

78 . A double stranded RNAi (dsRNA) comprising a sense strand complementary to an antisense strand, wherein said antisense strand comprises a region of complementarity to an ALAS1 RNA transcript, wherein each strand has about 14 to about 30 nucleotides, wherein said dsRNA is represented by formula (III):
sense:5′ n p -N a -(XXX) i -N b -YYY-N b -(ZZZ) j -N a - n q 3′
antisense:3′ n p ′-N a ′-(X′X′X′) k -N b ′-Y′Y′Y′-N b ′-(Z′Z′Z′) l -N a ′- n q ′5′  (III)
wherein: i, j, k, and 1 are each independently 0 or 1; p, p′, q, and q′ are each independently 0-6; each N a and N a ′ independently represents an oligonucleotide sequence comprising 0-25 nucleotides which are either modified or unmodified or combinations thereof, each sequence comprising at least two differently modified nucleotides; each N b and N b ′ independently represents an oligonucleotide sequence comprising 0-10 nucleotides which are either modified or unmodified or combinations thereof; each n p , n p ′, n q , and n q ′ independently represents an overhang nucleotide; XXX, YYY, ZZZ, X′X′X′, Y′Y′Y′, and Z′Z′Z′ each independently represent one motif of three identical modifications on three consecutive nucleotides; modifications on N b differ from the modification on Y and modifications on N b ′ differ from the modification on Y′.

79 .- 154 . (canceled)

Assignees

• Alnylam Pharmaceuticals Inc
• Icahn School Med Mount Sinai

Inventors

• Bettencourt Brian
• Fitzgerald Kevin
• Querbes William
• Desnick Robert J
• Yasuda Makiko

Classifications

• A61P3/00

  Drugs for disorders of the metabolism (of the blood or the extracellular fluid A61P7/00) · CPC title

• A61P25/08

  Antiepileptics; Anticonvulsants · CPC title

• A61P25/00

  Drugs for disorders of the nervous system · CPC title

• A61K47/549

  Sugars, nucleosides, nucleotides or nucleic acids · CPC title

• C12N15/63

  Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression · CPC title