Compositions and method for inhibiting hepcidin antimicrobial peptide (HAMP) or HAMP-related gene expression
View Patent ↗The invention relates to lipid formulated double-stranded ribonucleic acid (dsRNA) targeting a hepcidin antimicrobial peptide (HAMP) and/or HAMP-related gene, and methods of using the dsRNA to inhibit expression of HAMP and/or HAMP-related genes.
1. A double-stranded ribonucleic acid (dsRNA) compound, comprising a sense strand and an antisense strand that are each equal to or less than 30 nucleotides in length, wherein the compound is targeted to a nucleic acid molecule encoding transferrin receptor 2(TRF2), wherein the compound specifically hybridizes with the nucleic acid molecule so that the abundance of TRF2 mRNA is inhibited by at least 20% relative to a PBS control, and wherein the antisense strand comprises at least 15 contiguous nucleotides from the nucleotide sequence of SEQ ID NO:38.
2. The dsRNA of claim 1 , wherein the nucleotide sequence of the sense strand comprises 15 or more contiguous nucleotides of SEQ ID NO:35 with a start position of position 239 of the TFR2 mRNA transcript and the nucleotide sequence of the antisense strand comprises 15 or more contiguous nucleotides of SEQ ID NO:38 with a start position complementary to position 239 of the TFR2 mRNA transcript.
3. The dsRNA of claim 1 , wherein the nucleotide sequence of the sense strand comprises 16, 17, 18, 19, 20, or 21 contiguous nucleotides of SEQ ID NO:35 with a start position of position 239 of the TFR2 mRNA transcript and the nucleotide sequence of the antisense strand comprises 16, 17, 18, 19, 20, or 21 contiguous nucleotides of SEQ ID NO:38 with a start position complementary to position 239 of the TFR2 mRNA transcript.
4. The dsRNA of claim 1 , wherein the sense strand comprises the nucleotide sequence set forth in SEQ ID NO:35 and the antisense strand comprises the nucleotide sequence set forth in SEQ ID NO:38.
5. The dsRNA of claim 1 , further comprising a phosphorothioate at the first internucleotide linkage at the 3′ end of the sense strand, the antisense strand or both the sense strand and the antisense strand.
6. The dsRNA of claim 1 , wherein the sense strand, the antisense strand or both the sense and antisense strand further comprises at least one 3′n-overhang wherein the 3′-overhang comprises from 1 to 6 nucleotides.
7. The dsRNA of claim 1 , wherein the dsRNA further comprises a non-nucleotide moiety.
8. The dsRNA of claim 1 , wherein said dsRNA further comprises at least one modified nucleotide.
9. The dsRNA of claim 1 , further comprising a 2′-modified nucleotide in the sense strand, r the antisense strand or both the sense and antisense strand.
10. The dsRNA of claim 8 , wherein at least one of said modified nucleotides is selected from the group consisting of: a 2′-O-methyl modified nucleotide, a nucleotide comprising a 5′-phosphorothioate group, and a terminal nucleotide linked to a cholesteryl derivative or dodecanoic acid bisdecylamide group.
11. The dsRNA of claim 8 , wherein the modified nucleotide is selected from the group consisting of: 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.
12. The dsRNA of claim 1 , wherein the dsRNA is formulated in a nucleic acid lipid particle formulation.
13. The dsRNA of claim 1 , wherein the nucleic acid lipid particle formulation comprises a cationic lipid, a non-cationic lipid, and a cholesterol/ polyethyleneglycol-lipid conjugate.
14. The dsRNA of claim 1 , wherein the dsRNA is selected from the group consisting of AD-52551, AD-52552, AD-52557, AD-52558, AD-52563, AD-52564, AD-52569, AD-52570, AD-52574, AD-52575, AD-52579, AD-52580, AD-52584, AD-52585, AD-52589, AD-52590, and AD-47826.
15. A cell comprising the dsRNA of claim 1 .
16. A vector encoding at least one of the antisense strand and the sense strand of the dsRNA of claim 1 .
17. A cell comprising the vector of claim 15 .
18. A pharmaceutical composition comprising the dsRNA of claim 1 and a pharmaceutically acceptable carrier.
19. The pharmaceutical composition of claim 17 further comprising a lipid formulation.
20. A method of inhibiting TFR2 expression in a cell, the method comprising:
(a) introducing into the cell the dsRNA of claim 1 and
(b) maintaining the cell produced in step (a) for a time sufficient to obtain degradation of the mRNA transcript of a TFR2 gene, thereby inhibiting expression of the TFR2 gene in the cell.
21. A method of treating a disorder associated with TFR2 expression comprising administering to a subject in need of such treatment a therapeutically effective amount of the dsRNA of claim 1 .
22. The dsRNA of claim 1 , wherein the sense strand consists of the nucleotide sequence set forth in SEQ ID NO:35 and the antisense strand consists of the nucleotide sequence set forth in SEQ ID NO:38.