Compositions and methods for increasing erythropoietin (EPO) production
View Patent ↗The invention relates to double-stranded ribonucleic acid (dsRNA) compositions targeting one or more EGLN genes, EGLN1, EGLN2 and/or EGLN3 and methods of using such dsRNA compositions to inhibit expression of these genes.
1. A double-stranded ribonucleic acid (dsRNA) for inhibiting expression of Eg1 nine homolog 1 (EGLN1), wherein said dsRNA comprises a sense strand and an antisense strand,
wherein the sense strand comprises at least 15 contiguous nucleotides or chemically modified nucleotides differing by no more than 3 nucleotides or chemically modified nucleotides from a first chemically modified sequence set forth in
cAAGGuAcGcAAuAAcuGudTsdT (SEQ ID NO: 88),
and the antisense strand comprises at least 15 contiguous nucleotides or chemically modified nucleotides differing by no more than 3 nucleotides or chemically modified nucleotides from a second chemically modified sequence set forth in
AcAGUuAUUGCGuACCUUGdTsdT (SEQ ID NO: 89),
wherein c is 2′-O-methylcytidine, u is 2′-O-methyluridine, dT is 2′-deoxythymidine, and s is phosphorothioate linkage.
2. The dsRNA of claim 1 , wherein the dsRNA comprises a region of complementarity between the sense and antisense strands, wherein the region is at least 17 nucleotides or chemically modified nucleotides in length.
3. The dsRNA of claim 2 , wherein the dsRNA comprises a region of complementarity between the sense and antisense strands, wherein the region is between 19 and 21 nucleotides or chemically modified nucleotides in length.
4. The dsRNA of claim 1 , wherein either one or both of the sense and antisense strands comprises a 3′ overhang of at least 1 nucleotide.
5. The dsRNA of claim 1 , wherein the dsRNA further comprises a ligand selected from the group consisting of a lipid, a lipid based molecule, a protein, an antibody, a peptide, a peptidomimetic, and a polymer.
6. The dsRNA of claim 5 , wherein the ligand is conjugated to the 3′ end of the sense strand of the dsRNA.
7. A pharmaceutical composition for inhibiting expression of EGLN1 comprising the dsRNA of claim 1 .
8. The dsRNA of claim 1 , wherein the sense strand comprises at least 15 contiguous nucleotides or chemically modified nucleotides of the first chemically modified sequence.
9. The dsRNA of claim 1 , wherein the sense strand comprises the first chemically modified sequence.
10. The dsRNA of claim 1 , wherein the antisense strand comprises at least 15 contiguous nucleotides or chemically modified nucleotides of the second chemically modified sequence.
11. The dsRNA of claim 1 , wherein the antisense strand comprises the second chemically modified sequence.
12. The dsRNA of claim 6 , wherein the ligand is a lipid or a lipid based ligand.
13. The dsRNA of claim 1 , wherein the dsRNA is encapsulated in a stable nucleic acid-lipid particle.
14. A composition comprising
(1) a first double-stranded ribonucleic acid (dsRNA) for inhibiting expression of Eg1 nine homolog 1 (EGLN1), wherein said dsRNA comprises a sense strand and an antisense strand, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence of SEQ ID NO: 26 and the antisense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence of SEQ ID NO: 27, and wherein the dsRNA comprises a modified nucleotide selected from the group consisting of: a nucleotide comprising a 5′-phosphorothioate group, a terminal nucleotide linked to a cholesteryl derivative, a terminal nucleotide linked to a dodecanoic acid bisdecylamide group, a 2′-deoxy-2′-fluoro modified nucleotide, a locked nucleotide, an abasic nucleotide, a 2′-amino-modified nucleotide, a 2′-alkyl-modified nucleotide, a morpholino nucleotide, and a nucleotide comprising a non-natural base; and
(2) a second dsRNA for inhibiting expression of EGLN2 or EGLN3.
15. The composition of claim 14 , further comprising an anti-cancer therapy.
16. The composition of claim 14 , further comprising an angiogenesis inhibitor.
17. The composition of claim 14 , wherein the second dsRNA inhibits expression of EGLN2.
18. The composition of claim 14 , wherein the second dsRNA inhibits expression of EGLN3.
19. A method of inhibiting EGLN1 expression in a cell, the method comprising: contacting the cell with the dsRNA of claim 1 , thereby inhibiting expression of EGLN1 in the cell.
20. A method of treating a disorder mediated by EGLN expression comprising administering to a human subject in need of such treatment a therapeutically effective amount of the dsRNA of claim 1 .
21. The method of claim 20 , wherein the human subject has anemia or a condition associated with anemia.
22. The method of claim 21 , wherein the anemia is selected from the group consisting of anemia due to B12 deficiency, anemia due to folate deficiency, anemia due to iron deficiency, hemolytic anemia, hemolytic anemia due to G-6-PD deficiency, idiopathic aplastic anemia, idiopathic autoimmune hemolytic anemia, immune hemolytic anemia, megaloblastic anemia, pernicious anemia, secondary aplastic anemia, and sickle cell anemia.
23. The method of claim 21 , wherein the condition associated with anemia is selected from the group consisting of pale skin, dizziness, fatigue, headaches, irritability, low body temperature, numb/cold hands or feet, rapid heartbeat, reduced erythropoietin, shortness of breath, weakness and chest pain.
24. The method of claim 20 , wherein the human subject has a disorder selected from the group consisting of hypoxia, a neurological condition, renal disease or failure, a cancer of the blood, a cancer of the bone, and a cancer of the marrow.
25. A method of increasing erythropoietin levels in a human subject, the method comprising administering to the human subject a therapeutically effective amount of the dsRNA of claim 1 .