Claims for Patent: 10,125,364

Summary for Patent: 10,125,364

Title:	Compositions and methods for inhibiting expression of the ALAS1 gene
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.
Inventor(s):	Bettencourt; Brian (Groton, MA), Fitzgerald; Kevin (Brookline, MA), Querbes; William (Cambridge, MA), Desnick; Robert J. (New York, NY), Yasuda; Makiko (New York, NY)
Assignee:	ALYNYLAM PHARMACEUTICALS, INC. (Cambridge, MA) ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI (New York, NY)
Application Number:	14/814,911
Patent Claims:	1. A double-stranded ribonucleic acid (dsRNA) for inhibiting expression of ALAS1, comprising: (i) an antisense strand that comprises the sequence of SEQ ID NO: 1296; (ii) a sense strand comprising at least 15 contiguous nucleotides from SEQ ID NO: 1295; and (iii) a duplex region of 15-30 base pairs in length. 2. The dsRNA of claim 1, wherein said dsRNA comprises at least one modified nucleotide. 3. The dsRNA of claim 1, wherein each of the sense strand and the antisense strand is 19-24 nucleotides in length. 4. The dsRNA of claim 1, wherein at least one strand comprises a 3' overhang of at least 1 nucleotide. 5. The dsRNA of claim 4, wherein the 3' overhang is 2 nucleotides in length. 6. The dsRNA of claim 1 further comprising a ligand. 7. The dsRNA of claim 6, wherein said ligand is a GalNAc ligand. 8. An isolated cell comprising the dsRNA of claim 1. 9. A pharmaceutical composition for inhibiting expression of an ALAS1 gene, the composition comprising the dsRNA of claim 1. 10. 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. 11. 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. 12. The method of claim 11, wherein the subject is at risk for developing, or is diagnosed with, a porphyria. 13. The method of claim 11, wherein the dsRNA is administered before, during, or after an acute attack of porphyria. 14. The method of claim 11, wherein the method decreases a level of a porphyrin or a porphyrin precursor in the subject. 15. 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. 16. A vector encoding at least one strand of a dsRNA of claim 1. 17. An isolated cell comprising the vector of claim 16. 18. The method of claim 11, 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. 19. The dsRNA of claim 7, wherein the GalNAc ligand is attached to the 3' end of the sense strand. 20. The dsRNA of claim 1, wherein the at least one modified nucleotide is chosen from a 2'-O-methyl modified nucleotide, 2'-fluoro modified nucleotide, or both. 21. The dsRNA of claim 19, wherein the GalNAc ligand is attached to the 3' end of the sense strand via a linker. 22. The dsRNA of claim 7, wherein the GalNAc ligand is ##STR00026## 23. The dsRNA of claim 21, wherein the ligand and linker has the structure of: ##STR00027## 24. The method of claim 11, wherein the porphyria is a hepatic porphyria selected from acute intermittent porphyria (AIP) hereditary coproporphyria (HCP), variegate porphyria (VP), ALA deyhdratase deficiency porphyria (ADP), and hepatoerythropoietic porphyria. 25. The method of claim 11, wherein the dsRNA is administered during a prodrome. 26. The method of claim 11, wherein the subject has an elevated level of ALA, PBG, or both ALA and PBG. 27. A method of treating a subject with an elevated level of ALA, PBG, or both ALA and PBG, the method comprising administering to the subject the dsRNA of claim 1. 28. The dsRNA of claim 5, wherein one end of the dsRNA is blunt-ended. 29. The dsRNA of claim 28, wherein the dsRNA comprises one or more phosphorothioate linkages. 30. The dsRNA of claim 1, which comprises 20 or more modified nucleotides chosen from 2'-O-methyl modified nucleotides and 2'-fluro modified nucleotides. 31. The dsRNA of claim 30, comprising 2'-O-methyl modified nucleotides and 2'-fluro modified nucleotides over the entire length of the sense and antisense strands. 32. A double-stranded ribonucleic acid (dsRNA) for inhibiting expression of ALAS1, comprising: (i) an antisense strand that comprises the sequence of SEQ ID NO: 1296; (ii) a sense strand comprising at least 15 contiguous nucleotides from SEQ ID NO: 1295; (iii) a duplex region of 15-30 base pairs in length; (iv) one strand comprising a 3' overhang of at least 1 nucleotide; (v) one end that is blunt-ended; and (vi) at least one modified nucleotide chosen from a 2'-O-methyl modified nucleotide, 2'-fluoro modified nucleotide, or both. 33. A double-stranded ribonucleic acid (dsRNA) for inhibiting expression of ALAS1, wherein the dsRNA comprises: (i) a sense strand comprising at least 15 contiguous nucleotides from SEQ ID NO: 1295; (ii) an antisense strand complementary to at least nucleotides 871-889 of SEQ ID NO: 1; and (iii) a ligand comprising one or more N-acetylgalactosamine (GalNAc) derivatives. 34. The dsRNA of claim 33, wherein the dsRNA comprises at least one modified nucleotide, wherein at least one of the 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, a 2'-amino-modified nucleotide, a 2'-alkyl-modified nucleotide, a morpholino nucleotide, a phosphoramidate, and a non-natural base comprising nucleotide. 35. The dsRNA of claim 33, wherein the GalNAc derivative structure is as shown below and is attached to the 3' end of the sense strand of the dsRNA ##STR00028## 36. The dsRNA of claim 33, comprising a duplex region of 15-30 base pairs in length.