Polynucleotide agents targeting aminolevulinic acid synthase-1 (alas1) and uses thereof

Abstract

The invention relates to polynucleotide agents, e.g., antisense polynucleotide agents, targeting the ALAS1 gene, and methods of using such agents to alter (e.g., inhibit) expression of ALAS1 and to treat ALAS1 associated diseases, e.g., porphyria.

1 . (canceled) 
     
     
         2 . (canceled) 
     
     
         3 . A single-stranded antisense polynucleotide agent for inhibiting expression of aminolevulinic acid synthase-1 (ALAS1), wherein the agent comprises at least 8 contiguous nucleotides differing by no more than 3 nucleotides from any one of the nucleotide sequences listed in Tables 3 and 4, and wherein at least one of the contiguous nucleotides is a modified nucleotide. 
     
     
         4 . The agent of  claim 3 , wherein substantially all of the nucleotides of the antisense polynucleotide agent are modified nucleotides. 
     
     
         5 . The agent of  claim 3 , which is 10 to 40 nucleotides in length; 10 to 30 nucleotides in length; 18 to 30 nucleotides in length; or 10 to 24 nucleotides in length. 
     
     
         6 . The agent of  claim 3 , wherein the modified nucleotide comprises a modified sugar moiety selected from the group consisting of: a 2′-O-methoxyethyl modified sugar moiety, a 2′-methoxy modified sugar moiety, a 2′-O-alkyl modified sugar moiety, and a bicyclic sugar moiety. 
     
     
         7 . The agent of  claim 3 , wherein the modified nucleotide is a 5-methylcytosine. 
     
     
         8 . The agent of  claim 3 , wherein the modified nucleotide comprises a modified internucleoside linkage. 
     
     
         9 . The agent of  claim 3 , comprising a plurality of 2′-deoxynucleotides flanked on each side by at least one nucleotide having a modified sugar moiety. 
     
     
         10 . The agent of  claim 9 , wherein the agent is a gapmer comprising a gap segment comprised of linked 2′-deoxynucleotides positioned between a 5′ and a 3′ wing segment. 
     
     
         11 . The agent of  claim 3 , comprising
 a gap segment consisting of linked deoxynucleotides;   a 5′-wing segment consisting of linked nucleotides;   a 3′-wing segment consisting of linked nucleotides;   wherein the gap segment is positioned between the 5′-wing segment and the 3′-wing segment and wherein each nucleotide of each wing segment comprises a modified sugar.   
     
     
         12 . The agent of  claim 11 , wherein the gap segment is ten 2′-deoxynucleotides in length and each of the wing segments is five nucleotides in length. 
     
     
         13 . The agent of  claim 3 , wherein the agent further comprises a ligand at the 3′-terminus of the agent. 
     
     
         14 . The agent of  claim 13 , wherein the ligand is an N-acetylgalactosamine (GalNAc) derivative. 
     
     
         15 . A pharmaceutical composition for inhibiting expression of a aminolevulinic acid synthase-1 (ALAS1) gene comprising the agent of  claim 3 . 
     
     
         16 . A pharmaceutical composition comprising the agent of  claim 3 , and a lipid formulation. 
     
     
         17 . A method of inhibiting aminolevulinic acid synthase-1 (ALAS1) expression in a cell, the method comprising:
 (a) contacting the cell with the agent of  claim 3 ; and   (b) maintaining the cell produced in step (a) for a time sufficient to obtain antisense inhibition of an ALAS1 gene, thereby inhibiting expression of the ALAS gene in the cell.   
     
     
         18 . The method of  claim 17 , wherein the cell is within a human subject. 
     
     
         19 . (canceled) 
     
     
         20 . A method of preventing at least one symptom in or treating a subject having an ALAS1-associated disease, the method comprising administering to the subject a prophylactically effective amount or therapeutically effective amount of the agent of  claim 3 , thereby preventing at least one symptom in or treating the subject. 
     
     
         21 . (canceled) 
     
     
         22 . (canceled) 
     
     
         23 . The method of  claim 20 , wherein the ALAS1-associated disease is porphyria. 
     
     
         24 . The method of  claim 23 , wherein the porphyria is selected from the group consisting of X-linked sideroblastic anemia (XLSA), ALA deyhdratase deficiency porphyria (Doss porphyria), acute intermittent porphyria (AIP), congenital erythropoietic porphyria (CEP), prophyria cutanea tarda (PCT), hereditary coproporphyria (coproporphyria, or HCP), variegate porphyria (VP), erythropoietic protoporphyria (EPP), or transient erythroporphyria of infancy, acute hepatic porphyria, hepatoerythropoietic porphyria, and dual porphyria. 
     
     
         25 . The method of  claim 23 , wherein the agent or the composition is administered to the subject after an acute attack of porphyria; or during an acute attack of porphyria.