Polynucleotide agents targeting patatin-like phospholipase domain containing 3 (pnpla3) and methods of use thereof

Abstract

The invention relates to polynucleotide agents targeting a Patatin-Like Phospholipase Domain Containing 3 (PNPLA3) gene, and methods of using such polynucleotide agents to inhibit expression of a PNPLA3 gene and methods of treating subjects having Nonalcoholic Fatty Liver Disease (NAFLD) and/or a PNPLA3-associated disorder.

1 . An antisense polynucleotide agent for inhibiting expression of a Patatin-Like Phospholipase Domain Containing 3 (PNPLA3) gene, wherein the agent comprises about 4 to about 50 contiguous nucleotides, wherein at least one of the contiguous nucleotides is a modified nucleotide, and wherein the nucleotide sequence of the agent is at least about 80% complementary over its entire length to the equivalent region of the nucleotide sequence of any one of SEQ ID NOs:1, 3, 5, 7, 9, and 11. 
     
     
         2 . The agent of  claim 1 , wherein the equivalent region is any one of the target regions of SEQ ID NO:1 provided in Tables 3 and 4. 
     
     
         3 . An antisense polynucleotide agent for inhibiting expression of a Patatin-Like Phospholipase Domain Containing 3 (PNPLA3) gene, 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 the agent is about 8 to about 50 nucleotides in length. 
     
     
         4 . The agent of  claim 1 , wherein substantially all of the nucleotides of the antisense polynucleotide agent are modified nucleotides; or wherein all of the nucleotides of the antisense polynucleotide agent are modified nucleotides. 
     
     
         5 . The agent of  claim 1 , which is 10 to 40 nucleotides in length; 10 to 30 nucleotides in length; 18 to 30 nucleotides in length; 10 to 24 nucleotides in length; 18 to 24 nucleotides in length; 20 nucleotides in length; or 14 nucleotides in length. 
     
     
         6 . The agent of  claim 1 , wherein the modified nucleotide comprises a modified sugar moiety selected from the group consisting of: a 2′-O-methoxyethyl modified sugar moiety, a 2′-O-alkyl modified sugar moiety, and a bicyclic sugar moiety. 
     
     
         7 . (canceled) 
     
     
         8 . The agent of  claim 1 , wherein the modified nucleotide comprises a modified internucleoside linkage. 
     
     
         9 . The agent of  claim 1 , 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 . An antisense polynucleotide agent for inhibiting expression of a Patatin-Like Phospholipase Domain Containing 3 (PNPLA3) gene, 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 moiety.   
     
     
         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 1 , wherein the agent further comprises a ligand conjugated at the 3′-terminus. 
     
     
         14 . The agent of  claim 13 , wherein the ligand is an N-acetylgalactosamine (GalNAc) derivative. 
     
     
         15 . A pharmaceutical composition for inhibiting expression of a Patatin-Like Phospholipase Domain Containing 3 (PNPLA3) gene, comprising the agent of  claim 1 . 
     
     
         16 . A pharmaceutical composition comprising the agent of  claim 1 , and a lipid formulation. 
     
     
         17 . A method of inhibiting expression of a Patatin-Like Phospholipase Domain Containing 3 (PNPLA3) gene in a cell, the method comprising:
 (a) contacting the cell with the agent of  claim 1  or a pharmaceutical composition of  claim 15 ; and   (b) maintaining the cell produced in step (a) for a time sufficient to obtain antisense inhibition of a PNPLA3 gene, thereby inhibiting expression of the PNPLA3 gene in the cell.   
     
     
         18 . The method of  claim 17 , wherein the cell is within a subject. 
     
     
         19 . The method of  claim 18 , wherein the subject is a human. 
     
     
         20 .- 23 . (canceled)