Compositions and Methods for Inhibiting Gene Expression of Alpha-1 AntiTrypsin

Abstract

The invention relates to a RNA interference triggers for inhibiting the expression of an AAT gene through the mechanism of RNA interference. The invention also relates to a pharmaceutical composition comprising the AAT RNAi trigger together with an excipient capable of improving delivery of the RNAi trigger to a liver cell in vivo. Delivery of the AAT RNAi trigger to liver cells in vivo provides for inhibition of AAT gene expression and treatment of alpha 1-antitrypsin deficiency and associated diseases.

1 .- 20 . (canceled) 
     
     
         21 . A method for inhibiting the expression of an AAT gene in a cell, a tissue, or an organism, the method comprising introducing into a cell, tissue, or organism a composition that comprises an RNAi trigger molecule, wherein the RNAi trigger molecule comprises a sense strand sequence that is 18-30 nucleotides in length and an antisense strand sequence that is 18-30 nucleotides in length, wherein:
 the antisense strand sequence and the sense strand sequence are substantially complementary over a core stretch of at least 18 consecutive nucleotides, and the antisense strand sequence comprises in order the nucleobase sequence of nucleotides 1-18 of any one of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, or SEQ ID NO:5.   
     
     
         22 .- 23 . (canceled) 
     
     
         24 . The method of  claim 21 , wherein the sense strand and the antisense strand of the RNAi trigger molecule each independently comprises at least one modified nucleotide, wherein the modified nucleotide is selected from the group consisting of: a 2′-O-methyl modified nucleotide, a nucleotide comprising a 5′-phosphorothioate group, a 2′-deoxy-2′-fluoro modified nucleotide, a 2′-deoxy-modified nucleotide, and a deoxythymidine. 
     
     
         25 . The method of  claim 24 , wherein the antisense strand of the RNAi trigger molecule comprises at least one modified nucleotide that is a UNA (2′,3′-seco RNA nucleotide mimic). 
     
     
         26 . The method of  claim 25 , wherein the UNA is located on the antisense strand of the RNAi trigger molecule at nucleotide position 6 or nucleotide position 7 from the 5′ terminal end of the anti sense strand. 
     
     
         27 . The method of  claim 24 , wherein a targeting moiety is conjugated to the sense strand of the RNAi trigger molecule. 
     
     
         28 . The method of  claim 25 , wherein a targeting moiety is conjugated to the sense strand of the RNAi trigger molecule. 
     
     
         29 . The method of  claim 26 , wherein a targeting moiety is conjugated to the sense strand of the RNAi trigger molecule. 
     
     
         30 . The method of  claim 24 , where the targeting moiety comprises a cholesteryl group. 
     
     
         31 . The method of  claim 24 , wherein the targeting moiety comprises N-acetyl-galactosamine. 
     
     
         32 . The method of  claim 21 , wherein the sense strand of RNAi trigger molecule comprises in order the nucleobase sequence of nucleotides 1-18 of any one of SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO: 10, SEQ ID NO: 11, or SEQ ID NO: 12. 
     
     
         33 . The method of  claim 32 , wherein the antisense strand and the sense strand of the RNAi trigger molecule form a duplex comprising the nucleotide sequence pairs selected from the group consisting of: SEQ ID NOs:50/62; SEQ ID NOs:50/63; SEQ ID NOs:53/67; SEQ ID NOs:54/69; SEQ ID NOs:55/70; SEQ ID NOs:56/75; SEQ ID NOs:56/77; SEQ ID NOs:51/63; SEQ ID NOs:52/63; SEQ ID NOs:57/77; and SEQ ID NOs:58/77. 
     
     
         34 . The method of  claim 33 , wherein the antisense strand and the sense strand of the RNAi trigger molecule form a duplex comprising the nucleotide sequence pairs selected from the group consisting of: SEQ ID NOs:51/63; SEQ ID NOs:52/63; SEQ ID NOs:57/77; and SEQ ID NOs:58/77. 
     
     
         35 . The method of  claim 33 , wherein inhibiting expression of AAT gene in an organism treats or manages a pathological condition or disease caused by alpha-1 antitrypsin deficiency. 
     
     
         36 . The method of  claim 35 , wherein the pathological condition and disease caused by alpha-1 antitrypsin deficiency is selected from the group consisting of: chronic hepatitis, cirrhosis, hepatocellular carcinoma, and fulminant hepatic failure. 
     
     
         37 . The method of  claim 34 , wherein inhibiting expression of AAT gene in an organism treats or manages a pathological condition or disease caused by alpha-1 antitrypsin deficiency. 
     
     
         38 . The method of  claim 37 , wherein the pathological condition and disease caused by alpha-1 antitrypsin deficiency is selected from the group consisting of: chronic hepatitis, cirrhosis, hepatocellular carcinoma, and fulminant hepatic failure. 
     
     
         39 . The method of  claim 21 , wherein the composition is capable of inhibiting expression of an AAT gene in vitro by at least 40% at a concentration of 0.1 nM of the RNAi trigger molecule.