Rnai therapy for hepatitis b virus infection

Abstract

Described are compositions and methods for inhibition of Hepatitis B virus gene expression. RNA interference (RNAi) triggers and RNAi trigger conjugates for inhibiting the expression of Hepatitis B virus gene are described. Pharmaceutical compositions comprising one or more HBV RNAi triggers optionally with one or more additional therapeutics are also described. Delivery of the described HBV RNAi triggers to infected liver in vivo provides for inhibition of HBV gene expression and treatment.

1 . A composition for inhibiting expression of a hepatitis B virus gene comprising:
 (a) MLP-(L-T) x  wherein
 MLP is a melittin-like peptide, 
 L-T has the structure represented by —CO—C(CH 3 )═C(T)-COOH or
 —CO—C(T)=C(CH 3 )—COOH, wherein T comprises a targeting ligand having affinity for the an asialoglycoprotein receptor, and 
 
 x is greater than 80% of the number of primary amines of a population of MLPs, and 
   (b) an HBV RNAi trigger comprising an antisense strand and a sense strand wherein the antisense strand comprises nucleotides 2-18 of any of SEQ ID NO: 1-102.   
     
     
         2 . The composition of  claim 1 , wherein the HBV RNAi trigger comprises a sense strand comprising nucleotides 2-18 of any of SEQ ID NO: 103-209. 
     
     
         3 . The composition of  claim 1 , wherein the HBV RNAi trigger comprises a sense strand covalently linked to a cholesteryl group via a TEG group or a C6 group. 
     
     
         4 - 6 . (canceled) 
     
     
         7 . The composition of  claim 1  wherein the MLP comprises the amino acid sequence of SEQ ID NO: 650, SEQ ID NO: 644, SEQ ID NO: 654, SEQ ID NO: 694, SEQ ID NO: 700, SEQ ID NO: 701, SEQ ID NO: 751, or SEQ ID NO: 755. 
     
     
         8 . (canceled) 
     
     
         9 . The composition of  claim 1  wherein T comprises N-acetylgalactosamine (GalNAc). 
     
     
         10 . The composition of  claim 9  wherein -(L-T) has the structure represented by: 
       
         
           
           
               
               
           
         
         wherein n=1. 
       
     
     
         11 . The composition of  claim 1  wherein at least one nucleotide of the HBV RNAi trigger is modified. 
     
     
         12 . The composition of  claim 11  wherein the antisense strand of the HBV RNAi trigger comprises the nucleotide sequence of any of SEQ ID NO: 210-414. 
     
     
         13 . The composition of  claim 12  wherein the sense strand of the HBV RNAi trigger comprises the nucleotide sequence of any of SEQ ID NO: 415-643. 
     
     
         14 - 29 . (canceled) 
     
     
         30 . An HBV RNAi trigger comprising a sense strand and an antisense strand, wherein the sense strand comprises a modified nucleotide sequence according to:
 (a) nucleotides 8-26 of SEQf5 ID NO: 550;   (b) nucleotides 5-23 of SEQ ID NO: 608;   (c) nucleotides 7-24 of SEQ ID NO: 624;   (d) nucleotides 1-19 of SEQ ID NO: 629;   (e) nucleotides 3-21 of SEQ ID NO: 630;   (f) nucleotides 3-20 of SEQ ID NO: 631;   (g) nucleotides 3-20 of SEQ ID NO: 632; or   (h) nucleotides 3-12 of SEQ ID NO: 634.   
     
     
         31 . The HBV RNAi trigger of  claim 30 , wherein the antisense strand comprises a nucleobase sequence according to:
 (a) nucleobases 1-18 of SEQ ID NO: 17;   (b) nucleobases 1-19 of SEQ ID NO: 75;   (c) nucleobases 1-19 of SEQ ID NO: 76;   (d) nucleobases 1-20 of SEQ ID NO: 92; or   (e) nucleobases 1-19 of SEQ ID NO: 95.   
     
     
         32 . The HBV RNAi trigger of  claim 30 , wherein the antisense strand comprises a modified nucleotide sequence according to:
 (a) nucleotides 1-19 of SEQ ID NO: 371;   (b) nucleotides 1-19 of SEQ ID NO: 372;   (c) nucleotides 1-19 of SEQ ID NO: 401;   (d) nucleotides 1-19 of SEQ ID NO: 402;   (e) nucleotides 2-19 of SEQ ID NO: 403; or   (f) nucleotides 2-19 of SEQ ID NO: 404.   
     
     
         33 . An HBV RNAi trigger comprising a sense strand and an antisense strand, wherein the antisense strand comprises a modified nucleotide sequence according to:
 (a) nucleotides 1-19 of SEQ ID NO: 370;   (b) nucleotides 1-19 of SEQ ID NO: 371;   (c) nucleotides 1-19 of SEQ ID NO: 372;   (d) nucleotides 1-19 of SEQ ID NO: 396;   (e) nucleotides 1-19 of SEQ ID NO: 401; or   (f) nucleotides 1-19 of SEQ ID NO: 402.   
     
     
         34 . The HBV RNAi trigger of  claim 33 , wherein the sense strand comprises a nucleobase sequence according to:
 (a) nucleobases 5-23 of SEQ ID NO: 179;   (c) nucleobases 7-24 of SEQ ID NO: 192;   (b) nucleobases 1-19 of SEQ ID NO: 196;   (c) nucleobases 1-21 of SEQ ID NO: 197;   (c) nucleobases 1-21 of SEQ ID NO: 198;   (c) nucleobases 1-19 of SEQ ID NO: 199;   (c) nucleobases 3-21 of SEQ ID NO: 200;   
     
     
         35 . The HBV RNAi trigger of  claim 33 , wherein the sense strand comprises a modified nucleotide sequence according to:
 (a) nucleotides 5-23 of SEQ ID NO: 608;   (b) nucleotides 7-24 of SEQ ID NO: 624;   (c) nucleotides 1-19 of SEQ ID NO: 629;   (d) nucleotides 3-21 of SEQ ID NO: 630;   (e) nucleotides 3-20 of SEQ ID NO: 631;   (f) nucleotides 3-20 of SEQ ID NO: 632; or   (g) nucleotides 3-12 of SEQ ID NO: 634.   
     
     
         36 . The HBV RNAi trigger of  claim 32 , wherein the sense strand of the HBV RNAi trigger is covalently linked to a cholesteryl group. 
     
     
         37 . The HBV RNAi trigger of  claim 32 , wherein the sense strand is conjugated to a galactose trimer. 
     
     
         38 . The HBV RNAi trigger of  claim 35 , wherein the sense strand of the HBV RNAi trigger is covalently linked to a cholesteryl group. 
     
     
         39 . The HBV RNAi trigger of  claim 35 , wherein the sense strand is conjugated to a galactose trimer. 
     
     
         40 . A method of treating a subject with an HBV infection, comprising administering to the subject an effective amount of an RNAi agent according to  claim 30 .