Compositions and Methods for Inhibiting Viral Replication

Abstract

The present invention relates to a double-stranded ribonucleic acid (dsRNA) having a nucleotide sequence which is less that 30 nucleotides in length and which is substantially identical to at least a part of a 3′-untranslated region (3′-UTR) of a (+) strand RNA virus, such as HCV, as well as pharmaceutical compositions comprising the dsRNA, together with a pharmaceutically acceptable carrier. The pharmaceutical compositions are useful for treating infections and diseases caused by the replication or activity of the (+) strand RNA virus, as well as methods for inhibiting viral replication.

1 . A double-stranded ribonucleic acid (dsRNA) for inhibiting the replication of a (+) strand RNA virus, wherein the dsRNA comprises a sense RNA strand comprising a nucleotide sequence which is substantially identical to at least a part of a 3′-untranslated region (3′-UTR) of the (+) strand RNA virus, and wherein the dsRNA comprises an antisense RNA strand which is complementary to at least a part of the sense RNA strand, and wherein each strand of the dsRNA is 21 to 24 nucleotides in length. 
     
     
         2 . The dsRNA of  claim 1 , wherein the dsRNA comprises a blunt end. 
     
     
         3 . The dsRNA of  claim 1 , wherein the dsRNA comprises two blunt ends. 
     
     
         4 . The dsRNA of  claim 1 , wherein the antisense RNA strand and the sense RNA strand comprise a 3′-terminus and a 5′-terminus, and wherein at least one of said RNA strands comprises a nucleotide overhang of 1 to 3 nucleotides in length. 
     
     
         5 . The dsRNA of  claim 4 , wherein the nucleotide overhang is two nucleotides in length. 
     
     
         6 . The dsRNA of  claim 4 , wherein the nucleotide overhang is on the 3′-terminus of the antisense RNA strand. 
     
     
         7 . The dsRNA of  claim 4 , wherein the dsRNA further comprises a first end and a second end, wherein the first end comprises the 3′-terminus of the antisense RNA strand and the 5′-terminus of the sense RNA strand, and wherein the second end comprises the 5′-terminus of the antisense RNA strand and the 3′-terminus of the sense RNA strand, wherein the first end comprises a nucleotide overhang on the 3′-terminus of the antisense RNA strand, and wherein the second end is blunt. 
     
     
         8 . The dsRNA of  claim 4 , wherein the antisense RNA strand is 23 nucleotides in length and comprises a 2-nucleotide overhang at the 3′-terminus, wherein the sense RNA strand is 23 nucleotides in length and comprises a 2-nucleotide overhang at the 3′-terminus, and wherein the dsRNA has a 21-nucleotide double-stranded region. 
     
     
         9 . A pharmaceutical composition for inhibiting the replication of a (+) strand RNA virus in an organism, comprising a dsRNA and a pharmaceutically acceptable carrier, wherein the dsRNA comprises a sense RNA strand comprising a nucleotide sequence which is substantially identical to at least a part of a 3′-untranslated region (3′-UTR) of the (+) strand RNA virus, and wherein the dsRNA comprises an antisense RNA strand which is complementary to at least a part of the sense RNA strand, and wherein each strand of the dsRNA is 21 to 24 nucleotides in length. 
     
     
         10 . The pharmaceutical composition of  claim 9 , wherein the dsRNA comprises a blunt end. 
     
     
         11 . The pharmaceutical composition of  claim 9 , wherein the dsRNA comprises two blunt ends. 
     
     
         12 . The pharmaceutical composition of  claim 9 , wherein the antisense RNA strand and the sense RNA strand comprise a 3′-terminus and a 5′-terminus, and wherein at least one of said RNA strands comprise a nucleotide overhang of 1 to 3 nucleotides in length. 
     
     
         13 . The pharmaceutical composition of  claim 12 , wherein the nucleotide overhang is two nucleotides in length. 
     
     
         14 . The pharmaceutical composition of  claim 12 , wherein the nucleotide overhang is on the 3′-terminus of the antisense RNA strand. 
     
     
         15 . The pharmaceutical composition of  claim 12 , wherein the dsRNA further comprises a first end and a second end, wherein the first end comprises the 3′-terminus of the antisense RNA strand and the 5′-terminus of the sense RNA strand, and wherein the second end comprises the 5′-terminus of the antisense RNA strand and the 3′-terminus of the sense RNA strand, wherein the first end comprises a nucleotide overhang on the 3′-terminus of the antisense RNA strand, and wherein the second end is blunt. 
     
     
         16 . The pharmaceutical composition of  claim 12 , wherein the antisense RNA strand is 23 nucleotides in length and comprises a 2-nucleotide overhang at the 3′-terminus, wherein the sense RNA strand is 23 nucleotides in length and comprises a 2-nucleotide overhang at the 3′-terminus, and wherein the dsRNA has a 21-nucleotide double-stranded region. 
     
     
         17 . The pharmaceutical composition of  claim 9 , wherein a dosage unit of dsRNA is less than 5 milligram (mg) of dsRNA per kg body weight of the mammal. 
     
     
         18 . The pharmaceutical composition of  claim 9 , wherein a dosage unit of dsRNA is in a range of 0.01 to 2.5 milligrams (mg), 0.1 to 200 micrograms (μg), 0.1 to 100 μg per kilogram body weight of the mammal. 
     
     
         19 . The pharmaceutical composition of  claim 9 , wherein a dosage unit of dsRNA is less than 25 μg per kilogram body weight of the mammal. 
     
     
         20 . The pharmaceutical composition of  claim 9 , wherein the pharmaceutically acceptable carrier is an aqueous solution. 
     
     
         21 . The pharmaceutical composition of  claim 20 , wherein the aqueous solution is phosphate buffered saline. 
     
     
         22 . The pharmaceutical composition of  claim 9 , wherein the pharmaceutically acceptable carrier comprises a micellar structure selected from the group consisting of a liposome, capsid, capsoid, polymeric nanocapsule, and polymeric microcapsule. 
     
     
         23 . The pharmaceutical composition of  claim 9 , which is formulated to be administered by inhalation, infusion, injection, or orally. 
     
     
         24 . The pharmaceutical composition of  claim 9 , which is formulated to be administered by intravenous or intraperitoneal injection.