Compositions and methods for inhibiting expression of mutant egfr gene

Abstract

The invention relates to a double-stranded ribonucleic acid (dsRNA) targeting a mutant Epidermal Growth Factor Receptor (EGFR), and methods of using the dsRNA to inhibit expression of mutant EGFR.

1 . A double-stranded ribonucleic acid (dsRNA), wherein said dsRNA comprises at least two sequences that are complementary to each other and wherein a sense strand comprises a first sequence and an antisense strand comprises a second sequence comprising a region of complementarity which is substantially complementary to at least a part of a mRNA encoding an delta-Epidermal Growth Factor Receptor (deltaEGFR), and wherein said region of complementarity is less than 30 nucleotides in length. 
     
     
         2 . The dsRNA of  claim 1 , wherein said dsRNA comprises at least one modified nucleotide. 
     
     
         3 . The dsRNA of  claim 2 , wherein at least one of said modified nucleotides is chosen from the group of: a 2′-O-methyl modified nucleotide, a nucleotide comprising a 5′-phosphorothioate group, and a terminal nucleotide linked to a cholesteryl derivative or dodecanoic acid bisdecylamide group. 
     
     
         4 . The dsRNA of  claim 2 , wherein said modified nucleotide is chosen from the group of: a 2′-deoxy-2′-fluoro modified nucleotide, a 2′-deoxy-modified nucleotide, a locked nucleotide, an abasic nucleotide, 2′-amino-modified nucleotide, 2′-alkyl-modified nucleotide, morpholino nucleotide, a phosphoramidate, and a non-natural base comprising nucleotide. 
     
     
         5 . The dsRNA of  claim 1 , wherein the region of complementary is at least 15 nucleotides in length or is between 19 and 21 nucleotides in length. 
     
     
         6 . (canceled) 
     
     
         7 . The dsRNA of  claim 1 , wherein the dsRNA comprises a sense strand consisting of a sense strand sequence selected from Tables 2 and 3, and an antisense strand consisting of an antisense sequence selected from Tables 2 and 3. 
     
     
         8 . The dsRNA of  claim 1 , wherein the sense strand and the antisense strand are also substantially complementary to at least a part of an mRNA encoding a wildtype Epidermal Gro The dsRNA of  claim 1 , wherein the sense strand comprises a nucleotide sequence of SEQ ID NO:2, and the antisense strand comprises a nucleotide sequence of SEQ ID NO:3. 
     
     
         9 . The dsRNA of  claim 1 , wherein the sense strand comprises a nucleotide sequence of SEQ ID NO:2, and the antisense strand comprises a nucleotide sequence of SEQ ID NO:3. 
     
     
         10 . A cell containing the dsRNA of  claim 1 . 
     
     
         11 . A pharmaceutical composition for inhibiting expression of a deltaEGFR gene comprising the dsRNA of  claim 1 . 
     
     
         12 . The pharmaceutical composition of  claim 11 , further comprising a dsRNA comprising at least two sequences that are complementary to each other and wherein a sense strand comprises a first sequence and an antisense strand comprises a second sequence which is substantially complementary to at least a part of an mRNA encoding an Interleukin-6 (IL6), and wherein said region of complementarity is less than 30 nucleotides in length. 
     
     
         13 . The pharmaceutical composition of  claim 12 , wherein the IL6 dsRNA comprises a sense strand comprising a sequence selected from Tables 5-8, and an antisense sequence comprising an antisense sequence selected from Tables 5-8. 
     
     
         14 . A method of inhibiting deltaEGFR expression in a cell, the method comprising:
 (a) introducing into the cell the dsRNA of  claim 1 ; and   (b) maintaining the cell produced in step (a) for a time sufficient to obtain degradation of the mRNA transcript of the deltaEGFR gene, thereby inhibiting expression of the deltaEGFR gene in the cell.   
     
     
         15 . A method of treating a disorder mediated by deltaEGFR expression comprising administering to a human in need of such treatment a therapeutically effective amount of the dsRNA of  claim 1 . 
     
     
         16 . The method of  claim 15 , wherein the human has cancer, a tumor, an astrocytic tumor, or a glioma. 
     
     
         17 . (canceled) 
     
     
         18 . (canceled) 
     
     
         19 . (canceled) 
     
     
         20 . The method of  claim 15 , further comprising administering a therapeutically effective amount of a dsRNA comprising at least two sequences that are complementary to each other and wherein a sense strand comprises a first sequence and an antisense strand comprises a second sequence which is substantially complementary to at least a part of an mRNA encoding an IL6, and wherein said region of complementarity is less than 30 nucleotides in length. 
     
     
         21 . A vector comprising a nucleotide sequence that encodes at least one strand of a dsRNA, wherein one of the strands of said dsRNA is substantially complementary to at least a part of an mRNA encoding a deltaEGFR and wherein said dsRNA is less than 30 base pairs in length. 
     
     
         22 . The vector of  claim 21 , wherein the region of complementarity is at least 15 nucleotides in length or is 19 to 21 nucleotides in length. 
     
     
         23 . (canceled) 
     
     
         24 . A cell comprising the vector of  claim 20 . 
     
     
         25 . A double-stranded ribonucleic acid (dsRNA), wherein said dsRNA comprises at least two sequences that are complementary to each other and wherein a sense strand comprises a first sequence and an antisense strand comprises a second sequence comprising a region of complementarity which is substantially complementary to at least a part of a mRNA encoding IL-6, and wherein said region of complementarity is less than 30 nucleotides in length. 
     
     
         26 . The dsRNA of  claim 25 , wherein the IL6 dsRNA comprises a sense strand comprising a sequence selected from Tables 5-8, and an antisense sequence comprising an antisense sequence selected from Tables 5-8. 
     
     
         27 . A method of treating a disorder mediated by IL-6 expression comprising administering to a human in need of such treatment a therapeutically effective amount of the dsRNA of  claim 25 .