Compositions and methods for inhibiting expression of the hamp gene

Abstract

The invention relates to a double-stranded ribonucleic acid (dsRNA) for inhibiting the expression of the HAMP gene (HAMP gene), comprising an antisense strand having a nucleotide sequence which is less that 30 nucleotides in length, generally 19-25 nucleotides in length, and which is substantially complementary to at least a part of the HAMP gene. The invention also relates to a pharmaceutical composition comprising the dsRNA together with a pharmaceutically acceptable carrier; methods for treating diseases caused by HAMP gene expression and the expression of the HAMP gene using the pharmaceutical composition.

1 . A double-stranded ribonucleic acid (dsRNA) for inhibiting the expression of a human HAMP gene in a cell, 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 HAMP, and wherein said region of complementarity is less than 30 nucleotides in length and wherein said dsRNA, upon contact with a cell expressing said HAMP, inhibits expression of said HAMP gene by at least 40%. 
     
     
         2 . The dsRNA of  claim 1 , wherein said first sequence is selected from the group consisting SEQ ID NOs:1-36 and said second sequence is selected from the group consisting of SEQ ID NOs:37-72. 
     
     
         3 . The dsRNA of  claim 1 , wherein said dsRNA comprises at least one modified nucleotide. 
     
     
         4 . The dsRNA of  claim 2 , wherein said dsRNA comprises at least one modified nucleotide. 
     
     
         5 . The dsRNA of  claim 4 , wherein said modified nucleotide 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. 
     
     
         6 . The dsRNA of  claim 4 , 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. 
     
     
         7 . The dsRNA of  claim 4 , wherein said first sequence is selected from the group consisting of SEQ ID NOs: 217-233 and said second sequence is selected from the group consisting of SEQ ID NOs:234-250. 
     
     
         8 . A cell comprising the dsRNA of  claim 1 . 
     
     
         9 . A pharmaceutical composition for inhibiting the expression of the HAMP gene in an organism, comprising a dsRNA and a pharmaceutically acceptable carrier, wherein the 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 HAMP, and wherein said region of complementarity is less than 30 nucleotides in length and wherein said dsRNA, upon contact with a cell expressing said HAMP, inhibits expression of said HAMP. 
     
     
         10 . The pharmaceutical composition of  claim 9 , wherein said first sequence of said dsRNA is selected from the group consisting of SEQ ID NOs:1-36 and 217-233 and said second sequence is selected from the group consisting of SEQ ID NOs:37-72 and 234-250. 
     
     
         11 . A method for inhibiting the expression of the HAMP gene in a cell, the method comprising:
 (a) introducing into the cell a double-stranded ribonucleic acid (dsRNA), wherein the 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 HAMP, and wherein said region of complementarity is less than 30 nucleotides in length and wherein said dsRNA, upon contact with a cell expressing said HAMP, inhibits expression of said HAMP gene by at least 40%; and   (b) maintaining the cell produced in step (a) for a time sufficient to obtain degradation of the mRNA transcript of the HAMP gene, thereby inhibiting expression of the HAMP gene in the cell.   
     
     
         12 . A method of treating, preventing or managing pathological processes which can be mediated by down regulating HAMP gene expression comprising administering to a patient in need of such treatment, prevention or management a therapeutically or prophylactically effective amount of a dsRNA, wherein the 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 HAMP, and wherein said region of complementarity is less than 30 nucleotides in length and wherein said dsRNA, upon contact with a cell expressing said HAMP, inhibits expression of said HAMP gene. 
     
     
         13 . A vector for inhibiting the expression of the HAMP gene in a cell, said vector comprising a regulatory sequence operably linked to 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 a mRNA encoding HAMP and wherein said dsRNA is less than 30 base pairs in length and wherein said dsRNA, upon contact with a cell expressing said HAMP, inhibits the expression of said HAMP gene by at least 40%. 
     
     
         14 . A cell comprising the vector of  claim 13 . 
     
     
         15 . A double-stranded ribonucleic acid (dsRNA) for reducing the expression level of a human HAMP gene in a cell, 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 HAMP, and wherein said dsRNA, upon contact with a cell expressing said HAMP, reduces the expression level of said HAMP gene. 
     
     
         16 . The dsRNA of  claim 15 , wherein said contact reduces the expression level of said HAMP gene by at least 40%. 
     
     
         17 . The dsRNA of  claim 15 , wherein said contact is performed in vitro at 30 nM or less. 
     
     
         18 . A pharmaceutical composition for reducing the expression level of the HAMP gene in an organism, comprising the dsRNA of  claim 15  and a pharmaceutically acceptable carrier. 
     
     
         19 . A method of treating an HAMP associated disorder comprising administering to a patient in need of such treatment, a therapeutically effective amount of a dsRNA of  claim 15 . 
     
     
         20 . A method of treating an HAMP-associated disorder comprising administering to a patient in need of such treatment, a therapeutically effective amount of a dsRNA of  claim 15 .