Hepcidin Binding Nucleic Acids

Abstract

The present invention is related to a nucleic acid capable of binding to hepcidin.

1 - 99 . (canceled) 
     
     
         100 . An L-nucleic acid that binds hepcidin, wherein the L-nucleic acid comprises in 5→43′ direction a first terminal stretch of nucleotides, a central stretch of nucleotides and a second terminal stretch of nucleotides, wherein the central stretch of nucleotides comprises
 5′ RKAUGGGAKUAAGUAAAUGAGGRGUWGGAGGAAR 3′ [SEQ II) NO: 182] or 
 5′ RKAUGGGAKAAGUAAAUGAGGRGUWGGAGGAAR 3′ [SEQ ID NO:183], and 
 
       the first terminal stretch of nucleotides comprises five to eight nucleotides, and the second terminal stretch of nucleotides comprises five to eight nucleotides. 
     
     
         101 . The L-nucleic acid according to  claim 100 , wherein the nucleic acid is an antagonist of hepcidin. 
     
     
         102 . The L-nucleic acid according to  claim 100 , wherein the nucleic acid is an inhibitor of the hepcidin-ferroportin system. 
     
     
         103 . The L-nucleic acid according to  claim 100 , wherein the central stretch of nucleotides is essential for binding to hepcidin. 
     
     
         104 . The L-nucleic acid according to  claim 100 , wherein the central stretch of nucleotides comprises 5′ RKAUGGGAKUAAGUAAAUGAGGRGUUGGAGGAAR 3′ [SEQ ID NO:213]. 
     
     
         105 . The L-nucleic acid according to  claim 100 , wherein the first terminal stretch of nucleotides and the second terminal stretch of nucleotides optionally hybridize with each other to form a double-stranded structure. 
     
     
         106 . The L-nucleic acid molecule according to  claim 100 , wherein the first terminal stretch of nucleotides comprises 5 X 1 X 2 X 3 SBSBC3′ and the second terminal stretch of nucleotides comprises 5′ GVBVBX 4 X 5 X 6  3′, wherein
 a) X 1  is A, X 2  is G, X 3  is B, X 4  is S, X 5  is C, and X 6 ; is U or 
 b) X 1  is absent, X 2  is G, X 3  is B, X 4  is S, X 5  is C, and X 6  is U or 
 c) X 1  is A, X 2  is G, X 3  is B, X 4  is S, X 5  is C, and X 6  is absent. 
 
     
     
         107 . The L-nucleic acid molecule according to  claim 100 , wherein the first terminal stretch of nucleotides comprises 5′ X 1 X 2 X 3 SBSBC3′ and the second terminal stretch of nucleotides comprises 5′ GVBVYX 4 X 5 X 6  3′, wherein
 a) X 1  is absent, X 2  is G, X 3  is B, X 4  is S, X 5  is C, and X 6  is absent or 
 b) X 1  is absent, X 2  is absent, X 3  is B, X 4  is S, X 5  is C, and X 6  is absent or 
 c) X 1  is absent, X 2  is G, X 3  is B, X 4  is S, X 5  is absent, and X 6  is absent. 
 
     
     
         108 . The L-nucleic acid molecule according to  claim 100 , wherein the first terminal stretch of nucleotides comprises 5′ X 1 X 2 X 3 SBSBC3′ and the second terminal stretch of nucleotides comprises 5′ GVBVYX 4 X 5 X 6  3′, wherein X 1  is absent, X 2  is absent, X 3  is B or absent, X 4  is S or absent, X 5  is absent, and X 6  is absent. 
     
     
         109 . The L-nucleic acid according to  claim 100 , wherein the nucleic acid comprises a nucleic acid sequence according to any one of SEQ ID NOs:115 to 119, SEQ ID NO:121, SEQ. ID NO:142, SEQ ID NO:144, SEQ ID NO:146, SEQ ID NO:148, SEQ ID NO:151, SEQ ID NO:152, SEQ ID NO:175 or SEQ ID NO:176. 
     
     
         110 . The L-nucleic acid according to  claim 100 , wherein hepcidin is selected from the group consisting of human hepcidin-25, human hepcidin-22, human hepcidin-20, monkey hepcidin-25, monkey hepcidin-22 and monkey hepcidin-20. 
     
     
         111 . The L-nucleic acid according to  claim 100 , wherein the L-nucleic acid binds human hepcidin-25. 
     
     
         112 . The L-nucleic acid according to  claim 100 , comprising a modification group. 
     
     
         113 . The nucleic acid according to  claim 112 , wherein the modification group is selected from the group consisting of linear polyethylene glycol (PEG), branched PEG, hydroxyethyl starch (HES), a peptide, a protein, a polysaccharide, a sterol, polyoxypropylene, polyoxyamidate and poly (2-hydroxyethyl)-L-glutamine. 
     
     
         114 . The L-nucleic acid according to  claim 112 , wherein the modification group comprises a straight or branched PEG. 
     
     
         115 . The L-nucleic acid according to  claim 114 , wherein the PEG is from about 20,000 to about 120,000 Da. 
     
     
         116 . The L-nucleic acid according to  claim 112 , wherein the modification group is a HES comprising a molecular weight of from about 10,000 to about 200,000 Da. 
     
     
         117 . The L-nucleic acid according to  claim 112 , wherein the modification group is coupled to the nucleic acid via a linker. 
     
     
         118 . The nucleic acid according, to  claim 112 , wherein the modification group is coupled to the 5′-terminal nucleotide and/or the 3′-terminal nucleotide of the nucleic acid. 
     
     
         119 . A pharmaceutical composition comprising the L-nucleic acid of  claim 100  and optionally a further constituent selected from the group consisting of pharmaceutically acceptable excipients, pharmaceutically acceptable carriers and pharmaceutically active agents. 
     
     
         120 . A method of treating a disease or condition associated with hepcidin in a subject suspected of having said disease or condition, wherein the method comprises exposing the subject to the L-nucleic acid of  claim 100 . 
     
     
         121 . The method according to  claim 120 , wherein the disease or condition is selected from the group consisting of anemia, hypoferremia, pica, a condition with elevated hepcidin level, a condition with elevated iron level and a condition with iron overload. 
     
     
         122 . The method according to  claim 121 , wherein the anemia is selected from the group consisting of sideroblastic anemia, hypochromic microcytic anemia, anemia caused by chronic disease and/or disorder, anemia caused by inflammation, anemia caused by genetic disorders, anemia caused by acute infections, anemia caused by mutation in genes of iron and in homeostasis and anemia caused by cancer treatment. 
     
     
         123 . A complex comprising the L-nucleic acid according to  claim 100  and hepcidin. 
     
     
         124 . A method for detecting hepcidin comprising exposing a sample suspected of comprising hepcidin with the L-nucleic acid according to  claim 100  and determining presence of complexes of said L-nucleic acid and hepcidin, wherein presence of complexes correlates with presence of hepcidin in said sample. 
     
     
         125 . A method for identifying an antagonist or an agonist of hepcidin comprising:
 providing a candidate antagonist and/or a candidate agonist of hepcidin,   providing the L-nucleic acid according to  claim 100 ,   providing a test system which provides a signal in the presence of an antagonist and/or an agonist of hepcidin, and   determining whether the candidate antagonist is an antagonist of hepcidin and/or whether the candidate agonist is an agonist of hepcidin.   
     
     
         126 . A method for the detection of the L-nucleic acid according to  claim 100  in a sample, wherein the method comprises the steps of:
 a) providing a sample containing the L-nucleic acid according to  claim 100 ; 
 b) providing a capture probe, wherein the capture probe is at least partially complementary to a first part of the L-nucleic acid according to any one of  claim 100 , and a detection probe, wherein the detection probe is at least partially complementary to a second part of the L-nucleic acid according to  claim 100 , or, alternatively, the capture probe is at least partially complementary to a second part of the L-nucleic acid according to  claim 100  and the detection probe is at least partially complementary to the first part of the L-nucleic acid according to  claim 100 ; 
 c) allowing the capture probe and the detection probe to react either simultaneously or in any order with the L-nucleic acid according to  claim 100  or part thereof to form a complex; 
 d) optionally detecting Whether or not the capture probe is hybridized to the L-nucleic acid according to  claim 100  provided in step a); and 
 e) detecting the complex formed in step c) consisting of the L-nucleic acid according to  claim 100  the capture probe and the detection probe.