Hepcidin, hepcidin antagonists and methods of use

Abstract

The invention relates to purified, correctly folded hepcidin, antibodies that bind hepcidin, and methods of making and using such materials. Also provide are methods of treated hepcidin-related disorders.

What is claimed is: 
     
         1 . An isolated monoclonal antibody which specifically binds to human hepcidin, wherein said human hepcidin consists of the amino acid sequence set forth in SEQ ID NO: 9 and has a conformation comprising four disulfide-bond loops formed between residues 7 and 23, 10 and 13, 11 and 19, and 14 to 22 as located in SEQ ID NO: 9. 
     
     
         2 - 132 . (canceled) 
     
     
         133 . A composition comprising the antibody of  claim 1  and pharmaceutically acceptable carrier, diluent or excipient. 
     
     
         134 . A method for detecting the presence of human hepcidin in a biological sample comprising: incubating the sample with the antibody of  claim 1  under conditions that allow binding of the monoclonal antibody to mature human hepcidin of SEQ ID NO: 9; and detecting the bound monoclonal antibody or bound hepcidin. 
     
     
         135 . (canceled) 
     
     
         136 . The method of  claim 134 , further comprising incubating the sample with a second monoclonal antibody that binds mature hepcidin of SEQ ID NO: 9. 
     
     
         137 - 171 . (canceled) 
     
     
         172 . An immunoassay kit for detecting human hepcidin in a biological sample, the kit comprising the antibody of  claim 1  and, optionally purified hepcidin as a standard. 
     
     
         173 . An immunoassay kit for detecting human hepcidin in a biological sample, comprising:
 (a) a capture agent comprising an antibody that specifically binds to human hepcidin consisting of the amino acid sequence set forth in SEQ ID NO: 9; and   (b) a detection reagent comprising the antibody of  claim 1  that binds to said human hepcidin at an epitope distinct from that recognized by the antibody in (a).   
     
     
         174 . (canceled) 
     
     
         175 . An immunoassay kit for detecting human hepcidin in a biological sample, comprising:
 (a) a detection reagent comprising an antibody that specifically binds to human hepcidin consisting of the amino acid sequence set forth in SEQ ID NO:9; and   (b) a capture reagent comprising the antibody of  claim 1  that binds to said human hepcidin at an epitope distinct from that recognized by the antibody in (a).   
     
     
         176 - 178 . (canceled) 
     
     
         179 . A method of diagnosing a hepcidin-related disorder, comprising:
 (a) contacting a biological sample from a human suspected of having said disorder with the antibody of  claim 1  under conditions that allow binding of the antibody to human hepcidin; and   (b) detecting and/or quantitating the hepcidin bound to the antibody,   wherein the amount of hepcidin in said sample, as quantitated in (b), above a threshold level indicates the presence of hepcidin-related disorder and below the threshold level indicates the absence of hepcidin-related disorder.   
     
     
         180 . (canceled) 
     
     
         181 . The method according to  claim 179 , wherein said disorder is selected from the group consisting of: anemia, sepsis, anemia of inflammation, anemia of cancer, chemotherapy induced anemia, chronic inflammatory anemia, congestive heart failure, end stage renal disorder, chronic kidney disease (stage I, II, III, IV or V), iron deficiency anemia, a disorder of iron homeostasis, ferroportin disease, hemochromatosis, diabetes, inflammation, rheumatoid arthritis, arteriosclerosis, tumors, vasculitis, systemic lupus erythematosus, hemoglobinopathies, and red cell disorders. 
     
     
         182 - 184 . (canceled) 
     
     
         185 . A method of monitoring a treatment in which a hepcidin antagonist is administered, comprising:
 (a) contacting a biological sample, from a human that has been administered a hepcidin antagonist, with the antibody of  claim 1  under conditions that allow binding of the antibody to human hepcidin; and   (b) detecting and/or quantitating the hepcidin bound to the antibody,   wherein the amount of hepcidin, as quantitated in (b), above a threshold level indicates that the dose of hepcidin antagonist is not therapeutically effective and below said threshold level indicates that the dose of hepcidin antagonist is therapeutically effective.   
     
     
         186 - 191 . (canceled) 
     
     
         192 . A vial or prefilled syringe comprising the composition of  claim 1 . 
     
     
         193 . A method of treating a mammal in need thereof comprising administering to said mammal (a) an antibody that binds human hepcidin consisting of SEQ ID NO: 9 with a K D  of about 10 −7  M or less, and (b) an erythropoiesis stimulator selected from the group consisting of erythropoietin, erythropoietin variants and antibodies that bind erythropoietin. 
     
     
         194 - 196 . (canceled) 
     
     
         197 . A method of treating a mammal that is hypo-responsive to therapy with an erythropoiesis stimulator, comprising administering an antibody that binds human hepcidin consisting of SEQ ID NO: 9 with a K D  of about 10 −7 M or less. 
     
     
         198 - 204 . (canceled) 
     
     
         205 . A kit for treating a disorder associated with elevated hepcidin levels or a disorder of iron homeostasis, comprising the antibody of  claim 1  and an erythropoiesis stimulator. 
     
     
         206 - 209 . (canceled) 
     
     
         210 . A kit for treating a disorder associated with elevated hepcidin levels or a disorder of iron homeostasis, comprising the antibody of  claim 1  and a label attached to or packaged with the container, the label describing use of the antibody with an erythropoiesis stimulator. 
     
     
         211 - 236 . (canceled) 
     
     
         237 . An isolated monoclonal antibody which specifically binds to human hepcidin, wherein said human hepcidin consists of the amino acid sequence set forth in SEQ ID NO: 9 and has a conformation comprising at least one disulfide-bond loop formed between residues 10 and 13 and 14 to 22 as located in SEQ ID NO: 9.