Analogues of hepcidin mimetics with improved in vivo half lives

Abstract

The present invention provides hepcidin analogues with improved in vivo half-lives, and related pharmaceutical compositions and methods of use thereof.

What is claimed is: 
     
         1 . A hepcidin analogue comprising a polypeptide sequence of Formula (I):
   X—Y  (I)
   or a pharmaceutically acceptable salt or solvate thereof, wherein:   X is a peptide sequence having the formula Xa:   
       
         
           
                 
                 
               
                     
                   (Xa) 
                 
                     
                   (SEQ ID NO: 1) 
                 
                     
                   X1-Thr-His-X4-Pro-X6-X7-X8-Phe-X10 
                 
             
                
                
                
               
            
           
         
         
           wherein 
           X1 is Asp, isoGlu or Ida; 
           X4 is Phe, Phe(4-F), Phe(4-CN), 4-BIP, Phe(4-OCH 3 ), Tyr, Phe(2,3-(OCH 3 ) 2 ), Phe(2,3-Cl 2 ), or Dpa; 
           X6 is Cys or Pen; 
           X7 is any amino acid; 
           X8 is Ile, Leu, Val, nLeu, Lys or Arg; and 
           X10 is Lys, Glu or absent; and 
         
         Y is absent or present; 
         wherein if Y is present, Y is a peptide sequence having the formula Ya: 
       
       
         
           
                 
                 
               
                     
                   (Ya) 
                 
                     
                   (SEQ ID NO: 2) 
                 
                     
                   Y1-Y2-Y3-Y4-Y5-Y6-Y7 
                 
             
                
                
                
               
            
           
         
         
           wherein 
           Y1 is amino acid 
           Y2 is any amino acid 
           Y3 is any amino acid 
           Y4 is any amino acid 
           Y5 is any amino acid; 
           Y6 is Cys or Pen; and 
           Y7 is Lys or absent; and 
         
         wherein the hepcidin analogue comprises a conjugated half-life extension moiety, wherein the half-life extension moiety is optionally conjugated via a linker moiety. 
       
     
     
         2 . The hepcidin analogue of  claim 1 , comprising one of more of the following:
 X1 is Asp;   X4 is Phe or Dpa;   X7 is Ile, Leu, Val, nLeu, or Lys;   X7 is Ile or Lys;   X8 is Lys or Arg;   Y1 is Pro or hPro;   Y1 is Pro;   Y2 is Arg or Lys;   Y2 is Arg;   Y3 is Ser;   Y4 is Lys, Arg or His;   Y4 is Lys; or   Y5 is Gly or Sar.   
     
     
         3 . The hepcidin analogue of  claim 1 , wherein:
 X1 is Asp;   X4 is Phe or Dpa;   X7 is Ile, Leu, Val, nLeu, or Lys;   X8 is Lys or Arg;   Y1 is Pro or hPro;   Y2 is Arg or Lys;   Y3 is Ser;   Y4 is Lys, Arg or His; and   Y5 is Gly or Sar.   
     
     
         4 . The hepcidin analogue of  claim 3 , wherein:
 X7 is Ile or Lys;   Y1 is Pro;   Y2 is Arg; and   Y4 is Lys.   
     
     
         5 . The hepcidin analogue of any one of  claims 1 - 4 , comprising a structure of Formula II:
   R 1 —X-L-Y—R 2   (II)
   or a pharmaceutically acceptable salt or solvate thereof, wherein:   R 1  is hydrogen, a C1-C6 alkyl, a C6-C12 aryl, a C6-C12 aryl C1-C6 alkyl, or a C1-C20 alkanoyl, and including PEGylated versions alone or as spacers of any of the foregoing;   R 2  is OH or NH 2 ;   X is a peptide sequence having the formula Xa;   L is absent, a bond, or a linker moiety; and   Y is absent or present;   provided that if Y is present, Y is a peptide having the formula Ya.   
     
     
         6 . The hepcidin analogue of any one of  claims 1 - 5 , wherein X or Y further comprise one to three additional amino acids at the N-terminus or C-terminus. 
     
     
         7 . The hepcidin analogue of any one of  claims 1 - 6 , wherein Y is present. 
     
     
         8 . The hepcidin analogue of  claim 7 , comprising a disulfide bond between X6 and Y6. 
     
     
         9 . The hepcidin analogue of  claim 7  or  claim 8 , wherein L is a bond. 
     
     
         10 . The hepcidin analogue of any one of  claims 7 - 9 , comprising a half-life extension moiety conjugated to a Lys at X8 or X10. 
     
     
         11 . The hepcidin analogue of any one of  claims 1 - 10 , wherein the hepcidin analogue comprises one of the following structures: 
       
         
           
           
               
               
           
         
       
     
     
         12 . The hepcidin analogue of any one of  claims 1 - 6 , wherein Y is absent. 
     
     
         13 . The hepcidin analogue of  claim 12 , wherein X10 is absent. 
     
     
         14 . A dimer comprising two hepcidin analogues of  claim 12  or  claim 13 , wherein the two polypeptide sequence of Formula I or two structures of Formula II are dimerized via a linker moiety. 
     
     
         15 . The dimer of  claim 14 , wherein the linker moiety is bound to the C-terminus of each hepcidin analogue. 
     
     
         16 . The dimer of  claim 14  or  claim 15 , wherein the half-life extension moiety is conjugated to the linker moiety. 
     
     
         17 . The hepcidin analogue of any one of  claims 1 - 13  or the dimer of any one of  claims 14 - 16 , wherein the linker moiety is selected from IsoGlu, Dapa, PEGn where n=1 to 25, PEG11(40 atoms), OEG, IsoGlu-Ahx, IsoGlu-OEG-OEG, IsoGlu-PEG5, IsoGlu-PEGn where n=1 to 25 βAla-PEG2, and βAla-PEG11(40 atoms). 
     
     
         18 . The hepcidin analogue or dimer of any one of  claims 1 - 17 , wherein the half-life extension moiety is selected from C12 (Lauric acid), C14 (Mysteric acid), C16 (Palmitic acid), C18 (Stearic acid, C20, C12 diacid, C14 diacid, C16 diacid, C18 diacid, C20 diacid, biotin, and isovaleric acid. 
     
     
         19 . The hepcidin analogue or dimer of any one of  claims 1 - 18 , wherein the half-life extension moiety is attached to a linker moiety that is attached to the peptide. 
     
     
         20 . The hepcidin analogue or dimer of any one of  claims 1 - 19 , wherein the half-life extension moiety increases the molecular weight of the hepcidin analogue by about 50 D to about 2 KD. 
     
     
         21 . The hepcidin analogue or dimer of any one of  claims 1 - 20 , wherein the half-life extension moiety increases serum half-life, enhances solubility, and/or improves bioavailability of the hepcidin analogue. 
     
     
         22 . The hepcidin analogue or dimer of any one of  claims 1 - 21 , comprising an isovaleric acid moiety conjugated to the N-terminal Asp residue. 
     
     
         23 . The hepcidin analogue or dimer of any one of  claims 1 - 22 , comprising an amidated C-terminal residue. 
     
     
         24 . The hepcidin analogue or dimer of any one of  claims 1 - 23 , comprises the sequence: 
       
         
           
                 
               
                   (SEQ ID NO: 6) 
                 
                   Asp-Thr-His-Phe-Pro-Cys-Ile-Lys-Phe-Glu-Pro-Arg- 
                 
                   Ser-Lys-Gly-Cys-Lys. 
                 
             
                
                
                
               
            
           
         
       
     
     
         25 . The hepcidin analogue or dimer of any one of  claims 1 - 23 , comprising the sequence: 
       
         
           
                 
               
                   (SEQ ID NO: 5) 
                 
                   Asp-Thr-His-Phe-Pro-Cys-Ile-Lys-Phe-Lys-Pro-Arg- 
                 
                   Ser-Lys-Gly-Cys-Lys. 
                 
             
                
                
                
               
            
           
         
       
     
     
         26 . A polynucleotide encoding a peptide of the hepcidin analogue of any one of  claims 1 - 25 . 
     
     
         27 . A vector comprising the polynucleotide of  claim 26 . 
     
     
         28 . A pharmaceutical composition comprising the hepcidin analogue or dimer of any one of  claims 1 - 25 , the polynucleotide of  claim 26 , or the vector of  claim 27 , and a pharmaceutically acceptable carrier, excipient or vehicle. 
     
     
         29 . A method of binding a ferroportin or inducing ferroportin internalization and degradation, comprising contacting the ferroportin with at least one hepcidin analogue or dimer of any one of  claims 1 - 25  or a composition of  claim 28 . 
     
     
         30 . A method for treating a disease of iron metabolism in a subject in need thereof comprising providing to the subject an effective amount of the pharmaceutical composition of  claim 28 . 
     
     
         31 . The method of  claim 30 , wherein the pharmaceutical composition is provided to the subject by an oral, intravenous, peritoneal, intradermal, subcutaneous, intramuscular, intrathecal, inhalation, vaporization, nebulization, sublingual, buccal, parenteral, rectal, vaginal, or topical route of administration. 
     
     
         32 . The method of  claim 31 , wherein the pharmaceutical composition is provided to the subject by an oral or subcutaneous route of administration. 
     
     
         33 . The method of any one of  claims 30 - 32 , wherein the disease of iron metabolism is an iron overload disease. 
     
     
         34 . The method of any one of  claims 30 - 33 , wherein the pharmaceutical composition is provided to the subject at most twice daily, at most once daily, at most once every two days, at most once a week, or at most once a month. 
     
     
         35 . The method of any one of  claims 30 - 34 , wherein the hepcidin analogue is provided to the subject at a dosage of about 1 mg to about 100 mg. 
     
     
         36 . A device comprising the pharmaceutical composition of  claim 28 , for delivery of the hepcidin analogue to a subject, optionally orally or subcutaneously. 
     
     
         37 . A kit comprising the pharmaceutical composition of  claim 28 , packaged with a reagent, a device, or an instructional material, or a combination thereof.