Mini-Hepcidin Peptides and Methods of Using Thereof

Abstract

Disclosed herein are peptides which exhibit hepcidin activity and methods of making and using thereof.

1 . An isolated peptide having the following structural formula
   A1-A2-A3-A4-A5-A6-A7-A8-A9-A10   
       wherein
 A1 is Asp, Glu, pyroglutamate, Gln, Asn, or an unnatural amino acid commonly used as a substitute thereof; 
 A2 is Thr, Ser, Val, Ala, or an unnatural amino acid commonly used as a substitute thereof; 
 A3 is His, Asn, Arg, or an unnatural amino acid commonly used as a substitute thereof; 
 A4 is Phe, Leu, Ile, Trp, Tyr, or an unnatural amino acid commonly used as a substitute thereof which includes cyclohexylalanine; 
 A5 is Pro, Ser, or an unnatural amino acid commonly used as a substitute thereof; 
 A6 is Ile, Leu, Val, or an unnatural amino acid commonly used as a substitute thereof; 
 A7 is Cys, Ser, Ala, or an unnatural amino acid commonly used as a substitute thereof which includes S-tertiary butyl-cysteine; 
 A8 is Ile, Leu, Thr, Val, Arg, or an unnatural amino acid commonly used as a substitute thereof; 
 A9 is Phe, Leu, Ile, Tyr, or an unnatural amino acid commonly used as a substitute thereof which includes cyclohexylalanine; and 
 A10 is Cys, Ser, Ala, or an unnatural amino acid commonly used as a substitute thereof; 
 wherein the carboxy-terminal amino acid is in amide or carboxy-form; 
 wherein at least one sulfhydryl amino acid is present as one of the amino acids in the sequence; and 
 wherein A1, A2, A3, A1 to A2, A1 to A3, A10, A9 to A10, A8 to A10, or a combination thereof are optionally absent. 
 
     
     
         2 . The peptide of  claim 1 , wherein
 A1 is D-Asp, D-Glu, D-pyroglutamate, D-Gln, D-Asn, bhAsp, Ida, or N-MeAsp;   A2 is D-Thr, D-Ser, D-Val, Tle, Inp, Chg, bhThr, or N-MeThr;   A3 is D-His, D-Asn, DArg, Dpa, (D)Dpa, or 2-aminoindan;   A4 is D-Phe, D-Leu, D-Ile, D-Trp, Phg, bhPhe, Dpa, Bip, 1Nal, bhDpa, Amc, PheF5, hPhe, Igl, or cyclohexylalanine;   A5 is D-Pro, D-Ser, Oic, bhPro, trans-4-PhPro, cis-4-PhPro, cis-5-PhPro, Idc;   A6 is D-Ile, D-Leu, Phg, Chg, Amc, bhIle, Ach, and MeIle;   A7 is D-Cys, D-Ser, D-Ala, Cys(S-tBut), homoC, Pen, (D)Pen, Dap(AcBr), and Inp;   A8 is D-Ile, D-Leu, D-Thr, D-Val, D-Arg, Chg, Dpa, bhIle, Ach, or MeIle;   A9 is D-Phe, D-Leu, D-Ile, PheF5, N-MePhe, benzylamide, bhPhe, Dpa, Bip, 1Nal, bhDpa, cyclohexylalanine; or   A10 is D-Cys, D-Ser, D-Ala;   
       or a combination thereof. 
     
     
         3 . The peptide of  claim 1 , wherein
 A1 is Ala, D-Ala, Cys, D-Cys, Phe, D-Phe, Asp or D-Asp linked to Cys or D-Cys, Phe or D-Phe linked to a PEG molecule linked to chenodeoxycholate, ursodeoxycholate, or palmitoyl, or Dpa or (D)Dpa linked to palmitoyl;   A2 is Ala, D-Ala, Cys, D-Cys, Pro, D-Pro, Gly, or D-Gly;   A3 is Ala, D-Ala, Cys, D-Cys, Dpa, Asp or D-Asp linked to Dpa or (D)Dpa;   A4 is Ala, D-Ala, Pro, or D-Pro;   A5 is Ala, D-Ala, Pro, D-Pro, Arg, D-Arg;   A6 is Ala, D-Ala, Phe, D-Phe, Arg, D-Arg, Cys, D-Cys;   A7 is His, or D-His;   A8 is Cys, or D-Cys; or   A9 is Phe or D-Phe linked to RA, Asp, D-Asp, Asp or D-Asp linked to RB, bhPhe linked to RC, or cysteamide, wherein RA is —CONH 2 —CH 2 —CH 2 —S, -D-Pro linked to Pro-Lys or Pro-Arg, -bhPro linked to Pro linked to Pro-Lys or Pro-Arg, -D-Pro linked to bhPro-Lys or bhPro-Arg, wherein RB is -PEG11-GYIPEAPRDGQAYVRKDGEWVLLSTFL, -(PEG11)-(GPHyp)10, and wherein RC is -D-Pro linked to Pro-Lys or Pro-Arg, -D-Pro linked to bhPro-Lys or bhPro-Arg;   
       or a combination thereof. 
     
     
         4 . The peptide of  claim 1 , wherein A1 is Asp; A2 is Thr; A3 is His; A4 is Phe; A5 is Pro; A6 is Ile; A7 is Ala; A8 is Ile; A9 is Phe; and A10 is Cys in amide form; wherein A1 or A1 to A2 are optionally absent. 
     
     
         5 . The peptide of  claim 1 , wherein A1 is Asp, A2 is Thr, A3 is His, A4 is Phe, A5 is Pro, A6 is Ile, A7 is Cys or an unnatural thiol amino acid, A8 is Ile, A9 is Phe in amide form, and A10 is absent. 
     
     
         6 . The peptide of  claim 1 , wherein A1 and A2 are absent, A3 is His, A4 is Phe, A5 is Pro, A6 is Ile, A7 is Cys or an unnatural thiol amino acid, A8 is Ile in amide form, and A9 and A10 are absent. 
     
     
         7 . The peptide of  claim 1 , wherein A1 and A2 are absent, A3 is His, A4 is Phe, A5 is Pro, A6 is Ile, A7 is Cys or an unnatural thiol amino acid in amide form, and A8 to A10 are absent. 
     
     
         8 . The peptide of  claim 1 , wherein the peptide is a cyclic peptide. 
     
     
         9 . The peptide of according to  claim 1 , wherein the sequence is retroinverted such that A1 is the C-terminus and A10 is the N-terminus. 
     
     
         10 . The peptide according to  claim 1 , wherein the peptide has an addition at the N-terminus, C-terminus, or both. 
     
     
         11 . The peptide according to  claim 1 , wherein the peptide is selected from the group consisting of: Hep3-8, Hep3-9, Hep1-8, Hep1-9, Hep1-10 C7A, Hep9F4A, Hep9C7-SStBut, (D)C, homoC, Pen, (D)Pen, Cyc-1, Pr10, Pr11, Pr12, riHep7ΔDT, Pr23, Pr24, Pr25, Pr27, Pr28, F4bhPhe, F4Dpa, F4Bip, F4 1Nal, F4bhDpa, F9bhPhe, F9Dpa, F9Bip, F91Nal, F9bhDpa, Pr39, Pr40, Pr41, Pr42, Pr43, Pr44, Pr45, Pr46, Pr13, Pr14, Pr15, Pr16, Pr17, Pr18, Pr19, Pr20, Pr21, Pr22, Pr-1, Pr-2, Pr-3, and Pr-4. 
     
     
         12 . The peptide according to  claim 1 , wherein the peptide exhibits hepcidin activity. 
     
     
         13 . The peptide according to  claim 1 , wherein the peptide binds ferroportin. 
     
     
         14 . A composition which comprises at least one peptide according to  claim 1 . 
     
     
         15 . A method of binding a ferroportin or inducing ferroportin internalization and degradation which comprises contacting the ferroportin with at least one peptide according to any one of  claim 1  or the composition according to  claim 14 . 
     
     
         16 . A method of treating a disease of iron metabolism in a subject which comprises administering at least one peptide according to  claim 1  or the composition according to  claim 14  to the subject. 
     
     
         17 . The method of  claim 16 , wherein the disease of iron metabolism is an iron overload disease. 
     
     
         18 . A kit comprising at least one peptide according to  claim 1  or the composition according to  claim 14  packaged together with a reagent, a device, instructional material, or a combination thereof. 
     
     
         19 . A complex comprising at least one peptide according to  claim 1  bound to a ferroportin or an antibody. 
     
     
         20 . (canceled)