Conjugated hepcidin mimetics

1. A hepcidin analogue comprising:

(a) a peptide according to Formula I:

R 1 -Asp-Thr-His-B1-B2-B3-B4-Xaa1-B6-Xaa2-J-Y1-Y2-R 2 (I)

or

(b) a peptide dimer comprising two peptides according to Formula I,

or a pharmaceutically acceptable salt thereof,

wherein:

R 1 is hydrogen, C 1 -C 6 alkyl, C 6 -C 12 aryl, C 6 -C 12 aryl-C 1 -C 6 alkyl, C 1 -C 20 alkanoyl, or C 1 -C 20 cycloalkanoyl;

R 2 is —NH 2 or —OH;

Xaa1 is B5; and

i) B5 is absent, Lys, D-Lys, Orn, homoSer, Gln, (D)Gln, Lys(Ac), Ile, Abu, Leu, Ala, D-Ala, bAla, or Nleu; and

ii) Xaa2 is B7(L1Z);

or

Xaa1 is B5(L1Z); and

i) B5 is Lys, D-Lys, Orn, homoSer, Gln, Lys(Ac), Ile, Abu, Leu, or Nleu; and

ii) Xaa2 is absent or B7;

each of B1 and B6 is independently a-MePhe, NMe-Phe, 2-Nal, 1-Nal, D-1-Nal, D-2-Nal, 3,3-diPhenylGly, Tic, Bip, Trp, bhTrp, hPhe,

substituted Phe, substituted bhPhe, substituted Trp, or substituted bhTrp;

B2 is Pro, D-Pro, bhPro, D-bhPro, NPC, or D-NPC;

B3 is Cys, homoCys, (D)Cys, a-MeCys, or Pen;

B4 is Ile, Val, Leu, or NLeu;

B7 is Glu, Dapa, Lys, D-Lys, homoLys, or a-Me-Lys;

L1 is absent, Dapa, D-Dapa, isoGlu, PEG, Ahx, isoGlu-PEG, PEG-Ahx, isoGlu-Ahx, or isoGlu-PEG-Ahx;

Z is a half-life extension moiety;

J is Pro, -Pro-Arg-, -Pro-Lys-, -Pro-(D)Lys-, -Pro-Arg-Ser-, -Pro-Arg-Ser-Lys-(SEQ ID NO:249), -Pro-Arg-Ser-Lys-Sar-(SEQ ID NO:250), -Pro-Arg-Ser-Lys-Gly-(SEQ ID NO:251), or absent;

Y1 is Cys, homoCys, (D)Cys, NMeCys, aMeCys, or Pen;

Y2 is an amino acid or absent;

substituted Phe is phenylalanine wherein phenyl is substituted with F, Cl, Br, I, OH, methoxy, dimethoxy, dichloro, dimethyl, difluoro, pentafluoro, allyloxy, azido, nitro, 4-carbamoyl-2,6-dimethyl, trifluoromethoxy, trifluoromethyl, phenoxy, benzyloxy, carbamoyl, t-Bu, carboxyl, CN, or guanidine;

substituted bhPhe is b-homophenylalanine wherein phenyl is substituted with F, Cl, Br, I, OH, methoxy, dimethoxy, dichloro, dimethyl, difluoro, pentafluoro, allyloxy, azido, nitro, 4-carbamoyl-2,6-dimethyl, trifluoromethoxy, trifluoromethyl, phenoxy, benzyloxy, carbamoyl, t-Bu, carboxyl, CN, or guanidine;

substituted Trp is N-methyl-L-tryptophan, a-methyltryptophan, or tryptophan substituted with F, Cl, OH, or t-Bu;

substituted bhTrp is N-methyl-L-b-homotryptophan, a-methyl-b-homotryptophan, or b-homotryptophan substituted with F, Cl, OH, or t-Bu;

Peg1 is —C(O)—CH 2 —CH 2 —O—CH 2 —CH 2 —NH—;

Peg2 is —C(O)—CH 2 —CH 2 —O—CH 2 —CH 2 —O—CH 2 —CH 2 —NH—;

Peg4 is —C(O)—CH 2 —CH 2 —(O—CH 2 —CH 2 ) 4 —NH—;

Peg8 is —C(O)—CH 2 —CH 2 —(O—CH 2 —CH 2 ) 8 —NH—; and

Peg11 is —C(O)—CH 2 —CH 2 —(O—CH 2 —CH 2 ) 11 —O—CH 2 —CH 2 —NH—;

wherein

i) the peptide of formula I is optionally PEGylated with Peg1, Peg2, Peg4, Peg8, or Peg11 on one or more R 1 , B1, B2, B3, B4, B5, B6, B7, J, Y1, Y2, or R 2 ; and

ii) the peptide is cyclized via a disulfide bond between B3 and Y1.

2. The hepcidin analogue according to claim 1 , comprising a peptide according to Formula A-I:

R 1 -Asp-Thr-His-B1-B2-B3-B4-B5-B6-B7(L1Z)-J-Y1-Y2-R 2 (A-I)

or a pharmaceutically acceptable salt thereof,

wherein:

R 1 is hydrogen, C 1 -C 6 alkyl, C 6 -C 12 aryl, C 6 -C 12 aryl-C 1 -C 6 alkyl, C 1 -C 20 alkanoyl, or C 1 -C 20 cycloalkanoyl;

R 2 is —NH 2 or —OH;

each of B1 and B6 is independently

a-MePhe, NMe-Phe,

2-Nal, 1-Nal, D-1-Nal, D-2-Nal, 3,3-diPhenylGly, Tic, Bip, Trp, bhTrp, hPhe, Tyr(Me), substituted Phe, substituted bhPhe, substituted Trp, or substituted bhTrp;

B2 is Pro, D-Pro, bhPro, D-bhPro, NPC, or D-NPC;

B3 is Cys, homoCys, (D)Cys, a-MeCys, or Pen;

B4 is Ile, Val, Leu, or NLeu;

B5 is absent, Lys, D-Lys, Orn, homoSer, Gln, (D)Gln, Lys(Ac), Ile, Abu, Leu, Ala, D-Ala, bAla, or Nleu;

B7 is Dapa, Lys, D-Lys, homoLys, or a-Me-Lys; and wherein L1 is attached to N ε of Lys, D-Lys, homoLys, or a-Me-Lys; or N β of Dapa;

L1 is absent, Dapa, D-Dapa, or isoGlu, PEG, Ahx, isoGlu-PEG, PEG-Ahx, isoGlu-Ahx, or isoGlu-PEG-Ahx;

Z is a half-life extension moiety, wherein the half-life extension moiety is C 10 -C 21 alkanoyl;

J is Pro, -Pro-Arg-, -Pro-Lys-, -Pro-(D)Lys-, -Pro-Arg-Ser-, -Pro-Arg-Ser-Lys-(SEQ ID NO:249), -Pro-Arg-Ser-Lys-Sar-(SEQ ID NO:250), -Pro-Arg-Ser-Lys-Gly-(SEQ ID NO:251), or absent;

Y1 is Cys, homoCys, (D)Cys, NMeCys, aMeCys, or Pen;

Y2 is an amino acid or absent;

substituted Phe is phenylalanine wherein phenyl is substituted with F, Cl, Br, I, OH, methoxy, dimethoxy, dichloro, dimethyl, difluoro, pentafluoro, allyloxy, azido, nitro, 4-carbamoyl-2,6-dimethyl, trifluoromethoxy, trifluoromethyl, phenoxy, benzyloxy, carbamoyl, t-Bu, carboxyl, CN, or guanidine;

substituted bhPhe is b-homophenylalanine wherein phenyl is substituted with F, Cl, Br, I, OH, methoxy, dimethoxy, dichloro, dimethyl, difluoro, pentafluoro, allyloxy, azido, nitro, 4-carbamoyl-2,6-dimethyl, trifluoromethoxy, trifluoromethyl, phenoxy, benzyloxy, carbamoyl, t-Bu, carboxyl, CN, or guanidine;

substituted Trp is N-methyl-L-tryptophan, a-methyltryptophan, or tryptophan substituted with F, Cl, OH, or t-Bu;

substituted bhTrp is N-methyl-L-b-homotryptophan, a-methyl-b-homotryptophan, or b-homotryptophan substituted with F, Cl, OH, or t-Bu; and PEG is as described in claim 1 ;

wherein

i) the peptide of formula I is optionally PEGylated on one or more R 1 , B1, B2, B3, B4, B5, B6, J, Y1, Y2, or R 2 ;

ii) the peptide is cyclized via a disulfide bond between B3 and Y1; and

iii) when B6 is Phe, then B5 is other than Lys.

3. The hepcidin analogue according to claim 1 , comprising a peptide according to Formula B-I:

R 1 -Asp-Thr-His-B1-B2-B3-B4-B5(L1Z)-B6-B7-J-Y1-Y2-R 2 (B-I)

or a pharmaceutically acceptable salt thereof,

wherein:

R 1 is hydrogen, C 1 -C 6 alkyl, C 6 -C 12 aryl, C 6 -C 12 aryl-C 1 -C 6 alkyl, C 1 -C 20 alkanoyl, or C 1 -C 20 cycloalkanoyl;

R 2 is —NH 2 or —OH;

each of B1 and B6 is independently

NMe-Phe,

2-Nal, 1-Nal, D-1-Nal, D-2-Nal, 3,3-diPhenylGly, Tic, Bip, Trp, bhTrp, hPhe, Tyr(Me), substituted Phe, substituted bhPhe, substituted Trp, or substituted bhTrp;

B2 is Pro, D-Pro, bhPro, D-bhPro, NPC, or D-NPC;

B3 is Cys, homoCys, (D)Cys, a-MeCys, or Pen;

B4 is Ile, Val, Leu, or NLeu;

B5 is Lys, D-Lys, Orn, homoSer, Gln, Lys(Ac), Ile, Abu, Leu, or Nleu;

B7 is Glu or absent;

L 1 is absent or isoGlu, PEG, Ahx, isoGlu-PEG, PEG-Ahx, isoGlu-Ahx, or isoGlu-PEG-Ahx;

Z is a half-life extension moiety, wherein the half-life extension moiety is C 10 -C 21 alkanoyl;

J is Pro, Arg, Gly, -Pro-Arg-, -Pro-Lys-, -Pro-(D)Lys-, -Pro-Arg-Ser-, -Pro-Arg-Ser-Lys-(SEQ ID NO:249), -Pro-Arg-Ser-Lys-Sar-(SEQ ID NO:250), -Pro-Arg-Ser-Lys-Gly-(SEQ ID NO:251), or absent;

Y1 is Cys, homoCys or Pen;

Y2 is an amino acid or absent;

substituted Phe is phenylalanine wherein phenyl is substituted with F, Cl, Br, I, OH, methoxy, dimethoxy, dichloro, dimethyl, difluoro, pentafluoro, allyloxy, azido, nitro, 4-carbamoyl-2,6-dimethyl, trifluoromethoxy, trifluoromethyl, phenoxy, benzyloxy, carbamoyl, t-Bu, carboxyl, CN, or guanidine;

substituted bhPhe is b-homophenylalanine wherein phenyl is substituted with F, Cl, Br, I, OH, methoxy, dimethoxy, dichloro, dimethyl, difluoro, pentafluoro, allyloxy, azido, nitro, 4-carbamoyl-2,6-dimethyl, trifluoromethoxy, trifluoromethyl, phenoxy, benzyloxy, carbamoyl, t-Bu, carboxyl, CN, or guanidine;

substituted Trp is N-methyl-L-tryptophan, a-methyltryptophan, or tryptophan substituted with F, Cl, OH, or t-Bu;

substituted b-hTrp is N-methyl-L-b-homotryptophan, a-methyl-b-homotryptophan, or b-homotryptophan substituted with F, Cl, OH, or t-Bu; and PEG is as described in claim 1 ;

wherein

i) the peptide of formula I is optionally PEGylated on one or more R 1 , B1, B2, B3, B4, B6, B7, J, Y1, Y2, or R 2 ; and

ii) the peptide is cyclized via a disulfide bond between B3 and Y1; and

iii) when Y1 is Cys, and Y2 is Lys, then J is Pro, Arg, Gly, -Pro-Arg-, -Pro-Arg-Ser-,-Pro-Arg-Ser-Lys-(SEQ ID NO:249), or absent.

4. The hepcidin analogue or pharmaceutically acceptable salt thereof according to claim 1 , wherein —J-Y1-Y2—is —Cys-, -Pro-Cys-, -Lys-Cys-, -(D)Lys-Cys-,-(D)Lys-Pen-,-Dap-Cys-, -Cys-(D)Lys-, -Dap-hCys-, -Pro-Arg-Cys-, -Pro-Arg-Ser-Cys-(SEQ ID NO:253), -Pro-Arg-Ser-Lys-Cys-(SEQ ID NO:254), or —Pro-Arg-Ser-Lys-Sar-Cys-(SEQ ID NO:255).

5. The hepcidin analogue or pharmaceutically acceptable salt thereof according to claim 1 , wherein L1 is a single bond, iso-Glu, Ahx, iso-Glu-Ahx, PEG, PEG-Ahx, iso-Glu-PEG-Ahx; and wherein PEG is PEG1, PEG2, PEG3, PEG4, or PEG11.

6. The hepcidin analogue or pharmaceutically acceptable salt thereof according to claim 1 , wherein Z is Palm.

7. The hepcidin analogue or pharmaceutically acceptable salt thereof according to claim 1 , wherein R 2 is NH 2 .

8. The hepcidin analogue or pharmaceutically acceptable salt thereof according to claim 1 , wherein R 2 is OH.

9. A pharmaceutical composition comprising the hepcidin analogue or pharmaceutically acceptable salt thereof of claim 1 and a pharmaceutically acceptable carrier, excipient, or vehicle.

10. 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 9 .