Modified Mini-Hepcidin Peptides and Methods of Using Thereof

• US 20140336110A1
• Filed: 12/06/2012
• Published: 11/13/2014
• Est. Priority Date: 12/05/2008
• Status: Active Grant

• Current Assignee: Regents of the University of California (University of California)
• Original Assignee: Regents of the University of California (University of California)
• Inventors: Ganz, Tomas, Nemeth, Elizabeta, Ruchala, Piotr P.
• Granted Patent: US 9,328,140 B2
• Time in Patent Office: Days
• US Class Current: 514/5.4
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 38/04 Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof enzyme inhibitors A61K38/005; gastrins A61K38/2207, somatostatins A61K38/31, melanotropins A61K38/34 ; protease inhibitors A61K38/55
• CPC Class Codes: A61P 1/06 Anti-spasmodics, e.g. drugs for colics, esophagic dyskinesia
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 19/08 for bone diseases, e.g. rachitism, Paget's disease
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/14 for treating abnormal movements, e.g. chorea, dyskinesia
• CPC Class Codes: A61P 25/16 Anti-Parkinson drugs
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 3/04 Anorexiants; Antiobesity agents
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 5/50 for increasing or potentiating the activity of insulin
• CPC Class Codes: A61P 7/00 Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: A61P 7/06 Antianaemics
• CPC Class Codes: A61P 9/04 Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C07K 14/47 from mammals
• CPC Class Codes: C07K 14/575 : Hormones derived from pro-opiomelanocortin, pro-enkephalin or pro-dynorphin C07K14/665, e.g. corticotropin C07K14/695
• CPC Class Codes: C07K 16/00 : Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies antibodies with enzymatic activity, e.g. abzymes C12N9/0002
• CPC Class Codes: C07K 7/06 : having 5 to 11 amino acids

Abstract

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

Claims

1. An isolated peptide having the following structural formula IA or IB

A1-A2-A3-A4-A5-A6-A7-A8-A9-A10 

 

IA

A10-A9-A8-A7-A6-A5-A4-A3-A2-A1 

 

IB

wherein

A1 is Asp, D-Asp, Glu, D-Glu, pyroglutamate, D-pyroglutamate, Gln, D-Gln, Asn, D-Asn, or an unnatural amino acid commonly used as a substitute thereof such as bhAsp, Ida, Ida(NHPal), and N-MeAsp, preferably Ida and N-MeAsp;

A2 is Thr, D-Thr, Ser, D-Ser, Val, D-Val, Ile, D-Ile, Ala, D-Ala or an unnatural amino acid commonly used as a substitute thereof such as Tle, Inp, Chg, bhThr, and N-MeThr;

A3 is His, D-His, Asn, D-Asn, Arg, D-Arg, or an unnatural amino acid commonly used as a substitute thereof such as L-His(π

-Me), D-His(π

-Me), L-His(τ

-Me), or D-His(τ

-Me);

A4 is Phe, D-Phe, Leu, D-Leu, Ile, D-Ile, Trp, D-Trp, Tyr, D-Tyr, or an unnatural amino acid commonly used as a substitute thereof such as Phg, bhPhe, Dpa, Bip, 1Nal, 2Nal, bhDpa, Amc, PheF5, hPhe, Igl, or cyclohexylalanine, preferably Dpa;

A5 is Pro, D-Pro, Ser, D-Ser, or an unnatural amino acid commonly used as a substitute thereof such as Oic, bhPro, trans-4-PhPro, cis-4-PhPro, cis-5-PhPro, and Idc, preferably bhPro;

A6 is Arg, D-Arg, Ile, D-Ile, Leu, D-Leu, Thr, D-Thr, Lys, D-Lys, Val, D-Val, or an unnatural amino acid commonly used as a substitute thereof such as D-Nω

,ω

-dimethyl-arginine, L-Nω

,ω

-dimethyl-arginine, D-homoarginine, L-homoarginine, D-norarginine, L-norarginine, citrulline, a modified Arg wherein the guanidinium group is modified or substituted, Norleucine, norvaline, bhIle, Ach, N-MeArg, and N-MeIle, preferably Arg;

A7 is Cys, D-Cys, Ser, D-Ser, Ala, D-Ala, or an unnatural amino acid commonly used as a substitute thereof such as Cys(S-tBut), homoCys, Pen, (D)Pen, preferably S-tertiary butyl-cysteine, Cys(S-S-Pal), Cys(S-S-cysteamine-Pal), Cys(S-S-Cys-NHPal), and Cys(S-S-Cys);

A8 is Arg, D-Arg, Ile, D-Ile, Leu, D-Leu, Thr, D-Thr, Lys, D-Lys, Val, D-Val, or an unnatural amino acid commonly used as a substitute thereof such as D-Nω

,ω

-dimethyl-arginine, L-Nω

,ω

-dimethyl-arginine, D-homoarginine, L-homoarginine, D-norarginine, L-norarginine, citrulline, a modified Arg wherein the guanidinium group is modified or substituted, Norleucine, norvaline, bhIle, Ach, N-MeArg, and N-MeIle, preferably Arg;

A9 is Phe, D-Phe, Leu, D-Leu, Ile, D-Ile, Tyr, D-Tyr, Trp, D-Trp, Phe-R^a, D-Phe-R^a, Dpa-R^a, D-Dpa-R^a, Trp-R^a, bhPhe-R^a, or an unnatural amino acid commonly used as a substitute thereof such as PheF5, N-MePhe, benzylamide, 2-aminoindane, bhPhe, Dpa, Bip, 1Nal, 2Nal, bhDpa, and cyclohexylalanine, which may or may not have R^a linked thereto, preferably bhPhe and bhPhe-R^a, wherein R^a is palmitoyl-PEG-, wherein PEG is PEG11 or miniPEG3, palmitoyl-PEG-PEG, wherein PEG is PEG11 or miniPEG3, butanoyl (C4)-PEG11-, octanoyl (C8, Caprylic)-PEG11-, palmitoyl (C16)-PEG11-, or tetracosanoyl (C24, Lignoceric)-PEG11-; and

A10 is Cys, D-Cys, Ser, D-Ser, Ala, D-Ala, or an unnatural amino acid such as Ida, Ida(NHPal)Ahx, and Ida(NBzl2)Ahx;

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, A1 to A2, A10, or a combination thereof are optionally absent, with the proviso that the peptide is not one of the peptides as set forth in Table 1.

2. The peptide according to claim 1, wherein the peptide contains at least one of the following:

a) A1=N-MeAsp, Ida, or Ida(NHPal);

b) A5=bhPro;

c) A6=D-Val, D-Leu, Lys, D-Lys, Arg, D-Arg, Ach, bhArg, or N-MeArg;

d) A7=Cys(S-S-Pal), Cys(S-S-cysteamine-Pal), Cys(S-S-Cys-NHPal), or Cys(S-S-Cys); and

/or

e) A8=D-Val, D-Leu, Lys, D-Lys, Arg, D-Arg, Ach, bhArg, or N-MeArg.

3. The peptide according to claim 1, wherein:

i) when A1 is Ida and A9 is Phe, then A10 is not Ahx-Ida(NHPal);

ii) when A1 is Ida, A9 is not bhPhe-R^b, wherein R^b is S-(palmityl)thioglycolic-PEG-;

iii) when A4 is D-Phe, A7 is not D-Cys(S-S-tBut) and A9 is not D-Trp-R^c, wherein R^c is Butanoyl-PEG11-, Octanoyl-PEG11-, Palmitoyl-PEG11-, or Tetracosanoyl-PEG11-; and

iv) when A1 is Ida and A9 is bhPhe-R^d, wherein R^d is palmitoyl-PEG-miniPEG3-, A6 and A8 are not both D-Arg or both bhArg.

4. The peptide according to claim 1, wherein

A1 is D-Asp, N-MeAsp, Ida, or Ida(NHPal);

A2 is Thr or D-Thr;

A3 is His or D-His;

A4 is Dpa or D-Dpa;

A5 is Pro, D-Pro, bhPro, or Oic;

A6 is Ile, D-Ile, Arg, D-Val, D-Leu, Ach, or N-MeArg;

A7 is Cys, D-Cys, Cys(S-S-Pal), Cys(S-S-cysteamine-Pal), Cys(S-S-Cys-NHPal), or Cys(S-S-Cys);

A8 is Ile, D-Ile, Arg, D-Val, D-Leu, Ach, or N-MeArg;

A9 is Phe, D-Phe, Dpa, D-Dpa, Trp, D-Trp, bhPhe, Phe-R^a, D-Phe-R^a, Dpa-R^a, D-Dpa-R^a, Trp-R^a, bhPhe-R^a, wherein R^a is palmitoyl-PEG-, wherein PEG is PEG11 or miniPEG3, palmitoyl-PEG-PEG, wherein PEG is PEG11 or miniPEG3, butanoyl (C4)-PEG11-, octanoyl (C8, Caprylic)-PEG11-, palmitoyl (C16)-PEG11-, or tetracosanoyl (C24, Lignoceric)-PEG11-; and

A10, if present, is Ida(NHPal)Ahx or Ida(NBzl2)Ahx.

5. The peptide of according to claim 1, wherein the peptide is selected from the group consisting of:

PR42′

, PR47, PR48, PR49, PR50, PR51, PR52, PR53, PR56, PR57, PR58, PR59, PR60, PR61, PR63, PR65, PR66, PR67, PR68, PR69, PR70, PR71, PR72, PR73, PR74, and PR82, preferably PR47, PR48, PR49, PR50, PR51, PR52, PR53, PR56, PR57, PR58, PR59, PR60, PR61, PR63, PR65, PR66, PR67, PR68, PR69, PR70, PR71, PR72, PR73, PR74, and PR82.

6. The peptide according to claim 1, wherein the peptide exhibits hepcidin activity.

7. The peptide according to claim 1, wherein the peptide binds ferroportin.

8. A composition which comprises at least one peptide according to claim 1.

9. A method of binding a ferroportin or inducing ferroportin internalization and degradation which comprises contacting the ferroportin with at least one peptide according to claim 1.

10. A method of treating a disease of iron metabolism in a subject which comprises administering at least one peptide according to claim 1 to the subject.

11. The method of claim 10, wherein the disease of iron metabolism is an iron overload disease.

12. A kit comprising at least one peptide according to claim 1 packaged together with a reagent, a device, instructional material, or a combination thereof.

13. A complex comprising at least one peptide according to claim 1 bound to a ferroportin or an antibody.

16. The method of claim 10, wherein the at least one peptide is administered at an effective daily dose as a single daily dose or as divided daily doses.

17. The method of claim 16, wherein the effective daily dose is about 10-500 μ

g/kg/day and the at least one peptide is provided in a formulation for subcutaneous injection.

18. The method of claim 16, wherein the effective daily dose is about 10-1000 μ

g/kg/day and the at least one peptide is provided in a formulation for oral, pulmonary, or mucosal administration.

14-15. -15. (canceled)