Stabilized Exendin-4 compounds

• US 7,544,657 B2
• Filed: 10/02/2003
• Issued: 06/09/2009
• Est. Priority Date: 10/02/2002
• Status: Expired due to Fees

• Current Assignee: Zealand Pharma A/S
• Original Assignee: Zealand Pharma A/S
• Inventors: Knott, David, Jepsen, Trine, Ebbehoj, Kirsten, Larsen, Bjarne Due, Knudsen, Carsten Boye
• Primary Examiner(s): Landsman; Robert
• Assistant Examiner(s): Chandra; Gyan
• Application Number:
  US10/529,858
• Publication Number:
  US 20060194719A1
• Time in Patent Office: 2,077 Days
• Field of Search: None
• US Class Current: 514/1.1
• 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: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 25/18 Antipsychotics, i.e. neuroleptics; Drugs for mania or schizophrenia
• 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 3/06 Antihyperlipidemics
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• 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 14/57563 Vasoactive intestinal peptide [VIP]; Related peptides

Abstract

The present invention disclosed compositions comprising a stabilized Exendin-4 (1-39) and related compounds. The invention describes stabilized Exendin-4 agonists that include at least one modified amino acid residue particularly at positions Gln 13, Met14, Trp25, or Asn28 of the Exendin-4 (1-39) molecule. Disclosed are preferred modifications of deaminated, hydrolyzed, oxidized, or isomerized reaction products of the specified amino acid residues corresponding to the same positions in the Exendin-4 molecule. The invention also relates to methods of making and using the stabilized Exendin compounds, such as for the treatment of diabetes.

Claims

1. A composition comprising an exendin-4 (1-39) analog comprising a sequence selected from the group consisting of:

[Asp²⁸]Exendin-4 (1-39) (SEQ ID NO;

83),

des Pro³⁶[Asp²⁸]Exendin-4 (1-39) (SEQ ID NO;

15),

des Pro³⁶, Pro³⁷[Asp²⁸]Exendin-4 (1-39) (SEQ ID NO;

108), and

des Pro³⁶, Pro³⁷, Pro³⁸[Asp²⁸]Exendin-4 (1-39) (SEQ ID NO;

28).

2. The composition of claim 1 further comprising at least one peptide sequence Z of 4-20 amino acid residues covalently bound to the exendin analog;

or a pharmaceutically acceptable salt or solvate thereof.

3. The composition of claim 2, wherein Z comprises between about 4 to about 20 Lys amino acid units.

4. The composition of claim 2, wherein Z comprises 6 Lys amino acid units.

5. The composition of claim 2, wherein the exendin-4 (1-39) analog and Z are bonded by a peptide bond.

6. The composition of claim 2, wherein Z is covalently bound to the exendin-4 (1-39) analog at the C-terminal carbonyl function.

7. A composition of claim 2, further comprising a pharmaceutically acceptable carrier.

8. The composition of claim 7, wherein the composition comprises a depot formulation, microspheres, liposomes or the composition includes a stabilized liquid formulation.

9. A method for treating disease states associated with elevated blood glucose levels, said method comprising administering a therapeutically effective amount of the composition of claim 2.

10. A method for regulation of blood glucose levels, the method comprising administering a therapeutically effective amount of the composition of claim 2.

11. A method of stimulating insulin release in a mammal comprising administering an effective insulinotropic amount of the composition of claim 2.

12. The composition of claim 1 further comprising the following group linked to the C-terminus of the analog:

-Lys₆-NH₂.

13. The composition of claim 1, wherein the exendin-4 (1-39) analog comprises a sequence selected from the group consisting of:

H-(Lys)₆-des Pro³⁶[Asp²⁸]Exendin-4 (1-39)-Lys₆-NH₂ (SEQ ID NO;

27),

H-(Lys)₆-des Pro³⁶, Pro³⁷, Pro³⁸[Asp²⁸]Exendin-4 (1-39)-NH₂ (SEQ ID NO;

29),

H-Asn-(Glu)₅des Pro³⁶, Pro³⁷, Pro³⁸[Asp²⁸]Exendin-4 (1-39)-NH₂ (SEQ ID NO;

30),

des Pro³⁶, Pro³⁷, Pro³⁸[Asp²⁸]Exendin-4 (1-39)-(Lys)₆-NH₂ (SEQ ID NO;

31),

H-(Lys)₆-des Pro³⁶, Pro³⁷, Pro³⁸[Asp²⁸]Exendin-4 (1-39)-(Lys)₆-NH₂ (SEQ ID NO;

32), and

H-Asn-(Glu)₅-des Pro³⁶, Pro³⁷, Pro³⁸[Asp²⁸]Exendin-4 (1-39)-(Lys)₆-NH₂ (SEQ ID NO;

33),

or a pharmaceutically acceptable salt or solvate thereof.

14. The composition of claim 13, said analog comprising the sequence H-(Lys)₆-des Pro³⁶[Asp²⁸]Exendin-4 (1-39)-Lys₆-NH₂ (SEQ ID NO:

27).

15. The composition of claim 13, said analog comprising the sequence H-(Lys)₆-des Pro³⁶, Pro³⁷, Pro³⁸[Asp²⁸]Exendin-4 (1-39)-NH₂ (SEQ ID NO:

29).

16. The composition of claim 13, said analog comprising the sequence H-Asn-(Glu)₅des Pro³⁶, Pro³⁷, Pro³⁸[Asp²⁸]Exendin-4 (1-39)-NH₂ (SEQ ID NO:

30).

17. The composition of claim 13, said analog comprising the sequence des Pro³⁶, Pro³⁷, Pro³⁸[Asp²⁸]Exendin-4 (1-39)-(Lys)₆-NH₂ (SEQ ID NO:

31).

18. The composition of claim 13, said analog comprising the sequence H-(Lys)₆-des Pro³⁶, Pro³⁷, Pro³⁸[Asp²⁸]Exendin-4 (1-39)-(Lys)₆-NH₂ (SEQ ID NO:

32).

19. The composition of claim 13, said analog comprising the sequence H-Asn-(Glu)₅-des Pro³⁶, Pro³⁷, Pro³⁸[Asp²⁸]Exendin-4 (1-39)-(Lys)₆-NH₂ (SEQ ID NO:

33).

20. The composition of claim 1, wherein the composition includes a mixture of L- and D-amino acid residues.

21. A composition of any one of claims 3-20, further comprising a pharmaceutically acceptable carrier.

22. The composition of claim 21, wherein the composition comprises a depot formulation, microspheres, or liposomes, or the composition includes a stabilized liquid formulation.

23. A method for treating diabetes type 1, the method comprising administering a therapeutically effective amount of the composition of claim 1.

24. A method for treating disease states associated with elevated blood glucose levels, said method comprising administering a therapeutically effective amount of the composition of claim 1.

25. A method for regulation of blood glucose levels, the method comprising administering a therapeutically effective amount of the composition of claim 1.

26. A method of stimulating insulin release in a mammal comprising administering an effective insulinotropic amount of the composition of claim 1.

27. The composition of claim 1, said sequence comprising [Asp²⁸]Exendin-4 (1-39) (SEQ ID NO:

83).

28. The composition of claim 1, said sequence comprising des Pro³⁶[Asp²⁸]Exendin-4 (1-39) (SEQ ID NO:

15).

29. The composition of claim 1, said analog comprising the sequence des Pro³⁶, Pro³⁷[Asp²⁸]Exendin-4 (1-39) (SEQ ID NO:

108).

30. The composition of claim 1, said analog comprising the sequence des Pro³⁶, Pro³⁷, Pro³⁸[Asp²⁸]Exendin-4 (1-39) (SEQ ID NO:

28).

31. The composition of claim 1, wherein the amino acid residues of said exendin-4 (1-39) analog are in the L configuration.

32. The composition of claim 1, wherein the amino acid residues of said exendin-4 (1-39) analog are in the D configuration.