Stabilized Exendin-4 compounds
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.
1. A composition comprising an exendin-4 (1-39) analog comprising a sequence selected from the group consisting of:
[Asp 28 ]Exendin-4 (1-39) (SEQ ID NO:83),
des Pro 36 [Asp 28 ]Exendin-4 (1-39) (SEQ ID NO:15),
des Pro 36 , Pro 37 [Asp 28 ]Exendin-4 (1-39) (SEQ ID NO:108), and
des Pro 36 , Pro 37 , Pro 38 [Asp 28 ]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: -Lys 6 -NH 2 .
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 Pro 36 [Asp 28 ]Exendin-4 (1-39)-Lys 6 -NH 2 (SEQ ID NO:27),
H-(Lys) 6 -des Pro 36 , Pro 37 , Pro 38 [Asp 28 ]Exendin-4 (1-39)-NH 2 (SEQ ID NO:29),
H-Asn-(Glu) 5 des Pro 36 , Pro 37 , Pro 38 [Asp 28 ]Exendin-4 (1-39)-NH 2 (SEQ ID NO:30),
des Pro 36 , Pro 37 , Pro 38 [Asp 28 ]Exendin-4 (1-39)-(Lys) 6 -NH 2 (SEQ ID NO:31),
H-(Lys) 6 -des Pro 36 , Pro 37 , Pro 38 [Asp 28 ]Exendin-4 (1-39)-(Lys) 6 -NH 2 (SEQ ID NO:32), and
H-Asn-(Glu) 5 -des Pro 36 , Pro 37 , Pro 38 [Asp 28 ]Exendin-4 (1-39)-(Lys) 6 -NH 2 (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 Pro 36 [Asp 28 ]Exendin-4 (1-39)-Lys 6 -NH 2 (SEQ ID NO:27).
15. The composition of claim 13 , said analog comprising the sequence H-(Lys) 6 -des Pro 36 , Pro 37 , Pro 38 [Asp 28 ]Exendin-4 (1-39)-NH 2 (SEQ ID NO:29).
16. The composition of claim 13 , said analog comprising the sequence H-Asn-(Glu) 5 des Pro 36 , Pro 37 , Pro 38 [Asp 28 ]Exendin-4 (1-39)-NH 2 (SEQ ID NO:30).
17. The composition of claim 13 , said analog comprising the sequence des Pro 36 , Pro 37 , Pro 38 [Asp 28 ]Exendin-4 (1-39)-(Lys) 6 -NH 2 (SEQ ID NO:31).
18. The composition of claim 13 , said analog comprising the sequence H-(Lys) 6 -des Pro 36 , Pro 37 , Pro 38 [Asp 28 ]Exendin-4 (1-39)-(Lys) 6 -NH 2 (SEQ ID NO:32).
19. The composition of claim 13 , said analog comprising the sequence H-Asn-(Glu) 5 -des Pro 36 , Pro 37 , Pro 38 [Asp 28 ]Exendin-4 (1-39)-(Lys) 6 -NH 2 (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 [Asp 28 ]Exendin-4 (1-39) (SEQ ID NO:83).
28. The composition of claim 1 , said sequence comprising des Pro 36 [Asp 28 ]Exendin-4 (1-39) (SEQ ID NO:15).
29. The composition of claim 1 , said analog comprising the sequence des Pro 36 , Pro 37 [Asp 28 ]Exendin-4 (1-39) (SEQ ID NO:108).
30. The composition of claim 1 , said analog comprising the sequence des Pro 36 , Pro 37 , Pro 38 [Asp 28 ]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.