Variants of adrenomedullin and calcitonin gene-related peptide and methods of use

Patent metadata

Field	Value
Publication number	US-10723778-B2
Application number	US-201916593336-A
Country	US
Kind code	B2
Filing date	Oct 4, 2019
Priority date	May 13, 2015
Publication date	Jul 28, 2020
Grant date	Jul 28, 2020

Abstract

Variant peptides of calcitonin gene-related peptide alpha (αCGRP), calcitonin gene-related peptide beta (βCGRP), and adrenomedullin (AM) are disclosed, wherein the variant peptides have high binding affinity and agonistic or antagonistic activity for at least one receptor complex of CLR:RAMP1, CLR:RAMP2, and CLR:RAMP3. Also disclosed are methods of use of the variant peptides in therapeutic treatments.

First claim

What is claimed is:

1. A variant peptide having binding affinity for at least one receptor complex of the group consisting of calcitonin receptor-like receptor-receptor activity-modifying protein 1 (CLR:RAMP1), calcitonin receptor-like receptor-receptor activity-modifying protein 2 (CLR:RAMP2), and calcitonin receptor-like receptor-receptor activity-modifying protein 3 (CLR:RAMP3), the variant peptide having an affinity K i for the receptor complex of less than 1 μM, and the variant peptide having a structure as represented by Formula III (SEQ ID NO:51):

(III)

F-V-P-T-X 5 -X 6 -G-X 8 -X 9 -X 10 -X 11

wherein:
X 5 is N, D, or W,
X 6 is V, T, or W,
X 8 is P or S,
X 9 is W, Y, or H,
X 10 is A, S, or G,
X 11 is F or Y, wherein the F or Y is amidated;
wherein the peptide is optionally extended at the N-terminus by all or any portion of amino acids 1-41 of SEQ ID NO:1, all or any portion of amino acids 1-26 of SEQ ID NO:31, all or any portion of amino acids 1-26 of SEQ ID NO:32, or all or any portion of amino acids 1-29 of SEQ ID NO:47.

2. The variant peptide of claim 1 , wherein the variant peptide is directly bound to a carrier molecule.

3. The variant peptide of claim 1 , wherein the variant peptide is bound to the carrier molecule by a linker molecule.

4. The variant peptide of claim 1 , further defined as consisting of up to 70 amino acids.

5. The variant peptide of claim 1 , further defined as having agonistic activity for the at least one receptor complex.

6. The variant peptide of claim 1 , further defined as having antagonistic activity for the at least one receptor complex.

7. The variant peptide of claim 1 , wherein the affinity K i is less than about 500 nM.

8. A method of treating a subject for a condition regulated by a calcitonin receptor-like receptor-receptor activity-modifying protein (CLR:RAMP) receptor complex, comprising the step of:
administering to the subject in need of such therapy an effective amount of the variant peptide of claim 1 .

9. A variant peptide having binding affinity for at least one receptor complex of the group consisting of calcitonin receptor-like receptor-receptor activity-modifying protein 1 (CLR:RAMP1), calcitonin receptor-like receptor-receptor activity-modifying protein 2 (CLR:RAMP2), and calcitonin receptor-like receptor-receptor activity-modifying protein 3 (CLR:RAMP3), the variant peptide having an affinity K i for the receptor complex of less than 1 μM, and the variant peptide having a structure as represented by Formula IV (SEQ ID NO:52):

(IV)

T-X 2 -X 3 -G-X 5 -X 6 -X 7 -X 8

wherein:
X 2 is N, D, or W,
X 3 is V, T, or W,
X 5 is P,
X 6 is W, Y, or H,
X 7 is A, S, or G,
X 8 is F or Y, and wherein the F or Y is amidated.

10. The variant peptide of claim 9 , wherein the variant peptide is directly bound to a carrier molecule.

11. The variant peptide of claim 9 , wherein the variant peptide is bound to the carrier molecule by a linker molecule.

12. The variant peptide of claim 9 , further defined as consisting of up to 70 amino acids.

13. The variant peptide of claim 9 , further defined as having agonistic activity for the at least one receptor complex.

14. The variant peptide of claim 9 , further defined as having antagonistic activity for the at least one receptor complex.

15. The variant peptide of claim 9 , wherein the affinity K i is less than about 500 nM.

16. A method of treating a subject for a condition regulated by a calcitonin receptor-like receptor-receptor activity-modifying protein (CLR:RAMP) receptor complex, comprising the step of:
administering to the subject in need of such therapy an effective amount of the variant peptide of claim 9 .

17. The variant peptide of claim 1 , wherein X 5 is D, X 6 is V, X 8 is P, X 9 is W, X 10 is S, and X 11 is F (SEQ ID NO:39).

18. The variant peptide of claim 1 , wherein X 5 is D, X 6 is V, X 8 is P, X 9 is W, X 10 is S, and X 11 is Y (SEQ ID NO:40).

19. The variant peptide of claim 9 , further comprising an N-terminal portion comprising all or any portion of amino acids 1-44 of SEQ ID NO:1, all or any portion of amino acids 1-29 of SEQ ID NO:31, all or any portion of amino acids 1-29 of SEQ ID NO:32, or all or any portion of amino acids 1-32 of SEQ ID NO:47.

20. The variant peptide of claim 9 , wherein X 2 is N.

21. The variant peptide of claim 9 , wherein X 2 is D.

22. The variant peptide of claim 9 , wherein X 3 is V.

23. The variant peptide of claim 9 , wherein X 3 is T.

24. The variant peptide of claim 9 , wherein X 6 is W.

25. The variant peptide of claim 9 , wherein X 7 is S.

26. The variant peptide of claim 9 , wherein X 8 is F.

27. The variant peptide of claim 9 , wherein X 8 is Y.

28. The variant peptide of claim 9 , wherein X 6 is W, and X 7 is S.

29. The variant peptide of claim 9 , wherein X 2 is D, X 3 is V, X 6 is W, X 7 is S, and X 8 is F (SEQ ID NO:45).

30. The variant peptide of claim 9 , wherein X 2 is D, X 3 is V, X 6 is W, X 7 is S, and X 8 is Y (SEQ ID NO:46).

31. A variant peptide having binding affinity for at least one receptor complex of the group consisting of calcitonin receptor-like receptor-receptor activity-modifying protein 1 (CLR:RAMP1), calcitonin receptor-like receptor-receptor activity-modifying protein 2 (CLR:RAMP2), and calcitonin receptor-like receptor-receptor activity-modifying protein 3 (CLR:RAMP3), the variant peptide having an affinity K i for the receptor complex of less than 1 μM, and the variant peptide having a structure as represented by Formula IV (SEQ ID NO:52):

(IV)

T-X 2 -X 3 -G-X 5 -X 6 -X 7 -X 8

wherein:
X 2 is N, D, or W,
X 3 is V, T, or W,
X 5 is P or S,
X 6 is W or Y,
X 7 is A, S, or G,
X 8 is F or Y, and wherein the F or Y is amidated.

32. The variant peptide of claim 31 , further comprising an N-terminal portion comprising all or any portion of amino acids 1-44 of SEQ ID NO:1, all or any portion of amino acids 1-29 of SEQ ID NO:31, all or any portion of amino acids 1-29 of SEQ ID NO:32, or all or any portion of amino acids 1-32 of SEQ ID NO:47.

33. The variant peptide of claim 31 , wherein X 2 is N.

34. The variant

Assignees

• Univ Oklahoma

Inventors

• Pioszak Augen A

Classifications

• A61K38/00

  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 title

• C07K14/575

  Hormones (derived from pro-opiomelanocortin, pro-enkephalin or pro-dynorphin C07K14/665, e.g. corticotropin C07K14/695) · CPC title

• C07K14/57527

  Calcitonin gene related peptide · CPC title

• C07K14/585 Primary

  Calcitonins · CPC title