Proline-locked stapled peptides and uses thereof

• US 9,617,309 B2
• Filed: 09/26/2013
• Issued: 04/11/2017
• Est. Priority Date: 09/26/2012
• Status: Active Grant

• Current Assignee: President and Fellows of Harvard College (Harvard Corporation)
• Original Assignee: President and Fellows of Harvard College (Harvard Corporation)
• Inventors: Verdine, Gregory L., Hayashi, Kazuhiro
• Primary Examiner(s): Lukton, David
• Application Number:
  US14/431,280
• Publication Number:
  US 20150239937A1
• Time in Patent Office: 1,293 Days
• Field of Search: 530317
• CPC Class Codes: C07D 207/16 Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
• CPC Class Codes: C07K 1/113 without change of the primary structure
• CPC Class Codes: C07K 7/56 the cyclisation not occurring through 2,4-diamino-butanoic acid

Abstract

The present invention provides a new type of alpha-helix nucleating cross-link (“staple”) formed by olefin metathesis of a proline derivative with an alkenyl side chain and another amino acid derivative with an alkenyl side chain. The proline derivatives as described herein have been found to be strong nucleators of alpha-helix formation. The invention also provides moieties for shielding the free amide N—H'"'"'s at the N-terminus of an alpha-helix, thereby further stabilizing the helix. The proline derivatives, precursors prior to cross-linking, and the cross-linked peptides are provided as well as methods of using and preparing these compounds and peptides.

Claims

1. A polypeptide of Formula (I):

2. A composition comprising a polypeptide of claim 1, and an excipient.

3. The polypeptide of claim 1 of the formula:

4. The polypeptide of claim 3 selected from any one of the formula:

5. The polypeptide of claim 1 of the formula:

6. The polypeptide of claim 5 selected from any one of the formula:

7. The polypeptide of claim 1, wherein p is 1.

8. The polypeptide of claim 1, wherein q is 0.

9. The polypeptide of claim 1, wherein p is 1 and q is 0.

10. The polypeptide of claim 1, wherein each instance of R^e is, independently, hydrogen, —

CH₃, —

CH₂OH, —

COOH, or —

CH₂COOH.

11. The polypeptide of claim 1, wherein each instance of R^e is —

CH₃.

12. The polypeptide of claim 1, wherein each R^d is hydrogen.

13. The polypeptide of claim 1, wherein each instance of y is, independently, 1 to 8.

14. The polypeptide of claim 1, wherein each instance of y is 2.

15. The polypeptide of claim 1, wherein R^a is substituted or unsubstituted acyl.

16. The polypeptide of claim 1, wherein R^a is hydrogen.

17. The polypeptide of claim 1, wherein R^b is —

OR^B.

18. The polypeptide of claim 1, wherein R^b is a —

OH.

19. The polypeptide of claim 1, wherein each instance of K is, independently, —

CH₂—

, —

(CH₂)₂—

, or —

(CH₂)₃—

.

20. The polypeptide of claim 1, wherein each instance of K is —

CH₂—

.

21. The polypeptide of claim 1, wherein each instance of L is, independently, —

CH₂—

, —

(CH₂)₂—

, or —

(CH₂)₃—

.

22. The polypeptide of claim 1, wherein each instance of L is —

CH₂—

.

23. The polypeptide of claim 1, wherein corresponds to a double bond.

24. The polypeptide of claim 1, wherein each instance of R^e is —

CH₃.

25. The polypeptide of claim 1, wherein:

K and L are —

CH₂—

;

R^d is —

H;

R^e is —

CH₃;

j is 1;

p is 1;

q is 0;

y is 2; and

corresponds to a double bond.

26. The polypeptide of claim 3, wherein:

p is 1;

q is 0;

j is 1;

y is 2;

K is —

CH₂—

, —

(CH₂)₂—

, —

(CH₂)₃—

, or —

O—

(CH₂)_1-3—

;

L is —

(CH₂)_1-6—

;

R^d is —

H;

and R^e is —

CH₃; and

corresponds to a double bond.

27. The polypeptide of claim 1, wherein:

each instance of K and L, is, independently, a bond;

substituted or unsubstituted alkylene;

or substituted or unsubstituted heteroalkylene;

R^a is hydrogen;

substituted or unsubstituted aliphatic;

substituted or unsubstituted heteroaliphatic;

substituted or unsubstituted aryl;

substituted or unsubstituted heteroaryl;

substituted or unsubstituted acyl;

a resin;

an amino protecting group;

a label optionally joined by a linker, wherein the linker is a group consisting of one or more combinations of substituted or unsubstituted alkylene;

substituted or unsubstituted alkenylene;

substituted or unsubstituted alkynylene;

substituted or unsubstituted heteroalkylene;

substituted or unsubstituted heteroalkenylene;

substituted or unsubstituted carbocyclene;

substituted or unsubstituted heterocyclene;

substituted or unsubstituted arylene;

or substituted or unsubstituted heteroarylene;

R^b is, —

R^B, —

OR^B, or —

N(R^B)₂, wherein each instance of R^B is, independently, hydrogen;

substituted or unsubstituted aliphatic;

substituted or unsubstituted heteroaliphatic;

substituted or unsubstituted aryl;

substituted or unsubstituted heteroaryl;

substituted or unsubstituted acyl;

a resin;

a suitable hydroxyl or amino protecting group;

or two R^B groups together form a substituted or unsubstituted 5- to 6-membered heterocyclic or heteroaromatic ring;

each instance of R^KL is, independently, hydrogen or substituted or unsubstituted aliphatic;

each instance of R^d is, independently, hydrogen;

substituted or unsubstituted aliphatic;

substituted or unsubstituted heteroaliphatic;

substituted or unsubstituted aryl;

substituted or unsubstituted heteroaryl;

substituted or unsubstituted acyl;

or R^d is an amino protecting group;

each instance of R^e is, independently, a suitable amino acid side chain;

hydrogen;

or substituted or unsubstituted aliphatic;

each instance of G is, independently, a natural or unnatural amino acid or a group of the formula;