Beta-catenin targeting peptides and uses thereof

• US 8,957,026 B2
• Filed: 09/22/2011
• Issued: 02/17/2015
• Est. Priority Date: 09/22/2010
• Status: Expired due to Fees

• Current Assignee: President and Fellows of Harvard College (Harvard Corporation)
• Original Assignee: President and Fellows of Harvard College (Harvard Corporation)
• Inventors: Verdine, Gregory L., Grossmann, Tom N., Yeh, Tsung-Han Johannes
• Primary Examiner(s): Gamett, Daniel C
• Application Number:
  US13/825,709
• Publication Number:
  US 20140005118A1
• Time in Patent Office: 1,244 Days
• Field of Search: None
• US Class Current: 514/19.3
• 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: C07K 1/113 without change of the primary structure
• CPC Class Codes: C07K 14/47 from mammals
• CPC Class Codes: C07K 14/4702 Regulators; Modulating activity
• CPC Class Codes: C07K 14/4703 Inhibitors; Suppressors
• CPC Class Codes: C07K 14/4705 stimulating, promoting or activating activity
• CPC Class Codes: C07K 7/08 having 12 to 20 amino acids gastrins C07K14/595; somatostatins C07K14/655; melanotropins C07K14/68

Abstract

The invention relates to β-catenin targeting peptides comprising an α-helical segment that are optionally stapled or stitched, and pharmaceutical compositions thereof. Uses of the inventive β-catenin targeting polypeptides including methods for treatment of disease, such as diseases associated with aberrant Wnt signaling, including cancer, are also provided.

Claims

1. A β

-catenin binding polypeptide comprising an α

-helical segment, wherein the polypeptide comprises the amino acid sequence;

X₁X₂X₃XL₄X₅Y₁X₆X₇X₈Y₂X₉X ₁₀X₁₁A ₁₂X₁₃X₁₄X₁₅,

wherein

X₁ is selected from the group consisting of R, K and no amino acid;

X₂ is selected from the group consisting of R, K, W and no amino acid;

X₃ is selected from the group consisting of W, Y and Q;

X₄ is selected from the group consisting of P, S, W, A, F and L;

X₅ is selected from the group consisting of E, Q, R and W;

X₆ is selected from the group consisting of I and L;

X₇ is selected from the group consisting of I and L;

X₈ is selected from the group consisting of D and N;

X₉ is selected from the group consisting of H and W;

X₁₀ is selected from the group consisting of V, W, L, M, F, and I;

X₁₁ is selected from the group consisting of Q, E, L, D, H, R, S, and V;

X₁₂ is selected from the group consisting of R, S, and K;

X₁₃ is selected from the group consisting of V, I, L, W, F and H;

X₁₄ is selected from the group consisting of M, Norleucine, I, L, W, F and H;

X₁₅ is selected from the group consisting of R, G, P, E, H, K, Q, and no amino acid; and

Y₁ is selected from the group S, S⁵, R⁵, and Y₂ is selected from the group E, S⁵, and R⁵.

2. The polypeptide of claim 1, wherein the amino acid of X₁, X₂, X₁₁, X₁₂, or X₁₅ is a positively charged amino acid and/or the amino acid of X₃, X₄, X₅, X₆, X₇, X₉, X₁₀, X₁₃ or X₁₄ is a bulky, hydrophobic amino acid.

3. The polypeptide of claim 1, wherein Y₁ and Y₂ are S⁵.

4. The polypeptide of claim 1 comprising a free N-terminus, an acetylated N-terminus, a linker molecule, a labeling moiety, an affinity tag, and/or a targeting moiety.

5. The polypeptide of claim 4, wherein the linker molecule is β

-Ala or PEG₁.

6. The polypeptide of claim 4, wherein the labeling moiety is FITC.

7. The polypeptide of claim 4, wherein the affinity tag is biotin, FLAG, 6×

His, or myc.

8. The polypeptide of claim 4, wherein the targeting moiety is integrin, antibody, or antibody fragment.

9. A pharmaceutical composition comprising the polypeptide of claim 1.

10. A method of treating a cancer that exhibits Wnt-signaling dependent growth in a subject, the method comprising administering to a subject the pharmaceutical composition of claim 9 in an amount sufficient to treat the cancer.

11. A method of modulating Wnt-mediated transcription, the method comprising contacting a cell with the polypeptide of claim 1.

12. A method to inhibit cell proliferation that is dependent on aberrant Wnt-signaling, the method comprising contacting the cell with a polypeptide of claim 1.

13. A β

-catenin binding polypeptide comprising an α

-helical segment, wherein the polypeptide comprises the amino acid sequence;

X₁X₂X₃X₄X₅X₆X₇X₈X₉X₁₀X₁₁X₁₂X₁₃X₁₄X₁₅X₁₆,

wherein

X₁ is selected from the group consisting of R, V, E, Q, T, K, W, and L;

X₂ is selected from the group consisting of W, S, L, N, Y, K, V, and Q;

X₃ is selected from the group consisting of P, L, S, W, A, and F;

X₄ is selected from the group consisting of E, Q, and W;

X₅ isS;

X₆ is I;

X₇ is L;

X₈ is D;

X₉ is selected from the group consisting of E and Q;

X₁₀ is H;

X₁₁ is selected from the group consisting of W, V, F, M, L, and I;

X₁₂ is selected from the group consisting of E, Q, H, D, V, and L;

X₁₃ is selected from the group consisting of R and K;

X₁₄ is selected from the group consisting of V and W;

X₁₅ is selected from the group consisting of M, W, F, and E; and

X₁₆ is selected from the group consisting of R, G, P, E, and Q, with the proviso that the sequence is not the wild-type sequence from Axin.

14. The polypeptide of claim 13, wherein

X₁X₂X₃X₄X₅X₆X₇X₈X₉X₁₀X₁₁X₁₂X₁₃X₁₄X₁₅X₁₆ is modified to induce and/or stabilize an α

-helical conformation.

15. The polypeptide of claim 14, wherein the amino acids positioned at i, i+4 and/or i, i+7 are cross-linked by a hydrocarbon cross-link.

16. The polypeptide of claim 14, wherein the modification is selected from the group consisting of a peptide staple, a lactam cross-link, a disulfide cross-link, α

-methylation, N-caps, and a hydrogen bond surrogate.

17. The polypeptide of claim 16, wherein the modification is a peptide staple.

18. The polypeptide of claim 17, wherein the amino acids positioned at i, i+4 and/or i, i+7 are stapled.

19. The polypeptide of claim 13, wherein the sequence is not ENPESILDEHVQRVMR (SEQ ID NO:

1) (amino acids 469-484 of rat axin).

20. A β

-catenin binding polypeptide comprising an amino acid sequence of any one of SEQ ID NOs.;

28-58.

21. An isolated polypeptide comprising an α

-helical segment, wherein the peptide binds β

-catenin and comprises an amino acid sequence selected form the group consisting of the sequences;

ENPES⁵ILDS⁵HVQRVM (SEQ ID NO;

14), PES⁵ILDS⁵HVQRVM (SEQ ID NO;

15), PES⁵ILDS⁵HVRRVMR (SEQ ID NO;

16), PQS⁵ILDS⁵HVRRVMR (SEQ ID NO;

17), PQS⁵ILDS⁵HVRRWMR (SEQ ID NO;

18), PQS⁵ILDS⁵HVRRVWR (SEQ ID NO;

19), RWPQS⁵ILDS⁵HVRRVWR (SEQ ID NO;

20), RRWPQS⁵ILDS⁵HVRRVWR (SEQ ID NO;

21), RWPRS⁵ILDS⁵HVRRVWR (SEQ ID NO;

22), RRWPRS⁵ILDS⁵HVRRVWR (SEQ ID NO;

23), RWPQS⁵ILDS⁵HVRRWNleR (SEQ ID NO;

24), RWPRS⁵ILDS⁵HVRRWNleR (SEQ ID NO;

25), RWPQS⁵ILDS⁵HVRRRWR (SEQ ID NO;

26), RWPRS⁵ILDS⁵HVRRRWR (SEQ ID NO;

27).