Trophinin-binding peptides and uses thereof

• US 8,097,588 B2
• Filed: 02/06/2008
• Issued: 01/17/2012
• Est. Priority Date: 02/08/2007
• Status: Active Grant

• Current Assignee: Sanford Burnham Prebys Medical Discovery Institute
• Original Assignee: Sanford-Burnham Medical Research Institute (Sanford Burnham Prebys Medical Discovery Institute)
• Inventors: Fukuda, Michiko
• Primary Examiner(s): Tsang, Cecilia J
• Assistant Examiner(s): Heard, Thomas
• Application Number:
  US12/026,810
• Publication Number:
  US 20080200369A1
• Time in Patent Office: 1,441 Days
• Field of Search: None
• US Class Current: 514/9.8
• 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 5/1008 the side chain containing 0 or 1 carbon atoms, i.e. Gly, Ala
• CPC Class Codes: C07K 7/06 having 5 to 11 amino acids

Abstract

Disclosed herein are compositions and methods useful for promoting sperm motility, promoting embryonic stem cell formation, promoting trophoblast formation, or promoting neuronal growth. The compositions and methods are based on peptide sequences that bind trophinin, inhibit bystin-mediated arrest of epidermal growth factor (EGF) receptor, and promotes EGF receptor autophosphorylation.

Claims

1. A method of promoting sperm motility, comprising contacting the sperm in vitro with an isolated polypeptide comprising one or more trophinin-binding segments, wherein at least one of the trophinin-binding segments comprises the amino acid sequence X₁X₂X₃X₄ (SEQ ID NO:

1), wherein

X₁ is G, A, L, I, T, or S;

X₂ is W or Y;

X₃ is R, K, or H; and

X₄ is Q or N,

wherein the polypeptide is 50 amino acids or less in length.

2. The method of claim 1, wherein the sperm is used for in vitro fertilization.

3. The method of claim 2, wherein fertilization is increased by at least about 10%, 20%, 30%, 40%, 50%, or higher.

7. The method of claim 1, wherein the at least one trophinin-binding segment comprises the amino acid sequence X₁X₂X₃X₄X₅ (SEQ ID NO:

3), wherein X₅ is S, A, V, or L.

8. The method of claim 1, wherein at least one trophinin-binding segment comprises the amino acid sequence GWRQ (SEQ ID NO:

2).

9. The method of claim 1, wherein the polypeptide selectively binds trophinin.

10. The method of claim 1, wherein each trophinin-binding segment independently consists of from 4 to 50 amino acids.

11. The method of claim 1, wherein at least one trophinin-binding segment comprises the amino acid sequence GWRQLTARVP (SEQ ID NO:

4).

12. The method of claim 1, wherein at least one trophinin-binding segment consists essentially of the amino acid sequence GWRQLTARVP (SEQ ID NO:

4).

13. The method of claim 1, wherein the peptide has a length of less than 20 residues.

14. The method of claim 1, wherein the polypeptide binds to the extracellular domain of trophinin.

15. The method of claim 1, wherein polypeptide inhibits the bystin-mediated arrest of epidermal growth factor (EGF) receptor and promotes EGF receptor autophosphorylation.

16. The method of claim 1, wherein the polypeptide comprises two or more trophinin-binding segments.

17. The method of claim 1, wherein the polypeptide comprises two or more amino acid branches.

18. The method of claim 17, wherein the polypeptide further comprises a peptidyl core of 2, 3, 4, 5, 6, 7, 8, 9, 10 or more branched lysine residues, wherein two or more of the trophinin-binding segments are linked to two or more branched lysine residues.

4. A method of promoting sperm motility, comprising contacting the sperm in vitro with an isolated polypeptide comprising one or more trophinin-binding segments, wherein at least one of the trophinin-binding segments comprises the amino acid sequence X₁X₂X₃X₄ (SEQ ID NO:

1), wherein

X₁ is G;

X₂ is W;

X₃ is R; and

X₄ is Q.

5. The method of claim 4, wherein the at least one of the trophinin-binding segments comprises the amino acid sequence GWRQLTARVP (SEQ ID NO:

4), SSLMLGGWRQ (SEQ ID NO;

19), or SLSVGWRQSG (SEQ ID NO;

20).

6. A method of promoting sperm motility, comprising administering to a subject an isolated polypeptide comprising one or more trophinin-binding segments, wherein at least one of the trophinin-binding segments comprises the amino acid sequence X₁X₂X₃X₄ (SEQ ID NO:

1), wherein

X₁ is G, A, L, I, T, or S;

X₂ is W or Y;

X₃ is R, K, or H; and

X₄ is Q or N,

wherein the polypeptide is 50 amino acids or less in length.