KDR and VEGF/KDR binding peptides and their use in diagnosis and therapy
View Patent ↗The present invention provides polypeptides, peptide dimer, and multimeric complexes comprising at least one binding moiety for KDR or VEGF/KDR complex, which have a variety of uses wherever treating, detecting, isolating or localizing angiogenesis is advantageous. Particularly disclosed are synthetic, isolated polypeptides capable of binding KDR or VEGF/KDR complex with high affinity (e.g., having a K D <1 μM), and dimer and multimeric constructs comprising these polypeptides.
1. A method for treating an ocular disease that involves angiogenesis, comprising administering to a subject in need thereof an effective amount of a VEGF/KDR-binding peptide, which comprises a motif of:
(i) Cys-X 2 -X 3 -X 4 -X 5 -X 6 -Tyr-Cys, wherein X 2 is Ala, Arg, Glu, Lys, or Ser; X 3 is Ala, Asp, Gln, Glu, Thr, or Val; X 4 is Asp or Glu; X 5 is Trp or Tyr; and X 6 is Thr or Tyr; or
(ii) Cys-X 2 -X 3 -X 4 -X 5 -Trp-Gly-Gly-X 9 -X 10 -Cys (SEQ ID NO:14), wherein X 2 is Ala, Phe, or Trp; X 3 is Glu or Lys; X 4 is Asp, Ser, Trp, or Tyr; X 5 is Phe, Pro, or Ser, X 9 is Gln or Glu; and X 10 is Ile, Phe, or Val;
wherein the VEGF/KDR-binding peptide is either monomeric or homomultimeric.
2. The method of claim 1 , wherein the ocular disease is selected from the group consisting of diabetic retinopathy, retinopathy of prematurity, macular degeneration, and corneal graft rejection.
3. The method of claim 1 , wherein the peptide has fewer than 25 amino acids.
4. The method of claim 1 , wherein the peptide comprises Cys-X 2 -X 3 -X 4 -X 5 -X 6 -Tyr-Cys, in which X 2 is Glu; X 3 is Asp or Glu; X 4 is Asp or Glu; X 5 is Trp or Tyr; and X 6 is Tyr.
5. The method of claim 4 , wherein the peptide comprises an amino acid sequence selected from the group consisting of:
(a)
AGPTWCEDDWYYCWLFGTK,
(SEQ ID NO: 493)
(b)
AGPTWCEDDWYYCWLFGTGGGK,
(SEQ ID NO: 277)
(c)
AGPTWCEDDWYYCWLFGTGGGGK,
(SEQ ID NO: 480)
(d)
AGPTWCEDDWYYCWLFGTGGGKK,
(SEQ ID NO: 373)
and
(e) a variant thereof that includes a truncation of one or more residues at the C-terminus or the N-terminus.
6. The method of claim 1 , wherein the peptide comprises Cys-X 2 -X 3 -X 4 -X 5 -Trp-Gly-Gly-X 9 -X 10 -Cys (SEQ ID NO:14), in which X 2 is Phe or Trp; X 3 is Glu; X 4 is Asp, Trp, or Tyr; X 5 is Ser, X 9 is Glu; and X 10 is Ile, Phe, or Val.
7. The method of claim 6 , wherein the peptide comprises an amino acid sequence selected from the group consisting of:
(iii)
GDSRVCWEDSWGGEVCFRYDP,
(SEQ ID NO: 374)
(iv)
GDSRVCWEDSWGGEVCFRYDPGGGK,
(SEQ ID NO: 294)
and
(v)
GDSRVCWEDSWGGEVCFRYGGGK.
(SEQ ID NO: 372)
8. The method of claim 1 , wherein the peptide is modified.
9. The method of claim 8 , wherein the peptide is modified at the N-terminus, the C-terminus, or both.
10. The method of claim 9 , wherein the peptide has an acetylated N-terminus, an amidated C-terminus, or both.
11. The method of claim 1 , wherein the VEGF/KDR-binding peptide is in a dimeric construct containing two copies of the same VEGF/KDR-binding peptide.
12. The method of claim 1 , wherein the peptide is conjugated with a therapeutic agent.
13. The method of claim 1 , wherein the therapeutic agent is an antibody or a fragment thereof.
14. The method of claim 12 , wherein the peptide and the therapeutic agent is connected via a spacer.
15. The method of claim 14 , wherein the spacer is a substituted or unsubstituted alkyl chain, a polyethylene glycol derivative, an amino acid spacer, a sugar spacer, or an aromatic spacer.