FUSION PROTEINS FOR INHIBITING ANGIOGENESIS
The present invention relates to a biologic that inhibits angiogenesis. In particular, the present invention relates to fusion proteins that inhibit the integrin activated pathway and one other angiogenic factor-activated pathway, the compositions of these fusion proteins, as well as methods for producing and using the same.
1 . A fusion protein comprising:
an integrin binding peptide comprising disintegrin and its integrin binding fragments;
other protein binding peptide targeting an angiogenic factor; and
a Fc domain;
wherein the integrin binding peptide has at least one mutation on or adjacent to a RGD motif.
2 - 4 . (canceled)
5 . The fusion protein according to claim 1 comprising the integrin binding peptide, the Fc domain and the other protein binding peptide from C-terminus to N-terminus, or from N-terminus to C terminus.
6 - 8 . (canceled)
9 . A nucleic acid encoding the fusion protein of claim 1 .
10 . A dimer of the fusion protein of claim 1 .
11 . A vector comprising a nucleotide sequence encoding the fusion protein of claim 1 .
12 . A method of producing a fusion protein, comprising culturing a host cell comprising a nucleic acid encoding the fusion protein of claim 1 under a condition that produces the fusion protein, and recovering the fusion protein produced by the host cell.
13 . A method of treating/preventing an angiogenic disease comprising administering an effective amount of the fusion protein of claim 1 to a subject in need thereof.
14 . The fusion protein of claim 1 further comprising a linker sequence between the integrin binding peptide and the other protein binding peptide or a signal peptide sequence upstream of the other protein binding peptide.
15 . (canceled)
16 . The fusion protein of claim 1 , wherein the angiogenic factor comprises Angiopoietin (ANG), Ephrin (Eph), Fibroblast Growth Factor (FGF), Neuropilin (NRP), Plasminogen Activators, urokinase-type plasminogen activator receptor (uPAR), Platelet-Derived Growth Factor (PDGF), Tumor Growth Factor beta (TGF-β), Vascular Endothelial Growth Factor (VEGF), Vascular Endothelial cadherin (VE-cadherin), Insulin-like Growth Factor (IGF), Connective-Tissue Growth Factor (CTGF), Tumor Necrosis Factor alpha (TNF-α), Interlukine 1 (IL-1), Interlukine 6 (IL-6), Granulocyte Macrophage Colony-Stimulating Factor (GM-CSF), and receptors thereof.
17 . A composition comprising the fusion protein according to claim 1 and a pharmaceutically acceptable adjuvant, carrier or excipient.
18 . The method of claim 13 , wherein the angiogenic disease comprises rheumatoid arthritis, inflammatory arthritis, osteoarthritis, ocular and systemic cancer, tumor related metastasis and invasion, systemic fibrosis diseases, ocular disease characterized by neovascularization or ischemia uveitis, retinitis pigmentosa, age-related macular degeneration (AMD), diabetic retinopathy and diabetic macular edema (DME).
19 . The fusion protein according to claim 16 , wherein the VEGF receptors comprise an Ig-like domains D1-D7 of the VEGF receptors.
20 . The fusion protein according to claim 16 , wherein VEGF receptors comprise i) an Ig-like domain D2 of a VEGFR1 and an Ig-like domain D3 of a VEGFR2; ii) the amino acid sequence of SEQ ID NO: 10, or iii) an amino acid sequence having at least 90% identity to SEQ ID NO: 10.
21 . The fusion protein of claim 1 further comprising a GS or G 9 linker between the Fc domain and the integrin binding peptide or the other protein binding peptide.
22 . The fusion protein of claim 1 , wherein the disintegrin has an amino acid sequence selected from a group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, and SEQ ID NO: 7, or amino acid sequence having at least 85% sequence identity to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, and SEQ ID NO: 7.
23 . The method of claim 18 , wherein the systemic fibrosis diseases include idiopathic lung fibrosis (IPF), nonalcoholic steatohepatitis (NASH) or liver fibrosis, diabetic nephropathy and/or kidney fibrosis, dermal fibrosis or keloid, wound healing, cardio-fibrosis, ischemia induced stroke.
24 . The method of claim 18 , wherein the ocular disease characterized by neovascularization or ischemia is choroidal neovascularization.
25 . The fusion protein according to claim 20 , wherein the fusion protein has an amino acid sequence of SEQ ID NO: 16.