VEGF-activated trail ligands
View Patent ↗The present invention provides fusion proteins comprising an extracellular domain of a VEGF receptor and a death ligand. The fusion proteins bind to VEGF and to death receptors on tumor cells thereby inhibiting VEGF activation of VEGF receptors and inducing apoptosis in the tumor cells. Fusion proteins of the present invention are useful for inducing apoptosis and cytotoxic effects in cells, treating cancer and diseases or disorders related to unregulated angiogenesis and/or vasculogenesis. Thus, this invention further provides methods for treating angiogenesis related diseases using the fusion proteins, polynucleotides encoding the fusion proteins, vectors containing the polynucleotides, pharmaceutical compositions and kits containing the fusion proteins or the polynucleotides encoding the fusion proteins.
1. A soluble fusion protein which binds to a death receptor and induces apoptosis, the fusion protein comprising:
(i) a soluble vascular endothelial growth factor receptor-1 (VEGFR-1) polypeptide that binds a vascular endothelial growth factor (VEGF) polypeptide, wherein the VEGFR-1 polypeptide comprises an amino acid sequence at least 95% identical to amino acid residues 129 to 230 of SEQ ID NO: 19; and
(ii) a soluble human tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) extracellular domain,
wherein the C-terminus of the VEGFR-1 polypeptide is linked to the N-terminus of the TRAIL extracellular domain.
2. The fusion protein of claim 1 , wherein the VEGFR-1 is a human VEGFR-1.
3. The fusion protein of claim 1 , further comprising:
(iii) an epitope tag.
4. The fusion protein of claim 3 , wherein the epitope tag comprises a FLAG-like tag or an HA tag.
5. The fusion protein of claim 4 , wherein the epitope tag can be cleaved off.
6. A method of inducing a death receptor-mediated pathway comprising the step of:
contacting a TRAIL receptor expressing cell with an effective amount of a fusion protein of claim 1 , wherein the amount of the fusion protein is effective to induce the TRAIL receptor-mediated pathway.
7. The method of claim 6 , which is practiced in vitro.
8. The method of claim 6 , which is practiced in vivo.
9. The method of claim 6 , wherein the cell is a cancer cell.
10. The method of claim 9 , wherein the cancer cell overexpresses VEGF.
11. The method of claim 9 , wherein the cancer cell is selected from the group consisting of breast cancer cell, prostate cancer cell, colon cancer cell, lung cancer cell, glioblastoma cell, and ovarian cancer cell.
12. The method of claim 6 , wherein the TRAIL receptor-mediated pathway is modulated in a disease selected from the group consisting of rheumatoid arthritis, psoriasis, and macular degeneration.
13. The method of claim 6 , further comprising the step of contacting the death receptor expressing cell with a chemotherapeutic agent.
14. The method of claim 13 , wherein the chemotherapeutic agent is selected from the group consisting of camptothecin, etoposide, bisindolylmaleimide VIII, cisplatin, taxol, doxorubicin, temozolomide, bortezomid, LY294002, and valproic acid.
15. A pharmaceutical composition comprising:
(i) the fusion protein of claim 1 ; and
(ii) a pharmaceutically acceptable excipient, carrier and/or diluent.