Long-acting VEGF inhibitors for intraocular neovascularization

1. An anti-VEGF agent comprising a polypeptide, the polypeptide comprising a VEGF binding moiety operatively linked to an Fc domain of an IgG immunoglobulin, wherein the VEGF binding moiety comprises:

an IgG-like domain 2 of VEGFR-1 comprising a sequence having at least 95% sequence identity to SEQ ID NO: 16 and

two IgG-like domains 3 of VEGFR-1, each comprising a sequence having at least 95% sequence identity to SEQ ID NO: 17; and

wherein the VEGF binding moiety and the Fc domain are linked, respectively, from the N-terminus of the polypeptide to the C-terminus of the polypeptide, for the treatment of a VEGF-related condition of the eye.

2. The anti-VEGF agent of claim 1 , wherein the VEGF binding moiety further comprises, at its N-terminus, an IgG-like domain 1 of VEGFR-1 comprising a sequence having at least 95% sequence identity to SEQ ID NO: 15.

3. The anti-VEGF agent of claim 2 , wherein the Fc domain of an IgG immunoglobulin is an Fc domain of a human IgG1 immunoglobulin.

4. The anti-VEGF agent of claim 3 , comprising a polypeptide wherein the IgG-like domain 1 of VEGFR-1, the IgG-like domain 2 of VEGFR-1, the two IgG-like domains 3 of VEGFR-1, and the human IgG1 Fc domain are linked, respectively, from the N-terminus of the polypeptide to the C-terminus of the polypeptide.

5. The anti-VEGF agent of claim 1 , wherein the anti-VEGF agent comprises amino acids 27 to 659 of SEQ ID NO: 5.

6. The anti-VEGF agent of claim 1 , wherein the anti-VEGF agent has a heparin binding ability greater than aflibercept.

7. The anti-VEGF agent of claim 1 , wherein the anti-VEGF agent has a vitreous binding ability greater than aflibercept.

8. The anti-VEGF agent of claim 1 , wherein the anti-VEGF agent has a vitreous bound VEGF-stimulated endothelial cell proliferation-inhibiting ability greater than aflibercept.

9. The anti-VEGF agent of claim 1 , wherein the anti-VEGF agent at a concentration of 5 nM inhibits VEGF-stimulated proliferation of bovine choroidal microvascular endothelial cells to a greater extent than aflibercept.

10. The anti-VEGF agent of claim 1 , wherein the anti-VEGF agent has a longer duration after intraocular injection compared to aflibercept.

11. The anti-VEGF agent of claim 1 , wherein 1 nM of the anti-VEGF agent inhibits VEGF-stimulated proliferation of bovine choroidal endothelial cells by greater than 80%.

12. The anti-VEGF agent of claim 1 , wherein 2.5 μg of the anti-VEGF agent inhibits choroid neovascularization by at least 30% when intravitreally injected into a mouse eye 14 days before inducing a choroid injury by laser photocoagulation using a diode laser and a slit lamp with a spot size of 50 μm, power of 180 mW, and exposure duration of 100 ms.

13. The anti-VEGF agent of claim 1 , wherein 2.5 μg of the anti-VEGF agent inhibits choroid neovascularization at least as potently as 25 μg of aflibercept when the anti-VEGF agent or the aflibercept is intravitreously injected 1 day before a choroid injury induced by laser photocoagulation using a diode laser and a slit lamp with a spot size of 50 μm, power of 180 mW, and exposure duration of 100 ms.

14. A method of treating a VEGF related condition in an eye of a subject comprising administering to the subject the anti-VEGF agent of claim 1 .

15. The method of claim 14 , wherein the administering comprises an intravitreal injection.

16. The method of claim 14 , wherein the VEGF related condition comprises neovascularization.

17. The method of claim 16 , wherein the neovascularization is choroidal neovascularization.

18. The method of claim 16 , wherein the neovascularization is retinal neovascularization.

19. The method of claim 14 , wherein the VEGF related condition is age-related macular degeneration.