Optimized variants of anti-VEGF antibodies and methods of treatment thereof by reducing or inhibiting angiogenesis

Patent metadata

Field	Value
Publication number	US-10072075-B2
Application number	US-201615274612-A
Country	US
Kind code	B2
Filing date	Sep 23, 2016
Priority date	Sep 23, 2015
Publication date	Sep 11, 2018
Grant date	Sep 11, 2018

Abstract

The present invention provides anti-VEGF antibodies and compositions that include anti-VEGF antibodies (e.g., antibody conjugates, fusion proteins, and polymeric formulations), and uses thereof, for example for treatment of disorders associated with pathological angiogenesis. The present invention also provides methods of identifying antibody variants with improved properties, for example, enhanced binding affinity, stability, pharmacokinetics, and/or expression.

First claim

What is claimed is:

1. An isolated antibody that specifically binds vascular endothelial growth factor (VEGF), wherein the antibody comprises the following six hypervariable regions (HVRs):
(a) an HVR-H1 comprising the amino acid sequence of DYWIH (SEQ ID NO: 1);
(b) an HVR-H2 comprising the amino acid sequence of GX 1 TPX 2 GGX 3 X 4 X 5 YX 6 DSVX 7 X 8 (SEQ ID NO: 2), wherein X 1 is Ile or His, X 2 is Ala or Arg, X 3 is Tyr or Lys, X 4 is Thr or Glu, X 5 is Arg, Tyr, Gln, or Glu, X 6 is Ala or Glu, X 7 is Lys or Glu, and X 8 is Gly or Glu;
(c) an HVR-H3 comprising the amino acid sequence of FVFFLPYAMDY (SEQ ID NO: 3);
(d) an HVR-L1 comprising the amino acid sequence of RASQX 1 VSTAVA (SEQ ID NO: 4), wherein X 1 is Asp or Arg;
(e) an HVR-L2 comprising the amino acid sequence of X 1 ASFLYS (SEQ ID NO: 5), wherein X 1 is Ser or Met; and
(f) an HVR-L3 comprising the amino acid sequence of X 1 QGYGX 2 PFT (SEQ ID NO: 6), wherein X 1 is Gln, Asn, or Thr and X 2 is Ala, Gln, or Arg.

2. The antibody of claim 1 , wherein the antibody comprises the following six HVRs:
(a) an HVR-H1 comprising the amino acid sequence of DYWIH (SEQ ID NO: 1);
(b) an HVR-H2 comprising the amino acid sequence of GITPAGGYTRYADSVKG (SEQ ID NO: 7), GITPAGGYEYYADSVKG (SEQ ID NO: 21), or GITPAGGYEYYADSVEG (SEQ ID NO: 22);
(c) an HVR-H3 comprising the amino acid sequence of FVFFLPYAMDY (SEQ ID NO: 3);
(d) an HVR-L1 comprising the amino acid sequence of RASQDVSTAVA (SEQ ID NO: 8);
(e) an HVR-L2 comprising the amino acid sequence of SASFLYS (SEQ ID NO: 9); and
(f) an HVR-L3 comprising the amino acid sequence of QQGYGAPFT (SEQ ID NO: 10) or QQGYGNPFT (SEQ ID NO: 23).

3. The antibody of claim 2 , wherein the antibody comprises the following six HVRs:
(a) an HVR-H1 comprising the amino acid sequence of DYWIH (SEQ ID NO: 1);
(b) an HVR-H2 comprising the amino acid sequence of GITPAGGYTRYADSVKG (SEQ ID NO: 7);
(c) an HVR-H3 comprising the amino acid sequence of FVFFLPYAMDY (SEQ ID NO: 3);
(d) an HVR-L1 comprising the amino acid sequence of RASQDVSTAVA (SEQ ID NO: 8);
(e) an HVR-L2 comprising the amino acid sequence of SASFLYS (SEQ ID NO: 9); and
(f) an HVR-L3 comprising the amino acid sequence of QQGYGAPFT (SEQ ID NO: 10).

4. The antibody of claim 3 , wherein the antibody further comprises the following heavy chain variable (VH) domain framework regions (FRs):
(a) an FR-H1 comprising the amino acid sequence of EVQLVESGGGLVQPGGSLRLSCAASGFTIS (SEQ ID NO: 13);
(b) an FR-H2 comprising the amino acid sequence of WVRQAPGKGLEWVA (SEQ ID NO: 14);
(c) an FR-H3 comprising the amino acid sequence of RFTISADTSKNTAYLQMRSLRAEDTAVYYCAR (SEQ ID NO: 15); and
(d) an FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 16).

5. The antibody of claim 4 , wherein the antibody further comprises the following light chain variable (VL) domain FRs:
(a) an FR-L1 comprising the amino acid sequence of DIQMTQSPSSLSASVGDRVTITC (SEQ ID NO: 17);
(b) an FR-L2 comprising the amino acid sequence of WYQQKPGKAPKLLIY (SEQ ID NO: 18);
(c) an FR-L3 comprising the amino acid sequence of GVPSRFSGSGSGTDFTLTISSLQPEDAATYYC (SEQ ID NO: 19); and
(d) an FR-L4 comprising the amino acid sequence of FGQGTKVEIK (SEQ ID NO: 20).

6. The antibody of claim 1 , wherein the antibody is capable of inhibiting the binding of VEGF to a VEGF receptor.

7. The antibody of claim 6 , wherein the VEGF receptor is VEGF receptor 1 (Flt-1) or VEGF receptor 2 (KDR).

8. The antibody of claim 1 , wherein the antibody binds human VEGF (hVEGF) with a Kd of about 2 nM or lower; has a melting temperature (Tm) of greater than about 83.5° C.; and/or has an isoelectric point (pI) of lower than 8.

9. The antibody of claim 1 , wherein the antibody is monoclonal, human, humanized, or chimeric.

10. The antibody of claim 1 , wherein the antibody is an antibody fragment that binds VEGF.

11. The antibody of claim 10 , wherein the antibody fragment is selected from the group consisting of Fab, Fab-C, Fab′-SH, Fv, scFv, and (Fab′) 2 fragments.

12. The antibody of claim 1 , wherein the antibody is a monospecific antibody.

13. The antibody of claim 1 , wherein the antibody is a multispecific antibody.

14. The antibody of claim 13 , wherein the multispecific antibody is a bispecific antibody.

15. An antibody conjugate comprising (i) the antibody of claim 1 and (ii) a hydrophilic polymer covalently attached to the antibody.

16. The antibody conjugate of claim 15 , wherein the hydrophilic polymer is a hyaluronic acid (HA) polymer or a polyethylene glycol (PEG) polymer.

17. The antibody conjugate of claim 16 , wherein the hydrophilic polymer is an HA polymer.

18. The antibody conjugate of claim 15 , wherein the antibody is an antibody fragment that binds VEGF.

19. The antibody conjugate of claim 18 , wherein the antibody fragment is selected from the group consisting of Fab, Fab-C, Fab′, Fab′-SH, Fv, scFv, and (Fab′) 2 fragments.

20. The antibody conjugate of claim 15 , wherein the antibody conjugate has a hydrodynamic radius between about 10 nm and about 60 nm.

21. The antibody conjugate of claim 15 , wherein the antibody conjugate has an ocular half-life that is increased relative to a reference antibody that is not covalently attached to the hydrophilic polymer.

22. The antibody conjugate of claim 15 , wherein the antibody is covalently attached to the polymer by a reversible prodrug linker.

23. The antibody conjugate of claim 22 , wherein the polymer is a hydrogel.

24. The antibody conjugate of claim 23 , wherein the hydrogel is a PEG-based hydrogel.

25. A fusion protein comprising the antibody of claim 1 covalently attached to an HA binding domain.

26. The antibody of claim 1 , wherein the antibody further comprises (a) a VH domain comprising an amino acid sequence having at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 11, 40, or 42; (b) a VL domain comprising an amino acid sequence having at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 12, 41, or 46; or (c) a VH domain as in (a) and a VL domain as in (b).

27. A pharmaceutical composition comprising the antibody of claim 1 .

28. A method of reducing or inhibiting angiogenesis in a subject having a disorder associated with pathological angiogenesis, comprising administering to the subject an effective amount of the antibody of claim 1 , thereby reducing or inhibiting angiogenesis in the subject.

29. A method for treating a disorder associated with pathological angiogenesis, the method comprising administering an effective amount of the antibody of claim 1 to a subject in need of such treatment.

30. The method of claim 29 , wherein the disorder associated with pathological angiogenesis is an ocular disorder or a cell proliferative disorder.

31. The method of claim 30 , wherein the ocular disorder is selected from the group consisting of AMD, macular degeneration, macular edema, DME (including focal, non-center DME and diffuse, center-involved DME), retinopathy, DR (including PDR, NPDR, and high-altitude DR), other ischemia-related retinopathies, ROP, RVO (including CRVO and BRVO forms), CNV (including myopic CNV), corneal neovascularization, a disease associated with corneal neovascularization, retinal neovascularization, a disease associated with retinal/choroidal

Assignees

• Genentech Inc

Inventors

• Koenig Patrick
• Lee Chingwei Vivian
• Rajagopal Karthikan
• Famili Amin
• Fuh Germaine

Classifications

• A61P3/10

  for hyperglycaemia, e.g. antidiabetics · CPC title

• A61P9/10

  for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis · CPC title

• A61P35/00

  Antineoplastic agents · CPC title

• A61P27/06

  Antiglaucoma agents or miotics · CPC title

• A61P27/02

  Ophthalmic agents · CPC title