HUMANIZED ANTI-EGFL7 ANTIBODIES AND METHODS USING SAME

• US 20130064822A1
• Filed: 11/02/2012
• Published: 03/14/2013
• Est. Priority Date: 05/08/2009
• Status: Active Grant

• Current Assignee: Genentech, Inc. (Roche Holding AG)
• Original Assignee: Genentech, Inc. (Roche Holding AG)
• Inventors: Ye, Weilan, Dennis, Mark S., Fredrickson, Jill
• Granted Patent: US 8,574,576 B2
• Time in Patent Office: Days
• US Class Current: 424/136.1
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 2039/507 Comprising a combination of two or more separate antibodies
• CPC Class Codes: A61K 31/282 Platinum compounds
• CPC Class Codes: A61K 31/337 having four-membered rings, e.g. taxol
• CPC Class Codes: A61K 31/513 having oxo groups directly attached to the heterocyclic ring, e.g. cytosine
• CPC Class Codes: A61K 31/519 ortho- or peri-condensed with heterocyclic rings
• CPC Class Codes: A61K 31/555 containing heavy metals, e.g. hemin, hematin, melarsoprol
• CPC Class Codes: A61K 31/56 Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
• CPC Class Codes: A61K 38/19 Cytokines; Lymphokines; Interferons
• CPC Class Codes: A61K 39/001131 Epidermal growth factor [EGF]
• CPC Class Codes: A61K 39/3955 against proteinaceous materials, e.g. enzymes, hormones, lymphokines
• CPC Class Codes: A61K 39/39558 against tumor tissues, cells, antigens
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61P 1/04 for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
• CPC Class Codes: A61P 1/08 for nausea, cinetosis or vertigo; Antiemetics
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 15/00 Drugs for genital or sexual disorders for disorders of sex hormones A61P5/24; Contraceptives
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 17/06 Antipsoriatics
• CPC Class Codes: A61P 19/02 : for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 21/00 : Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 27/02 : Ophthalmic agents
• CPC Class Codes: A61P 27/04 : Artificial tears; Irrigation solutions
• CPC Class Codes: A61P 27/06 : Antiglaucoma agents or miotics
• CPC Class Codes: A61P 29/00 : Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 3/00 : Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 3/10 : for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 31/00 : Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 31/04 : Antibacterial agents
• CPC Class Codes: A61P 35/00 : Antineoplastic agents
• CPC Class Codes: A61P 37/04 : Immunostimulants
• CPC Class Codes: A61P 37/06 : Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 43/00 : Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 5/14 : of the thyroid hormones, e.g. T3, T4
• CPC Class Codes: A61P 9/00 : Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/10 : for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C07K 16/18 : against material from animals or humans
• CPC Class Codes: C07K 16/22 : against growth factors ; against growth regulators
• CPC Class Codes: C07K 16/46 : Hybrid immunoglobulins hybrids of an immunoglobulin with a peptide not being an immunoglobulin C07K19/00
• CPC Class Codes: C07K 16/468 : Immunoglobulins having two or more different antigen binding sites, e.g. multifunctional antibodies
• CPC Class Codes: C07K 2317/24 : containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/56 : variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C07K 2317/565 : Complementarity determining region [CDR]
• CPC Class Codes: C07K 2317/567 : Framework region [FR]
• CPC Class Codes: C07K 2317/73 : Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
• CPC Class Codes: C07K 2317/92 : Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value

Abstract

The present invention concerns antibodies to EGFL7 and the uses of same.

Claims

1. A method of reducing or inhibiting perfusion and permeability of a tumor in a subject, the method comprising administering to an individual in need of such treatment an effective amount of an anti-EGFL7 antibody, the antibody comprising the following hypervariable region (HVR) sequences:

(i) HVR-L1 comprising KX₁SX₂SX₃DYX₄GDSYX₅S, wherein X₁ is A or R;

X₂ is H or Q;

X₃ is G or V;

X₄ is selected from the group consisting of D, L, R, S, and W; and

X₅ is M or V (SEQ ID NO;

     210);

(ii) HVR-L2 comprising GASX₁X₂EX₃, wherein X₁ is N or Y;

X₂ is selected from the group consisting of L, R and Y; and

X₃ is Q or S (SEQ ID NO;

     211);

(iii) HVR-L3 comprising QQNNEX₁PX₂T, wherein X₁ is D or E; and

X₂ is F or Y (SEQ ID NO;

     212);

(iv) HVR-H1 comprising GX₁X₂X₃X₄TYGX₅S, wherein X₁ is H or V;

X₂ is R or T;

X₃ is selected from the group consisting of F, G, R, and S;

X₄ is selected from the group consisting of D, G, R, and T; and

X₅ is M or Y (SEQ ID NO;

     213);

(v) HVR-H2 comprising GWINX₁X₂SGVPTX₃AX₄X₅X₆X₇X₈, wherein X₁ is selected from the group consisting of I, M, T, and W;

X₂ is H or R;

X₃ is selected from group consisting of I, M, T, and Y;

X₄ is D or H;

X₅ is selected from group consisting of D, M and T;

X₆ is F or Y;

X₇ is K or S; and

X₈ is G or R (SEQ ID NO;

214, and

(vi) HVR-H3 comprising AX₁LGSX₂AVDX₃, wherein X₁ is N or R;

X₂ is selected from the group consisting of C, R, H, and Y; and

X₃ is A or Y (SEQ ID NO;

     215).

3. The method of claim 1, wherein HVR-L1 comprises the amino acid sequence KASQSVDYSGDSYMS (SEQ ID NO:

234), HVR-L2 comprises the amino acid sequence GASYRES (SEQ ID NO;

     235), HVR-L3 comprises the amino acid sequence QQNNEEPYT (SEQ ID NO;

     236), HVR-H1 comprises the amino acid sequence GHTFTTYGMS (SEQ ID NO;

     34), HVR-H2 comprises the amino acid sequence GWINTHSGVPTYADDFKG (SEQ ID NO;

     35), and HVR-H3 comprises the amino acid sequence ARLGSYAVDY (SEQ ID NO;

     237).

4. The method of claim 1, wherein the heavy chain comprises the following framework sequences:

FR-H1 comprises EX₁QLVESGGGLVQPGGSLRLSCAAS, wherein X₁ is I or V (SEQ ID NO;

     216);

FR-H2 comprises WVRQAPGKGLEWX₁, wherein X₁ is I, M, or V (SEQ ID NO;

     217);

FR-H3 comprises

RFTX₁SX₂DX₃SX₄X₅TX₆YLQMNSLRAEDTAVYX₇CAR, wherein X₁ is F or I;

X₂ is L or R;

X₃ is N or T, X₄ is selected from the group consisting of A, E, K and T;

X₅ is N or S;

X₆ is selected from the group consisting of A, L, M, T and V; and

X₇ is F or Y (SEQ ID NO;

     218); and

FR-H4 comprises WGQGTLVTVSS (SEQ ID NO;

     219).

5. The method of claim 4, wherein the heavy chain comprises the following framework sequences:

FR-H1 comprises EVQLVESGGGLVQPGGSLRLSCAAS (SEQ ID NO;

     197);

FR-H2 comprises WVRQAPGKGLEWV (SEQ ID NO;

     198);

FR-H3 comprises RFTISX₁DNSKNTX₂YLQMNSLRAEDTAVYYCAR, wherein X₁ L or R;

X₂ is selected from the group consisting of A, L, M, T and V (SEQ ID NO;

     220); and

FR-H4 comprises WGQGTLVTVSS (SEQ ID NO;

     200).

6. The method of claim 4, wherein the heavy chain comprises the following framework sequences:

FR-H1 comprises EIQLVESGGGLVQPGGSLRLSCAAS (SEQ ID NO;

     238);

FR-H2 comprises WVRQAPGKGLEWM (SEQ ID NO;

     239);

FR-H3 comprises RFTISX₁DNSKSTX₂YLQMNSLRAEDTAVYFCAR, wherein X₁ L or R;

X₂ is selected from the group consisting of A, L, M, T and V (SEQ ID NO;

     240); and

FR-H4 comprises WGQGTLVTVSS (SEQ ID NO;

     219).

7. The method of claim 1, wherein the light chain comprises the following framework sequences:

FR-L1 comprises DIQMTQSPSSLSASVGDRVTITC (SEQ ID NO;

     201), FR-L2 comprises WYQQKPGKAPKLLIY (SEQ ID NO;

     202), FR-L3 comprises GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC (SEQ ID NO;

     203), FR-L4 comprises FGQGTKVEIK (SEQ ID NO;

     221) or FGQGTKVEIKR (SEQ ID NO;

     204).

8. The method of claim 1, wherein the light chain comprises the variable domain sequence of 4F11.v17 as shown in FIG. 15 (SEQ ID NO:

82) or the variable domain sequence of 4F11.v22 as shown in FIG. 15 (SEQ ID NO;

     83).

9. The method of claim 1, wherein the heavy chain comprises the variable domain sequence of 4F11.v17 as shown in FIG. 16 (SEQ ID NO:

84) or the variable domain sequence of 4F11.v22 as shown in FIG. 16 (SEQ ID NO;

     85).

10. The method of claim 1, wherein the light chain comprises the variable domain sequence of 4F11.v17 as shown in FIG. 15 (SEQ ID NO:

82) and the heavy chain comprises the variable domain sequence of 4F11.v17 as shown in FIG. 16 (SEQ ID NO;

     84).

11. The method of claim 1, wherein the light chain comprises the variable domain sequence of 4F11.v22 as shown in FIG. 15 (SEQ ID NO:

83) and the heavy chain comprises the variable domain sequence of 4F11.v22 as shown in FIG. 16 (SEQ ID NO;

     85).

12. The method of claim 1, wherein at least a portion of the framework sequence is a human consensus framework sequence.

13. The method of claim 12, comprising human x subgroup 1 consensus framework sequence or heavy chain human subgroup III consensus framework sequence.

26. The method of claim 1 or 14, wherein said antibody is a bispecific antibody.

27. The method of claim 26, wherein said bispecific antibody binds to vascular endothelial growth factor (VEGF).

28. The method of claim 1 or 14, wherein the tumor is selected from the group consisting of breast, colorectal, lung, esophageal, bladder, ovarian, pancreatic, and hepatocellular tumor.

29. The method of claim 28, wherein the tumor is a colorectal or lung tumor.

30. The method of claim 1 or 14, further comprising administering to the individual an effective amount of a second medicament, wherein the anti-EGFL7 antibody is a first medicament.

31. The method of claim 30, wherein the second medicament is another antibody, a chemotherapeutic agent, a cytotoxic agent, an anti-angiogenic agent, an immunosuppressive agent, a prodrug, a cytokine, a cytokine antagonist, cytotoxic radiotherapy, a corticosteroid, an anti-emetic, a cancer vaccine, an analgesic, or a growth-inhibitory agent.

32. The method of claim 31, wherein the second medicament is an anti-VEGF antibody.

33. The method of claim 32, where the second medicament is bevacizumab or ranibizumab.

34. The method of claim 30, further comprising administering to the individual an effective amount of a further medicament, wherein the anti-EGFL7 antibody is a first medicament and an anti-VEGF antibody is the second medicament.

35. The method of claim 34, wherein the further medicament is a chemotherapeutic agent.

36. The method of claim 35, wherein the chemotherapeutic agent is FOLFOX.

37. The method of claim 35, wherein the chemotherapeutic agent is carboplatin and paclitaxel.

2. A method of reducing or inhibiting perfusion and permeability of a tumor in a subject, the method comprising administering to an individual in need of such treatment an effective amount of an anti-EGFL7 antibody, the antibody comprising the following HVR sequences:

HVR-L1 comprising an amino acid sequence selected from SEQ ID NOs;

37-43, HVR-L2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs;

44-47, HVR-L3 comprising the amino acid sequence SEQ ID NO;

48, HVR-H1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs;

49-57, HVR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs;

58-73, and HVR-H3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs;

74-77.

14. A method of reducing or inhibiting perfusion and permeability of a tumor in a subject, the method comprising administering to an individual in need of such treatment an effective amount of an anti-EGFL7 antibody, the antibody comprising a variable domain comprising the following HVR sequences:

(i) HVR-L1 comprising X₁X₂X₃X₄X₅X₆VX₇X₈X₉X₁₀ITYLX₁₁, wherein X₁ is selected from the group consisting of L, Q, R, S, and T;

X₂ is selected from the group consisting of P, T, and W;

X₃ is H or S;

X₄ is D or Q;

X₅ is G or S;

X₆ is L or V;

X₇ is H or P;

X₈ is selected from the group consisting of I, L, P, T, and Y;

X₉ is selected from the group consisting of N, Q or S;

X₁₀ is selected from the group consisting of A, G, and S; and

X₁₁ is G or H (SEQ ID NO;

     222);

(ii) HVR-L2 comprising RVSNX₁X₂S, wherein X₁ is D or R; and

X₂ is selected from the group consisting of A, G, F, I, and T (SEQ ID NO;

     223);

(iii) HVR-L3 comprising X₁QSX₂X₃VPLT, wherein X₁ is selected from the group consisting of A, G, I, K, L, N, T, and V;

X₂ is C or T; and

X₃ is F or H (SEQ ID NO;

     224);

(iv) HVR-H1 comprising GYX₁X₂X₃DX₄YX₅N, wherein X₁ is N or T;

X₂ is F or V;

X₃ is selected from the group consisting of I, M, R, and S;

X₄ is selected from the group consisting of Y, Q, and K; and

X₅ is I or M (SEQ ID NO;

     225);

(v) HVR-H2 comprising GDINX₁X₂X₃X₄X₅X₆HX₇X₈X₉X₁₀X₁₁X₁₂X₁₃, wherein X₁ is selected from the group consisting of A, L, N, and P;

X₂ is selected from the group consisting of D, L, and R;

X₃ is selected from the group consisting of G, K, N, R, S, and Y;

X₄ is G or S;

X₅ is selected from the group consisting of G, I, K, R, S, T, and V;

X₆ is selected from the group consisting of G, R, and T;

X₇ is selected from the group consisting of I, V, L, and Y;

X₈ is N or S;

X₉ is selected from the group consisting of A, N, and Q;

X₁₀ is K or V;

X₁₁ is F or Q;

X₁₂ is K or T; and

X₁₃ is selected from the group consisting of G, H, R, and S (SEQ ID NO;

     226); and

(vi) HVR-H3 comprising X₁REGVYHX₂YDDYAX₃DY, wherein X₁ is selected from the group consisting of A, N, and T;

X₂ is D or P; and

X₃ is M or W (SEQ ID NO;

     227).

16. The method of claim 14, wherein HVR-L1 comprises the amino acid sequence RTSQSLVHINAITYLH (SEQ ID NO:

241), HVR-L2 comprises the amino acid sequence RVSNRFS (SEQ ID NO;

     101), HVR-L3 comprises the amino acid sequence GQSTHVPLT (SEQ ID NO;

     131), HVR-H1 comprises the amino acid sequence GYTFIDYYMN (SEQ ID NO;

     103), HVR-H2 comprises the amino acid sequence GDINLDNSGTHYNQKFKG (SEQ ID NO;

     242), and HVR-H3 comprises the amino acid sequence AREGVYHDYDDYAMDY (SEQ ID NO;

     105).

17. The method of claim 14, wherein the heavy chain comprises the following framework sequences:

FR-H1 comprises EVQLVESGGGLVQPGGSLRLSCAAS (SEQ ID NO;

     197);

FR-H2 comprises WVRQAPGKGLEWX₁, wherein X₁ is I or V (SEQ ID NO;

     228);

FR-H3 comprises RX₁TX₂SX₃DX₄SX₅X₆TX₇YX₈QMNSLRAEDTAVYYC, wherein X₁ is F or V;

X₂ is I or L;

X₃ is selected from the group consisting of L, R, and V;

X₄ is K or N;

X₅ is selected from the group consisting of K, N, R, and S;

X₆ is N or S;

X₇ is selected from the group consisting of A, L, and V; and

X₈ is L or M (SEQ ID NO;

     229); and

FR-H4 comprises WGQGTLVTVSS (SEQ ID NO;

     200).

18. The method of claim 17, wherein the heavy chain comprises the following framework sequences:

FR-H1 comprises EVQLVESGGGLVQPGGSLRLSCAAS (SEQ ID NO;

     197);

FR-H2 comprises WVRQAPGKGLEWV (SEQ ID NO;

     198);

FR-H3 comprises RFTISRDX₁SKNTX₂YLQMNSLRAEDTAVYYCAR, wherein X₁ is N or K; and

X₂ is selected from the group consisting of A, L, and V (SEQ ID NO;

     230); and

FR-H4 comprises WGQGTLVTVSS (SEQ ID NO;

     200).

19. The method of claim 14, wherein the light chain comprises the following framework sequences:

FR-L1 comprises DIQMTQSPSSLSASVGDRVTITC (SEQ ID NO;

     201), FR-L2 comprises WYQQKPGKAPKLLIY (SEQ ID NO;

     202), FR-L3 comprises GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC (SEQ ID NO;

     203), FR-L4 comprises FGQGTKVEIK (SEQ ID NO;

     221) or FGQGTKVEIKR (SEQ ID NO;

     204).

20. The method of claim 14, wherein the light chain comprises the variable domain sequence of 18F7.v6 as shown in FIG. 27 (SEQ ID NO:

193) or the variable domain sequence of 18F7.v6k as shown in FIG. 27 (SEQ ID NO;

     194).

21. The method of claim 14, wherein the heavy chain comprises the variable domain sequence of 18F7.v6 as shown in FIG. 28 (SEQ ID NO:

195) or the variable domain sequence of 18F7.v6k as shown in FIG. 28 (SEQ ID NO;

     196).

22. The method of claim 14, wherein the light chain comprises the variable domain sequence of 18F7.v6 as shown in FIG. 27 (SEQ ID NO:

193) and the heavy chain comprises the variable domain sequence of 18F7.v6 as shown in FIG. 28 (SEQ ID NO;

     195).

23. The method of claim 14, wherein the light chain comprises the variable domain sequence of 18F7.v6k as shown in FIG. 27 (SEQ ID NO:

194) and the heavy chain comprises the variable domain sequence of 18F7.v6k as shown in FIG. 28 (SEQ ID NO;

     196).

24. The method of claim 14, wherein at least a portion of the framework sequence is a human consensus framework sequence.

25. The method of claim 24, comprising human κ

subgroup 1 consensus framework sequence or heavy chain human subgroup III consensus framework sequence.

15. A method of reducing or inhibiting perfusion and permeability of a tumor in a subject, the method comprising administering to an individual in need of such treatment an effective amount of an anti-EGFL7 antibody, the antibody comprising the following HVR sequences:

HVR-L1 comprising an amino acid sequence selected from SEQ ID NOs;

106-124 and 243-246, HVR-L2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs;

125-129, HVR-L3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs;

130-145, HVR-H1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs;

146-153, HVR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs;

154-187 and 247, and HVR-H3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs;

188-192.