Humanized anti-ox40 antibodies and uses thereof

Patent metadata

Field	Value
Publication number	US-2016137740-A1
Application number	US-201514877547-A
Country	US
Kind code	A1
Filing date	Oct 7, 2015
Priority date	Oct 10, 2014
Publication date	May 19, 2016
Grant date	—

Abstract

The disclosure provides humanized anti-OX40 antibodies. Also provided are methods of making such antibodies, and methods of use, e.g., treatment of cancer.

First claim

What is claimed is:

1 . An antibody or antigen-binding fragment thereof comprising a humanized heavy chain variable region (VH) and a humanized light chain variable region (VL);
wherein the VH comprises an amino acid sequence with the formula:
HFW1-HCDR1-HFW2-HCDR2-HFW3-HCDR3-HFW4,
wherein HFW1 is SEQ ID NO: 6 or SEQ ID NO: 7, HCDR1 is SEQ ID NO: 8, HFW2 is SEQ ID NO: 9, HCDR2 is SEQ ID NO: 14, SEQ ID NO: 15 or SEQ ID NO: 16, HFW3 is SEQ ID NO: 17, HCDR3 is SEQ ID NO: 25, SEQ ID NO: 26, or SEQ ID NO: 27, and HFW4 is SEQ ID NO: 28; wherein the VL comprises the amino acid sequence SEQ ID NO: 29 or SEQ ID NO: 32; and wherein the antibody or fragment thereof can specifically bind to human OX40.

2 . The antibody or fragment thereof of claim 1 , wherein the amino acid sequence of HFW2 is SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, or SEQ ID NO: 13.

3 . The antibody or fragment thereof of claim 1 , wherein the amino acid sequence of HFW3 is SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, or SEQ ID NO: 24.

4 . The antibody or fragment thereof of claim 1 , wherein the VH comprises the amino acid sequence SEQ ID NO: 33, SEQ ID NO: 35, SEQ ID NO: 37, SEQ ID NO: 39, SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, SEQ ID NO: 55, SEQ ID NO: 57, SEQ ID NO: 59, SEQ ID NO: 61, SEQ ID NO: 63, SEQ ID NO: 65, or SEQ ID NO: 67.

5 . The antibody or fragment thereof of claim 1 , wherein the VL comprises the amino acid sequence SEQ ID NO: 29 and the VH comprises the amino acid sequence SEQ ID NO: 59.

6 . The antibody or fragment thereof of claim 1 further comprising a light chain constant region or fragment thereof fused to the C-terminus of the VL.

7 . The antibody or fragment thereof of claim 6 , wherein the light chain constant region is a human kappa constant region.

8 . The antibody or fragment thereof of claim 1 , further comprising a heavy chain constant region or fragment thereof fused to the C-terminus of the VH.

9 . The antibody or fragment thereof of claim 8 , wherein the heavy chain constant region is a human IgG1 constant region, a human IgG4P constant region, a human IgG1TM constant region or a murine IgG1 constant region.

10 . The antibody or fragment thereof of claim 9 , wherein the heavy chain constant region is a human IgG1 constant region.

11 . The antibody or fragment thereof of claim 1 comprising the heavy chain amino acid sequence SEQ ID NO: 71 and the light chain amino acid sequence SEQ ID NO: 30.

12 . The antibody or fragment of claim 1 , wherein the antigen-binding fragment is an Fv fragment, an Fab fragment, an F(ab′)2 fragment, an Fab′ fragment, a dsFv fragment, an scFv fragment, or an sc(Fv)2 fragment, or any combination thereof.

13 . The antibody or fragment thereof of claim 1 , which can specifically bind to cynomolgus monkey, or rhesus monkey OX40.

14 . The antibody or fragment thereof of claim 13 , which can specifically bind to OX40 as expressed on Jurkat cells, primary activated CD4 + or CD8 + T cells from human, cynomolgus monkey, rhesus monkey, or any combination thereof.

15 . The antibody or fragment thereof of claim 1 , which does not bind to murine or rat OX40.

16 . The antibody or fragment thereof of claim 1 , which does not cross react with related Tumor Necrosis Factor Receptor Superfamily (TNFRSF) proteins.

17 . The antibody or fragment thereof of claim 14 , wherein the binding affinity for human OX40 expressed on primary activated human CD4 + T cells is about 250 pM to about 370 pM as measured by flow cytometry.

18 . The antibody or fragment thereof of claim 17 , wherein the binding affinity is about 312 pM.

19 . The antibody or fragment thereof of claim 17 , which can achieve 20% receptor occupancy on primary activated human CD4 + T cells (EC 20 ) at about 63 to about 93 pM, 50% receptor occupancy on primary activated human CD4 + T cells (EC 50 ) at about 250 to about 370 pM, and 90% receptor occupancy on primary activated human CD4 + T cells (EC 90 ) at about 2290 to about 3330 pM as measured by flow cytometry.

20 . The antibody or fragment thereof of claim 19 , wherein EC 20 is about 78 pM, EC 50 is about 312 pM, and EC 90 is about 2810 pM.

21 . The antibody or fragment thereof of claim 14 , wherein the binding affinity for human OX40 expressed on OX40-overexpressing Jurkat cells is about 250 pM to about 600 pM as measured by flow cytometry.

22 . The antibody or fragment thereof of claim 21 , wherein the binding affinity is about 424 pM.

23 . The antibody or fragment thereof of claim 21 , which can achieve EC 20 on OX40-overexpressing Jurkat cells at about 60 to about 150 pM, EC 50 on OX40-overexpressing Jurkat cells at about 250 to about 600 pM, and EC 90 on OX40-overexpressing Jurkat cells at about 2260 to about 4380 pM as measured by flow cytometry.

24 . The antibody or fragment thereof of claim 23 , wherein EC 20 is about 106 pM, EC 50 is about 424 pM, and EC 90 is about 3820 pM.

25 . The antibody or fragment thereof of claim 14 , wherein the binding affinity for cynomolgus monkey OX40 expressed on primary activated cynomolgus monkey CD4 + T cells is about 340 pM to about 820 pM as measured by flow cytometry.

26 . The antibody or fragment thereof of claim 25 , wherein the binding affinity is about 580 pM.

27 . The antibody or fragment thereof of claim 14 , wherein the binding affinity for rhesus monkey OX40 expressed on primary activated rhesus monkey CD4 + T cells is about 130 pM to about 600 pM as measured by flow cytometry.

28 . The antibody or fragment thereof of claim 27 , wherein the binding affinity is about 370 pM.

29 . The antibody or fragment thereof of claim 1 , which can induce dose-dependent proliferation of activated CD4 + T cells and dose-dependent cytokine release in primary activated human CD4 + T cells in a plate-based assay.

30 . The antibody or fragment thereof of claim 29 , wherein a 20% maximal proliferation response (EC 20 ) can be achieved in primary activated human CD4 + T cells at an antibody concentration of about 14 pM to about 28 pM, a 50% maximal proliferation response (EC 50 ) can be achieved in primary activated human CD4 + T cells at an antibody concentration of about 0.3 pM to about 130 pM, and a 90% maximal proliferation response (EC 90 ) can be achieved in primary activated human CD4 + T cells at an antibody concentration of about 50 pM to about 90 pM, all as measured by flow cytometry.

31 . The antibody or fragment thereof of claim 30 , wherein EC 20 is about 21 pM, EC 50 is about 28 pM, and EC 90 is about 72 pM.

32 . The antibody or fragment thereof of claim 29 , wherein the cytokine is IFNγ, TNFα, IL-5, IL-10, IL-2, IL-4, IL-13, IL-8, IL-12 p70, IL-1β, or any combination thereof.

33 . The antibody or fragment thereof of claim 32 wherein the cytokine is IFNγ, TNFα, IL-5, IL-10, IL-13, or any combination thereof.

34 . The antibody or fragment thereof of claim 1 , which can achieve CD4 + T cell proliferation and cytokine release in primary activated cynomolgus monkey CD4 + T cells and in primary activated rhesus monkey CD4 + T cells.

35 . The antibody or fragment

Assignees

• Medimmune Llc

Inventors

• Hammond Scott A
• Oberst Michael
• Du Qun
• Damschroder Melissa

Classifications

• A61P35/00

  Antineoplastic agents · CPC title

• C07K2317/24

  containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered · CPC title

• C07K2317/51

  Complete heavy chain or Fd fragment, i.e. VH + CH1 · CPC title

• C07K16/2878 Primary

  against the NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95 · CPC title

• C07K2317/515

  Complete light chain, i.e. VL + CL · CPC title