Anti-WT1/HLA bi-specific antibody

Patent metadata

Field	Value
Publication number	US-10239952-B2
Application number	US-201415034782-A
Country	US
Kind code	B2
Filing date	Nov 7, 2014
Priority date	Apr 1, 2011
Publication date	Mar 26, 2019
Grant date	Mar 26, 2019

Abstract

Disclosed herein is a bi-specific form of a T cell receptor mimic (TCRm) mAb with reactivity to human immune effector cell antigen and a WT1 peptide/HLA-A epitope. This antibody selectively bound to leukemias and solid tumor cells expressing WT1 and HLA-A as well as activated resting human T cells to release interferon-(IFN-γ) and to kill the target cancer cells in vitro. Importantly, the antibody mediated autologous T cell proliferation and directed potent cytotoxicity against fresh ovarian cancer cells. Therapeutic activity in vivo of the antibody was demonstrated in NOD SCID SCID Yc*(NSG) mice with three different human cancers expressing WT1/HLA-A2 including disseminated Ph+ acute lymphocytic leukemia (ALL), disseminated acute myeloid leukemia, and peritoneal mesothelioma. In both of the leukemia xenograft models, mice that received the antibody and T cells also showed longer survival and delayed limb paralysis. Also provided are methods for stimulating a primary T cell response comprising stimulating cytotoxic T cells against a first tumor antigen and a secondary T cell response comprising stimulating effector T cells and/or memory T cells against a first tumor antigen and/or against a second tumor antigen using the bi-specific antibodies described herein.

First claim

The invention claimed is:

1. A recombinant antibody comprising:
(i) a first antigen-binding portion comprising:
(A) a heavy chain (HC) variable region comprising HC-CDR1, HC-CDR2 and HC-CDR3, and a light chain (LC) variable region comprising LC-CDR1, LC-CDR2 and LC-CDR3, wherein:
(1) the HC-CDR1, HC-CDR2 and HC-CDR3 comprises amino acid sequences set forth in SEQ ID NOs: 2, 3, and 4, respectively, and the LC-CDR1, LC-CDR2 and LC-CDR3 comprises amino acid sequences set forth in SEQ ID NOs: 8, 9, and 10, respectively;
(2) the HC-CDR1, HC-CDR2 and HC-CDR3 comprises amino acid sequences set forth in SEQ ID NOs: 20, 21, and 22, respectively, and the LC-CDR1, LC-CDR2 and LC-CDR3 comprises amino acid sequences set forth in SEQ ID NOs: 26, 27, and 28, respectively;
(3) the HC-CDR1, HC-CDR2 and HC-CDR3 comprises amino acid sequences set forth in SEQ ID NOs: 38, 39, and 40, respectively, and the LC-CDR1, LC-CDR2 and LC-CDR3 comprises amino acid sequences set forth in SEQ ID NOs: 44, 45, and 46, respectively;
(4) the HC-CDR1, HC-CDR2 and HC-CDR3 comprises amino acid sequences set forth in SEQ ID NOs: 56, 57, and 58, respectively, and the LC-CDR1, LC-CDR2 and LC-CDR3 comprises amino acid sequences set forth in SEQ ID NOs: 62, 63, and 64, respectively;
(5) the HC-CDR1, HC-CDR2 and HC-CDR3 comprises amino acid sequences set forth in SEQ ID NOs: 92, 93, and 94, respectively, and the LC-CDR1, LC-CDR2 and LC-CDR3 comprises amino acid sequences set forth in SEQ ID NOs: 80, 81, and 82, respectively; or
(5) the HC-CDR1, HC-CDR2 and HC-CDR3 comprises amino acid sequences set forth in SEQ ID NOs: 74, 75, and 76, respectively, and the LC-CDR1, LC-CDR2 and LC-CDR3 comprises amino acid sequences set forth in SEQ ID NOs: 98, 99, and 100, respectively;

(B) a V H and a V L comprising first and second amino acid sequences as set forth in SEQ ID NOs: 14 and 16, 32 and 34, 50 and 52, 68 and 70, 86 and 88, or 104 and 106; or
(C) an scFv comprising an amino acid sequence as set forth in SEQ ID NO: 18, 36, 54, 72, 90, or 108; and

(ii) a second antigen-binding portion comprising an amino acid sequence set forth in SEQ ID NO: 111, 112, or 113.

2. The recombinant antibody of claim 1 , wherein said first antigen-binding portion and/or said second antigen-binding portion is an antibody fragment selected from the group consisting of a Fab fragment; a monovalent fragment consisting of the VL, VH, CL and CH1 domains; a F(ab)2 fragment; a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; a Fd fragment consisting of the VH and CH1 domains; a Fv fragment consisting of the VL and VH domains of a single arm of an antibody; a dAb fragment; an isolated CDR; and a scFv.

3. The recombinant antibody of claim 2 , wherein said first antigen-binding portion and/or said second antigen-binding portion is a scFv.

4. The recombinant antibody of claim 1 , wherein said first antigen-binding portion specifically binds to WT1/HLA and said second antigen-binding portion specifically binds to an immune effector cell surface antigen.

5. The recombinant antibody of claim 4 , wherein said immune effector cell is selected from the group consisting natural killer (NK) cells, macrophages, and T cells.

6. The recombinant antibody of claim 4 , wherein said immune effector cell is a CD3 + cell and the recombinant antibody specifically binds to CD3.

7. The recombinant antibody of claim 1 , comprising the amino acid sequence set forth in SEQ ID NO: 110.

8. A pharmaceutical composition comprising the recombinant antibody of claim 1 .

Assignees

• Memorial Sloan Kettering Cancer Center
• Eureka Therapeutics Inc

Inventors

• Scheinberg David
• Xiang Jingyi
• Dao Tao
• Yan Su
• Liu Cheng

Classifications

• C07K2317/622

  Single chain antibody (scFv) · CPC title

• C07K2317/34

  Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues · CPC title

• C07K2317/73

  Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation · CPC title

• C07K16/32 Primary

  against translation products of oncogenes · CPC title

• C07K16/2809

  against the T-cell receptor (TcR)-CD3 complex · CPC title