Bispecific antibodies to ror1 and cd3

Abstract

There is described bispecific antibodies which selectively bind to Receptor Tyrosine Kinase Like Orphan Receptor 1 (ROR1) and the CD3 subunit of the T-Cell Receptor (TCR), their production and their use. Also described is the use of the bispecific antibodies in the treatment of cancer.

1 . A bispecific antibody molecule comprising a first antigen binding domain which selectively binds to Receptor Tyrosine Kinase Like Orphan Receptor 1 (ROR1), wherein the first antigen binding domain binds to an epitope of ROR1 comprising amino acid Gln-261; and
 a second antigen binding domain which selectively binds to the CD3 subunit of the T-Cell Receptor (TCR).   
     
     
         2 . The antibody molecule of  claim 1 , wherein the first antigen binding domain comprises a light chain variable domain and a heavy chain variable domain, wherein the light chain variable domain comprises a light chain complementarity determining region (LCDR)1, an LCDR2 and an LCDR3, wherein LCDR1 comprises the amino acid sequence set forth in SEQ ID NO: 16; LCDR2 comprises the amino acid sequence set forth in SEQ ID NO: 18; and LCDR3 comprises the amino acid sequence set forth in SEQ ID NO: 20; and wherein the heavy chain variable domain comprises a heavy chain complementarity determining region (HCDR)1, an HCDR2 and an HCDR3, wherein HCDR1 comprises the amino acid sequence set forth in SEQ ID NO: 23; HCDR2 comprises the amino acid sequence set forth in SEQ ID NO: 25; and HCDR3 comprises the amino acid sequence set forth in SEQ ID NO: 57; wherein the sequence of each complementarity determining region may differ from the given sequence at up to two amino acid positions. 
     
     
         3 . The antibody molecule of  claim 2 , wherein the sequence of each CDR may differ from the given sequence at one amino acid position. 
     
     
         4 . The antibody molecule of any preceding claim, wherein LCDR1 comprises the amino acid sequence set forth in SEQ ID NO: 16; LCDR2 comprises the amino acid sequence set forth in SEQ ID NO: 18; and LCDR3 comprises the amino acid sequence set forth in SEQ ID NO: 20; and wherein HCDR1 comprises the amino acid sequence set forth in SEQ ID NO: 23; HCDR2 comprises the amino acid sequence set forth in SEQ ID NO: 25; and HCDR3 comprises the amino acid sequence set forth in SEQ ID NO: 57. 
     
     
         5 . The antibody molecule of any preceding claim, wherein HCDR3 comprises an amino acid sequence selected from any of the sequences set forth in SEQ ID NOs: 27, 36, 44 and 49. 
     
     
         6 . The antibody molecule of any preceding claim, wherein the first antigen binding domain has a light chain variable domain which comprises a Light Chain Framework Region (LCFR)1 comprising the amino acid sequence as set forth as one of SEQ ID NOs: 15, 29, 50 and 53; an LCFR2 comprising the amino acid sequence as set forth as one of SEQ ID NOs: 17, 30, 38 and 46; an LCFR3 comprising the amino acid sequence as set forth as one of SEQ ID NOs: 19, 31, 39, 47 and 54; and an LCFR4 comprising the amino acid sequence as set forth as one of SEQ ID NOs: 21, 32 and 40, wherein the sequence of each framework region may differ from the given sequence at up to five amino acid positions. 
     
     
         7 . The antibody molecule of any preceding claim, wherein the first antigen binding domain has a light chain variable domain which comprises an LCFR1 comprising the amino acid sequence as set forth as one of SEQ ID NOs: 29, 50 and 53; an LCFR2 comprising the amino acid sequence as set forth as one of SEQ ID NOs: 30, 38 and 46; an LCFR3 comprising the amino acid sequence as set forth as one of SEQ ID NOs: 31, 39, 47 and 54; and an LCFR4 comprising the amino acid sequence as set forth as one of SEQ ID NOs: 32 and 40, wherein the sequence of each framework region may differ from the given sequence at up to five amino acid positions. 
     
     
         8 . The antibody molecule of any preceding claim, wherein the first antigen binding domain has a heavy chain variable domain which comprises a Heavy Chain Framework Region (HCFR)1 comprising the amino acid sequence as set forth as one of SEQ ID NOs: 22, 33, 41 and 55; an HCFR2 comprising the amino acid sequence as set forth as one of SEQ ID NOs: 24, 34, 42 and 51; an HCFR3 comprising the amino acid sequence as set forth as one of SEQ ID NOs: 26, 35, 43, 48, 52 and 56; and an HCFR4 comprising the amino acid sequence as set forth as one of SEQ ID NOs: 28, 37 and 45, wherein the sequence of each framework region may differ from the given sequence at up to five amino acid positions. 
     
     
         9 . The antibody molecule of any preceding claim, wherein the first antigen binding domain has a heavy chain variable domain which comprises an HCFR1 comprising the amino acid sequence as set forth as one of SEQ ID NOs: 33, 41 and 55; an HCFR2 comprising the amino acid sequence as set forth as one of SEQ ID NOs: 34, 42 and 51; an HCFR3 comprising the amino acid sequence as set forth as one of SEQ ID NOs: 35, 43, 48, 52 and 56; and an HCFR4 comprising the amino acid sequence as set forth as one of SEQ ID NOs: 37 and 45, wherein the sequence of each framework region may differ from the given sequence at up to five amino acid positions. 
     
     
         10 . The antibody molecule of any preceding claim, wherein the light chain variable domain of the first antigen binding domain comprises the amino acid sequence as set forth as one of SEQ ID NOs: 3, 4, 5, 6, 7 and 8, or a sequence having at least 90% identity thereto. 
     
     
         11 . The antibody molecule of any preceding claim, wherein the light chain variable domain of the first antigen binding domain comprises the amino acid sequence as set forth as one of SEQ ID NOs: 4, 5, 6, 7 and 8, or a sequence having at least 90% identity thereto. 
     
     
         12 . The antibody molecule of any preceding claim, wherein the heavy chain variable domain of the first antigen binding domain comprises the amino acid sequence as set forth as one of SEQ ID NOs: 9, 10, 11, 12, 13 and 14, or a sequence having at least 90% identity thereto. 
     
     
         13 . The antibody molecule of any preceding claim, wherein the heavy chain variable domain of the first antigen binding domain comprises the amino acid sequence as set forth as one of SEQ ID NOs: 10, 11, 12, 13 and 14, or a sequence having at least 90% identity thereto. 
     
     
         14 . The antibody molecule of any preceding claim, wherein the light chain variable domain of the first antigen binding domain comprises the amino acid sequence as set forth as one of SEQ ID NOs: 4, 5, 6, 7 and 8, or a sequence having at least 90% identity thereto; and
 the heavy chain variable domain of the first antigen binding domain comprises the amino acid sequence as set forth as one of SEQ ID NOs: 10, 11, 12, 13 and 14, or a sequence having at least 90% identity thereto.   
     
     
         15 . The antibody molecule of any preceding claim, wherein the light chain variable domain of the first antigen binding domain comprises the amino acid sequence as set forth as SEQ ID NO: 4 or a sequence having at least 90% identity thereto, and the heavy chain variable domain of the first antigen binding domain comprises the amino acid sequence as set forth as one of SEQ ID NOs: 10, 12 and 13 or a sequence having at least 90% identity thereto. 
     
     
         16 . The antibody molecule of any preceding claim, wherein for the first antigen binding domain:
 (a) the light chain variable domain comprises the amino acid sequence as set forth as SEQ ID NO: 4 and the heavy chain variable domain comprises the amino acid sequence as set forth as SEQ ID NO: 10;   (b) the light chain variable domain comprises the amino acid sequence as set forth as SEQ ID NO: 5 and the heavy chain variable domain comprises the amino acid sequence as set forth as SEQ ID NO: 11;   (c) the light chain variable domain comprises the amino acid sequence as set forth as SEQ ID NO: 6 and the heavy chain variable domain comprises the amino acid sequence as set forth as SEQ ID NO: 12;   (d) the light chain variable domain comprises the amino acid sequence as set forth as SEQ ID NO: 7 and the heavy chain variable domain comprises the amino acid sequence as set forth as SEQ ID NO: 13; or   (e) the light chain variable domain comprises the amino acid sequence as set forth as SEQ ID NO: 8 and the heavy chain variable domain comprises the amino acid sequence as set forth as SEQ ID NO: 14,   wherein each light chain variable domain and heavy chain variable domain above may have at least 90% identity to the amino acid sequence set forth above.   
     
     
         17 . The antibody molecule of  claim 1 , wherein the first antigen binding domain comprises a light chain variable domain and a heavy chain variable domain, wherein the light chain variable domain comprises a light chain complementarity determining region (LCDR)1, an LCDR2 and an LCDR3, wherein LCDR1 comprises the amino acid sequence set forth in SEQ ID NO: 58; LCDR2 comprises the amino acid sequence set forth in SEQ ID NO: 59; and LCDR3 comprises the amino acid sequence set forth in SEQ ID NO: 20; and wherein the heavy chain variable domain comprises a heavy chain complementarity determining region (HCDR)1, an HCDR2 and an HCDR3, wherein HCDR1 comprises the amino acid sequence set forth in SEQ ID NO: 60; HCDR2 comprises the amino acid sequence set forth in SEQ ID NO: 61; and HCDR3 comprises the amino acid sequence set forth in SEQ ID NO: 62; wherein the sequence of each complementarity determining region may differ from the given sequence at up to two amino acid positions. 
     
     
         18 . The antibody molecule of  claim 17 , wherein for the first antigen binding domain, the light chain variable domain comprises the amino acid sequence as set forth as one of SEQ ID NOs: 3, 4, 5, 6, 7 and 8, and the heavy chain variable domain comprises the amino acid sequence as set forth as one of SEQ ID NOs: 9, 10, 11, 12, 13 and 14,
 wherein the light chain variable domain and heavy chain variable domain above may have at least 90% identity to the amino acid sequence set forth above.   
     
     
         19 . The antibody molecule of  claim 17  or  claim 18 , wherein for the first antigen binding domain:
 (a) the light chain variable domain comprises the amino acid sequence as set forth as SEQ ID NO: 3 and the heavy chain variable domain comprises the amino acid sequence as set forth as SEQ ID NO: 9; 
 (b) the light chain variable domain comprises the amino acid sequence as set forth as SEQ ID NO: 4 and the heavy chain variable domain comprises the amino acid sequence as set forth as SEQ ID NO: 10; 
 (c) the light chain variable domain comprises the amino acid sequence as set forth as SEQ ID NO: 5 and the heavy chain variable domain comprises the amino acid sequence as set forth as SEQ ID NO: 11; 
 (d) the light chain variable domain comprises the amino acid sequence as set forth as SEQ ID NO: 6 and the heavy chain variable domain comprises the amino acid sequence as set forth as SEQ ID NO: 12; 
 (e) the light chain variable domain comprises the amino acid sequence as set forth as SEQ ID NO: 7 and the heavy chain variable domain comprises the amino acid sequence as set forth as SEQ ID NO: 13; or 
 (f) the light chain variable domain comprises the amino acid sequence as set forth as SEQ ID NO: 8 and the heavy chain variable domain comprises the amino acid sequence as set forth as SEQ ID NO: 14, 
 wherein the light chain variable domain and heavy chain variable domain above may have at least 90% identity to the amino acid sequence set forth above. 
 
     
     
         20 . The antibody molecule of any preceding claim, wherein the second antigen binding domain comprises a light chain variable domain and a heavy chain variable domain, wherein the light chain variable domain comprises the amino acid sequence as set forth as one of SEQ ID NOs: 63, 64, 65, 66, 67 and 68, or a sequence having at least 90% identity thereto, and wherein the heavy chain variable domain comprises the amino acid sequence as set forth as one of SEQ ID NOs: 69, 70, 71, 72, 73 and 74, or a sequence having at least 90% identity thereto. 
     
     
         21 . The antibody molecule of any preceding claim, wherein the first antigen binding domain is a scFv antibody. 
     
     
         22 . The antibody molecule of any preceding claim, wherein the second antigen binding domain is a scFv antibody. 
     
     
         23 . The antibody molecule of any preceding claim, wherein the first and second antigen binding domains are scFv antibodies covalently linked by a peptide linker. 
     
     
         24 . The antibody molecule of any preceding claim, wherein the antibody molecule comprises the sequence of SEQ ID NO. 75 or 76, or a sequence having at least 90% sequence identity thereto. 
     
     
         25 . An isolated nucleic acid molecule encoding the antibody molecule of any one of  claims 1  to  24 . 
     
     
         26 . The isolated nucleic acid molecule of  claim 25 , operably linked to a promoter. 
     
     
         27 . An expression vector comprising the isolated nucleic acid molecule of  claim 25  or  claim 26 . 
     
     
         28 . An isolated host cell transformed with the nucleic acid molecule of  claim 25  or  26 , or the vector of  claim 27 . 
     
     
         29 . The antibody molecule of any one of  claims 1  to  24  or the nucleic acid molecule of  claim 25  for use in the treatment of cancer. 
     
     
         30 . The use of an antibody molecule of any one of  claims 1  to  24  or the nucleic acid molecule of  claim 25  in the manufacture of a medicament for the treatment of cancer. 
     
     
         31 . A method for treating cancer comprising administering to a subject an antibody molecule according to any one of  claims 1  to  24 , the nucleic acid molecule of  claim 25  or the vector of  claim 26 . 
     
     
         32 . The use of  claim 29  or  claim 30 , or the method of  claim 31 , wherein the cancer is leukaemia, pancreatic cancer, prostate cancer, colon cancer, bladder cancer, ovarian cancer, glioblastoma, testicular cancer, uterine cancer, adrenal cancer, breast cancer, lung cancer, melanoma, neuroblastoma, sarcoma or renal cancer.