# US 11,306,142 B2 · Bispecific antibodies to ROR1 and CD3 > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 11,306,142 B2 - Document ID: US11306142B2 - Status: Granted - Kind: B2 - Application number: 16/628,404 - Filed: 2018-07-05 - Published: 2020-05-21 - Granted: 2022-04-19 - Earliest claimed: 2017-07-05 - Assignees: UCL Business PLC - Applicants: UCL Business LTD - Inventors: Amit Nathwani; Satyen Gohil; Marco Della Peruta - Web page: https://patentgrep.com/patent/US11306142B2 ## CPC - C07K 16/2803: {against the immunoglobulin superfamily} - C07K 16/2809: {against the T-cell receptor (TcR)-CD3 complex} - A61P 35/00: Antineoplastic agents - A61K 2039/505: {comprising antibodies} - C07K 2317/24: containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered - C07K 2317/31: multispecific - C07K 2317/33: Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity - C07K 2317/34: Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues - C07K 2317/622: Single chain antibody (scFv) - C07K 2317/73: Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) ## 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. ## Claims ### Claim 1 A bispecific antibody comprising a first antigen binding domain which selectively binds to Receptor Tyrosine Kinase Like Orphan Receptor 1 (ROR1); and a second antigen binding domain which selectively binds to the CD3 subunit of the T-Cell Receptor (TCR); 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; and 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 a LCDR1, an LCDR2 and an LCDR3 as shown in any one of SEQ ID NOs: 63-68; and wherein the heavy chain variable domain comprises a HCDR1, an HCDR2 and an HCDR3 as shown in any one of SEQ ID NOs: 69-74. ### Claim 2 (depends on 1) The antibody of claim 1, wherein HCDR3 of the first antigen binding domain comprises an amino acid sequence selected from any of the sequences set forth in SEQ ID NOs: 27, 36, 44 and 49. ### Claim 3 (depends on 1) The antibody of claim 1, wherein: (a) 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; or (b) 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. ### Claim 4 (depends on 1) The antibody of claim 1, wherein: (a) 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; or (b) 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. ### Claim 5 (depends on 1) The antibody of claim 1, wherein: (a) 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; (b) 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; (c) 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; or (d) 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. ### Claim 6 (depends on 1) The antibody of claim 1, wherein: (a) 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; or (b) 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; or (c) for the first antigen binding domain: (i) 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; (ii) 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; (iii) 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; (iv) 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 (v) 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. ### Claim 7 A bispecific antibody comprising a first antigen binding domain which selectively binds to Receptor Tyrosine Kinase Like Orphan Receptor 1 (ROR1); and a second antigen binding domain which selectively binds to the CD3 subunit of the T-Cell Receptor (TCR), 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 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; and, wherein for the second antigen binding domain: (a) the light chain variable domain comprises the amino acid sequence as set forth as SEQ ID NO: 63 and the heavy chain variable domain comprises the amino acid sequence as set forth as SEQ ID NO: 69; (b) the light chain variable domain comprises the amino acid sequence as set forth as SEQ ID NO: 64 and the heavy chain variable domain comprises the amino acid sequence as set forth as SEQ ID NO: 70; (c) the light chain variable domain comprises the amino acid sequence as set forth as SEQ ID NO: 65 and the heavy chain variable domain comprises the amino acid sequence as set forth as SEQ ID NO: 71; (d) the light chain variable domain comprises the amino acid sequence as set forth as SEQ ID NO: 66 and the heavy chain variable domain comprises the amino acid sequence as set forth as SEQ ID NO: 72; (e) the light chain variable domain comprises the amino acid sequence as set forth as SEQ ID NO: 67 and the heavy chain variable domain comprises the amino acid sequence as set forth as SEQ ID NO: 73; or (f) the light chain variable domain comprises the amino acid sequence as set forth as SEQ ID NO: 68 and the heavy chain variable domain comprises the amino acid sequence as set forth as SEQ ID NO: 74. ### Claim 8 (depends on 1) The antibody of claim 1, wherein the light chain variable domain of the second antigen binding 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 of the second antigen binding 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. ### Claim 9 (depends on 1, 7) The antibody of claim 1 or 7, wherein: (a) the first antigen binding domain is a scFv antibody; (b) the second antigen binding domain is a scFv antibody; or (c) the first and second antigen binding domains are scFv antibodies covalently linked by a peptide linker. ### Claim 10 (depends on 1, 7) The antibody of claim 1 or 7, wherein the antibody comprises the sequence of SEQ ID NO. 75 or 76, or a sequence having at least 90% sequence identity thereto. ### Claim 11 (depends on 1, 7) The antibody of claim 1 or 7, wherein: (i) the antibody is an IgG or an IgM, and/or (ii) wherein the antibody is an IgG1 or an IgG2. ### Claim 12 (depends on 1, 7) A pharmaceutical composition comprising the antibody of claim 1 or 7, and a pharmaceutically acceptable carrier. ### Claim 13 (depends on 1, 7) An isolated nucleic acid encoding the antibody of claim 1 or 7. ### Claim 14 (depends on 1, 7) An isolated host cell transformed with a nucleic acid encoding the antibody of claim 1 or 7. ### Claim 15 (depends on 1, 7) A method for treating a ROR1-expressing cancer, the method comprising administering to a subject in need thereof, a therapeutically effective amount of an antibody of claim 1 or 7, and thereby treating the ROR1-expressing cancer. ### Claim 16 (depends on 15) The method of claim 15, wherein the ROR1-expressing 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.