# US 12,600,801 B2 · Single-domain antibodies that bind ROR1 > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 12,600,801 B2 - Document ID: US12600801B2 - Status: Granted - Kind: B2 - Application number: 18/350,617 - Filed: 2023-07-11 - Published: 2024-03-21 - Granted: 2026-04-14 - Earliest claimed: 2022-07-11 - Assignees: Fred Hutchinson Cancer Center - Applicants: Fred Hutchinson Cancer Center - Inventors: Jason Price; Colin E. Correnti; James M. Olson - Web page: https://patentgrep.com/patent/US12600801B2 ## CPC - C07K 16/2809: {against the T-cell receptor (TcR)-CD3 complex} - C07K 16/468: {Immunoglobulins having two or more different antigen binding sites, e.g. multifunctional antibodies} - C07K 16/30: from tumour cells - C07K 16/2803: {against the immunoglobulin superfamily} - C07K 2317/22: from camelids, e.g. camel, llama or dromedary - C07K 2317/24: containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered - C07K 2317/31: multispecific - C07K 2317/52: Constant or Fc region; Isotype - C07K 2317/565: Complementarity determining region [CDR] - C07K 2317/569: Single domain, e.g. dAb, sdAb, VHH, VNAR or nanobody® - C07K 2317/73: Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation - C07K 2317/77: Internalization into the cell - 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) ## Abstract Single-domain antibodies that bind receptor tyrosine kinase (ROR1) are described. The single-domain antibodies can be used for multiple purposes including in research, imaging, diagnosis, and treatment of ROR1-related conditions. The disclosed single-domain antibodies can be used as anti-cancer therapeutics such as antibody-drug conjugates, multi-domain binding molecules, or recombinant receptors or can be used as cancer imaging/diagnostic agents. ## Claims ### Claim 1 A single-domain antibody that binds receptor tyrosine kinase (ROR1), the single domain antibody comprising a set of complementarity determining regions (CDRs) comprising: (i) a CDR1 having the sequence as set forth in SEQ ID NO: 6, a CDR2 having the sequence as set forth in SEQ ID NO: 7, and a CDR3 having the sequence as set forth in SEQ ID NO: 8 according to IMGT; (ii) a CDR1 having the sequence as set forth in SEQ ID NO: 9, a CDR2 having the sequence as set forth in SEQ ID NO: 10, and a CDR3 having the sequence as set forth in SEQ ID NO: 11 according to Kabat; (iii) a CDR1 having the sequence as set forth in SEQ ID NO: 12, a CDR2 having the sequence as set forth in SEQ ID NO: 13, and a CDR3 having the sequence as set forth in SEQ ID NO: 11 according to Chothia; (iv) a CDR1 having the sequence as set forth in SEQ ID NO: 14, a CDR2 having the sequence as set forth in SEQ ID NO: 15, and a CDR3 having the sequence as set forth in SEQ ID NO: 8 according to North; (v) a CDR1 having the sequence as set forth in SEQ ID NO: 16, a CDR2 having the sequence as set forth in SEQ ID NO: 17, and a CDR3 having the sequence as set forth in SEQ ID NO: 18 according to Contact; (vi) a CDR1 having the sequence as set forth in SEQ ID NO: 19, a CDR2 having the sequence as set forth in SEQ ID NO: 20, and a CDR3 having the sequence as set forth in SEQ ID NO: 21 according to IMGT; (vii) a CDR1 having the sequence as set forth in SEQ ID NO: 22, a CDR2 having the sequence as set forth in SEQ ID NO: 23, and a CDR3 having the sequence as set forth in SEQ ID NO: 24 according to Kabat; (viii) a CDR1 having the sequence as set forth in SEQ ID NO: 25, a CDR2 having the sequence as set forth in SEQ ID NO: 26, and a CDR3 having the sequence as set forth in SEQ ID NO: 24 according to Chothia; (ix) a CDR1 having the sequence as set forth in SEQ ID NO: 27, a CDR2 having the sequence as set forth in SEQ ID NO: 28, and a CDR3 having the sequence as set forth in SEQ ID NO: 21 according to North; (x) a CDR1 having the sequence as set forth in SEQ ID NO: 29, a CDR2 having the sequence as set forth in SEQ ID NO: 30, and a CDR3 having the sequence as set forth in SEQ ID NO: 31 according to Contact; (xi) a CDR1 having the sequence as set forth in SEQ ID NO: 32, a CDR2 having the sequence as set forth in SEQ ID NO: 20, and a CDR3 having the sequence as set forth in SEQ ID NO: 33 according to IMGT; (xii) a CDR1 having the sequence as set forth in SEQ ID NO: 34, a CDR2 having the sequence as set forth in SEQ ID NO: 23, and a CDR3 having the sequence as set forth in SEQ ID NO: 24 according to Kabat; (xiii) a CDR1 having the sequence as set forth in SEQ ID NO: 35, a CDR2 having the sequence as set forth in SEQ ID NO: 26, and a CDR3 having the sequence as set forth in SEQ ID NO: 33 according to Chothia; (xiv) a CDR1 having the sequence as set forth in SEQ ID NO: 36, a CDR2 having the sequence as set forth in SEQ ID NO: 28, and a CDR3 having the sequence as set forth in SEQ ID NO: 33 according to North; (XV) a CDR1 having the sequence as set forth in SEQ ID NO: 29, a CDR2 having the sequence as set forth in SEQ ID NO: 30, and a CDR3 having the sequence as set forth in SEQ ID NO: 37 according to Contact; (xvi) a CDR1 having the sequence as set forth in SEQ ID NO: 32, a CDR2 having the sequence as set forth in SEQ ID NO: 20, and a CDR3 having the sequence as set forth in SEQ ID NO: 21 according to IMGT; (xvii) a CDR1 having the sequence as set forth in SEQ ID NO: 35, a CDR2 having the sequence as set forth in SEQ ID NO: 26, and a CDR3 having the sequence as set forth in SEQ ID NO: 24 according to Chothia; or (xviii) a CDR1 having the sequence as set forth in SEQ ID NO: 36, a CDR2 having the sequence as set forth in SEQ ID NO: 28, and a CDR3 having the sequence as set forth in SEQ ID NO: 21 according to North wherein the single-domain antibody is not linked to an IgM Fc region or multimerizing fragment thereof. ### Claim 2 (depends on 1) The single-domain antibody of claim 1, wherein the single-domain antibody comprises a sequence having at least 90% sequence identity to the sequence as set forth in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, or SEQ ID NO: 5. ### Claim 3 (depends on 1) The single-domain antibody of claim 1, wherein the single-domain antibody comprises the sequence as set forth in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, or SEQ ID NO: 5. ### Claim 4 (depends on 1) A heavy chain only antibody (HcAb) comprising the single-domain antibody of claim 1 linked to an IgG, IgA, IgD, or IgE Fc region of an antibody. ### Claim 5 (depends on 1) A multi-domain binding molecule comprising at least two binding domains wherein at least one binding domain comprises a single-domain antibody of claim 1 and wherein the multi-domain binding molecule does not comprise an IgM Fc region or multimerizing fragment thereof. ### Claim 6 (depends on 5) The multi-domain binding molecule of claim 5, wherein the multi-domain binding molecule comprises a CD19 binding domain. ### Claim 7 (depends on 6) The multi-domain binding molecule of claim 6, wherein the CD19 binding domain comprises an anti-CD19 scFv. ### Claim 8 (depends on 6) The multi-domain binding molecule of claim 6, wherein the CD19 binding domain comprises a variable heavy chain sequencing comprising a CDRH1 having the sequence as set forth in SEQ ID NO: 66, a CDRH2 having the sequence as set forth in SEQ ID NO: 67, and a CDRH3 having the sequence as set forth in SEQ ID NO: 68; and a variable light chain sequence comprising a CDRL1 having the sequence as set forth in SEQ ID NO: 63, a CDRL2 having the sequence as set forth in SEQ ID NO: 64, and a CDRL3 having the sequence as set forth in SEQ ID NO: 65; or a variable heavy chain sequence comprising a CDRH1 having the sequence as set forth in SEQ ID NO: 72, a CDRH2 having the sequence as set forth in SEQ ID NO: 73, and a CDRH3 having the sequence as set forth in SEQ ID NO: 74; and a variable light chain sequence comprising a CDRL1 having the sequence as set forth in SEQ ID NO: 69, a CDRL2 having the sequence as set forth in SEQ ID NO: 70, and a CDRL3 having the sequence as set forth in SEQ ID NO: 71. ### Claim 9 (depends on 5) The multi-domain binding molecule of claim 5, wherein the multi-domain binding molecule comprises an immune cell engaging molecule. ### Claim 10 (depends on 9) The multi-domain binding molecule of claim 9, wherein the immune cell engaging molecule activates a B cell, T cell, natural killer (NK) cell, or macrophage. ### Claim 11 (depends on 10) The multi-domain binding molecule of claim 10, wherein the T cell is a CD3 T cell, a CD4 T cell, a CD8 T cell, a central memory T cell, an effector memory T cell, and/or a naïve T cell. ### Claim 12 (depends on 9) The multi-domain binding molecule of claim 9, wherein a binding domain of the immune cell engaging molecule binds CD3, CD28, CD8, NKG2D, CD8, CD16, KIR2DL4, KIR2DS1, KIR2DS2, KIR3DS1, NKG2C, NKG2E, NKG2D, NKp30, NKp44, NKp46, NKp80, DNAM-1, CD11b, CD11c, CD64, CD68, CD119, CD163, CD206, CD209, F4/80, IFGR2, Toll-like receptors 1-9, IL-4Rα, or MARCO. ### Claim 13 (depends on 9) The multi-domain binding molecule of claim 9, wherein a binding domain of the immune cell engaging molecule binds CD3. ### Claim 14 (depends on 13) The multi-domain binding molecule of claim 13, wherein the CD3 binding domain is derived from the OKT3 antibody. ### Claim 15 (depends on 13) The multi-domain binding molecule of claim 13, wherein the binding domain that binds CD3 has a variable heavy chain sequence comprising a CDRH1 having the sequence as set forth in SEQ ID NO: 82, a CDRH2 having the sequence as set forth in SEQ ID NO: 83, and a CDRH3 having the sequence as set forth in SEQ ID NO: 84; and a variable light chain sequence comprising a CDRL1 having the sequence as set forth in SEQ ID NO: 79, a CDRL2 having the sequence as set forth in SEQ ID NO: 80, and a CDRL3 having the sequence as set forth in SEQ ID NO: 81. ### Claim 16 (depends on 9) The multi-domain binding molecule of claim 9, wherein a binding domain of the immune cell engaging molecule binds CD28. ### Claim 17 (depends on 16) The multi-domain binding molecule of claim 16, wherein the CD28 binding domain is derived from the TGN1412 antibody. ### Claim 18 (depends on 5) The multi-domain binding molecule of claim 5, wherein the single-domain antibody is linked to a first Fc region of the multi-domain binding molecule and a binding domain that binds CD3 is linked to the second Fc region of the multi-domain binding molecule. ### Claim 19 (depends on 1) A method of treating a subject in need thereof having an ROR1-related cancer, the method comprising administering a therapeutically effective amount of the single-domain antibody of claim 1 to the subject thereby treating the subject in need thereof having the ROR1-related cancer. ### Claim 20 (depends on 19) The method of claim 19, wherein the ROR1-related cancer comprises chronic lymphocytic leukemia (CLL), hairy cell leukemia (HCL), mantle cell lymphoma (MCL), marginal zone lymphoma (MZL), diffuse large B cell lymphoma (DLBCL), follicular lymphoma (FL), multiple myeloma (MM), acute lymphocytic leukemia (ALL), acute myeloid leukemia (AML), chronic myeloid leukemia (CML), breast cancer, ovarian cancer, pancreatic cancer, lung cancer, or neuroblastoma.