# US 11,077,178 B2 · Chimeric antigen receptor (CAR) that targets chemokine receptor CCR4 and its use > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 11,077,178 B2 - Document ID: US11077178B2 - Status: Granted - Kind: B2 - Application number: 16/334,724 - Filed: 2017-09-20 - Published: 2019-08-01 - Granted: 2021-08-03 - Earliest claimed: 2016-09-21 - Assignees: The United States of America, as represented by the Secretary, Department of Health and Human Services; The United States Government as represented by the Department of Veterans Affairs - Applicants: THE UNITED STATES OF AMERICA, as represented by the Secretary, Department of Health and Human Services; The United States Government as represented by the Department of Veterans Affairs - Inventors: Liyanage Parakrama Perera; Thomas Alexander Waldmann; Kevin Charles Conlon; Pin-Yu Perera - Web page: https://patentgrep.com/patent/US11077178B2 ## CPC - C07K 16/2866: {against receptors for cytokines, lymphokines, interferons} - A61K 31/675: having nitrogen as a ring hetero atom, e.g. pyridoxal phosphate - A61K 31/7076: containing purines, e.g. adenosine, adenylic acid - A61K 40/11: T-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer [LAK] cells - A61K 40/31: Chimeric antigen receptors [CAR] - A61K 40/42: Cancer antigens - A61K 40/4236: {Chemokines} - A61K 40/4239: {Cell cycle regulated proteins, e.g. cyclin, CDC, CDK or INK-CCR} - A61P 35/00: Antineoplastic agents - A61P 35/02: specific for leukemia - C07K 14/7051: {T-cell receptor (TcR)-CD3 complex} - C07K 14/70517: {CD8} - C07K 14/70578: {NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95 (NGF-receptor C07K14/71, TNF-receptor C07K14/7151)} - A61K 2239/48: Blood cells, e.g. leukemia or lymphoma - C07K 2317/24: containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered - C07K 2317/622: Single chain antibody (scFv) - C07K 2319/03: containing a transmembrane segment - C07K 2319/33: fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies - 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) - 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) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS ## Abstract A chimeric antigen receptor is disclosed that includes: (a) an scFv comprising a light chain variable domain (VL) and a heavy chain variable domain (VH), wherein the scFv specifically binds to CCR4; (b) a hinge and transmembrane domain from CD8; (c) an intracellular 4-1BB signaling domain; and (d) an intracellular CD3 zeta signaling domain, wherein (a)-(d) are in N to C terminal order. Uses of the chimeric antigen receptor, such as for treating a malignancy, are also disclosed. ## Claims ### Claim 1 A method for treating a subject with a malignancy that produces CCR4 mRNA, comprising: transducing CD3+ T cells and/or natural killer cells from the subject with a nucleic acid molecule encoding a chimeric antigen receptor, wherein the nucleic acid molecule comprises the nucleic acid sequence of SEQ ID NO: 11, and wherein the chimeric antigen receptor comprises: (a) an extracellular scFv comprising a light chain variable domain (VL) and a heavy chain variable domain (VH), wherein the scFv specifically binds to CCR4, wherein the VH comprises a heavy chain complementarity determining region (HCDR)1, an HCDR2 and an HCDR3, wherein the HCDR1 comprises amino acids 26-33 of SEQ ID NO: 1, the HCDR2 comprises amino acids 51-58 of SEQ ID NO: 1, and the HCDR3 comprises amino acids 99-107 of SEQ ID NO: 1, and wherein the VL comprises a light chain complementarity determining region (LCDR)1, a LCDR2 and a LCDR3, wherein the LCDR1 comprises amino acids 27-38 of SEQ ID NO: 2, the LCDR2 comprises amino acids 56-61 of SEQ ID NO: 2, and the LCDR3 comprises amino acids 95-102 of SEQ ID NO: 2; (b) a hinge and transmembrane domain from CD8; (c) an intracellular 4-1BB signaling domain; (d) an intracellular CD3 zeta signaling domain, wherein (a)-(d) are in N-terminal to C-terminal order, and wherein the chimeric antigen receptor further comprises an immunoglobulin signal sequence comprising the amino acid sequence of SEQ ID NO: 4; to produce autologous transduced cells that express the chimeric antigen receptor; and administering to the subject a therapeutically effective amount of the autologous transduced cells that express the chimeric antigen receptor, thereby treating the malignancy that produces CCR4 mRNA in the subject. ### Claim 2 (depends on 1) The method of claim 1, wherein the malignancy is adult T cell leukemia, cutaneous T cell lymphoma, anaplastic large cell lymphoma, Hodgkin's lymphoma, peripheral T cell lymphoma or a diffuse large B cell lymphoma. ### Claim 3 (depends on 2) The method of claim 2, wherein the adult T cell leukemia is chronic adult T cell leukemia, acute adult T cell leukemia, smoldering T cell leukemia, or lymphomatous T cell leukemia. ### Claim 4 (depends on 1) The method of claim 1, wherein the hinge and the transmembrane domain from CD8 comprise the amino acid sequence set forth as SEQ ID NO: 7. ### Claim 5 (depends on 1) The method of claim 1, wherein the intracellular 4-1BB signaling domain comprises the amino acid sequence set forth as SEQ ID NO: 8. ### Claim 6 (depends on 1) The method of claim 1, wherein the CD3 zeta signaling domain comprises the amino acid sequence set forth as SEQ ID NO: 9. ### Claim 7 (depends on 1) The method of claim 1, comprising a nucleic acid molecule encoding the VL that is at least 95% identical to SEQ ID NO: 12. ### Claim 8 (depends on 1) The method of claim 1, wherein the nucleic acid molecule encoding the chimeric antigen receptor comprises a nucleic acid sequence encoding the VH that is at least 95% identical to SEQ ID NO: 13. ### Claim 9 (depends on 8) The method of claim 8, wherein the nucleic acid sequence encoding the VH is SEQ ID NO: 13, and wherein the nucleic acid molecule comprises a nucleic acid sequence encoding the VL, and wherein the nucleic acid sequence encoding the VL is SEQ ID NO: 12. ### Claim 10 (depends on 1) The method of claim 1, wherein the CD8 transmembrane domain and hinge are encoded by a nucleic acid sequence at least 80% identical to SEQ ID NO: 15, wherein the encoded protein functions as a CD8 transmembrane domain and hinge. ### Claim 11 (depends on 10) The method of claim 10, wherein the nucleic acid molecule encoding the chimeric antigen receptor comprises the nucleic acid sequence of SEQ ID NO: 15. ### Claim 12 (depends on 1) The method of claim 1, wherein the 4-1BB signaling domain is encoded by a nucleic acid sequence at least 80% identical to SEQ ID NO: 16, wherein the encoded protein functions as a 4-1BB signaling domain. ### Claim 13 (depends on 1) The method of claim 1, wherein the CD3 Zeta signaling domain is encoded by nucleic acid sequence at least 80% identical to SEQ ID NO: 17, wherein the encoded protein functions as a CD3 Zeta signaling domain. ### Claim 14 (depends on 13) The method of claim 13, wherein the nucleic acid molecule encoding the chimeric antigen receptor comprises the nucleic acid sequence of SEQ ID NO: 17. ### Claim 15 (depends on 1) The method of claim 1, wherein the nucleic acid molecule encoding the chimeric antigen receptor is codon-optimized for expression in human cells. ### Claim 16 (depends on 1) The method of claim 1, wherein the nucleic acid molecule encoding the chimeric antigen receptor comprises the nucleic acid sequence of SEQ ID NO: 18. ### Claim 17 (depends on 1) The method of claim 1, comprising transducing the CD3+ T cells and/or natural killer cells from the subject with an expression vector comprising the nucleic acid molecule encoding the chimeric antigen receptor, wherein the vector is a viral vector. ### Claim 18 (depends on 17) The method of claim 17, wherein the viral vector is a lentiviral vector or a gamma retroviral vector. ### Claim 19 (depends on 1) The method of claim 1, wherein the CD3+ T cells are autologous to the subject. ### Claim 20 (depends on 1) The method of claim 1, wherein the CD3+ T cells comprise CD3+CD4+ T cells and/or CD3+CD8+ T cells. ### Claim 21 (depends on 1) The method of claim 1, wherein the method reduces the number of Foxp3+CD4+ Treg cells in the subject. ### Claim 22 (depends on 1) The method of claim 1, wherein the subject is human. ### Claim 23 (depends on 1) The method of claim 1, wherein the malignancy is a solid tumor. ### Claim 24 (depends on 1) The method of claim 1, wherein the malignancy is a lymphoid malignancy. ### Claim 25 (depends on 1) The method of claim 1, wherein the method reduces the number of Foxp3+CD4+ Treg cells a tumor microenvironment in the subject. ### Claim 26 (depends on 1) The method of claim 1, wherein the nucleic acid molecule encoding the chimeric antigen receptor is RNA. ### Claim 27 (depends on 1) The method of claim 1, wherein the method comprises transducing CD3+ T cells from the subject with the nucleic acid molecule encoding the chimeric antigen receptor, and wherein the method further comprises: stimulating the CD3+ cells from the subject with an antibody that specifically binds CD3 and an antibody that specifically binds CD28. ### Claim 28 A method for treating a subject with a malignancy that produces CCR4 mRNA, comprising: transducing CD3+ T cells and/or natural killer cells from the subject with a nucleic acid molecule encoding a chimeric antigen receptor, wherein the chimeric antigen receptor comprises the amino acid sequence set forth as SEQ ID NO: 10 to produce autologous transduced cells that express the chimeric antigen receptor; and administering to the subject a therapeutically effective amount of the autologous transduced cells that express the chimeric antigen receptor, thereby treating the malignancy that produces CCR4 mRNA in the subject. ### Claim 29 (depends on 28) The method claim 28, wherein the nucleic acid molecule encoding the chimeric antigen receptor comprises the nucleic acid sequence of SEQ ID NO: 16. ### Claim 30 A method for treating a subject with a malignancy that produces CCR4 mRNA, comprising: transducing CD3+ T cells and/or natural killer cells from the subject with a nucleic acid molecule encoding a chimeric antigen receptor, to produce autologous transduced cells that express the chimeric antigen receptor, wherein the nucleic acid molecule comprises the nucleic acid sequence of SEQ ID NO: 14, and wherein the chimeric antigen receptor comprises: (a) an extracellular scFv comprising a light chain variable domain (VL) and a heavy chain variable domain (VH), wherein the scFv specifically binds to CCR4, wherein the VH comprises a heavy chain complementarity determining region (HCDR)1, an HCDR2 and an HCDR3, wherein the HCDR1 comprises amino acids 26-33 of SEQ ID NO: 1, the HCDR2 comprises amino acids 51-58 of SEQ ID NO: 1, and the HCDR3 comprises amino acids 99-107 of SEQ ID NO: 1, and wherein the VL comprises a light chain complementarity determining region (LCDR)1, a LCDR2 and a LCDR3, wherein the LCDR1 comprises amino acids 27-38 of SEQ ID NO: 2, the LCDR2 comprises amino acids 56-61 of SEQ ID NO: 2, and the LCDR3 comprises amino acids 95-102 of SEQ ID NO: 2, wherein scFv comprises a linker, and (b) a hinge and transmembrane domain from CD8; (c) an intracellular 4-1BB signaling domain; (d) an intracellular CD3 zeta signaling domain, wherein (a)-(d) are in N-terminal to C-terminal order; and administering to the subject a therapeutically effective amount of the autologous transduced cells that express the chimeric antigen receptor, thereby treating the malignancy that produces CCR4 mRNA in the subject.