# US 11,026,976 B2 · Nucleic acid molecules encoding chimeric antigen receptors comprising a CD20 binding domain > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 11,026,976 B2 - Document ID: US11026976B2 - Status: Granted - Kind: B2 - Application number: 16/664,223 - Filed: 2019-10-25 - Published: 2020-04-16 - Granted: 2021-06-08 - Earliest claimed: 2016-10-07 - Assignees: Novartis AG; The Trustees of the University of Pennsylvania - Applicants: Novartis AG; The Trustees of the University of Pennsylvania - Inventors: Barbara Brannetti; Jennifer Brogdon; Boris Engels; Brian Walter Granda; Lu Huang; Ming Lei; Na Li; Jimin Zhang; Carla Guimaraes; Saar Gill; Marco Ruella; Regina M. Young - Web page: https://patentgrep.com/patent/US11026976B2 ## CPC - A61K 38/1774: {Immunoglobulin superfamily (e.g. CD2, CD4, CD8, ICAM molecules, B7 molecules, Fc-receptors, MHC-molecules)} - 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 40/4211: {CD19 or B4} - A61K 40/4212: {CD22, BL-CAM, siglec-2 or sialic acid binding Ig-related lectin 2} - A61K 40/4221: {CD20} - A61P 35/00: Antineoplastic agents - C07K 14/7051: {T-cell receptor (TcR)-CD3 complex} - C07K 16/2803: {against the immunoglobulin superfamily} - C07K 16/2887: {against CD20} - C07K 16/2896: {against molecules with a "CD"-designation, not provided for elsewhere} - C12N 5/0636: {T lymphocytes} - C12N 15/85: for animal cells - A61K 2039/505: {comprising antibodies} - A61K 2039/804: {Blood cells [leukemia, lymphoma]} - A61K 2239/31: characterized by the route of administration - A61K 2239/38: characterised by the dose, timing or administration schedule - 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 2317/31: multispecific - 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 - 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 - 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) - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) ## Abstract The invention provides compositions and methods for treating diseases associated with expression of CD20 or CD22. The invention also relates to chimeric antigen receptor (CAR) specific to CD20 or CD22, vectors encoding the same, and recombinant T or natural killer (NK) cells comprising the CD20 CAR or CD22 CAR. The invention also includes methods of administering a genetically modified T cell or NK cell expressing a CAR that comprises a CD20 or CD22 binding domain. ## Claims ### Claim 1 An isolated chimeric antigen receptor (CAR) molecule comprising a CD20 binding domain, a transmembrane domain, and an intracellular signaling domain, wherein the CD20 binding domain comprises a light chain complementarity determining region 1 (LCDR1), light chain complementarity determining region 2 (LCDR2), light chain complementarity determining region 3 (LCDR3) heavy chain complementarity determining region 1 (HCDR1), heavy chain complementarity determining region 2 (HCDR2), and heavy chain complementarity determining region 3 (HCDR3), wherein the LCDR1, LCDR2, LCDR3, HCDR1, HCDR2, and HCDR3 comprise: (i) SEQ ID NOs: 147, 148, 149, 136, 137, and 138, respectively; (ii) SEQ ID NOs: 150, 151, 152, 139, 140, and 141, respectively; (iii) SEQ ID NOs: 153, 154, 155, 142, 143, and 144, respectively; (iv) SEQ ID NOs: 929, 930, 931, 926, 927, and 928, respectively; (v) SEQ ID NOs: 228, 229, 230, 217, 218, and 219, respectively; (vi) SEQ ID NOs: 231, 232, 233, 220, 221, and 222, respectively; (vii) SEQ ID NOs: 234, 235, 236, 223, 224, and 225, respectively; or (viii) SEQ ID NOs: 944, 945, 988, 941, 942, and 943, respectively. ### Claim 2 (depends on 1) The isolated CAR molecule of claim 1, wherein the CAR comprises a light chain variable region (VL) and a heavy chain variable region (VH), wherein: (i) the VL comprises the amino acid sequence of SEQ ID NO: 156, 129, 439, 237, 264, 291, 318, or 443 or a sequence with 95% identity thereto; (ii) the VH comprises the amino acid sequence of SEQ ID NO: 145, 118, 437, 226, 253, 280, 307, or 441 or an amino acid sequence with at least 95% identity thereto; or (iii) the VL and VH comprise: (a) SEQ ID NOs: 156 and 145, respectively; (b) SEQ ID NOs: 129 and 118 respectively; (c) SEQ ID NOs: 439 and 437, respectively; (d) SEQ ID NOs: 237 and 226, respectively; (e) SEQ ID NOs: 264 and 253, respectively; (f) SEQ ID NOs: 291 and 280, respectively; (g) SEQ ID NOs: 318 and 307, respectively; or (h) SEQ ID NOs: 443 and 441, respectively. ### Claim 3 (depends on 1) The isolated CAR molecule of claim 1, wherein the CD20 binding domain is an scFv. ### Claim 4 (depends on 1) The isolated CAR molecule of claim 1, wherein the CD20 binding domain comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 159, SEQ ID NO: 240, SEQ ID NO: 132, SEQ ID NO: 267, SEQ ID NO: 294, SEQ ID NO: 321, or an amino acid sequence with at least 95% identity thereto. ### Claim 5 (depends on 1) The isolated CAR molecule of claim 1, wherein the transmembrane domain comprises a transmembrane domain of a protein selected from the group consisting of the alpha, beta or zeta chain of the T-cell receptor, CD28, CD3 epsilon, CD45, CD4, CDS, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD123, CD134, CD137 and CD154. ### Claim 6 (depends on 1) The isolated CAR molecule of claim 1, wherein the transmembrane domain comprises the amino acid sequence of SEQ ID NO: 801 or an amino acid sequence with at least 95% identity thereto. ### Claim 7 (depends on 1) The isolated CAR molecule of claim 1, wherein the CD20 binding domain is connected to the transmembrane domain by a hinge region, wherein the hinge region comprises the amino acid sequence of SEQ ID NO: 799 or SEQ ID NO: 814, or an amino acid sequence with at least 95% identity thereto. ### Claim 8 (depends on 1) The isolated CAR molecule of claim 1, wherein the intracellular signaling domain comprises a costimulatory domain, wherein the costimulatory domain comprises a functional signaling domain of a protein selected from the group consisting of OX40, CD2, CD27, CD28, CDS, ICAM-1, LFA-1 (CD11a/CD18), ICOS (CD278) and 4-1BB (CD137). ### Claim 9 (depends on 1) The isolated CAR molecule of claim 1, wherein the intracellular signaling domain comprises a costimulatory domain, wherein the costimulatory domain comprises the amino acid sequence of SEQ ID NO: 803, or an amino acid sequence with at least 95% identity thereto. ### Claim 10 (depends on 1) The isolated CAR molecule of claim 1, wherein the intracellular signaling domain comprises a primary signaling domain, wherein the primary signaling domain comprises a functional signaling domain of CD3 zeta. ### Claim 11 (depends on 1) The isolated CAR molecule of claim 1, wherein the intracellular signaling domain comprises a primary signaling domain, wherein the primary signaling domain comprises the amino acid sequence of SEQ ID NO: 805 or SEQ ID NO: 807, or an amino acid sequence with at least 95% identity thereto. ### Claim 12 (depends on 1) The isolated CAR molecule of claim 1, wherein the intracellular signaling domain comprises a functional signaling domain of 4-1BB and/or a functional signaling domain of CD3 zeta. ### Claim 13 (depends on 1) The isolated CAR molecule of claim 1, wherein: (i) the intracellular signaling domain comprises the amino acid sequence of SEQ ID NO: 803 and/or the amino acid sequence of SEQ ID NO: 805; (ii) the intracellular signaling domain comprises the amino acid sequence of SEQ ID NO:803 and/or the amino acid sequence of SEQ ID NO: 807; (iii) the intracellular signaling domain comprises the amino acid sequence of SEQ ID NO: 803, or an amino acid sequence with at least 95% identity thereto, and/or the amino acid sequence of SEQ ID NO: 805 or SEQ ID NO: 807, or an amino acid sequence with at least 95% identity thereto; or (iv) the intracellular signaling domain comprises the amino acid sequence of SEQ ID NO: 803 and the amino acid sequence of SEQ ID NO: 805 or SEQ ID NO: 807, wherein the sequences comprising the intracellular signaling domain are expressed in the same frame and as a single polypeptide chain. ### Claim 14 (depends on 1) The isolated CAR molecule of claim 1, further comprising a leader sequence, wherein the leader sequence comprises the amino acid sequence of SEQ ID NO: 797, or an amino acid sequence with at least 95% identity thereto. ### Claim 15 (depends on 1) The isolated CAR molecule of claim 1, comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 161, SEQ ID NO: 242, SEQ ID NO: 134, SEQ ID NO: 269, SEQ ID NO: 296, SEQ ID NO: 323, or an amino acid sequence with at least 95% identity thereto, wherein the CAR molecule comprises or does not comprise a signal peptide of SEQ ID NO: 797. ### Claim 16 (depends on 1) An isolated polypeptide molecule comprising the CAR molecule of claim 1 and a second CAR molecule that binds a B-cell antigen. ### Claim 17 (depends on 16) The isolated polypeptide molecule of claim 16, wherein the second CAR molecule binds to CD19, CD22, CD10, CD34, CD123, FLT-3, ROR-1, CD79b, CD79a, or CD179b. ### Claim 18 (depends on 1, 16) The isolated polypeptide molecule of claim 16, further comprising a cleavable peptide between the CAR molecule of claim 1 and the second CAR molecule. ### Claim 19 (depends on 16) The isolated CAR of claim 16, wherein the B-cell antigen is CD19 or CD22. ### Claim 20 (depends on 16) The isolated CAR molecule of claim 16, wherein the CAR molecule that binds a B-cell antigen binds CD19 and comprises: (i) a HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 comprising SEQ ID NOs: 773, 774, 775, 776, 777, and 778, respectively; (ii) a scFv comprising the amino acid sequence of SEQ ID NO: 765; or (iii) a CAR comprising the amino acid sequence of SEQ ID NO: 764, wherein the CAR molecule comprises or does not comprise a signal peptide comprising the amino acid sequence of SEQ ID NO: 797. ### Claim 21 (depends on 16) The isolated CAR molecule of claim 16, wherein the CAR molecule that binds a B-cell antigen binds CD19 and comprises: (i) a HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 comprising SEQ ID NOs: 785, 786, 787, 788, 789, and 790, respectively; (ii) a scFv comprising the amino acid sequence of SEQ ID NO: 1047; or (iii) a CAR comprising the amino acid sequence of SEQ ID NO: 1049, wherein the CAR molecule comprises or does not comprise a signal peptide comprising the amino acid sequence of SEQ ID NO: 797. ### Claim 22 (depends on 16) The isolated CAR molecule of claim 16, wherein the CAR molecule that binds a B-cell antigen binds CD22 and comprises: (i) a HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 comprising: (a) SEQ ID NOs: 719, 720, 721, 730, 731, and 732, respectively; (b) SEQ ID NOs: 722, 723, 724, 733, 734, and 735, respectively; (c) SEQ ID NOs: 725, 726, 727, 736, 737, and 738, respectively; or (d) SEQ ID NOs: 1036, 1037, 1038, 1039, 1040, and 1041, respectively; (ii) a light chain variable region (VL) and a heavy chain variable region (VH), wherein: (a) the VL comprises the amino acid sequence of SEQ ID NO: 840 or 739; (b) the VH comprises the amino acid sequence of SEQ ID NO: 839 or 728; or (c) the VL and VH comprise: (a) SEQ ID NOs: 840 and 839, respectively; or (b) SEQ ID NOs: 739 and 728, respectively; (iii) a scFv comprising the amino acid sequence of SEQ ID NO: 742, 835, 836, or 837; or (iv) a CAR comprising the amino acid sequence of SEQ ID NO: 744, wherein the CAR molecule comprises or does not comprise a signal peptide comprising the amino acid sequence of SEQ ID NO: 797. ### Claim 23 (depends on 22) The isolated CAR molecule of claim 22, wherein: (i) the VH and VL sequences are connected directly without a linker; (ii) the VH and VL sequences are connected via a (Gly4-Ser)n linker, wherein n is 0, 1, 2, 3, 4, 5, or 6; (iii) the VH and VL sequences are connected via a linker, wherein the linker comprises the amino acid sequence of SEQ ID NO: 834; or (iv) the VH and VL sequences are connected via a linker, wherein the linker comprises the amino acid sequence of SEQ ID NO: 741.