# US 12,594,321 B2 · Bispecific chimeric antigen receptor that binds CD19 and CD20, encoding nucleic acid molecules thereof and methods of use thereof to treat cancer > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 12,594,321 B2 - Document ID: US12594321B2 - Status: Granted - Kind: B2 - Application number: 18/529,915 - Filed: 2023-12-05 - Published: 2024-08-01 - Granted: 2026-04-07 - 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; Ming Lei; Jimin Zhang - Web page: https://patentgrep.com/patent/US12594321B2 ## 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 A bispecific chimeric antigen receptor (CAR) molecule comprising: (a) first antigen binding domain that binds to CD20 comprising a heavy chain variable domain (VH) comprising an amino acid sequence of SEQ ID NO: 512, a linker comprising an amino acid sequence of SEQ ID NO: 541, and a light chain variable domain (VL) comprising an amino acid sequence of SEQ ID NO: 514; (b) a (Gly4-Ser)5 linker; (c) a second antigen binding domain that binds to CD19 comprising an amino acid sequence of SEQ ID NO: 771; (d) a CD8 hinge comprising an amino acid sequence of SEQ ID NO: 799; (e) a CD8 transmembrane domain comprising an amino acid sequence of SEQ ID NO: 801; (f) a 4-1BB costimulatory domain comprising an amino acid sequence of SEQ ID NO: 803; and (g) a CD3z intracellular signaling domain comprising an amino acid sequence of SEQ ID NO: 807. ### Claim 2 (depends on 1) The bispecific CAR molecule of claim 1, wherein the CAR molecule comprises two scFvs. ### Claim 3 (depends on 1) A nucleic acid molecule encoding the bispecific CAR molecule of claim 1. ### Claim 4 (depends on 3) A vector comprising the nucleic acid molecule of claim 3. ### Claim 5 (depends on 4) The vector of claim 4, wherein the vector: (a) is selected from the group consisting of a DNA, a RNA, a plasmid, a lentivirus vector, adenoviral vector, and a retrovirus vector; and/or (b) is an in vitro transcribed vector. ### Claim 6 (depends on 4) The vector of claim 4, wherein the vector comprises a single promoter. ### Claim 7 (depends on 6) The vector of claim 6, wherein the single promoter is an EF-1a promoter. ### Claim 8 (depends on 7) The vector of claim 7, wherein the EF-1a promoter comprises the nucleic acid sequence of SEQ ID NO: 833. ### Claim 9 (depends on 3) A cell comprising the nucleic acid molecule of claim 3. ### Claim 10 (depends on 9) The cell of claim 9, wherein the cell is: (a) a human T cell or NK cell, optionally wherein the T cell is a CD8+ T cell; and/or (b) an autologous T cell or NK cell or an allogeneic T cell or NK cell. ### Claim 11 (depends on 3) A method of making a cell, comprising transducing a T cell or an NK cell with the nucleic acid molecule of claim 3. ### Claim 12 (depends on 3) A method of generating a population of RNA-engineered cells comprising introducing an in vitro transcribed RNA or synthetic RNA into a cell, wherein the RNA comprises the nucleic acid molecule of claim 3. ### Claim 13 A population of immune effector cells, comprising a bispecific chimeric antigen receptor (CAR) molecule comprising: (a) first antigen binding domain that binds to CD20 comprising a heavy chain variable domain (VH) comprising an amino acid sequence of SEQ ID NO: 512, a linker comprising an amino acid sequence of SEQ ID NO: 541, and a light chain variable domain (VL) comprising an amino acid sequence of SEQ ID NO: 514; (b) a (Gly4-Ser)5 linker; (c) a second antigen binding domain that binds to CD19 comprising an amino acid sequence of SEQ ID NO: 771; (d) a CD8 hinge comprising an amino acid sequence of SEQ ID NO: 799; (e) a CD8 transmembrane domain comprising an amino acid sequence of SEQ ID NO: 801; (f) a 4-1BB costimulatory domain comprising an amino acid sequence of SEQ ID NO: 803; and (g) a CD3z intracellular signaling domain comprising an amino acid sequence of SEQ ID NO: 807. ### Claim 14 (depends on 13) The population of immune effector cells of claim 13, wherein the population comprises autologous T cells. ### Claim 15 (depends on 3) A method of treating cancer, wherein said method comprises administering to the mammal an effective amount of a population of T cells or NK cells comprising a nucleic acid molecule of claim 3. ### Claim 16 (depends on 15) The method of claim 15, wherein the mammal has a cancer, and wherein the cancer is acute myeloid leukemia (AML), B-cell acute lymphoblastic leukemia (BALL), small lymphocytic lymphoma (SLL), acute lymphoblastic leukemia (ALL), chronic myelogenous leukemia (CML), chronic lymphocytic leukemia (CLL), mantle cell lymphoma (MCL), B cell prolymphocytic leukemia, blastic plasmacytoid dendritic cell neoplasm, Burkitt's lymphoma, diffuse large B cell lymphoma (DLBCL), follicular lymphoma, hairy cell leukemia, small cell-lymphoma, large cell-follicular lymphoma, a malignant lymphoproliferative condition, MALT lymphoma, Marginal zone lymphoma, multiple myeloma, myelodysplasia, or myelodysplastic syndrome, myeloproliferative neoplasm, non-Hodgkin's lymphoma, Hodgkin's lymphoma, plasmablastic lymphoma, plasmacytoid dendritic cell neoplasm, Waldenstrom macroglobulinemia, preleukemia, or a combination thereof. ### Claim 17 (depends on 15) The method of claim 15, wherein the cell is administered in combination with: (i) an agent that increases the efficacy of the cell, (ii) an agent that ameliorates one or more side effects associated with administration of the cell, or (iii) an agent that treats a cancer. ### Claim 18 (depends on 13) A method of treating diffuse large B cell lymphoma (DLBCL) in a subject in need thereof, the method comprising administering the population of immune effector cells of claim 13 to the subject, wherein the population of immune effector cells comprises T cells or NK cells.