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EP-4737481-A2 - CHIMERIC ANTIGEN RECEPTOR AND CAR-T CELLS THAT BIND BCMA

EP4737481A2EP 4737481 A2EP4737481 A2EP 4737481A2EP-4737481-A2

Abstract

The invention relates to an isolated chimeric antigen receptor polypeptide (CAR), wherein the CAR comprises an extracellular antigen-binding domain, comprising an antibody or antibody fragment that binds a B Cell Maturation Antigen (BCMA) polypeptide. The CAR preferably binds an epitope comprising one or more amino acids of residues 13 to 32 of the N-terminus of human BCMA. The invention further relates to a nucleic acid molecule encoding the CAR of the invention, a genetically modified immune cell, preferably a T cell, expressing the CAR of the invention and the use of said cell in the treatment of a medical disorder associated with the presence of pathogenic B cells, such as a disease of plasma cells, memory B cells and/or mature B cells, in particular multiple myeloma, non-Hodgkin's lymphoma or autoantibody-dependent autoimmune diseases.

Inventors

  • REHM, Armin
  • HÖPKEN, Uta Elisabeth
  • BLUHM, Julia
  • UCKERT, WOLFGANG
  • KIEBACK, ELISA
  • MARINO, Stephen

Assignees

  • Max-Delbrück-Centrum für Molekulare Medizin in der Helmholtz-Gemeinschaft

Dates

Publication Date
20260506
Application Date
20170607

Claims (15)

  1. A chimeric antigen receptor (CAR) polypeptide, wherein the CAR comprises: i. an extracellular antigen-binding domain that binds a B Cell Maturation Antigen (BCMA) polypeptide, ii. a transmembrane domain, and iii. an intracellular domain, and wherein said antigen-binding domain comprises a variable heavy chain (VH), said VH having at least 80% sequence identity to SEQ ID NO: 11, and comprising: - a heavy chain complementary determining region 1 (H-CDR1) comprising SEQ ID NO: 34 (RYWX 1 S), wherein X 1 is I, F, or M; - a heavy chain complementary determining region 2 (H-CDR2) comprising amino acids 50-67 of SEQ ID NO: 53 (EINPZ 2 SSTINYAPSLKX 11 X 12 ), wherein Z 2 is S, N, or D; X 11 is D or G; and X 12 is K or R; and - a heavy chain complementary determining region 3 (H-CDR3) comprising SEQ ID NO: 36 (SLYX 4 DYGDAX 5 DYW), wherein X 4 is Y, and X 5 is Y or M; and wherein said antigen-binding domain comprises a variable light chain (VL), said VL having at least 80% sequence identity to SEQ ID NO: 12, and comprising: - a light chain complementary determining region 1 (L-CDR1) comprising SEQ ID NO: 37 (KASQSVX 1 X 2 NVA), wherein X 1 X 2 is ES or DS; - a light chain complementary determining region 2 (L-CDR2) comprising SEQ ID NO 29 (SASLRFS), and - a light chain complementary determining region 3 (L-CDR3) comprising SEQ ID NO 30 (QQYNNYPLTFG).
  2. The chimeric antigen receptor (CAR) polypeptide according to the preceding claim, wherein the VH has at least 90% sequence identity to SEQ ID NO: 11, and wherein the VL has at least 90% sequence identity to SEQ ID NO: 12.
  3. The chimeric antigen receptor (CAR) polypeptide according to any one of the preceding claims, comprising the following sequences: i. H-CDR1: RYWFS (SEQ ID NO. 25) or RYWMS, ii. H-CDR2: EINPSSSTINYAPSLKDK (SEQ ID NO. 26) or EINPDSSTINYAPSLKDK, iii. H-CDR3: SLYYDYGDAYDYW (SEQ ID NO. 27) or SLYYDYGDAMDYW, iv. L-CDR1: KASQSVESNVA (SEQ ID NO. 28) or KASQSVDSNVA, v. L-CDR2: SASLRFS (SEQ ID NO. 29), and vi. L-CDR3: QQYNNYPLTFG (SEQ ID NO. 30).
  4. The chimeric antigen receptor (CAR) polypeptide according to any one of the preceding claims, wherein (a) the VH comprises the sequence of SEQ ID NO: 41, and the H-CDR1, H-CDR2, and H-CDR3 of claim 1, and (b) the VL comprises the sequence of SEQ ID NO: 50, and the L-CDR1, L-CDR2, and L-CDR3 of claim 1.
  5. The chimeric antigen receptor (CAR) polypeptide according to claim 1, wherein the VH domain comprises at least W33, E50, L99, Y100, Y101 and A106 of SEQ ID NO 11, and wherein the VL domain comprises at least S31, A34, F49 or Y49, S50, L53, F55, S67, Q89, Y91, Y94 and L96 of SEQ ID NO 12.
  6. The chimeric antigen receptor (CAR) polypeptide according to claim 1, wherein the VH domain comprises at least the CDR sequences of SEQ NO 25-27, and wherein the VL domain comprises at least the CDR sequences of SEQ ID NO 28-30.
  7. The chimeric antigen receptor (CAR) polypeptide according to any one of the preceding claims, comprising the VH and VL domains according to SEQ ID NO 11 and SEQ ID NO 12, respectively.
  8. The chimeric antigen receptor (CAR) polypeptide according to any one of the preceding claims, wherein when said CAR is expressed in a genetically modified immune cell, preferably a T lymphocyte, said immune cell binds BCMA on the surface of a non-Hodgkin's lymphoma (B-NHL) via said CAR and is activated, thereby inducing cytotoxic activity against said B-NHL, wherein the B-NHL preferably is JeKo-1, DOHH-2, SU-DHL4, JVM-3 and/or MEC-1 cell lines.
  9. The chimeric antigen receptor (CAR) polypeptide according to any one of the preceding claims, wherein the extracellular antigen-binding domain comprises: i. a linker polypeptide positioned between the VH and VL domains; and/or ii. a spacer polypeptide positioned between the extracellular antigen-binding domain and the transmembrane domain.
  10. The chimeric antigen receptor (CAR) polypeptide according to the preceding claim, wherein the linker is selected from a Whitlow or Gly-Ser linker, or linkers with at least 80% sequence identity to SEQ ID NO: 13 or 14, and/or wherein the spacer is selected from an IgG1-CD28 spacer of SEQ ID NO: 15, an IgG1Δ - 4-1BB spacer of SEQ ID NO: 16, an IgG4 (Hi-CH2-CH3) spacer of SEQ ID NO: 17, an IgG4 (Hi-CH3) spacer of SEQ ID NO: 18, an IgG4 (Hi) spacer of SEQ ID NO: 19, or a spacer with at least 80% sequence identity to any one of SEQ ID NO: 15 to 19.
  11. The chimeric antigen receptor (CAR) polypeptide according to any one of the preceding claims i. wherein the transmembrane domain is selected from (i) that of CD8α, comprising SEQ ID NO: 20, or (ii) that of CD28, comprising SEQ ID NO: 21, or (iii) a transmembrane domain with at least 80% sequence identity to SEQ ID NO: 20 or 21, ii. wherein the intracellular domain comprises (i) a co-stimulatory signaling domain of 4-1BB comprising SEQ ID NO: 22, or (ii) a co-stimulatory signaling domain of CD28 comprising SEQ ID NO: 23, or (iii) a co-stimulatory signaling domain with at least 80% sequence identity to SEQ ID NO: 22 or 23, iii. comprising a signaling domain, wherein said signaling domain is selected from (i) a signaling domain of CD3zeta comprising SEQ ID NO: 24, or (ii) a signaling domain with at least 80% sequence identity to SEQ ID NO: 24, and/or iv. comprising a tandem co-stimulatory signaling domain, comprising (i) a 4-1BB co-stimulatory signaling domain comprising SEQ ID NO: 22, and (ii) a CD28 co-stimulatory signaling domain comprising SEQ ID NO: 23, and (iii) a signaling/activation domain of CD3zeta comprising SEQ ID NO: 24.
  12. A nucleic acid molecule encoding a chimeric antigen receptor (CAR) polypeptide according to any one of the preceding claims.
  13. A cell comprising a nucleic acid molecule encoding a CAR according to any one of claims 1 to 11.
  14. The cell according to the preceding claim, wherein the cell is an immune effector cell, a progenitor of an effector cell, or a cell that can be induced to differentiate into an immune effector cell, or wherein the cell is a T cell or an NK cell, wherein the T cell or NK cell comprises BCMA-specific cytotoxicity upon activation through the CAR binding of BCMA.
  15. The cell according to claims 13 or 14 for use in the treatment of multiple myeloma, non-Hodgkin's lymphoma, an autoantibody-dependent autoimmune disease, systemic lupus erythematosus (SLE) or rheumatoid arthritis.

Description

The invention relates to an isolated chimeric antigen receptor polypeptide (CAR), wherein the CAR comprises an extracellular antigen-binding domain, comprising an antibody or antibody fragment that binds a B Cell Maturation Antigen (BCMA) polypeptide. The CAR preferably binds an epitope comprising one or more amino acids of residues 13 to 32 of the N-terminus of human BCMA. The invention further relates to a nucleic acid molecule encoding the CAR of the invention, a genetically modified immune cell, preferably a T cell, expressing the CAR of the invention and the use of said cell in the treatment of a medical disorder associated with the presence of pathogenic B cells, such as a disease of plasma cells, memory B cells and/or mature B cells, in particular multiple myeloma, non-Hodgkin's lymphoma or autoantibody-dependent autoimmune diseases. BACKGROUND OF THE INVENTION In cancer immunotherapy, adoptive transfer of T cells (ATT) genetically modified to recognize tumor-specific or tumor-associated antigens is a promising approach in order to eradicate tumor and tumor stem cells. Thus, in contrast to traditional chemo-, radiation- and surgical therapies, tumor recurrence can be potentially avoided. Moreover, novel pathway-selective drugs often allow for excellent tumor control, but the disease course usually switches to a chronic phase without definite tumor elimination. The advent of genetically modified T cells that express CARs has proven a tremendous success in B cell lymphoma/leukemia treatment, despite the fact that patients were heavily pre-treated and had previously received several lines of chemotherapies, antibody therapies or even autologous/allogeneic bone marrow transplantations. Thus, ATT with CAR-T cells was used successfully as salvage therapy. CARs are synthetic, engineered immunoglobulin-derived receptors that can recognize surface antigens in an MHC-independent fashion. Unlike TCRs, CARs have a broader range of affinities that can engage the target antigen without necessarily exhibiting cross-reactivity. The target antigens must be surface-deposited and can include tumor-associated proteins, carbohydrates or even glycolipids. Another advantage of CAR-T cells is their rapid generation by transduction of autologous T cells, which can be either of CD4+ or CD8+ origin. CARs can be produced "off-the-shelf" and their targets are typically broadly expressed (>90%) in a defined tumor entity, as shown for CD19+ B-cell leukemias and lymphomas. It has been suggested that CAR T cells act as a "living drug" that could be maintained even after a single T cell infusion. A strong medical demand exists for the chimeric antigen receptor (CAR)-T cell product described herein. Firstly, multiple myeloma is an incurable B cell non-Hodgkin lymphoma (B-NHL) which is derived from a malignantly transformed plasma cell clone. As a peculiarity, tumor cells localize predominantly to the bone marrow. This disease is the most frequent tumor of bone and bone marrow, has a 10-year survival rate of 50% among intensely treated younger patients, and is responsible for 2% of annual deaths from cancer. The incidence rate is 5/100.000, and the median age at diagnosis is 70 years, indicating that in many patients' co-morbidities exist that preclude intense and prolonged chemotherapies. The standard of care is chemotherapy, either alone or in combination with autologous stem cell transplantation, immunomodulatory drugs, local irradiation, proteasome inhibitors, and for very few patients allogeneic stem cell transplantation is applicable. Despite intense treatments with the aforementioned modalities, the disease usually relapses and after multiple lines of therapies, secondary resistance develops. Secondly, the much larger group of classical B-NHL contain diverse entities of neoplasias derived from B lymphocytes that usually home to secondary lymphatic organs such as diffuse large B cell lymphoma (DLBCL), follicular lymphoma (FL), and a subgroup of chronic lymphocytic leukemia (CLL). While the total incidence rate of all NHL is about 10-12/100.000 (>85% of B cell origin), most of them are diseases of adults with a substantial increase in the elderly. The demographic development would predict that total numbers will increase due to aging of Western societies. Clinically, B-NHL are heterogeneous and can be distinguished by an aggressive and indolent course. Substantial progress has been made over the last 15 years in the treatment of B-NHL, the standard of care is combined antibody/chemotherapy, either alone or in combination with autologous stem cell transplantation, immunomodulatory drugs, irradiation, proteasome inhibitors, signaling pathway inhibitors, and for very few patients allogeneic stem cell transplantation applies. Because in many B-NHL entities median age at diagnosis is >55-60 years, co-morbidities also exist that preclude intense and extended chemotherapies or even allogeneic bone marrow transplantations. The advent of a