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JP-2026076334-A - Composition and method for treating cancer using a chimeric antigen receptor targeting glypican 3

JP2026076334AJP 2026076334 AJP2026076334 AJP 2026076334AJP-2026076334-A

Abstract

[Challenge] To provide a novel chimeric antigen receptor (CAR) T-cell therapy that effectively treats cancer while minimizing the risk of dangerous inflammatory responses such as cytokine release syndrome. [Solution] An isolated nucleic acid sequence encoding CAR, wherein the CAR includes an antigen-binding domain specific to glypican 3 (GPC3), the antigen-binding domain has an equilibrium dissociation constant (KD) of approximately 100 nanomoles (nM) or less, and the CAR construct provides an isolated nucleic acid sequence that does not induce cytokine production in GPC3-expressing cells. [Selection Diagram] Figure 1A

Inventors

  • ギルブレス，ライアン
  • ムーディー，ゴードン
  • ジャルディーノ トーチア，マリア，レティジア
  • ワン，チオン
  • マム，ジョン

Assignees

  • メディミューン，エルエルシー

Dates

Publication Date
20260511
Application Date
20260217
Priority Date
20191220

Claims (20)

  1. An isolated nucleic acid sequence encoding a chimeric antigen receptor (CAR), wherein the CAR includes an antigen-binding domain specific to glypican 3 (GPC3), the antigen-binding domain has an equilibrium dissociation constant ( KD ) of approximately 100 nanomoles (nM) or less, and the construct of the CAR is an isolated nucleic acid sequence that does not induce cytokine production in GPC3 cells.
  2. The isolated nucleic acid sequence according to claim 1, wherein the antigen-binding domain of the encoded CAR comprises an antibody or its antigen-binding fragment.
  3. The isolated nucleic acid sequence according to claim 2, wherein the antigen-binding domain of the encoded CAR is a Fab or a single-chain variable region fragment (scFv).
  4. The isolated nucleic acid sequence according to claim 3, wherein the antigen-binding domain is an scFv containing the nucleic acid sequence of SEQ ID NO: 33 or SEQ ID NO: 34.
  5. An isolated nucleic acid sequence according to any one of claims 1 to 4, further encoding a transmembrane domain, a co-stimulatory domain, and a signaling domain.
  6. The isolated nucleic acid sequence according to claim 5, wherein the encoded transmembrane domain includes a CD28 transmembrane domain.
  7. The isolated nucleic acid sequence according to claim 5, wherein the encoded co-stimulatory domain comprises one or more of the following: CD28, 4-1BB, CD3 zeta, OX-40, ICOS, CD27, GITR, and MyD88/CD40 co-stimulatory domains.
  8. The isolated nucleic acid sequence according to claim 5, wherein the encoded co-stimulatory domain comprises one or more CD28, 4-1BB, and CD3 zeta co-stimulatory domains.
  9. The isolated nucleic acid sequence according to claim 5, wherein the encoded signal domain includes a sequence encoding the CSFR2 signal peptide.
  10. An isolated nucleic acid sequence according to any one of claims 1 to 9, further encoding a hinge/spacer domain.
  11. The isolated nucleic acid sequence according to claim 10, wherein the encoded hinge/spacer domain is an IgG4P hinge/spacer.
  12. The isolated nucleic acid sequence according to claim 1, wherein the nucleic acid sequence includes SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, or SEQ ID NO: 26.
  13. An anti-GPC3 chimeric antigen receptor (CAR) comprising an antigen-binding domain, wherein the antigen-binding domain comprises an antibody, Fab, or scFv comprising a heavy chain variable region (VH) and a light chain variable region (VL); The VH comprises CDR1 containing the amino acid sequence of SEQ ID NO: 37, CDR2 containing the amino acid sequence of SEQ ID NO: 38, and CDR3 containing the amino acid sequence of SEQ ID NO: 39; and the VL comprises CDR1 containing the amino acid sequence of SEQ ID NO: 40 or SEQ ID NO: 43, CDR2 containing the amino acid sequence of SEQ ID NO: 41 or SEQ ID NO: 44, and CDR3 containing the amino acid sequence of SEQ ID NO: 42 or SEQ ID NO: 45, an anti-GPC3 CAR.
  14. The anti-GPC3 CAR according to claim 13, wherein the VH comprises the amino acid sequence of SEQ ID NO: 27 or SEQ ID NO: 29.
  15. The anti-GPC3 CAR according to claim 13, wherein the VL comprises the amino acid sequence of SEQ ID NO: 28 or SEQ ID NO: 30.
  16. The anti-GPC3 CAR according to any one of claims 13 to 15, further comprising a transmembrane domain, a co-stimulatory domain, and a signaling domain.
  17. The anti-GPC3 CAR according to claim 16, wherein the CAR comprises the amino acid sequence of SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, or SEQ ID NO: 25.
  18. A vector comprising a nucleic acid sequence encoding a chimeric antigen receptor (CAR), wherein the nucleic acid sequence comprises SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 26, SEQ ID NO: 33, or SEQ ID NO: 34.
  19. A cell containing the vector according to claim 18.
  20. Cells comprising a nucleic acid sequence encoding a chimeric antigen receptor (CAR), wherein the CAR includes an antigen-binding domain specific to glypican 3 (GPC3), the antigen-binding domain has an equilibrium dissociation constant ( KD ) of approximately 100 nanomoles (nM) or less, and the CAR construct does not induce cytokine production in GPC3 cells.

Description

This disclosure relates to the treatment of cancer using chimeric antigen receptor T cells. 1. Chimeric Antigen Receptor T-Cell Therapy Chimeric antigen receptor (CAR) T-cell therapy is a specific form of cell-based immunotherapy that uses genetically modified T cells to fight cancer. In CAR T-cell therapy, T cells are collected from the patient's blood, ex vivo engineered to express CARs containing both antigen-binding and T-cell activation domains, expanded into a larger population, and administered to the patient. CAR T cells act as living drugs, binding to cancer cells and causing their destruction. When successful, the effects of CAR T-cell therapy tend to be long-lasting, as evidenced by the detection of persistent and expanding CAR T cells in patients long after clinical remission. 2. Structure and Function of CARs The antigen-binding domain of a CAR is an extracellular region that targets surface antigens on tumor cells. Suitable target antigens may be proteins, phosphorylated proteins, peptide-MHCs, carbohydrates, or glycolipid molecules. Ideal target antigens are widely expressed on tumor cells to enable targeting at a high rate in cancer cells. Ideal candidate target antigens are also typically expressed minimally in normal tissues, limiting off-tumor and on-target toxicity. The antigen-binding domain of a CAR contains a targeting moiety, such as an antibody single-chain variable region fragment (scFv), which is directed towards the target antigen. The T cell activation domain of a CAR is located intracellularly and activates T cells in response to its antigen-binding domain, which interacts with its target antigen. The T cell activation domain can contain one or more costimulatory domains, which are intracellular domains of known activated T cell receptors. Since costimulatory domains have different effects on the dynamics, cytotoxic function, and safety profile of CAR T cells, the selection and positioning of costimulatory domains within the CAR construct influences the function and fate of CAR T cells. The extracellular antigen-binding domain and intracellular T-cell activation domain of CAR are linked by a transmembrane domain, a hinge, and an optional spacer region. The hinge domain is a short peptide fragment that provides conformational freedom to facilitate binding to the target antigen on tumor cells. It can be used alone or in conjunction with a spacer domain that causes the scFv to protrude from the T-cell surface. The optimal length of the spacer depends on the proximity of the binding epitope to the cell surface. CAR T therapy targeting B lymphocyte antigen CD19 (Kymriah®, Novartis) is considered promising for pediatric acute lymphoblastic leukemia, while CAR T therapy targeting B cell maturation antigen ("bb2121," a joint research project between Celgene® and Bluebirdbio®) is considered promising for relapsed/refractory multiple myeloma. More recent data suggest that the CAR approach may also be effective against solid tumors. GD2 CAR natural killer T cell (NKT) therapy has shown activity in neuroblastoma (Heczey A, et al. Invariant NKT cells with chimeric antibody receptor provide a novel platform for safe and effective cancer immunotherapy. Blood; 124(18): 2824-33, 2014), and pembrolizumab-mediated mesothelin CAR T cells have shown antitumor effects against mesothelioma. However, further targets for treating solid tumors are needed. 3. Challenges of CAR T-cell therapy Unfortunately, the complexity of CAR T-cell-based therapy can lead to undesirable and dangerous consequences. Off-tumor effects such as neurotoxicity and acute respiratory distress syndrome are potential adverse effects of CAR T-cell therapy and can be fatal. Cytokine release syndrome (CRS) is the most common acute toxicity associated with CAR T cells. CRS occurs when lymphocytes become highly activated and release excessive amounts of inflammatory cytokines. In CRS patients, elevated levels of interleukin-2, interleukin-6, interleukin-1 beta, GM-CSF, and/or C-reactive protein in the serum are sometimes observed when these factors are measured. CRS is classified by severity and diagnosed as one of four grades (mild to severe), with more severe cases clinically characterized by high fever, hypotension, hypoxia, and/or multi-organ toxicity in the patient. One study reported that 92% of patients with acute lymphoblastic leukemia treated with anti-CD19 CAR T-cell therapy experienced CRS, and 50% of these patients developed grade 3–4 symptoms. Therefore, further CAR T-cell-based therapies are needed to enhance the capabilities of effective cancer treatment. However, new CAR T-cell therapies must be devised that effectively treat cancer while minimizing the risk of dangerous inflammatory responses such as CRS. GPC3 expression in cancer and normal tissue. 1A. Anti-GPC3 antibody staining in hepatocellular carcinoma (HCC), non-small cell lung cancer (NSCLC), and ovarian cancer. 1B. Results of immunohistochemical staining (IHC