Methods of treating cancer or tumors

Abstract

The present invention provides a method for treating, preventing, or delaying the progression of cancer and / or tumor in a subject, the method comprising administering to the subject a therapeutic regimen comprising an effective amount of a PD-1 axis-binding antagonist and a population of modified immunoresponsive cells that express or display a heterologous TCR. The present invention also provides a method for enhancing immune function in a subject with cancer and / or tumor, the method comprising administering to the subject a therapeutic regimen comprising an effective amount of a PD-1 axis-binding antagonist and a population of modified immunoresponsive cells that express or display a heterologous TCR.

Description

[Technical Field]

[0001] The present invention relates to the treatment of cancer or tumors by co-administration of a PD-1 axis-coupled antagonist, such as an anti-PD-1 or anti-PD-L1 antibody, and immune-responsive cells, such as T cells that present a heterologous T cell receptor (TCR) or chimeric antigen receptor (CAR). [Background technology]

[0002] Immune tolerance, T cell exhaustion, and functional suppression in the tumor microenvironment are challenges to cancer immunotherapy, and one goal for successful cancer treatment is to explore therapies that aim to disrupt tolerance in the tumor microenvironment and enhance and maintain T cell activation.

[0003] The antitumor immune response is initiated by the release and uptake of tumor antigens by dendritic cells (DCs), which process the antigens and present them via MHC to T cells that present antigen-specific T cell receptors. This stimulation of T cells leads to their activation and proliferation. An effective antitumor immune response requires the maintenance of an activated T cell response to effectively eliminate the tumor. The problem persists that the tumor microenvironment is immunosuppressive due to the tumor cells themselves as well as the suppressive immune population of myeloid suppressor cells (MDSCs) and regulatory T cells (Tregs), and that immune tolerance can arise from the suppression of any of the key stages of the antitumor immune response, namely, dendritic cells; antigen presentation; major histocompatibility complex; T cell priming, signaling, activation or transport, or T cell permeation into the tumor; T cell receptors; MDSCs, Tregs, and cancer-associated fibroblasts, which promote high levels of inhibitory ligands and cytokines. This problem can lead to patients not responding to cancer treatment with immune effectors, such as antitumor monoclonal antibodies, chimeric antigen receptor (CAR) T cells, or adoptive transplantation of TCR-modified T cells, or to patients relapsing after treatment despite having measurable antibodies or transduced T cells in their circulating blood. Certain cancers, such as esophageal cancer, gastric cancer, liver cancer, bile duct cancer, pancreatic cancer, head and neck cancer, lung cancer, ovarian cancer, and breast cancer, appear to present specific challenges in this regard.

[0004] In addition, primary and acquired resistance pose a significant challenge to effective cancer and tumor treatment with checkpoint inhibitors, such as PD-1 axis-coupled antagonist therapy with anti-PD-1 or PD-L1 antibodies. Various mechanisms are thought to underlie such resistance, including tumor microenvironment suppressors, low tumor immunogenicity, tumor PD1 resistance, T cell infiltration or exclusion mechanisms, tumor resistance to interferon, and further upmodulation of checkpoint inhibitor receptors by T cells. Some cancers appear to be highly resistant to checkpoint inhibitor therapy, and patients who exhibit partial responses, non-responses, and acquired resistance to checkpoint inhibitor therapy represent a significant problem in cancer treatment.

[0005] Therefore, there is a need to develop therapies that address primary and acquired resistance to cancer and tumor checkpoint inhibitor therapy, as well as therapies that improve T cell function, enhance T cell survival and efficacy, activate the immune system, and / or overcome the locally immunosuppressive tumor microenvironment.

[0006] The present invention relates to a combination therapy for cancer and / or tumors comprising a PD-1 axis-linked antagonist and a population of modified immune-responsive cells expressing or presenting a heterologous TCR or CAR, wherein this combination therapy results in an enhancement of the immune response against cancer or tumors. [Overview of the project]

[0007] The present invention relates to a combination therapy for cancer and / or tumors comprising a PD-1 axis-linked antagonist and a population of modified immune-responsive cells expressing or presenting a heterologous TCR or CAR. Preferably, the combination therapy results in an enhancement of the immune response against cancer or tumor.

[0008] The present invention provides a method for treating, preventing, or delaying the progression of cancer and / or tumors in a subject, comprising the step of administering an effective amount of a PD-1 axis-binding antagonist and a population of modified immune-responsive cells expressing or presenting a heterologous TCR or CAR to the subject. The present invention also provides a method for enhancing the immune response against cancer and / or tumors in a subject, comprising the step of administering an effective amount of a PD-1 axis-binding antagonist and a population of modified immune-responsive cells expressing or presenting a heterologous TCR or CAR to the subject.

[0009] Furthermore, the present invention provides a PD-1 axis-coupled antagonist for use in combination with a population of modified immune-responsive cells expressing or presenting a heterologous TCR or CAR, for use in the treatment, prevention, or delay of cancer and / or tumor progression in a subject, or for use in enhancing the immune response against cancer and / or tumors.

[0010] In the present invention, there is further provided the use of a PD-1 axis-binding antagonist in the manufacture of a medicament for the treatment, prevention or delay of the exacerbation of cancer and / or tumor in a subject, or for enhancing the immune response against cancer and / or tumor, in combination with a population of modified immune-responsive cells expressing or presenting a heterologous TCR or CAR.

[0011] The present invention also provides herein a manufacturing process of one or more medicaments for the treatment, prevention or delay of the exacerbation of cancer and / or tumor in a subject, or for enhancing the immune response against cancer or tumor, which comprises the use of a PD-1 axis-binding antagonist and a population of modified immune-responsive cells expressing or presenting a heterologous TCR or CAR.

[0012] There is also provided a method for treating, preventing or delaying the exacerbation of cancer and / or tumor in a subject, which method comprises the step of applying to the subject a treatment regimen comprising an effective amount of a PD-1 axis-binding antagonist and a population of modified immune-responsive cells expressing or presenting a heterologous TCR or CAR.

[0013] The present invention also provides a PD-1 axis-binding antagonist for use in a method for treating, preventing or delaying the exacerbation of cancer and / or tumor in a subject, the method comprising the step of applying to the subject a treatment regimen comprising an effective amount of the PD-1 axis-binding antagonist and a population of modified immune-responsive cells expressing or presenting a heterologous TCR or CAR.

[0014] There is also provided a method for enhancing the immune response against cancer and / or tumor in a subject, which method comprises the step of applying to the subject a treatment regimen comprising an effective amount of a PD-1 axis-binding antagonist and a population of modified immune-responsive cells expressing or presenting a heterologous TCR or CAR.

[0015] Also provided is a PD-1 axis-binding antagonist for use in a method for enhancing an immune response against cancer and / or tumors in a subject, the method comprising applying to the subject a treatment regimen comprising an effective amount of the PD-1 axis-binding antagonist and a population of modified immunoreactive cells that express or present a heterologous TCR or CAR.

[0016] According to the present invention, treatment results in complete response, partial response or disease stabilization in the subject, and / or a sustained response in the subject either during or after treatment cessation, optionally improved as compared to before application of the treatment or as compared to treatment with the PD-1 axis-binding antagonist alone or with the modified immunoreactive cells alone.

[0017] PD-1 axis-binding antagonist According to the present invention, the PD-1 axis-binding antagonist is a molecule that inhibits the interaction between a PD-1 axis-binding partner and its cognate binding partner or reduces, prevents or inhibits signal transduction resulting from the interaction between PD-1 and this binding partner, e.g., one or more of PD-L1, PD-L2. The PD-1 axis-binding antagonist can reduce, inhibit or prevent the PD1 axis-mediated inhibitory effect on T cells or overcome T cell dysfunction or exhaustion, e.g., resulting in restoring or enhancing T cell function, e.g., as demonstrated by the ability to T cell proliferation, cytokine production, killing of target cells, activation, CD28 signal transduction, infiltration into tumors, recognition and binding of dendritic cell-presented antigen, and / or production of interferon.

[0018] According to the present invention, a PD-1 axis-binding antagonist can be an antagonist in the sense that it antagonizes, blocks, inhibits, or reduces the biological activity of the antigen to which it binds, for example, a PD-1-related target. Therefore, by antagonizing, blocking, inhibiting, or reducing PD-1-mediated signaling, a PD-1 axis-binding antagonist can restore functional responses by T cells, such as T cell proliferation, cytokine production, and target cell killing, thereby rescuing T cells from a dysfunctional state to an antigen-stimulated state and enhancing T cell function by overcoming T cell exhaustion, anergy, dysfunction, or tumor immunity, enhancing tumor immunogenicity, or improving sustained responses.

[0019] According to the present invention, the PD-1 axially coupled antagonist may be (a) a PD-1 antagonist and / or a binding antagonist, (b) a PD-L1 antagonist and / or a binding antagonist, or (c) a PD-L2 antagonist and / or a binding antagonist. The PD-1 axially coupled antagonist may bind to PD-1 and / or SEQ ID NO: 1, or (b) PD-L1 and / or SEQ ID NO: 2.

[0020] According to the present invention, a PD-1 axis-binding antagonist can (a) inhibit the binding of PD-1 to its ligand-binding partner, and (b) inhibit the binding of PD-L1 to its ligand-binding partner. Therefore, if a PD-1 axis-binding antagonist is a PD-1-binding antagonist, it can inhibit the binding of PD-1 to (a) PD-L1, (b) PD-L2, or (c) one or more of both PD-L1 and PD-L2. PD-1-binding antagonists may include anti-PD-1 antibodies, antigen-binding fragments, immunoadhesins, fusion proteins, and oligopeptides that inhibit, reduce, prevent, or interfere with signaling due to the interaction between PD-1 and PD-L1 and / or PD-L2. PD-1-binding antagonists can make dysfunctional T cells less dysfunctional by reducing negative costimulatory signals mediated by or through cell surface proteins expressed on T lymphocytes and / or by PD-1-mediated signaling, thereby enhancing T cell effector function in response to antigens and / or enhancing T cell function by, for example, overcoming T cell exhaustion, anergy, dysfunction, tumor immunity, enhancing tumor immunogenicity, and / or improving sustained responses.

[0021] Therefore, if a PD-1 axis-binding antagonist is a PD-L1-binding antagonist, it may inhibit the binding of PD-L1 to one or more of its binding partners, such as (a) PD1, (b) CD80, and (c) B7-1, and / or reduce, inhibit, prevent, or interfere with signaling resulting from the interaction between PD-L1 and one or more of these binding partners. PD-L1-binding antagonists may include anti-PD-L1 antibodies, antigen-binding fragments, immunoadhesins, fusion proteins, or oligopeptides, and / or reduce negative costimulatory signals mediated by or through cell surface proteins expressed on T lymphocytes and / or mediated by PD-L1-mediated signaling, thereby making dysfunctional T cells less dysfunctional, thereby enhancing the T cell effector response to antigens, and / or enhancing T cell function by, for example, overcoming T cell exhaustion, anergy, dysfunction, tumor immunity, enhancing tumor immunogenicity, or improving sustained responses.

[0022] Therefore, if a PD-1 axis-binding antagonist is also a PD-L2-binding antagonist, it may bind to PD-L2 and / or to SEQ ID NO: 3, and / or inhibit the binding of PD-L2 to one or more of its binding partners, e.g., PD-1, and / or reduce, inhibit, prevent, or interfere with signaling resulting from the interaction between PD-L2 and one or more of these binding partners. A PD-L2-binding antagonist may include an anti-PD-L2 antibody, an antigen-binding fragment, an immunoadhesin, a fusion protein, or an oligopeptide, and / or reduce negative costimulatory signals mediated by or through cell surface proteins expressed on T lymphocytes and / or mediated by PD-L2-mediated signaling, thereby making dysfunctional T cells less dysfunctional and thereby enhancing the T cell effector response to the antigen.

[0023] According to the present invention, the PD-1 axis-binding antagonist, PD-1-binding antagonist, or PD-L1-binding antagonist may be an antibody, and here, optionally, (a) The anti-PD-L1 antibody inhibits the binding between PD-L1 and PD-1 and / or between PD-L1 and B7-1, (b) Anti-PD-L1 antibodies inhibit PD-L1 on the surface of cancer cells from transmitting signals to intracellular pathways, (c) The anti-PD-1 antibody inhibits the binding between PD-L1 and PD-1 and / or between PD-L2 and PD-1, (d) Anti-PD-1 antibodies inhibit PD-1 on the surface of T cells from transmitting signals to the intracellular pathway. As mentioned above, the intracellular pathway may be the intracellular pathway of T cells, and this stimulation can cause T cells to become dysfunctional, thereby reducing the T cell effector function that responds to antigens.

[0024] According to the present invention, the PD-L1-binding antagonist may be selected from (a) durvalumab, Imfinzi, or MEDI4736, (b) atezolizumab, Tecentriq, or MPDL3280A, (c) avelumab, Bavencio, or MSB0010718C, or (d) MDX-1105, or BMS-936559.

[0025] Therefore, PD-L1 binding antagonists are (a) Heavy chain containing SEQ ID NO: 4, and light chain containing SEQ ID NO: 5, or their variable regions or their CDRs, (b) Heavy chain containing SEQ ID NO: 6, and light chain containing SEQ ID NO: 7, or their variable regions or their CDRs, (c) The heavy chain containing SEQ ID NO: 8, and the light chain containing SEQ ID NO: 9, or their variable regions or their CDRs, (d) The heavy chain containing SEQ ID NO: 10, and the light chain containing SEQ ID NO: 11, or their variable regions or their CDRs, (e) Any antigen-binding region from (a) to (d) It may be an antibody containing [a specific component].

[0026] According to the present invention, the PD-1 conjugated antagonist may be selected from (a) pembrolizumab, Keytruda, lambrolizumab, or MK-3475, (b) semiprimab, libutayo, or REGN-2810, or (c) BMS / ONO, nivolumab, Opdivo, ONO-4538, BMS-936558, or MDX1106.

[0027] Therefore, PD-1 binding antagonists are (a) The heavy chain containing SEQ ID NO: 12, and the light chain containing SEQ ID NO: 13, or their variable regions or their CDRs, (b) The heavy chain containing SEQ ID NO: 14, and the light chain containing SEQ ID NO: 15, or their variable regions or their CDRs, (c) The heavy chain containing SEQ ID NO: 16, and the light chain containing SEQ ID NO: 17, or their variable regions or their CDRs, (d) Any antigen-binding region from (a) to (c) It may be an antibody containing [a specific component].

[0028] According to the present invention, the PD-1 axis-conjugated antagonist, PD-1 or PD-L1 or PD-L2-conjugated antagonist may be an antibody, for example, a monoclonal, human or humanized full-length antibody or its antigen-binding fragment, e.g., Fv, Fab, Fab', Fab'-SH, F(ab')2; diabody; linear antibody; single-chain antibody molecule or scFv. The antibody isotype may be selected from any of the five classes of immunoglobulins having heavy chains named alpha, delta, epsilon, gamma, and mu (M), respectively: IgA, IgD, IgE, IgG, and IgM. The gamma and alpha class antibodies may be any of the subclasses: IgG1, IgG2A, IgG2B, IgG3, IgG4, IgAl, and IgGA2.

[0029] According to the present invention, a PD-1 axis-binding antagonist, which may be a PD-1 binding, PD-L1 binding antagonist, or PD-L2 binding antagonist, may be an immunoadhesin. The immunoadhesin may contain an adhesin domain including a binding site for a receptor or ligand, for example, together with an immunoglobulin constant domain derived from any isotype such as IgG-1, IgG-2, IgG2A, IgG2B, IgG-3 or IgG-4 subtype, IgA, IgA-1, IgA-2, IgE, IgD or IgM, and / or may contain a hinge, CH2 and CH3 or hinge, CH1, CH2 and CH3 region of an immunoglobulin molecule. Therefore, immunoadhesins may be polypeptides comprising an extracellular or PD-1 binding moiety of PD-L1 or PD-L2, or an extracellular or PD-L1 or PD-L2 binding moiety of PD-1, for example, PD-L1ECD-Fc, PD-L2ECD-Fc, and PD-1ECD-Fc, fused to the constant domain of an immunoglobulin sequence.

[0030] Immune-responsive cells According to the present invention, modified immune-responsive cells may be lymphoid cells, including B, T, or natural killer (NK) cells. Modified immune-responsive cells may be lymphoid cells, including T cells, natural killer T (NKT) cells, and their precursors, including embryonic stem cells and pluripotent stem cells (e.g., those that can differentiate into lymphoid cells). T cells may be lymphocytes that mature in the thymus, are the main cause of cell-mediated immunity, and are also involved in the adaptive immune system. According to the present invention, T cells may include, but are not limited to, helper T cells, cytotoxic T cells, memory T cells (including central memory T cells, stem cell-like memory T cells (or stem-like memory T cells), and two types of effector memory T cells, e.g., TEM cells and TEMRA cells), regulatory T cells (also known as suppressor T cells), natural killer T cells, mucosa-associated invariant T cells, and gamma-delta T cells. Cytotoxic T cells (CTLs or killer T cells) are a subset of T lymphocytes capable of inducing the death of infected somatic or tumor cells. The subject's own T cells can be genetically modified to target specific antigens via the introduction of a TCR. Preferably, the modified immune-responsive cells are T cells, optionally CD4 + T cells or CD8 + These are T cells. Therefore, modified immune-responsive cells may be T cells, optionally CD4+ T cells or CD8+ T cells, or modified immune-responsive cells may be a population of modified T cells, optionally CD4+ T cells or CD8+ T cells, or a mixed population of CD4+ T cells and CD8+ T cells.

[0031] Heterogeneous TCR / CAR According to the present invention, modified immune-responsive cells may express heterologous T cell receptors (TCRs) or heterologous chimeric antigen receptors (CARs). Upon binding to an antigen, modified immune-responsive cells may exhibit T cell effector function and / or cytolytic activity against cells possessing the antigen, and / or may proliferate and / or divide. In certain embodiments, modified immune-responsive cells containing TCRs exhibit equivalent or better therapeutic efficacy compared to cells containing chimeric antigen receptors (CARs) targeting the same cancer and / or tumor antigen and / or peptide (antigen peptide). Activated modified immune-responsive cells containing TCRs or CARs may secrete antitumor cytokines, including but not limited to TNF-alpha, IFNγ, and IL2.

[0032] According to the present invention, modified immune-responsive cells may include nucleic acids, constructs, or vectors encoding heterologous T cell receptors (TCRs) or heterologous chimeric antigen receptors (CARs), or heterologous nucleic acids, constructs, or vectors. Optionally, the TCR may be an affinity-enhanced TCR, such as a specific peptide-enhanced affinity receptor (SPEAR) TCR.

[0033] The terms "exogenous" or "exogenous" refer to polypeptides or nucleic acids that are foreign to a particular biological system, e.g., cells or host cells, and that do not naturally exist in that system, and which can be introduced into the system artificially or by recombinant means. Thus, the expression of heterologous TCRs or CARs can alter the immunogenicity specificity of T cells so that these T cells recognize or exhibit enhanced recognition of one or more tumor or cancer antigens and / or peptides present on the surface of cancer cells in an individual with cancer. T cell modification and subsequent proliferation can be carried out in vitro and / or ex vivo.

[0034] Cancer antigens According to the present invention, cancer and / or tumor antigens or their peptide antigens may be cancer testicular antigen, NY-ESO-1, MART-1 (melanoma antigen recognized by T cells), WT1 (Wilms tumor 1), gp100 (glycoprotein 100), tyrosinase, PRAME (antigen preferentially expressed in melanoma), p53, HPV-E6 / HPV-E7 (human papillomavirus), HBV,TRAIL, DR4, thyroglobulin, TGFBII frameshift antigen, LAGE-1A, KRAS, CMV (cytomegalovirus), CEA (carcinoembryonic antigen), AFP (alpha-fetoprotein), MAGE-A1, MAGE-A2, MAGE-A3, MAGE-A4, MAGE-A6, MAGE-A8 and MAGE-A9, MAGE-A10 or MAGE-A12 or their peptide antigens. According to the present invention, preferably, the tumor antigen is MAGE-A4, for example, SEQ ID NO: 36 or this peptide antigen. Preferably, the cancer and / or tumor antigen peptide has the amino acid sequence GVYDGREHTV of SEQ ID NO: 18.

[0035] Co-stimulatory ligand According to the present invention, modified immune-responsive cells may further comprise at least one, optionally, one, two, three, or four exogenous or recombinant (e.g., transduction of this costimulatory ligand into the cell) costimulatory ligands. Modified immune-responsive cells may co-express a TCR or CAR and at least one exogenous costimulatory ligand. Interactions between the TCR or CAR and at least one exogenous costimulatory ligand may result in non-antigen-specific signaling and cell activation. Costimulatory ligands include, but are not limited to, members of the tumor necrosis factor (TNF) superfamily and immunoglobulin (Ig) superfamily ligands. TNF is a cytokine involved in systemic inflammation and stimulates the acute phase response. The TNF superfamily members include, but are not limited to, nerve growth factor (NGF), CD40L (CD40L) / CD154, CD137L / 4-1BBL, TNF-alpha, CD134L / OX40L / CD252, CD27L / CD70, Fas ligand (FasL), CD30L / CD153, tumor necrosis factor beta (TNFP) / lymphotoxin-alpha (LTa), lymphotoxin-beta (TTb), CD257 / B-cell activator (BAFF) / Blys / THANK / Tall-I, glucocorticoid-inducible TNF receptor ligand (GITRL), and TNF-associated apoptosis-inducing ligand (TRAIL), LIGHT (TNFSF14). The immunoglobulin (Ig) superfamily is a large group of cell surface and soluble proteins involved in cell recognition, binding, or adhesion processes. Such proteins share structural features with immunoglobulins, and they possess an immunoglobulin domain (fold). Immunoglobulin superfamily ligands include, but are not limited to, CD80 and CD86, both ligands for CD28. In certain embodiments, at least one co-stimulatory ligand is selected from the group consisting of 4-1BBL, CD275, CD80, CD86, CD70, OX40L, CD48, TNFRSF14, and combinations thereof.At least one exogenous or recombinant costimulatory ligand may be 4-1BBL or CD80, preferably at least one exogenous or recombinant costimulatory ligand is 4-1BBL. Modified immune-responsive cells may contain two exogenous or recombinant costimulatory ligands, preferably the two exogenous or recombinant costimulatory ligands are 4-1BBL and CD80.

[0036] Modified immune-responsive cells may contain at least one exogenous or recombinant (e.g., transduction of this construct into cells) construct that overcomes the immunosuppressive tumor microenvironment. Such constructs may, but are not limited to, cyclic AMP phosphodiesterase and dominant-negative transforming growth factor beta (TGF beta) receptor II. Modified immune-responsive cells, modified T cells, or populations of modified T cells may be modified to release cytokines that have a positive effect on the cytolytic activity of these cells. Such cytokines may, but are not limited to, interleukin-7, interleukin-15, and interleukin-21.

[0037] Specific binding TCR / CAR According to the present invention, modified immune-responsive cells, such as modified T cells, can be modified to express heterologous TCRs or CARs that bind to or specifically bind to tumor cells and / or tissues, and / or cancer cells and / or tissues, of subjects, patients, or cancer patients suffering from a disease or cancerous condition. Subjects, patients, or cancer patients can subsequently be treated with modified immune-responsive cells or modified T cells or populations thereof according to the present invention. Cancer patients suitable for treatment according to the present invention using modified immune-responsive cells or modified T cells can be identified by a method comprising the steps of obtaining a sample of tumor and / or cancer cells from an individual or subject having a tumor and / or cancer, and identifying the tumor and / or cancer cells as cells that bind to TCRs or CARs expressed by modified immune-responsive cells.

[0038] According to the present invention, heterologous TCRs or CARs bind to or specifically bind to cancer and / or tumor antigens or their peptide antigens. According to the present invention, heterologous TCRs or CARs bind to or specifically bind to cancer and / or tumor antigens or their peptide antigens that are associated with cancer symptoms and / or presented by tumors or cancer cells or tissues.

[0039] The strength of binding between heterologous TCRs or CARs and specific target cancer and / or tumor antigens or their peptide antigens is described by specificity, which can be described by the dissociation constant, Kd, ​​and rate between bound and unbound states in the receptor-ligand system. In addition, the less difference there is between the bindable cancer and / or tumor antigens or their peptide antigens, and the heterologous TCRs or CARs, the higher the binding specificity.

[0040] According to the present invention, heterologous TCRs or CARs can bind to 10, 9, 8, 7, 6, 5, 4, 3, or fewer than 2 different cancer and / or tumor antigens or their peptide antigens, for example, to a single cancer and / or tumor antigen or their peptide antigen.

[0041] According to the present invention, the heterologous TCR or CAR may bind with a dissociation constant of 0.01 μM to 100 μM, 0.01 μM to 50 μM, 0.01 μM to 20 μM, 0.05 μM to 20 μM, or 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1 μM, 0.15 μM, 0.2 μM, 0.25 μM, 0.3 μM, 0.35 μM, 0.4 μM, 0.45 μM, 0.5 μM, 0.55 μM, 0.6 μM, 0.65 μM, 0.7 μM, 0.75 μM, 0.8 μM, 0.85 μM, 0.9 μM, 0.95 μM, 1.0 μM, 1.5 μM, 2.0 μM, 2.5 μM, 3.0 μM, 3.5 μM, 4.0 μM, 4.5 μM, 5.0 μM, 5.5 μM, 6.0 μM, 6.5 μM, 7.0 μM, 7.5 μM, 8.0 μM, 8.5 μM, 9.0 μM, 9.5 μM, 10.0 μM, or a dissociation constant of 10 μM to 1000 μM, 10 μM to 500 μM, 50 μM to 500 μM or 10, 20, 30, 40, 50, 60, 70, 80, 90, 100 μM, 150 μM, 200 μM, 250 μM, 300 μM, 350 μM, 400 μM, 450 μM, 500 μM, and is optionally measured by surface plasmon resonance, optionally at 25°C, optionally at a pH of 6.5 to 6.9 or 7.0 to 7.5. Dissociation constant, K D or k off / k on is determined by experimentally measuring the dissociation rate constant k off and the association rate constant k on The TCR dissociation constant may be measured using a soluble form of TCR, where the TCR includes a TCR alpha chain variable domain and a TCR beta chain variable domain. Thus, the heterologous TCR or CAR for use according to the present invention efficiently and / or with high affinity binds to an HLA presenting GVYDGREHTV, optionally to a peptide presenting molecule, e.g., HLA and, e.g., HLA-A * 02 or HLA-A *It is possible to bind with dissociation constants of, for example, 0.01 μM to 100 μM, such as 50 μM, 100 μM, 200 μM, 500 μM, preferably 0.05 μM to 20.0 μM, with or without the formation of a complex with 0201 or with the formation of a complex with a peptide-presenting molecule.

[0042] According to the present invention, modified immune-responsive cells, for example, modified T cells, may optionally include heterologous TCRs or CARs that are associated with cancer symptoms and / or can bind, specifically bind, and / or bind with high affinity to cancer and / or tumor antigens or their peptide antigens presented by tumors or cancer cells or tissues, and optionally the cancer and / or tumor antigens or their peptide antigens may include peptide-presenting molecules, for example, HLA and, for example, HLA-A * 02 or HLA-A * It is recognized by heterologous TCRs or CARs, either in conjunction with the formation of a complex with O201, or without presentation involving the formation of a complex with a peptide-presenting molecule, such as HLA.

[0043] According to the present invention, xenogeneic T cell receptors (TCRs) or CARs, and modified immune-responsive cells containing xenogeneic T cell receptors (TCRs) or CARs may have the property of binding to cancer and / or tumor antigens or their peptide antigens endogenously expressed on the surface of tumor cells, wherein, optionally, the binding is independent of the presentation of cell surface antigens as a complex with peptide-presenting or antigen-presenting molecules, such as major histocompatibility complex (MHC) or human leukocyte antigen (HLA) or major histocompatibility complex class-associated protein (MR) 1.

[0044] According to the present invention, the TCR or CAR binding may optionally be specific to one cancer and / or tumor antigen, e.g., MAGE protein, e.g., MAGE A4, or its peptide antigen, compared to closely related cancer and / or tumor antigens or their peptide antigen sequences. The closely related cancer and / or tumor antigens or their peptide antigen sequences may be sequences of similar or identical length and / or have similar or identical numbers of amino acid residues. The closely related peptide antigen sequences may share 50, 60, 70, or 80-90% identity, preferably 80-90% identity, and / or differ by 1, 2, 3, or 4 amino acid residues. The closely related peptide sequence may be derived from the polypeptide sequence GVYDGREHTV of Sequence ID No. 18.

[0045] Binding affinity can be determined by equilibrium methods (e.g., enzyme-linked immunosorbent assay (ELISA) or radioimmunoassay (RIA)) or kinetics (e.g., BIACORE® analysis). Binding activity is, for example, the sum of the strengths of mutual binding between two molecules at multiple sites, taking into account the binding titer of their interaction. According to the present invention, improved affinity and / or binding activity to cancer and / or tumor antigens or their peptide antigens, or cancer and / or tumor antigens or their peptide antigens presented by tumors or cancer cells or tissues and recognized by heterologous TCRs or CARs, can be demonstrated by immune-responsive cells compared to immune-responsive cells lacking or possessing a different heterologous TCR or CAR.

[0046] Selective binding TCR / CAR According to the present invention, heterologous TCRs or CARs may selectively bind to cancer and / or tumor antigens or their peptide antigens, and optionally, are associated with cancer symptoms and / or are presented by tumors or cancer cells or tissues, wherein optionally, the cancer and / or tumor antigens or their peptide antigens may optionally be peptide-presenting molecules, such as HLA and, for example, HLA-A *02 or HLA-A * It is recognized by heterologous TCRs or CARs, either in conjunction with the formation of a complex with O201 or without presentation in conjunction with the formation of a complex with a peptide-presenting molecule or HLA, and is preferably expressed by tumor cells or cancer cells or tissues.

[0047] Selective binding refers to the binding of a heterologous TCR or CAR to one cancer and / or tumor antigen or its peptide antigen with higher affinity compared to another antigen. Selective binding is expressed by the equilibrium constant for the substitution of another ligand antigen by one ligand antigen with the heterologous TCR or CAR.

[0048] Specific / selective binding of TCRs / CARs According to the present invention, the binding of a heterologous TCR or CAR is selective and / or specific to cancer and / or tumor antigens or their peptide antigens, which may be cancer testicular antigens, NY-ESO-1, MART-1 (melanoma antigen recognized by T cells), WT1 (Wilms tumor 1), gp100 (glycoprotein 100), tyrosinase, PRAME (antigen preferentially expressed in melanoma), p53, HPV-E6 / HPV-E7 (human papillomavirus), HBV,TRAIL, DR4, thyroglobulin, TGFBII frameshift antigen, LAGE-1A, KRAS, CMV (cytomegalovirus), CEA (carcinoembryonic antigen), AFP (alpha-fetoprotein), MAGE-A1, MAGE-A2, MAGE-A3, MAGE-A4, MAGE-A6, MAGE-A8 and MAGE-A9, MAGE-A10 or MAGE-A12 or their peptide antigens. Preferably, the tumor antigen is MAGE-A4 or this peptide antigen.

[0049] According to the present invention, heterologous TCRs or CARs may bind to and / or specifically to and / or selectively to peptide-presenting molecules, such as cancer and / or tumor antigens or HLAs that present or exhibit these peptide antigens, i.e., peptide fragments of cancer and / or tumor antigens (pHLAs), where the HLAs all correspond to MHC class I (A, B, and C) which are HLA class 1 or specific alleles thereof, or the HLAs correspond to MHC class II (DP, DM, DO, DQ, and DR) or specific alleles thereof, preferably the HLAs are class 1, and preferably the alleles are HLA-A2 or HLA-A * 02 or HLA-A2+ or _HLA-A * O2-positive HLA, preferably HLA- * It is 0201. Alternatively, heterologous TCRs or CARs may bind and / or specifically and / or selectively to cancer and / or tumor antigens or their peptide antigens that are not presented or exhibited by HLA.

[0050] Preferably, heterologous TCRs or CARs are not spontaneously expressed by immune-responsive cells (i.e., the TCR or CAR is exogenous or heterologous). Heterologous TCRs may include αβTCR heterodimers. Heterologous TCRs or CARs may be recombinant, synthetic, or artificial TCRs or CARs, i.e., TCRs that do not exist in nature. For example, heterologous TCRs may be modified to enhance their affinity or binding activity to specific cancer and / or tumor antigens or their peptide antigens (i.e., affinity-enhancing TCRs or specific peptide-enhancing affinity receptor (SPEAR) TCRs). Affinity-enhancing TCRs or (SPEAR) TCRs may include one or more mutations compared to naturally occurring TCRs, for example, one or more mutations in the high-frequency variable complementarity-determining regions (CDRs) of the variable regions of the TCRα and β chains. Such mutations may, optionally, enhance the affinity of the TCR to MHCs exhibiting peptide fragments of tumor antigens when expressed by tumors and / or cancer cells. Suitable methods for generating affinity-enhanced or mature TCRs include screening libraries of TCR mutants using phage or yeast displays, and are well known in the art (see, for example, Robbins et al J Immunol (2008) 180(9):6116; San Miguel et al (2015) Cancer Cell 28 (3) 281-283; Schmitt et al (2013) Blood 122 348-256; Jiang et al (2015) Cancer Discovery 5 901). Preferred affinity-enhanced TCRs may bind to tumor or cancer cells expressing MAGE family tumor antigens, e.g., MAGE A4 or this peptide antigen, e.g., sequence GVYDGREHTV of SEQ ID NO: 18.

[0051] According to the present invention, a heterologous TCR may be a MAGE A4 TCR that comprises the α-chain reference amino acid sequence of SEQ ID NO: 21 or a variant thereof and the β-chain reference amino acid sequence of SEQ ID NO: 23 or a variant thereof. The variant may have an amino acid sequence having at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity with respect to the reference amino acid sequence. The TCR may be encoded by the α-chain reference nucleotide sequence of SEQ ID NO: 22 or a variant thereof and the β-chain reference nucleotide sequence of SEQ ID NO: 24 or a variant thereof. The variant may have a nucleotide sequence having at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity with respect to the reference nucleotide sequence.

[0052] According to the present invention, the TCR may include a TCR alpha chain variable domain and a TCR beta chain variable domain, where, (i) The alpha-chain variable domain contains a CDR having the sequence of VSPFSN(αCDR1) of SEQ ID NO: 27 or amino acids 48-53 of SEQ ID NO: 21, LTFSEN(αCDR2) of SEQ ID NO: 28 or amino acids 71-76 of SEQ ID NO: 21, and CVVSGGTDSWGKLQF(αCDR3) of SEQ ID NO: 29 or amino acids 111-125 of SEQ ID NO: 21. (ii) The beta-chain variable domain is the sequence of KGHDR (βCDR1) of SEQ ID NO: 30 or amino acids 46-50 of SEQ ID NO: 23, SFDVKD (βCDR2) of SEQ ID NO: 31 or amino acids 68-73 of SEQ ID NO: 23, and CATSGQGAYEEQFF (βCDR3) of SEQ ID NO: 32 or amino acids 110-123 of SEQ ID NO: 23. Or include CDRs having sequences with at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity to these, and optionally, sequences having 100% sequence identity to these.

[0053] Therefore, the TCR may include an alpha-chain variable domain containing an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to the sequence of amino acid residues 1-136 of SEQ ID NO: 25 or SEQ ID NO: 22, and / or a beta-chain variable domain containing an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to the sequence of amino acid residues 1-133 of SEQ ID NO: 26 or SEQ ID NO: 23.

[0054] The term “precursor TCR” is used herein to refer to the TCR comprising the MAGE A4TCRα chain and MAGE A4TCRβ chain of Sequence ID No. 21 and 23, respectively. It is desirable to have a mutated or modified TCR compared to the precursor TCR that has equal, equivalent or higher affinity and / or equal, equivalent or slower dissociation rate to the peptide-HLA complex than the precursor TCR. According to the present invention, the heterologous TCR may have two or more mutations present in the alpha chain variable domain and / or the beta chain variable domain compared to the precursor TCR, and may be represented as a “modified TCR” or “mutated TCR”. Such mutations may improve binding affinity and / or specificity and / or selectivity and / or binding activity to MAGE A4 or this peptide antigen. In certain embodiments, there may be one, two, three, four, five, six, seven, or eight mutations in the alpha chain variable domain, e.g., four or eight mutations, and / or one, two, three, four, or five mutations in the beta chain variable domain, e.g., five mutations. In some embodiments, the α-chain variable domain of the TCR of the present invention may include an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the amino acid residue sequence of SEQ ID NO: 25. In some embodiments, the β-chain variable domain of the TCR of the present invention may include an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the amino acid residue sequence of SEQ ID NO: 26.

[0055] According to the present invention, the TCR has an alpha chain variable domain in which the amino acid sequence of amino acid residues 1-136 of SEQ ID NO: 25 or SEQ ID NO: 21, or the amino acid residues 1-47, 54-70, 77-110 and 126-136 thereof, are at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% of the respective sequences of amino acid residues 1-47, 54-70, 77-110 and 126-136 of SEQ ID NO: 25. The TCR may contain an amino acid sequence having % identity, and / or each of amino acid residues 48-53, 71-76, and 111-125, CDR1, CDR2, and CDR3 having at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to each of the sequences of amino acid residues 48-53, 71-76, and 111-125, CDR1, CDR2, and CDR3 of SEQ ID NO: 25.

[0056] According to the present invention, the TCR has an alpha chain variable domain, (i) The amino acid residues 1-47 may (a) have at least 70%, 75%, 80%, 85%, 90%, or 95% identity with the sequence of amino acid residues 1-47 of SEQ ID NO: 25, or (b) have one, two, or three amino acid residues that are inserted or deleted compared to residues 1-47 of SEQ ID NO: 25. (ii) Amino acid residues 48-53 are CDR1, which is VSPFSN of SEQ ID NO: 27 or amino acids 48-53 of SEQ ID NO: 25. (iii) The amino acid residues 54-70 may (a) have at least 70%, 75%, 80%, 85%, 90%, or 95% identity with the sequence of amino acid residues 54-70 of SEQ ID NO: 25, or (b) have one, two, or three amino acid residues that have been inserted or deleted compared to the sequence of amino acid residues 54-70 of SEQ ID NO: 25. (iv) Amino acid residues 71-76 may be LTFSEN of SEQ ID NO: 28 or amino acids 71-76 of SEQ ID NO: 25, which are CDR2. (v) The amino acid residues 77-110 may have at least 70%, 75%, 80%, 85%, 90%, or 95% identity with the sequence of amino acid residues 77-110 of SEQ ID NO: 25, or may have one, two, or three insertions, deletions, or substitutions compared to the sequence of amino acid residues 77-110 of SEQ ID NO: 25. (vi) Amino acids 111-125 may be CDR3, such as CVVSGGTDSWGKLQF of SEQ ID NO: 29 or amino acids 111-125 of SEQ ID NO: 25. (vii) The amino acid residues 126-136 may have at least 70%, 75%, 80%, 85%, 90%, or 95% identity with the sequence of amino acid residues 126-136 of SEQ ID NO: 25, or may have one, two, or three insertions, deletions, or substitutions compared to the sequence of amino acid residues 126-136 of SEQ ID NO: 25. May contain sequences and TCRs.

[0057] According to the present invention, a TCR may include a TCR in which the beta-chain variable domain contains an amino acid sequence in which amino acid residues 1-45, 51-67, 74-109, and 124-133 have at least 70%, 75%, 80%, 85%, 90%, or 95% identity with each of the amino acid residues 1-45, 51-67, 74-109, and 124-133 of SEQ ID NO: 26, and amino acid residues 46-50, 68-73, and 110-123 have at least 70%, 75%, 80%, 85%, 90%, or 95% identity with each of the amino acid residues 46-50, 68-73, and 110-123 of SEQ ID NO: 26, CDR1, CDR2, and CDR3.

[0058] According to the present invention, the TCR has a beta chain variable domain, (i) The amino acid residues 1-45 may (a) have at least 70%, 75%, 80%, 85%, 90%, or 95% identity with the sequence of amino acid residues 1-45 of SEQ ID NO: 26, or (b) have one, two, or three amino acid residues that have been inserted or deleted compared to residues 1-45 of SEQ ID NO: 26. (ii) Amino acid residues 46-50 are KGHDR of SEQ ID NO: 30, which is CDR1, or amino acids 46-50 of SEQ ID NO: 26. (iii) The amino acid residues 51-67 may (a) have at least 70%, 75%, 80%, 85%, 90%, or 95% identity with the sequence of amino acid residues 51-67 of SEQ ID NO: 26, or (b) have one, two, or three amino acid residues that have been inserted or deleted compared to the sequence of amino acid residues 51-67 of SEQ ID NO: 26. (iv) Amino acid residues 68-73 may be SFDVKD of SEQ ID NO: 31 or amino acids 68-73 of SEQ ID NO: 26, which are CDR2. (v) The amino acid residues 74-109 may have at least 70%, 75%, 80%, 85%, 90%, or 95% identity with the sequence of amino acid residues 74-109 of SEQ ID NO: 26, or they may have one, two, or three insertions, deletions, or substitutions compared to the sequence of amino acid residues 74-109 of SEQ ID NO: 26. (vi) Amino acids 110-123 may be CDR3, CATSGQGAYEEQFF of SEQ ID NO: 32 or amino acids 110-123 of SEQ ID NO: 26. (vii) The amino acid residues 124-133 may have at least 70%, 75%, 80%, 85%, 90%, or 95% identity with the sequence of amino acid residues 124-133 of SEQ ID NO: 26, or may have one, two, or three insertions, deletions, or substitutions compared to the sequence of amino acid residues 124-133 of SEQ ID NO: 26. May contain sequences and TCRs.

[0059] The identity of amino acid and nucleotide sequences is generally determined according to the GAP algorithm (GCG Wisconsin Package®, Accelrys, San Diego, CA). GAP uses the Needleman-Bunsch algorithm (J. Mol. Biol. (48): 444-453 (1970)) to align two complete sequences, thereby maximizing the number of fits and minimizing the number of gaps. Generally, default parameters are used, with a gap generation penalty of 12 and a gap elongation penalty of 4. While the use of GAP may be preferred, other algorithms may also be used, such as BLAST, psiBLAST, or TBLASTN (using the method in Altschul et al. (1990) J. Mol. Biol. 215: 405-410), FASTA (using the method in Pearson and Lipman (1988) PNAS USA 85: 2444-2448), or the Smith-Waterman algorithm (Smith and Waterman (1981) J. Mol Biol. 147: 195-197), and default parameters are generally used.

[0060] A particular amino acid sequence variant may differ from the reference sequence by the insertion, addition, substitution, or deletion of one amino acid, two, three, four, five to ten, ten to twenty, or twenty to thirty amino acids. In some embodiments, the variant sequence may include a reference sequence having one, two, three, four, five, six, seven, eight, nine, ten, or more residues that have been inserted, deleted, or substituted. For example, up to 15, up to 20, up to 30, or up to 40 residues may be inserted, deleted, or substituted.

[0061] In some preferred embodiments, the variant may differ from the reference sequence by one, two, three, four, five, six, seven, eight, nine, ten or more conservative substitutions. Conservative substitutions include amino acid substitutions with different amino acids having similar properties. For example, an aliphatic residue may be replaced with another aliphatic residue, a nonpolar residue with another nonpolar residue, an acidic residue with another acidic residue, a basic residue with another basic residue, a polar residue with another polar residue, or an aromatic residue with another aromatic residue. Conservative substitutions include, for example, the following groups: (i) Alanine and glycine, (ii) Glutamic acid, aspartic acid, glutamine and asparagine, (iii) Arginine and lysine, (iv) Asparagine, glutamine, glutamic acid and aspartic acid, (v) Isoleucine, leucine and valine, (vi) Phenylalanine, tyrosine and tryptophan, (vii) Serine, threonine, and cysteine It can be present between amino acids in the following context.

[0062] CD8α coreceptor According to the present invention, a population of modified immune-responsive cells expressing or presenting a heterologous TCR or CAR may further express or present a heterologous coreceptor. The heterologous coreceptor may be a CD8 coreceptor. The CD8 coreceptor may include a dimer, i.e., a pair of CD8 chains, comprising CD8-α and CD8-β chains or CD8-α and CD8-α chains. Preferably, the CD8 coreceptor is a CD8αα coreceptor comprising CD8-α and CD8-α chains. The CD8α coreceptor may include an amino acid sequence having at least 80% identity with SEQ ID NO: 19, SEQ ID NO: 19, or a variant thereof. The CD8α coreceptor may be a homodimer.

[0063] The CD8 coreceptor binds to class 1 MHC and enhances TCR signaling. According to the present invention, the CD8 coreceptor may include or be a variant of the reference amino acid sequence of SEQ ID NO: 19. The variant may have an amino acid sequence having at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity to the reference amino acid sequence of SEQ ID NO: 19. The CD8 coreceptor may be encoded by or be a variant of the reference nucleotide sequence of SEQ ID NO: 20. The variant may have a nucleotide sequence that has at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity with respect to the reference nucleotide sequence of Sequence ID No. 20.

[0064] According to the present invention, heterologous CD8 coreceptors have an Ig-like V-type domain, (i) CDR1 is VLLSNPTSG of SEQ ID NO: 33 or amino acids 45-53 of SEQ ID NO: 19, (ii) CDR2, YLSQNKPK of SEQ ID NO. 34 or amino acids 72-79 of SEQ ID NO. 19, (iii) LSNSIM of SEQ ID NO: 35, which is CDR3, or the sequence of amino acids 80-117 of SEQ ID NO: 19, Alternatively, it may include a CD8 coreceptor comprising a CDR having a sequence having at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity.

[0065] According to the present invention, a heterologous CD8 coreceptor may include an amino acid sequence in which residues 22-135 of the amino acid sequence of SEQ ID NO: 19, or its amino acid residues 22-44, 54-71, 80-117, and 124-135, have at least 70%, 75%, 80%, 85%, 90%, or 95% identity with the respective sequences of amino acid residues 22-44, 54-71, 80-117, 124-135, CDR1, CDR2, and CDR3 of SEQ ID NO: 19, and amino acid residues 45-53, 72-79, and 118-123 have at least 70%, 75%, 80%, 85%, 90%, or 95% identity with the respective sequences of amino acid residues 45-53, 72-79, and 118-123 of SEQ ID NO: 19, or the Ig-like V-type domain may include a CD8 coreceptor containing these sequences.

[0066] According to the present invention, the CD8 coreceptor is (i) The amino acid residues 22-44 may (a) have at least 70%, 75%, 80%, 85%, 90%, or 95% identity with the sequence of amino acid residues 22-44 of SEQ ID NO: 19, or (b) have one, two, or three amino acid residues that have been inserted or deleted compared to residues 22-44 of SEQ ID NO: 19. (ii) Amino acid residues 45-53 are VLLSNPTSG of SEQ ID NO: 33 or amino acids 45-53 of SEQ ID NO: 19, (iii) The amino acid residues 54-71 may (a) have at least 70%, 75%, 80%, 85%, 90%, or 95% identity with the sequence of amino acid residues 54-71 of SEQ ID NO: 19, or (b) have one, two, or three amino acid residues that have been inserted or deleted compared to the sequence of amino acid residues 54-71 of SEQ ID NO: 19. (iv) Amino acid residues 72-79 may be YLSQNKPK of SEQ ID NO: 34 or amino acids 72-79 of SEQ ID NO: 19, (v) The amino acid residues 80-117 have at least 70%, 75%, 80%, 85%, 90%, or 95% identity with the sequence of amino acid residues 80-117 of SEQ ID NO: 19, or may have one, two, or three insertions, deletions, or substitutions compared to the sequence of amino acid residues 80-117 of SEQ ID NO: 19. (vi) Amino acids 118-123 may be LSNSIM of SEQ ID NO: 35 or amino acids 80-117 of SEQ ID NO: 19, which are CDR3. (vii) The amino acid residues 124-135 may have at least 70%, 75%, 80%, 85%, 90%, or 95% identity with the sequence of amino acid residues 124-135 of SEQ ID NO: 19, or may have one, two, or three insertions, deletions, or substitutions compared to the sequence of amino acid residues 124-135 of SEQ ID NO: 19. The sequence may be included, or this Ig-like type V domain may contain a CD8 coreceptor that includes these sequences.

[0067] When expressed by modified immune-responsive cells expressing heterologous CD8 coreceptors, if optionally presented on HLA, improved affinity and / or binding activity and / or T cell activation can be demonstrated compared to modified immune-responsive cells that do not express heterologous CD8 coreceptors, as can be determined by assays disclosed herein relating to or against stimulation by antigen peptides, tumors, or cancer antigens. Heterologous CD8 on modified immune-responsive cells may interact with or specifically bind to MHC, which may be class I or class II, preferably class I major histocompatibility complex (MHC), HLA-I molecule, or MHC class I HLA-A / B2M dimer, and preferably CD8-α interacts with the α3 portion (residues 223-229) of class I MHC, preferably via the IgV-like domain of CD8. Therefore, heterologous CD8 selectively enhances the TCR binding of immune-responsive cells to HLA and / or antigen peptides bound or presented by HLApMHCI or pHLA on the surface of antigen-presenting cells, dendritic cells, and / or tumor or cancer cells, or tumor or cancer tissue, compared to immune-responsive cells lacking heterologous CD8. Therefore, heterologous CD8 selectively enhances the dissociation rate (k) of cell (TCR) / peptide-major histocompatibility complex class I (pMHCI) interactions of immune-responsive cells on the surface of antigen-presenting cells, dendritic cells, and / or tumor or cancer cells, or tumor or cancer tissue. off The presence of heterologous CD8 may be enhanced or increased compared to cells lacking heterologous CD8, which may also lead to an enhanced or increased half-life, thereby potentially resulting in improved ligation affinity and / or binding activity. Heterologous CD8 may enhance the organization of TCRs on the surface of immune-responsive cells, enabling synergistic action in pHLA binding, potentially leading to improved therapeutic binding activity. Therefore, heterologous CD8 coreceptor-modified immune-responsive cells may bind to or interact with LCK (lymphocyte-specific protein tyrosine kinase) in a zinc-dependent manner, leading to the activation of transcription factors such as NFAT, NF-κB, and AP-1.

[0068] According to the present invention, modified immune-responsive cells may optionally enhance or increase CD40L expression, cytokine production, cytotoxic activity, induction of dendritic cell maturation, or induction of dendritic cell cytokine production in response to tumor or cancer cells or tissues, compared to immune-responsive cells lacking heterologous CD8 coreceptors.

[0069] therapeutic action According to the present invention and the methods and uses thereof, T cell function is enhanced by, for example, increased secretion of γ-interferon by CD8+ T cells, increased T cell proliferation, increased internal signaling, increased antigen responsiveness, increased secretion of cytokines and / or interferon, increased target cell killing, increased T cell activation, increased CD28 signaling, improved ability of T cells to infiltrate tumors, and improved ability to recognize and bind dendritic cell-presented antigens, compared to levels before treatment or intervention, or compared to treatment with either a PD-1 axis-linked antagonist alone or modified immune-responsive cells expressing or presenting a heterologous TCR alone, by at least 10%, or 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 120%, 150%, 200%, or more.

[0070] According to the present invention, as well as the methods and uses thereof, tumor immunity or evasion of immune recognition by tumors is attenuated, as measured by tumor binding, tumor reduction, and tumor removal, resulting in improved tumor recognition and attack by the immune system, thereby enabling the treatment of tumor immunity. Accordingly, the present invention provides a treatment for tumor immunity and / or a treatment for tumor immunity that is enhanced by at least 10%, or 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 120%, 150%, 200%, or more, compared to pre-treatment or pre-intervention levels, or compared to treatment with either a PD-1 axis-linked antagonist alone or modified immune-responsive cells expressing or presenting a heterologous TCR alone, as measured by one or more of the following: tumor binding, tumor reduction, and tumor removal.

[0071] According to the present invention and its methods and uses, an improvement or enhancement of oncoimmunogenicity, as measured by the ability to induce an immune response in response to a tumor or tumor antigen, is achieved, for example, by increasing cytokine and / or interferon secretion, increasing T cell proliferation, antigen responsiveness, target cell killing, T cell activation, CD28 signaling, the ability of T cells to infiltrate tumors, or the ability to recognize and bind dendritic cell-presented antigens, compared to pre-treatment or pre-intervention levels, or compared to treatment with either a PD-1 axis-binding antagonist alone or modified immune-responsive cells expressing or presenting a heterologous TCR alone, by at least 10%, or 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 120%, 150%, 200%, or more, as judged by one or more of the following: increased cytokine and / or interferon secretion, increased T cell proliferation, antigen responsiveness, target cell killing, T cell activation, CD28 signaling, the ability of T cells to infiltrate tumors, or improved ability to recognize and bind dendritic cell-presented antigens, compared to pre-treatment or pre-intervention levels, or compared to treatment with either a PD-1 axis-binding antagonist alone or modified immune-responsive cells expressing or presenting a heterologous TCR alone.

[0072] According to the present invention, as well as the methods and uses thereof, an improved sustained response is obtained, which is preferably enhanced by at least 10%, or 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 120%, 150%, 200%, or more, compared to levels before treatment or intervention, or compared to treatment with either a PD-1 axis-linked antagonist alone or modified immune-responsive cells expressing or presenting a heterologous TCR alone, for example, by reducing tumor growth or tumor growth rate, or maintaining tumor size after discontinuation of treatment, as determined by one or more measurements of tumor size or tumor number. For example, the sustained response may have a duration of at least the same as the duration of treatment, or at least 1.5, 2.0, 2.5, or 3.0 times the duration of treatment or more.

[0073] In the context of T cell activity, the term “dysfunction” refers to a state of reduced immune responsiveness to antigen stimulation, including T cell exhaustion and / or anergy, where T cells can recognize and bind to antigens, but their effectiveness in exacerbating the immune response or fighting tumor growth is diminished. Dysfunctional T cells demonstrate impaired ability to translate antigen recognition into downstream T cell effector functions, such as proliferation, cytokine and interferon production, or target cell killing, and / or appear to be unresponsive or unresponsive to antigen recognition, as is characteristic of T cell dysfunction disorders. “T cell dysfunction disorders” may be associated with inappropriate increases in PD-1-mediated T cell signaling, T cells with reduced ability to generate proliferation and / or cytokine and / or cytolytic activity, T cell anergy, and tumor immunity.

[0074] "T cell exhaustion" includes a state of T cell dysfunction due to persistent TCR signaling as part of the response to cancer, preventing an optimal response to the tumor. Exhaustion may be found to act either through intracellular negative regulatory (co-stimulatory) pathways (e.g., PD-1, PD-1 axis, B7-H3, B7-H4) or extracellular negative regulatory pathways (immunomodulatory cytokines). T cell exhaustion is characterized by insufficient effector function, persistent expression of inhibitory receptors, and altered transcriptional activity distinct from that of functional effector or memory T cells. T cell anergy, even in the context of co-stimulation, frequently arises from impaired T cell receptor-mediated signaling and unresponsiveness to antigen stimulation, resulting in such T cells failing to undergo clonal proliferation and / or acquire effector function.

[0075] Administration According to the present invention, PD-1 axis-linked antagonists and modified immune-responsive cells can be administered separately, over time, or simultaneously.

[0076] therefore, (a) PD-1 axis-linked antagonists may be administered before, concurrently with, or after modified immune-responsive cells, or (b) PD-1 axis-coupled antagonists may be administered before and concurrently with modified immune-responsive cells, or (c) PD-1 axis-linked antagonists may be administered before and after modified immune-responsive cells, or (d) PD-1 axis-linked antagonists may be administered concurrently with and after modified immune-responsive cells, or (e) PD-1 axis-coupled antagonists may be administered after modified immune-responsive cells, or (f) PD-1 axis-linked antagonists may be administered before, concurrently with, and after modified immune-responsive cells.

[0077] For example, the first administration of a PD-1 axis-coupled antagonist after administration of modified immune-responsive cells may be at any one of the following time points: 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 days after administration of the modified immune-responsive cells, preferably 14-16, 17-21, or 22-26 days later, preferably 17-22 days later, preferably 17, 21, or 22 days later.

[0078] According to the present invention, PD-1 axis-linked antagonists and / or modified immune-responsive cells may be administered sequentially, optionally as a single dose, or intermittently, optionally as two or more doses.

[0079] According to the present invention, modified immune-responsive cells can be administered as a single dose. Modified immune-responsive cells can be administered in any one of the following doses: approximately 500 million to approximately 1 billion cells, approximately 2 billion cells, approximately 3 billion cells, approximately 4 billion cells, approximately 5 billion cells, approximately 6 billion cells, approximately 7 billion cells, approximately 8 billion cells, approximately 9 billion cells, approximately 10 billion cells, approximately 11 billion cells, approximately 12 billion cells, approximately 13 billion cells, approximately 14 billion cells, approximately 15 billion cells, approximately 16 billion cells, approximately 17 billion cells, approximately 18 billion cells, approximately 19 billion cells, approximately 20 billion cells, or approximately 21 billion cells. Modified immune-responsive cells may be administered in doses of approximately 100 million to 200 million cells, 300 million to 400 million cells, 500 million to 600 million cells, 700 million to 800 million cells, or 900 million to 1 billion cells, or optionally, 500 million to 1 billion cells, 2 billion to 5 billion cells, or 6 billion to 10 billion cells.

[0080] According to the present invention, PD-1 axis-linked antagonists and / or modified immune-responsive cells can be administered intravenously, intramuscularly, subcutaneously, topically, orally, percutaneously, intraperitoneally, intraorbitally, by transplantation, by inhalation, intraarachnoidally, intraventricularly or nasally, or by intravenous infusion. Preferably, PD-1 axis-linked antagonists and / or modified immune-responsive cells can be administered intravenously, i.e., by intravenous infusion.

[0081] According to the present invention, a PD-1 axially coupled antagonist may be administered in any of the following doses: approximately 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, or 60 mg / kg. PD-1 axis-coupled antagonists can be administered in one of the following doses: approximately 0.5 mg / kg to approximately 1.5 mg / kg, approximately 1 mg / kg to approximately 2 mg / kg, approximately 3 mg / kg to approximately 5 mg / kg, approximately 6 mg / kg to approximately 9 mg / kg, approximately 10 mg / kg to approximately 15 mg / kg, approximately 16 mg / kg to approximately 20 mg / kg, approximately 20 mg / kg to approximately 25 mg / kg, approximately 1 mg / kg to approximately 9 mg / kg, or approximately 10 mg / kg to approximately 20 mg / kg. According to the present invention, the PD-1 axial coupling antagonist is approximately 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370 The PD-1 axis-coupled antagonist may be administered in doses of 380, 390, 400, 410, 420, 430, 440, 450, 460, 470, 480, 490, 500, 510, 520, 530, 540, 550, 560, 570, 580, 590, 600, 650, 700, 750, 800, 850, 900, 950, and 1000 mg, preferably in doses of 200, 250, or 300 mg. According to the present invention, the PD-1 axis-coupled antagonist may be administered in a fixed dose, or the dose may vary, for example, when two or more doses exist or in an administration cycle.

[0082] According to the present invention, the PD-1 axis coupled antagonist is (a) Single dose in each of one or more administration cycles, (b) One or more doses in each of the one or more administration cycles, (c) A single dose on day 1 of each of one or more administration cycles, (d) One or more doses in each of one or more dosing cycles, including the dose on day 1 of each of one or more dosing cycles, (e) One or more doses in each of one or more dosing cycles, where at least one dose is on day 1 of each cycle. It can be administered as such.

[0083] According to the present invention, a PD-1 axially coupled antagonist can be administered in an administration cycle, where the administration cycle may be 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 days, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, or 12 months. Therefore, the administration cycle may be 10-12 days, 11-13 days, 14-17 days, 18-21 days, 22-24 days, 24-27 days, 28-30 days, or 31 days, 1 week, 2 weeks, 3 weeks, 4 weeks, or 1 month, 2 months, 6 months, preferably 3 weeks or 21 days.

[0084] According to the present invention, the PD-1 axis-coupled antagonist can be administered before the administration of modified immune-responsive cells as one or more doses in each of one or more administration cycles, and after the administration of modified immune-responsive cells as one or more doses in each of one or more administration cycles, where optionally, at least one dose may be on day 1 of each cycle. Alternatively, the PD-1 axis-coupled antagonist can be administered before the administration of modified immune-responsive cells as one or more doses in each of one or more administration cycles, simultaneously with the administration of modified immune-responsive cells (optionally, as a single dose), and after the administration of modified immune-responsive cells as one or more doses in each of one or more administration cycles, where optionally, at least one dose may be on day 1 of each cycle. Alternatively, the PD-1 axis-coupled antagonist can be administered after the administration of modified immune-responsive cells as one or more doses in each of one or more administration cycles, where optionally, at least one dose may be on day 1 of each cycle.

[0085] According to the present invention, the PD-1 axis-coupled antagonist can be administered as a single dose in each of one or more administration cycles before administration of modified immune-responsive cells, and as a single dose in each of one or more administration cycles after administration of modified immune-responsive cells, where optionally, the single dose may be on day 1 of each cycle. Alternatively, the PD-1 axis-coupled antagonist can be administered as a single dose in each of one or more administration cycles before administration of modified immune-responsive cells, simultaneously with administration of modified immune-responsive cells (optionally, modified immune-responsive cells are administered as a single dose), and as a single dose in each of one or more administration cycles after administration of modified immune-responsive cells, where optionally, the single dose may be on day 1 of each cycle. Alternatively, the PD-1 axis-coupled antagonist may be administered as a single dose in each of one or more administration cycles after administration of modified immune-responsive cells (optionally, the modified immune-responsive cells are administered as a single dose), and optionally, the single dose may be on day 1 of each cycle. According to the present invention, the PD-1 axis-coupled antagonist may be administered as a fixed single dose of approximately 200 mg on day 1 of each of one or more administration cycles, where the administration cycle is 21 days, and / or the modified immune-responsive cells are administered as a single dose of approximately 5 billion to approximately 10 billion cells, and optionally, the first dose of the PD-1 axis-coupled antagonist after administration of modified immune-responsive cells may be on day 17, day 21, or day 22 after administration of the immune-responsive cells.

[0086] According to the present invention, a PD-1 axis-linked antagonist can be administered for a specific period of time, and the administration cycle of the PD-1 axis-linked antagonist can also be administered for a specific period of time, for example, for a specific period after administration of modified immune-responsive cells. The specific period may be any of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, or 31 months, preferably 24 months.

[0087] The method according to the present invention is (a) Administer a PD-1 axis-coupled antagonist in one or more doses in each of one or more administration cycles prior to administration of modified immune-responsive cells, optionally, at least one dose being on day 1 of each cycle. (b) The disease state is determined after administration of a PD-1 axis-binding antagonist but before administration of modified immune-responsive cells, and compared with the state before administration of the PD-1 axis-binding antagonist. If disease stabilization or disease exacerbation is determined, (c) A step comprising administering modified immune-responsive cells, and administering a PD-1 axis-coupled antagonist in one or more doses in each of one or more administration cycles, after or concurrently with and after administration of modified immune-responsive cells, wherein optionally, at least one dose may be on day 1 of each cycle, and optionally, the PD-1 axis-coupled antagonist may be administered for a specific period of time. This may include the following: Furthermore, optionally, the initial dose of the PD-1 axis-coupled antagonist after administration of modified immune-responsive cells may be on day 17, day 21, or day 22 after administration of the immune-responsive cells.

[0088] The method according to the present invention is (a) Administer a PD-1 axis-coupled antagonist in one or more doses in each of one or more administration cycles prior to administration of modified immune-responsive cells, optionally, at least one dose being on day 1 of each cycle. (b) The disease state is determined after administration of a PD-1 axis-binding antagonist but before administration of modified immune-responsive cells, and compared with the state before administration of the PD-1 axis-binding antagonist. If a complete response or partial response is determined, (c) Administer a PD-1 axis-coupled antagonist in one or more doses in each of one or more administration cycles, without administering modified immune-responsive cells, optionally at least one dose on day 1 of each cycle, and optionally administer the PD-1 axis-coupled antagonist for a specific period or the shorter of the period for determining disease progression, and then optionally perform the step in step (c) for determining disease stability or disease progression. (d) The step of administering modified immune-responsive cells, and administering a PD-1 axis-coupled antagonist in one or more doses in each of one or more administration cycles, after or concurrently with and after administration of modified immune-responsive cells, wherein at least one dose may be on day 1 of each cycle, and further optionally, the PD-1 axis-coupled antagonist may be administered for a specific period of time. This may include the following: Furthermore, optionally, the initial dose of the PD-1 axis-coupled antagonist after administration of modified immune-responsive cells may be on day 17, day 21, or day 22 after administration of the immune-responsive cells.

[0089] According to the present invention, "complete response" (CR) is determined when all target lesions or tumors are evaluated or measured as having disappeared. "Partial response" (PR) is determined when, for example, the sum of the measured lengths of the target lesions or tumors decreases by at least 30% compared to a control or pre-treatment comparison. "Disease progression" (PD) is determined when, for example, the sum of the measured lengths of the target lesions or tumors increases by at least 20% compared to a control or pre-treatment comparison since the start of treatment or the presence of one or more new lesions. "Stable disease" (SD) is determined when, since the start of treatment, the sum of the lengths of the target lesions or tumors does not decrease or decline sufficiently to be classified as a PR, nor does it increase sufficiently to be classified as a PD, compared to the minimum SLD.

[0090] According to the present invention, cancer may be recurrent cancer, refractory cancer, recurrent cancer, locally recurrent cancer, or metastatic cancer, unresectable or locally limited cancer, cancer for which there are no surgical or radiotherapy options, or inoperable cancer, or any combination thereof. The subject may have recurrent cancer, refractory cancer, recurrent cancer, locally recurrent cancer, or metastatic cancer, or locally limited or inoperable cancer, or any combination thereof.

[0091] According to the present invention, cancer and / or tumors may express MAGE protein, peptide, antigen or this peptide antigen and / or PD-L1 or PD-L2 [optionally, CPS ≥ 1], and optionally express MAGE-A4 protein, peptide, antigen or this peptide antigen and / or PD-L1 or PD-L2 [optionally, CPS ≥ 1].

[0092] According to the present invention, cancers include lung cancer, non-small cell lung cancer (NSCLC), metastatic or advanced NSCLC, squamous cell NSCLC, adenocarcinoma NSCLC, adenosquamous cell NSCLC, large cell NSCLC, ovarian cancer, gastric cancer, urothelial carcinoma, esophageal cancer, gastroesophageal junction (EGJ) cancer, melanoma, bladder cancer, head and neck cancer, head and neck squamous cell carcinoma (HNSCC), oral cancer, oropharyngeal cancer, hypopharyngeal cancer, pharyngeal cancer, laryngeal cancer, tonsil cancer, tongue cancer, soft palate cancer, pharyngeal cancer, synovial sarcoma, and myxoid round cell carcinoma. Selection may be made from hyaluroliposarcoma (MRCLS), where, optionally, the cancer or tumor expresses the MAGE protein, peptide, antigen or this peptide antigen and / or PD-L1 or PD-L2 [optionally, CPS ≥ 1], optionally, the MAGE-A4 protein, peptide, antigen or this peptide antigen and / or PD-L1 or PD-L2 [optionally, CPS ≥ 1], where CPS means composite positive score.

[0093] According to the present invention, cancer may be selected from any one of the following: breast cancer, metastatic breast cancer, liver cancer, renal cell carcinoma, synovial sarcoma, urothelial carcinoma or tumor, pancreatic cancer, colorectal cancer, metastatic stomach cancer, metastatic gastric cancer, metastatic liver cancer, metastatic ovarian cancer, metastatic pancreatic cancer, metastatic colorectal cancer, metastatic lung cancer, colorectal cancer or adenocarcinoma, lung cancer or adenocarcinoma, pancreatic cancer or adenocarcinoma, myxoid adenoma, pancreatic ductal carcinoma, or hematological malignancy, wherein optionally, the cancer or tumor expresses MAGE protein, peptide, antigen or this peptide antigen and / or PD-L1 or PD-L2 [optionally, CPS ≥ 1], and optionally, expresses MAGE-A4 protein, peptide, antigen or this peptide antigen and / or PD-L1 or PD-L2 [optionally, CPS ≥ 1]. According to the present invention, the cancer may be recurrent or metastatic HNSCC that has disease progression during or after platinum-containing chemotherapy.

[0094] The present invention further provides a method or use for subjects who have not received pre-cancer treatment, or who have received pre-cancer treatment and / or are unresponsive to pre-cancer treatment, or whose disease has worsened during or after pre-cancer treatment.

[0095] According to the present invention, pre-treatment may include one or more systemic and / or local therapies, such as surgery, radiotherapy, cryotherapy, laser therapy, topical therapy, chemotherapy, hormone therapy, targeted drugs, or immunotherapy. Therefore, pre-treatment may include one or more local therapies, such as surgery, radiotherapy, cryotherapy, laser therapy, or topical therapy, and / or one or more systemic therapies, such as chemotherapy, hormone therapy, targeted drugs, or immunotherapy.

[0096] According to the present invention, pretreatment may include a PD-1 axis-binding antagonist, a PD-L1-binding antagonist, or a PD-1-binding antagonist. Therefore, pretreatment is (a) Anti-PD-L1 antibodies that inhibit the binding between PD-L1 and PD-1 and / or between PD-L1 and B7-1, (b) Anti-PD-L1 antibodies that inhibit PD-L1 on the surface of cancer cells from transmitting signals to intracellular pathways. (c) Anti-PD-1 antibodies that inhibit the binding between PD-L1 and PD-1 and / or between PD-L2 and PD-1. (d) Anti-PD-1 antibodies that inhibit PD-1 on the surface of T cells from transmitting signals to intracellular pathways. (e)(i) durvalumab, imfinzi or MEDI4736, (ii) Atezolizumab, Tecentriq or MPDL3280A, (iii) Avelumab, Bavencio or MSB0010718C (iv) MDX-1105, BMS-936559 A PD-L1-binding antagonist selected from the following, (f)(i) Pembrolizumab, Keytruda, lambrolizumab or MK-3475, (ii) Semiprimab, ributayo or REGN-2810, (iii) BMS / ONO, nivolumab, Opdivo, ONO-4538, BMS-936558, or MDX1106 Selected PD-1 binding antagonist It may include any of the following:

[0097] According to the present invention, the pretreatment may include an epidermal growth factor receptor antagonist, optionally cetuximab. According to the present invention, if the pretreatment includes chemotherapy, it may include one or more platinum compounds, optionally selected from lipoplatin, cisplatin, carboplatin, oxaliplatin, nedaplatin, triplatin tetranitrate, phenanthriplatin, satraplatin, and picoplatin. In addition, or / or if the pretreatment includes chemotherapy, it may include one or more chemotherapeutic agents selected from methotrexate, capecitabine, taxane, anthracycline, paclitaxel, docetaxel, paclitaxel protein-binding particles, doxorubicin, epirubicin, 5-fluorouracil, cyclophosphamide, afatinib, vincristine, etoposide, or combinations thereof. In addition, if prior treatment includes chemotherapy, this may include one or more chemotherapeutic agents selected from FEC: 5-fluorouracil, epirubicin, cyclophosphamide; FAC: 5-fluorouracil, doxorubicin, cyclophosphamide; AC: doxorubicin, cyclophosphamide; EC: epirubicin, cyclophosphamide.

[0098] According to the present invention, the subject does not need to have received repeated prior treatments within 12 months or less since the last treatment, or within 6 months or less since the last treatment.

[0099] According to the present invention, subjects may be receiving a PD-1 axially coupled antagonist, such as pembrolizumab, semiprimab, nivolumab, or Opdivo, or may be untreated, or may have received one, two, three, four, five, six, or seven prior doses of a PD-1 axially coupled antagonist before treatment. According to the present invention, subjects may have received one, two, or three prior lines of platinum therapy before treatment, or may have had up to one prior line of platinum therapy before treatment.

[0100] According to the present invention, the subject does not need to have received any prior adjuvant therapy (postoperative radiation and / or chemotherapy) repeatedly within 12 months or less since the last treatment, or repeatedly within 6 months or less since the last treatment.

[0101] According to the present invention, by treatment, (a) Survival without progression, (b) Time until exacerbation, (c) Duration of response, (d) overall survival; (e) Objective effectiveness or objective effectiveness, (f) Total response or total response rate, (g) Partial response or partial response rate, (h) Complete response or complete response rate, (i) Disease stability rate or median disease stability, (j) Median progression-free survival, (k) median time to exacerbation, (l) median duration of response, or (m) Median overall survival, (n) Median objective response or median objective response rate, (o) Median overall response rate or median overall response rate, (p) median partial response or median partial response rate, (q) Median complete response or median complete response, (r) Median disease stability rate or median disease stability One or more of these factors are prolonged, improved, or effectively prolonged or effectively improved compared to treatment with a PD-1 axis-coupled antagonist alone or modified immune-responsive cells alone.

[0102] According to the present invention, by treatment, (a) Best Overall Response (BOR), (b) Time to Response (TTR), (c) Duration of Response (DoR), (d) Duration of Disease Stability (DoSD), (e) Progression-Free Survival (PFS), or (f) Overall Survival (OS) are prolonged, improved, or effectively prolonged or effectively improved compared to treatment with a PD-1 axis-linked antagonist alone or modified immune-responsive cells alone.

[0103] Best overall response (BOR) can be defined as the best response recorded from the date of T-cell infusion to disease progression. Time to confirmed response (TTR) can be defined as the period from T-cell infusion to the start date of confirmed response. Duration of response (DoR) can be defined as the period from the start date of confirmed response to the date of disease progression (PD), disease progression (or death). Duration of disease stability (DoSD) can be defined as the period from T-cell infusion to the date of PD, disease progression (or death). Progression-free survival (PFS) can be defined as the interval from T-cell infusion to the earliest date of disease progression, based on RECIST v1.1 or death by any cause. Overall survival (OS) can be defined as the period from T-cell infusion to death by any cause.

[0104] Progression-free survival (PFS) refers to the time from treatment (or randomization) to the first disease progression or death. Time to progression (TTP) is equivalent to PFS, but does not count patients who die from causes other than the cancer or tumor being treated. Duration of response (DoR) is the length of time that cancer, tumor, or lesion continues to respond to treatment without growing or spreading. According to the present invention, DoR, TTP, and PFS can be evaluated using the Criteria for Evaluation of Treatment Response in Solid Tumors (RECIST) or as a determination of progression using the CA-125 level.

[0105] According to the present invention, PFS and / or TTP and / or DoR or the median thereof may be, or can be extended or improved by, at least 2 weeks, 3 weeks, 1 month, 2 months, 2.3 months, 2.5 months, 2.9 months, 3 months, 3.5 months, 3.8 months, 4 months, 4.5 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 16 months, 18 months, 20 months, 22 months, 2 years, 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, or 10 years compared to treatment with a PD-1 axis-linked antagonist alone (control) or modified immune-responsive cells alone (control). In one embodiment, PFS and / or TTP and / or DoR or the median thereof are extended by about 2.9 to 3.8 months compared to the control. In one embodiment, PFS and / or TTP and / or DoR or the median thereof are extended by at least about 3.8 months compared to the control. In another embodiment, PFS and / or TTP and / or DoR or their median are extended by approximately 2.3 months, and in one embodiment, PFS and / or TTP and / or DoR or their median are extended by approximately 6 months compared to the control.

[0106] "Overall survival" refers to subjects who maintain life for a specified period. According to the present invention, overall survival or this median may be about 6 months, about 1 year, about 1.5 years, about 2 years, about 3 years, about 4 years, about 5 years, about 6 years, about 7 years, about 8 years, about 9 years, or about 10 years from the start of the method or treatment of the present invention or from the first diagnosis, or may be improved or extended, and the events used for survival analysis may be death from any cause. "Survival" refers to subjects who maintain life and includes progression-free survival (PFS) and overall survival (OS). "Overall survival" is the length of time from the date of diagnosis of the disease, tumor and / or cancer or the start of treatment in which a subject diagnosed with the disease is still alive. Survival can be estimated by the Kaplan-Meier method, and any difference in survival is calculated using a stratified log-rank test. "Extended survival" or "increased likelihood of survival" means an increase in PFS and / or OS in the treated subjects compared to treatment with PD-1 axis-coupled antagonist alone (control) or modified immune-responsive cells alone (control). According to the present invention, overall survival or survival may be, or can be extended or improved, at least 1 month, 2 months, 2.3 months, 2.5 months, 2.9 months, 3 months, 3.5 months, 3.8 months, 4 months, 4.5 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 16 months, 18 months, 20 months, 22 months, 2 years, 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, or 10 years, compared to treatment with, for example, a PD-1 axis-linked antagonist alone (control) or modified immune-responsive cells alone (control).

[0107] "Objective response rate" (ObRR) is the percentage of subjects having a predetermined amount of tumor size reduction, optionally determined by the sum of the target lesions or tumor longest diameters (SLD) over the shortest time. "Overall response rate" (ORR) is defined as the percentage of subjects having a partial or complete response to treatment and does not include disease stability. ORR is generally defined as the sum of complete responses (CR) and partial responses (PR) over a specific period. According to the present invention, ObRR and / or ORR and / or PR and / or CR and / or SD may be at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95% compared to treatment with a PD-1 axis-coupled antagonist alone (control) or modified immune-responsive cells alone (control), for example.

[0108] According to the present invention, the method may further include the step of determining the expression level of a biomarker in a sample derived from a subject, wherein the level of the biomarker is compared to a reference level to determine the likelihood of the subject responding to treatment or the level of response of the subject to treatment, wherein the sample is obtained either before, during, or after treatment. The reference level may be the level of the subject before treatment or a level associated with the presence or absence of cancer. The biomarker may be a T-effector-related gene, e.g., CD8A, perforin (PRF1), granzyme A (GZMA), granzyme B (GZMB), interferon-γ (IFN-γ), CXCL9, or CXCL10. The biomarker may be an activated stroma-related gene, e.g., transforming growth factor β (TGF-β), fibroblast-activating protein (FAP), podoplanin (PDPN), collagen gene, or biglycan (BGN). The biomarker may be a myeloid suppressor cell-related gene, such as CD68, CD163, FOXP3, or androgen regulatory gene 1. Alternatively, the biomarker may be PD-L1, CD8, or an androgen receptor (AR) gene.

[0109] The present invention further provides a method for enhancing immune function in subjects having cancer and / or tumors, comprising the step of applying to the subject a therapeutic regime comprising an effective amount of a PD-1 axis-binding antagonist and a population of modified immune-responsive cells expressing or presenting a heterologous TCR, for example, as previously described herein in relation to the method of treatment and aspects, embodiments, and features relating thereto.

[0110] Therefore, in the present invention, (a) The priming, activation, proliferation, and / or cytolytic activity of CD8 T cells in an individual is enhanced compared to before the application of treatment, or compared to treatment with PD-1 axis-coupled antagonist alone or modified immune-responsive cells alone. (b) The number of CD8 T cells increased compared to before the application of treatment, or compared to treatment with PD-1 binding antagonist alone or modified immune-responsive cells alone. (c) In the subject, the expression of MHC class I antigen is selectively increased by cancer and / or tumor cells compared to before the application of treatment, or compared to treatment with PD-1 axis-binding antagonist alone or modified immune-responsive cells alone, and, optionally, the expression of MHC class I antigen is not increased by the subject PBMC cells. (d) In the subject, maturation and activity are enhanced by antigen-presenting cells compared to before treatment or compared to treatment with PD-1 axis-linked antagonist alone or modified immune-responsive cells alone, and optionally, the antigen-presenting cells are dendritic cells. (e) Serum levels of IL-10 and / or IL-8 in the individual are reduced compared to before the application of treatment, or compared to treatment with PD-1 axis-coupled antagonist alone or modified immune-responsive cells alone. (f) In the cancer and / or tumor of the target, the level of T cell infiltration is elevated compared to before the application of treatment or compared to treatment with PD-1 axis-binding antagonist alone or modified immune-responsive cells alone. (g) The target T cells reduce the level of T cell PD-1 expression compared to before treatment, or compared to treatment with PD-1 axis-coupled antagonist alone or modified immune-responsive cells alone. It also provides methods to enhance immune function.

[0111] Therefore, (a) CD8 T cell activation was compared to before the application of treatment, or to treatment with PD-1 axis-bound antagonist alone or modified immune-responsive cells alone, compared to gamma IFN. + (b) Maturation of antigen-presenting cells may be characterized by an increased frequency of CD8 T cells and / or enhanced cytolytic activity, and (b) maturation of CD83 + (c) activation of antigen-presenting cells may be characterized by an increased frequency of dendritic cells, (d) CD8 T cells may be antigen-specific CD8 T cells.

[0112] According to the present invention, (a) A kit comprising a PD-1 axis-linked antagonist and a package insert containing instructions for using the PD-1 axis-linked antagonist in combination with a population of modified immune-responsive cells expressing or presenting a heterologous TCR to treat or delay cancer and / or tumor progression in a subject. (b) A kit comprising a PD-1 axis-linked antagonist and a population of modified immune-responsive cells expressing or presenting a heterologous TCR, and a package insert containing instructions for using the PD-1 axis-linked antagonist and the population of modified immune-responsive cells expressing or presenting a heterologous TCR to treat or delay cancer and / or tumor progression in a subject. (c) A kit comprising a population of modified immune-responsive cells expressing or presenting a heterologous TCR, and a PD-1 axis-binding antagonist and a kit comprising instructions for using the population of modified immune-responsive cells expressing or presenting a heterologous TCR in combination with the PD-1 axis-binding antagonist to treat or delay cancer and / or tumor progression in a subject. The available PD-1 axis-conjugated antagonists are, optionally, anti-PD-L1 antibodies, anti-PD-1 antibodies, and anti-PD-1 immunoadhesins.

[0113] According to the present invention, a subject, i.e., a subject having cancer and / or tumor, may be PD-L1 expression positive (PD-L1+) and / or HLA-A2 expression positive (HLA-A2+) and / or MAGE-A4 expression positive (MAGE-A4+). According to the present invention, subject (a) is HLA-A in any allele * 02:05 Not positive, (b) HLA-A * 02:07 (and A within the antigen-binding domain) * Even in alleles that have the same protein sequence as 02:07, any A * 02 Nullaller (named with the suffix "N", for example, A * 02:32N) is the only HLA-A * 02 as an allele (for example, HLA allele A) * 02:04 and A * (c) Subjects with 02:07 are not eligible, nor do they have CNS transfer.

[0114] The present invention will be further described by reference to the following drawings and embodiments. [Brief explanation of the drawing]

[0115] [Figure 1] This figure shows the increase in CD3 and PD-L1 detected in the responder's tumor after injection. [Figure 2] This figure shows the increase in CD3 and PD-L1 in non-responders after injection. [Figure 3] This figure shows that PD-1 expression is upregulated by MAGE-A4SPEAR T cells upon stimulation compared to non-transduced (NTD) T cells. [Figure 4] This figure shows that PD-1 expression is upregulated upon stimulation by MAGE-A4+CD8SPEAR T cells compared to non-transduced (NTD) T cells. [Figure 5]This figure shows that PD-L1 is expressed in A375 cells (A375.GFP MAGE-A4+ melanoma cell line) and is upregulated by IFN-γ. [Figure 6] This figure shows a scheme for a pre-activation assay that stimulates and upregulates PD-1 expression on SPEAR T cells (xenogeneic MAGE-A4TCR or xenogeneic MAGE-A4TCR + xenogeneic CD8-expressing T cells). [Figure 7] This figure shows that PD-1 is upregulated by pre-activation on A2M4SPEAR T cells. [Figure 8] This figure shows the supernatant of A2M4 cultures in (A) initial stimulation and (B) restimulation with and without anti-PD-1 antibody. IFN-gamma ELISA data are shown here (n=6 small donors). [Figure 9] This diagram shows the scheme for combination therapy with A2M4SPEAR T cells and a PD-1 axis-binding antagonist (pembrolizumab) for cancer patients. [Modes for carrying out the invention]

[0116] array Sequence ID 1, PD1 - Human programmed cell death protein (Homo sapiens)

[0117] [ka]

[0118] Sequence ID No. 2, PD1L1-Human Programmed Cell Death Ligand 1 (Human (Homo sapiens))

[0119] [ka]

[0120] Sequence ID 3, PD1L2-Human Programmed Cell Death Ligand 1 (Human (Homo sapiens))

[0121] [ka]

[0122] Sequence ID 4, durvalumab heavy chain sequence

[0123] [ka]

[0124] Sequence ID 5, durvalumab light chain sequence

[0125] [ka]

[0126] Sequence ID 6, atezolizumab heavy chain sequence

[0127] [ka]

[0128] Sequence ID 7, atezolizumab light chain sequence

[0129] [ka]

[0130] Sequence ID 8, avelumab heavy chain sequence

[0131] [ka]

[0132] Sequence ID 9, avelumab light chain sequence

[0133] [ka]

[0134] Sequence ID 10, MDX1105 heavy chain sequence

[0135] [ka]

[0136] Sequence ID 11, MDX1105 light chain sequence

[0137] [ka]

[0138] Sequence ID 12, pembrolizumab-DB09037 > heavy chain sequence

[0139] [ka]

[0140] Sequence ID 13, pembrolizumab light chain sequence

[0141] [ka]

[0142] Sequence ID 14, nivolumab heavy chain sequence

[0143] [ka]

[0144] Sequence ID 15, nivolumab light chain sequence

[0145] [ka]

[0146] Sequence ID 16, semiprimab heavy chain sequence

[0147] [ka]

[0148] Sequence ID 17, semiprimab light chain sequence

[0149] [ka]

[0150] Sequence ID No. 18, MAGE A4 peptide GVYDGREHTV

[0151] Sequence ID 19, (CD8α)CDR is in bold and underlined, signal sequence is in italics and underlined.

[0152] [ka]

[0153] Sequence ID 20, (CD8α)

[0154] [ka]

[0155] Sequence ID 21, (MAGE A4TCRα chain)CDR is in bold and underlined.

[0156] [ka]

[0157] Sequence ID 22, (MAGE A4TCRα chain coding sequence)

[0158] [ka]

[0159] Sequence ID 23, (MAGE A4TCRβ chain)CDR is in bold and underlined.

[0160] [ka]

[0161] Sequence ID 24, (MAGE A4TCRβ-chain coding sequence)

[0162] [ka]

[0163] Sequence ID 25, (MAGE A4TCRα chain variable region) 136AA-CDR is in bold and underlined. MKKHLTTFLVILWLYFYRGNGKNQVEQSPQSLIILEGKNCTLQCNYTVSPFSNLRWYKQDTGRGPVSLTILTFSENTKSNGRYTATLDADTKQSSLHITASQLSDSASYICVVSGGTDSWGKLQFGAGTQVVVTPD

[0164] Sequence ID No. 26, (MAGE A4TCRβ chain variable region) 133AA-CDR is in bold and underlined. MASLLFFCGAFYLLGTGSMDADVTQTPRNRITKTGKRIMLECSQTKGHDRMYWYRQDPGLGLRLIYYSFDVKDINKGEISDGYSVSRQAQAKFSLSLESAIPNQTALYFCATSGQGAYEEQFFGPGTRLTVLE

[0165] Sequence ID 27, CDR1MAGE A4TCRα chain (residues 48-53) VSFSSN

[0166] Sequence ID 28, CDR2MAGE A4TCRα chain (residues 71-76) LTFSEN

[0167] Sequence ID 29, CDR3MAGE A4TCRα chain (residues 111-125) CVVSGGTDSWGKLQF

[0168] Sequence ID 30, CDR1MAGE A4TCRβ chain (residues 46-50) KGHDR

[0169] Sequence ID 31, CDR2MAGE A4TCRβ chain (residues 68-73) SFDVKD

[0170] Sequence ID 32, CDR3MAGE A4TCRβ chain (residues 110-123) CATSGQGAYEEQFF

[0171] Sequence ID 33, CDR1CD8α (residues 45-53) VLLSNPTSG

[0172] Sequence ID 34, CDR2CD8α (residues 72-79) YLSQNKPK

[0173] Sequence ID 35, CDR3CD8α (residues 118-123) LSNSIM

[0174] Sequence ID 36, MAGE A4

[0175] [ka] [Examples]

[0176] [Example 1] Using tissue biopsy samples taken from subjects with synovial sarcoma (a tumor expressing MAGE A4) before and after T cell (SPEAR T cells, i.e., target T cells modified using a heterologous TCR specific to the MAGE A4 cancer-testicular antigen) injection, the inventors demonstrated the absence of PD-L1 expression in the tissue before injection. The patients were responsive to SPEAR T cell therapy. After SPEAR T cell injection, measured levels of CD3 (part of the T cell receptor (TCR) complex on mature T lymphocytes) expressed in tissues that signal T cell infiltration were elevated, and genetically modified T cells were present. In addition, induction of PD-L1 in tumors associated with T cell infiltration was observed, Figure 1. Data from biopsies derived from ovarian cancer patients unresponsive to SPEAR T-cell therapy demonstrated that PD-L1 expression was positive in pre-infusion biopsies, and that after SPEAR T-cell infusion, T-cell / genetically modified T-cell infiltration (indicated by CD3) increased, and a strong, associated induction of PD-L1 was observed in the tumor, as shown in Figure 2.

[0177] The inventors demonstrated that exposure of genetically modified T cells expressing xenogeneic TCRs to MAGE-A4 upregulated PD-1 expression in response to stimulation with the target antigen MAGE-A4 compared to non-transduced (NTD) T cells lacking xenogeneic TCRs, Figure 3. The same effect was observed in genetically modified T cells expressing xenogeneic MAGE-A4 and xenogeneic CD8, Figure 4. The data indicate that the activity of modified T cells can be enhanced by blocking the PD1 / PD-L1 interaction. The inventors also demonstrated that PD-L1 is upregulated in response to interferon by cancer cell lines, Figure 5. The data in Figure 5 demonstrate that what is clinically observed in patients is also observed in in vitro cancer cell assay systems. As part of the immune response to cancer, T cells produce interferon with antitumor effects via the JAK / STAT pathway of target cells. In patients, the tumor's PD-L1 upregulation response to interferon represents part of the tumor cell's immune evasion adaptation.

[0178] The inventors further investigated the effects of PD-1 blockade on cytotoxicity and effector cytokine production by gene-modified T cells (SPEAR T cells) expressing heterologous TCRs against MAGE-A4, using pre-stimulation protocols and ELISA assays for cytokines. T cell pre-activation was performed as shown in Figure 6. Here, gene-modified T cells expressing heterologous TCRs against MAGE-A4 (including gene-modified T cells expressing both heterologous MAGE-A4 and heterologous CD8) were targeted at tumor cells (irradiated A375MAGE-A4). + Melanoma cells were stimulated, cultured, and isolated from cancer cells, then restimulated in the presence or absence of PD-1 blockade with an anti-PD-1 antibody. The data in Figure 7 show that PD-1 expression on pre-activated T cells increased across both CD4+ and CD8+ cell populations over a 7-day initial stimulation period.

[0179] Using a pre-stimulation model, the inventors showed that in both modified MAGE-A4TCR and modified MAGE-A4TCR+CD8 T cells, IFN gamma, IL-2, and granzyme B are produced during initial stimulation, but this ability is lost after restimulation of this pre-activation step. However, in both modified MAGE-A4TCR and modified MAGE-A4TCR+CD8 T cells, restimulation in the presence of an anti-PD-1 antibody partially restores this cytokine activity, repairing IFN gamma and granzyme B production, but not IL-2 production. Figure 8 shows data for MAGE-A4TCR and IFN gamma. In conclusion, the inventors demonstrated this ability to rescue exhausted modified T cell cytokine function when used in combination with an anti-PD-1 antibody.

[0180] [Example 2] in vivo tumor model The inventors found that in an NSG strain mouse A375 (melanoma) xenograft model, transduction (ADP-A2M4) MAGE-A4TCR T cells 1×10 6This combination was further investigated using intraperitoneal q4d (twice daily for 4 days) administration of the anti-PD-1 antibody pembrolizumab (Keytruda) at a dose of 10 mg / kg, and its effect on tumor burden was recorded according to the following protocol. 72 tumor-bearing mice were randomized into 9 groups of n=8 and treated as shown in Table 1 below.

[0181] [Table 1]

[0182] All animals in groups 1 and 2 will receive either an isotype control (group 1) or pembrolizumab (group 2) via intravenous injection (IP) starting on day 0 of Q4D (i.e., days 0, 4, 8, 12, 16, and 20) (maximum IP dose of 10 ml / kg). No other treatments will be applied (day 0 will be the day for T cell administration, as in groups 3-9).

[0183] All animals in groups 3 and 4 will receive a single dose of NTD T cells via IV at the recommended dose of 5 ml / kg on day 0. No other treatments will be applied to animals in group 3. In addition, animals in group 4 will receive pembrolizumab via IV from day 1 (i.e., days 1, 5, 9, 13, and 17) on Q4D (maximum dose via IV of 10 ml / kg).

[0184] All animals in groups 5, 6, 7, 8, and 9 received a single dose of T cells at the recommended dose of 5 ml / kg via IV on day 0, and groups 5, 6, 7, 8, and 9 received a second treatment as described below. - Group 5: Isotype controls were administered via IP (intravenous injection) from day 1 of Q4D (i.e., days 1, 5, 9, 13, 17, 21, 25, 29, 33, 37, 41, 45, 49, 53, and 57) (maximum dose via IP: 10 ml / kg). - Group 6: Pembrolizumab administered via intravenous injection (IP) from day -1 of Q4D (i.e., days -1, 3, 7, 11, 15, 19, 23, 27, 31, 35, 39, 43, 47, 51, 55, and 59) (maximum dose via IP: 10 ml / kg) - Group 7: Pembrolizumab administered via intravenous injection (IP) starting on day 7 of Q4D (i.e., days 7, 11, 15, 19, 23, 27, 31, 35, 39, 43, 47, 51, 55, and 59) (maximum dose via IP: 10 ml / kg) - Group 8: Pembrolizumab administered via intravenous injection (IP) starting on day 14 of Q4D (i.e., days 14, 18, 22, 26, 30, 34, 38, 42, 46, 50, 54, and 58) (maximum dose via IP: 10 ml / kg) - Group 9: Pembrolizumab administered via intravenous injection (IP) from day 21 of Q4D (i.e., days 21, 25, 29, 33, 37, 41, 45, 49, 53, and 57) (maximum dose via IP: 10 ml / kg)

[0185] In all cases, tumor volume is measured three times a week using calipers from the start of treatment, and the animals are weighed three times a week.

[0186] [Example 3] clinical research The study will be designed to investigate combination therapy with pembrolizumab and ADP-A2M4 (SPEAR T cells modified to express a MAGE-A4 specific TCR) for the treatment of patients with relapsed or metastatic HNSCC. Participants will either have not previously received systemic therapy for metastatic disease, or will have experienced disease progression during or after platinum-containing chemotherapy. Disease may be histologically or cytogenetically confirmed, and / or measurable according to RECIST v1.1.

[0187] The trial is a single-arm phase II trial (n=10 patients) in patients with advanced / recurrent head and neck squamous cell carcinoma (HNSCC) who have not received any checkpoint inhibitor treatment, and who have either received 0-1 prior-line systemic therapy including platinum for metastatic disease, or who are checkpoint inhibitor-free at pretreatment screening and are scheduled to initiate pembrolizumab treatment for advanced disease, or who have recently initiated pembrolizumab treatment (receiving 1-2 doses). Patients are defined as PD-L1+, HLA-A2+, and MAGE-A4 expression: ≥30% of cells are 2+ / 3+ by IHC [i.e., the tumor shows MAGE-A4 expression defined as ≥30% of tumor cells being ≥2+ by IHC (immunohistochemical testing)]. The primary endpoint of measurement is ORR according to RECIST v1.1. The standard historical response rate for pembrolizumab monotherapy, a care PD-1 therapy for advanced / recurrent head and neck squamous cell carcinoma (HNSCC), is 19%.

[0188] Prior to initiating pembrolizumab treatment, subjects undergo apheresis to obtain a T cell population, which is then transduced with the MAGE A4 antigen (specifically, the specific MAGE A4 antigen peptide SEQ ID NO: 18) ADP-A2M4 "SPEAR" TCR to proliferate the cells and cryopreserve them for later use. Subsequently, subjects are treated with up to three doses of pembrolizumab (200 mg IV infusion over 30 minutes every three weeks), followed by a scan of the disease status, e.g., as measurable by RECIST v1.1. Subjects experiencing disease exacerbation or disease stabilization are treated with a lymphocyte depletion therapy regimen, such as fludarabine (30 mg / m²). 2 (4 days at a rate of 600 mg / m²) and cyclophosphamide (600 mg / m²). 2 / days for 3 days) and SPEAR T cell infusion: Proceed to administration of non-escalating doses of 1 to 10 billion transduced cells of ADP-A2M4 and continuation of pembrolizumab (200 mg IV infusion over 30 minutes every 3 weeks) until either disease progression or the shorter of 24 months. Subjects who are scanned and determined to have a complete or partial response will not receive lymphocyte depletion and cell infusion, but will continue pembrolizumab (200 mg IV infusion over 30 minutes every 3 weeks) until either disease progression or the shorter of 24 months, and then may receive lymphocyte depletion, cell infusion, and combination administration of pembrolizumab. Optionally, apheresis is performed before starting pembrolizumab. The first dose of pembrolizumab after cell infusion can be on day 17 or 22 after cell infusion.

[0189] In this study, the exclusion criteria for subjects were: (a) HLA-A * 02:05 positive in any allele, (b) HLA-A * 02:07 (and alleles having the same protein sequence as A * 02:07 within the antigen-binding domain), or any A * 02 null allele (named with the suffix "N", e.g., A * 02:32N) as the only HLA-A * 02 allele (e.g., a subject having HLA alleles A * 02:04 and A * 02:07 is eligible), (c) CNS metastasis, (d) any prior checkpoint inhibitor therapy, (e) any prior cell therapy, (f) if receiving chemotherapy, the washout period required before leukapheresis or including 3 weeks of LD.

[0190] The primary endpoint of measurement is the ORR according to RECISTv1.1. The treatment scheme is shown in Figure 9. The inventions described in the original claims of this application are listed below. [Invention 1] A method for treating, preventing or delaying cancer and / or tumor progression in a subject, comprising the step of applying a therapeutic regime to the subject comprising an effective amount of a PD-1 axis-binding antagonist and a population of modified immune-responsive cells expressing or presenting a heterologous TCR. [Invention 2] The aforementioned PD-1 axis coupled antagonist, (a) PD-1 binding antagonist, (b) PD-L1 binding antagonist, (c) PD-L2 binding antagonist A method according to Invention 1, selected from the group consisting of the following. [Invention 3] The method according to Invention 2, wherein the PD-1 axial coupling antagonist (a) binds to PD-1 and / or SEQ ID NO: 1, or (b) binds to PD-L1 and / or SEQ ID NO: 2. [Invention 4] (a) The PD-1 binding antagonist inhibits the binding of PD-1 to its ligand-binding partner, (b) The PD-L1 binding antagonist inhibits the binding of PD-L1 to its ligand-binding partner. The method according to Invention 3. [Invention 5] The aforementioned PD-1 binding antagonist, (a) PD-L1, (b) PD-L2, (c) Both PD-L1 and PD-L2 The method according to Invention 4, which inhibits the binding of PD-1 to one or more of the following. [Invention 6] The aforementioned PD-L1 binding antagonist, (a) PD1, (b) CD80, (c)B7-1 The method according to Invention 4, which inhibits the binding of PD-L1 to one or more of the following. [Invention 7] The aforementioned PD-1 or PD-L1 binding antagonist is an antibody, and can be optionally selected. (a) The anti-PD-L1 antibody inhibits the binding between PD-L1 and PD-1 and / or between PD-L1 and B7-1, (b) The anti-PD-L1 antibody inhibits PD-L1 on the surface of cancer cells from transmitting signals to intracellular pathways, (c) The anti-PD-1 antibody inhibits the binding between PD-L1 and PD-1 and / or between PD-L2 and PD-1, (d) The anti-PD-1 antibody inhibits PD-1 on the surface of T cells from transmitting signals to intracellular pathways. The method according to any one of inventions 3 to 6. [Invention 8] The aforementioned PD-L1 binding antagonist, (a) durvalumab, imfinzi or MEDI4736, (b) Atezolizumab, Tecentriq or MPDL3280A, (c) Avelumab, Bavencio or MSB0010718C, (d) MDX-1105, BMS-936559 A method according to invention 7, selected from the above. [Invention 9] The aforementioned PD-1 binding antagonist, (a) Pembrolizumab, Keytruda, Lambrolizumab or MK-3475, (b) Semiprimab, ributayo or REGN-2810, (c) BMS / ONO, nivolumab, Opdivo, ONO-4538, BMS-936558 or MDX1106 A method according to invention 7, selected from the above. [Invention 10] The aforementioned PD-L1 binding antagonist, (a) Heavy chain containing SEQ ID NO: 4, and light chain containing SEQ ID NO: 5, or their variable regions or their CDRs, (b) Heavy chain containing SEQ ID NO: 6, and light chain containing SEQ ID NO: 7, or their variable regions or their CDRs, (c) The heavy chain containing SEQ ID NO: 8, and the light chain containing SEQ ID NO: 9, or their variable regions or their CDRs, (d) Heavy chain containing SEQ ID NO: 10, and light chain containing SEQ ID NO: 11, or their variable regions or their CDRs The method according to Invention 8, wherein the antibody contains an antibody. [Invention 11] The aforementioned PD-1 binding antagonist, (a) The heavy chain containing SEQ ID NO: 12, and the light chain containing SEQ ID NO: 13, or their variable regions or their CDRs, (b) The heavy chain containing SEQ ID NO: 14, and the light chain containing SEQ ID NO: 15, or their variable regions or their CDRs, (c) Heavy chain containing SEQ ID NO: 16, and light chain containing SEQ ID NO: 17, or their variable regions or their CDRs The method according to Invention 9, wherein the antibody contains an antibody. [Invention 12] The method according to invention 10 or 11, wherein the antibody is a monoclonal, human or humanized antibody, and is a full-length antibody or an antigen-binding fragment thereof, Fv, Fab, Fab', Fab'-SH, F(ab')2; a diabody, a linear antibody; a single-chain antibody molecule, scFv. [Invention 13] The method according to Invention 2, wherein the PD-1 axis-coupled antagonist is a PD-L2-coupled antagonist and / or binds to Sequence ID No. 3. [Invention 14] The method according to Invention 13, wherein the PD-L2-binding antagonist is an antibody. [Invention 15] The method according to Invention 13, wherein the PD-L2 binding antagonist is an immunoadhesin. [Invention 16] The method according to any of the above inventions, wherein the heterologous TCR binds to or specifically binds to cancer and / or tumor antigens or their peptide antigens. [Invention 17] The method according to any of the above inventions, wherein the heterologous TCR binds to or specifically binds to cancer and / or tumor antigens or their peptide antigens that are associated with cancer symptoms and / or presented by a tumor or cancer cells or tissue. [Invention 18] The aforementioned cancer and / or tumor antigen or its peptide antigen forms a complex with a peptide-presenting molecule, optionally with major histocompatibility complex (MHC) or human leukocyte antigen (HLA), optionally with class I or class II, and the peptide is HLA-A2 or HLA-A * 02, or HLA-A * The method according to invention 16 or 17, which forms a complex with 0201. [Invention 19] The method according to any one of inventions 16 to 18, wherein the cancer and / or tumor antigen or a peptide antigen thereof is a cancer testicular antigen. [Invention 20] The method according to any one of inventions 16 to 19, wherein the cancer and / or tumor antigen or its peptide antigen is selected from any of NY-ESO-1, MART-1 (melanoma antigen recognized by T cells), WT1 (Wilms tumor 1), gp100 (glycoprotein 100), tyrosinase, PRAME (antigen preferentially expressed in melanoma), p53, HPV-E6 / HPV-E7 (human papillomavirus), HBV,TRAIL, DR4, thyroglobulin, TGFBII frameshift antigen, LAGE-1A, KRAS, CMV (cytomegalovirus), CEA (carcinoembryonic antigen), AFP (alpha-fetoprotein), MAGE-A1, MAGE-A2, MAGE-A3, MAGE-A4, MAGE-A6, MAGE-A8 and MAGE-A9, MAGE-A10 or MAGE-A12 or their peptide antigens. [Invention 21] The method according to any one of inventions 16 to 20, wherein the cancer and / or tumor antigen or the peptide antigen thereof is MAGE-A4 or this peptide, preferably sequence GVYDGREHTV, i.e., sequence number 18. [Invention 22] The method according to any one of inventions 16 to 21, wherein the heterologous TCR specifically and / or selectively binds to the cancer and / or tumor antigen or its peptide antigen and / or the peptide-presenting molecule and / or the complex thereof. [Invention 23] The heterogeneous TCR comprises a TCR alpha chain variable domain and a TCR beta chain variable domain, (i) The alpha chain variable domain includes a CDR having the sequence of VSPFSN(αCDR1) of SEQ ID NO: 27 or amino acids 48-53 of SEQ ID NO: 21, LTFSEN(αCDR2) of SEQ ID NO: 28 or amino acids 71-76 of SEQ ID NO: 21, and CVVSGGTDSWGKLQF(αCDR3) of SEQ ID NO: 29 or amino acids 111-125 of SEQ ID NO: 21. (ii) The beta chain variable domain is the sequence of KGHDR(βCDR1) of SEQ ID NO: 30 or amino acids 46-50 of SEQ ID NO: 23, SFDVKD(βCDR2) of SEQ ID NO: 31 or amino acids 68-73 of SEQ ID NO: 23, and CATSGQGAYEEQFF(βCDR3) of SEQ ID NO: 32 or amino acids 110-123 of SEQ ID NO: 23, Or a method according to any of the above inventions, comprising a CDR having a sequence having at least 80% sequence identity to these. [Invention 24] The method according to any one of the above inventions, wherein the heterogeneous TCR comprises a TCR in which the alpha chain variable domain comprises an amino acid sequence having at least 80% identity to SEQ ID NO: 25, and / or the beta chain variable domain comprises an amino acid sequence having at least 80% identity to SEQ ID NO: 26. [Invention 25] The method according to any one of the inventions, wherein a population of modified immune-responsive cells expressing or presenting a heterologous TCR further expresses or presents a heterologous coreceptor, and optionally the coreceptor is a CD8 coreceptor. [Invention 26] The method according to Invention 25, wherein the heterogeneous CD8 coreceptor is a heterodimer or homodimer, a CD8αb heterodimer or a CD8αα homodimer. [Discussion 27] The aforementioned heterologous CD8 coreceptor, (a) CDR1 having at least 80% sequence identity to the amino acid sequence VLLSNPTSG of SEQ ID NO: 33, CDR2 having at least 80% sequence identity to the amino acid sequence YLSQNKPK of SEQ ID NO: 34, and CDR3 having at least 80% sequence identity to the amino acid sequence LSNSIM of SEQ ID NO: 35. (b) CDR1 is the amino acid sequence VLLSNPTSG of SEQ ID NO: 33, CDR2 is the amino acid sequence YLSQNKPK of SEQ ID NO: 34, and CDR3 is the amino acid sequence LSNSIM of SEQ ID NO: 35 (c) an amino acid sequence having at least 80% sequence identity to amino acid numbers 22-235 of SEQ ID NO: 19, or (d) Amino acid sequences that have 100% sequence identity with amino acid numbers 22-235 of the sequence of SEQ ID NO: 19 A method according to invention 25 or 26, including the method described in invention 25 or 26. [Invention 28] The method according to any one of the inventions, wherein a population of modified immune-responsive cells expressing or presenting a heterologous TCR further expresses or presents a heterologous costimulatory ligand, optionally 4-1BBL or CD80. [Invention 29] The modified immune-responsive cells are (a) B cells, T cells, or natural killer (NK) cells, or (b) T cells, optionally selected, CD4 + T cells or CD8 + A method according to any one of Inventions 1 to 28, wherein the cell is a T cell. [Invention 30] The method according to any one of inventions 1 to 29, wherein the PD-1 axis-linked antagonist and modified immune-responsive cells are administered separately, over time, or simultaneously. [Invention 31] (a) Administer the PD-1 axis-coupled antagonist before, simultaneously with, or after the modified immune-responsive cells, (b) Administer the PD-1 axis-coupled antagonist before and after the modified immune-responsive cells, or (c) The PD-1 axis-linked antagonist is administered simultaneously with and after the modified immune-responsive cells, (d) The PD-1 axis-coupled antagonist is administered before and concurrently with the modified immune-responsive cells, (e) Administer the PD-1 axis-coupled antagonist after the modified immune-responsive cells, or (f) Administer the PD-1 axis-linked antagonist before, concurrently with, and after the modified immune-responsive cells. A method according to any one of inventions 1 to 30. [Invention 32] A method according to any one of inventions 1 to 31, comprising administering the PD-1 axis-coupled antagonist and / or the modified immune-responsive cells continuously or intermittently. [Invention 33] The method according to any one of inventions 1 to 31, wherein the modified immune-responsive cells are administered as a single dose. [Invention 34] The method according to any one of the inventions, wherein the modified immune-responsive cells are administered in a dose of approximately 500 million to 1 billion cells, approximately 2 billion to 5 billion cells, or approximately 6 billion to 10 billion cells. [Invention 35] The aforementioned PD-1 axial coupling antagonist (a) Approximately 1-9 or approximately 10-20 mg / kg (b) Approximately 3-5 mg / kg (c) Approximately 50-200 mg or approximately 300-500 mg, (d) Approximately 200mg The method according to any of the above inventions, wherein the dose is administered, and optionally, the dose is a fixed dose. [Invention 36] The aforementioned PD-1 axial coupling antagonist (a) Single dose in each of one or more administration cycles, (b) One or more doses in each of the one or more administration cycles, (c) A single dose on day 1 of each of one or more administration cycles, (d) One or more doses in each of one or more dosing cycles, where at least one dose is on day 1 of each cycle. A method according to any of the above inventions, administered as such. [Invention 37] The aforementioned administration cycle (a) 14-17, 18-21, 22-24, 24-27, 28-30 or 31, (b) 1 week, 2 weeks, 3 weeks, 4 weeks or 1 month The method according to any of the above inventions. [Invention 38] The method according to any one of the inventions, wherein the initial dose of the PD-1 axis-coupled antagonist after administration of the modified immune-responsive cells is administered on day 17, day 21, or day 22 after administration of the immune-responsive cells. [Invention 39] The method according to any one of the inventions, wherein the PD-1 axis-coupled antagonist is administered in one or more doses in each of one or more administration cycles before administration of the modified immune-responsive cells, and is administered in one or more doses in each of one or more administration cycles after administration of the modified immune-responsive cells. [Invention 40] (a) The PD-1 axis-coupled antagonist is administered in one or more doses in each of one or more administration cycles prior to the administration of the modified immune-responsive cells. (b) The disease state is assessed by comparing it with the state before administration of the PD-1 axis-coupled antagonist, and if it is determined that the disease is stable or has worsened, (c) Administer modified immune-responsive cells, and administer the PD-1 axis-binding antagonist in one or more doses in each of one or more administration cycles after the administration of the modified immune-responsive cells. The method described in Invention 39. [Invention 41] The method according to the 40th invention, wherein in step (c), the PD-1 axis-coupled antagonist is administered for a specific period of time. [Invention 42] (a) The PD-1 axis-coupled antagonist is administered in one or more doses in each of one or more administration cycles prior to the administration of the modified immune-responsive cells. (b) The disease state is determined by comparing it with the state before administration of the PD-1 axis-coupled antagonist, and if a complete response or partial response is determined, (c) The PD-1 axis-coupled antagonist is administered in one or more doses in each of one or more administration cycles without administering the modified immune-responsive cells, and then optionally, in step (c), it is determined whether the disease is stable or worsening. (d) Administer modified immune-responsive cells, and administer the PD-1 axis-coupled antagonist in one or more doses in each of one or more administration cycles after or concurrently with and after the administration of the modified immune-responsive cells. The method described in Invention 39. [Invention 43] The method according to Invention 42, wherein in step (c) or (d), the PD-1 axis-coupled antagonist is administered for a specific period or for a period shorter than the period for determining disease exacerbation. [Invention 44] The method according to invention 41 or 43, wherein the aforementioned specific period is 24 months. [Invention 45] The method according to any of the above inventions, wherein the PD-1 axis-coupled antagonist and / or modified immune-responsive cells are administered intravenously, i.e., by intravenous infusion. [Invention 46] The method according to any of the inventions, wherein the cancer is a recurrent cancer, or refractory cancer, or a recurring cancer, or a locally recurring cancer, or a metastatic cancer, or an unresectable or locally limited cancer, or a cancer for which there are no surgical or radiotherapy options, or an inoperable cancer. [Invention 47] The method of any of the above inventions, wherein the subject has recurrent cancer, refractory cancer, or recurring cancer, or locally recurrent cancer, metastatic cancer, or locally limited or inoperable cancer. [Invention 48] The aforementioned cancers include lung cancer, non-small cell lung cancer (NSCLC), metastatic or advanced NSCLC, squamous cell NSCLC, adenocarcinoma NSCLC, adenosquamous cell NSCLC, large cell NSCLC, ovarian cancer, gastric cancer, urothelial carcinoma, esophageal cancer, gastroesophageal junction (EGJ) cancer, melanoma, bladder cancer, head and neck cancer, head and neck squamous cell carcinoma (HNSCC), oral cancer, oropharyngeal cancer, hypopharyngeal cancer, pharyngeal cancer, laryngeal cancer, tonsil cancer, tongue cancer, soft palate cancer, pharyngeal cancer, The method according to any of the above inventions, wherein a cancer or tumor is selected from synovial sarcoma or myxoid round cell liposarcoma (MRCLS), and optionally, the cancer or tumor expresses MAGE antigen or this peptide antigen and / or PD-L1 or PD-L2 [optionally, CPS ≥ 1], or optionally, MAGE-A4 antigen or this peptide antigen and / or PD-L1 or PD-L2 [optionally, CPS ≥ 1]. [Invention 49] The cancer is selected from one of the following: breast cancer, metastatic breast cancer, liver cancer, renal cell carcinoma, synovial sarcoma, urothelial carcinoma or tumor, pancreatic cancer, colorectal cancer, metastatic stomach cancer, metastatic gastric cancer, metastatic liver cancer, metastatic ovarian cancer, metastatic pancreatic cancer, metastatic colorectal cancer, metastatic lung cancer, colorectal cancer or adenocarcinoma, lung cancer or adenocarcinoma, pancreatic cancer or adenocarcinoma, mucinous adenoma, pancreatic ductal carcinoma, or hematological malignancy, and optionally, the cancer or tumor expresses MAGE antigen or this peptide antigen and / or PD-L1 or PD-L2 [optionally, CPS ≥ 1], and optionally, expresses MAGE-A4 antigen or this peptide antigen and / or PD-L1 or PD-L2 [optionally, CPS ≥ 1], and the method according to any of the above inventions. [Invention 50] The method according to any of the above inventions, wherein the subject has not received prior cancer treatment. [Invention 51] The method according to any one of Inventions 1 to 49, wherein the subject has received pre-cancer treatment and / or is unresponsive to pre-cancer treatment. [Invention 52] The method according to Invention 51, wherein the prior treatment includes, optionally, one or more of local therapies, surgery, radiotherapy, cryotherapy, laser therapy, and topical therapies, and / or systemic therapies, such as one or more of chemotherapy, hormone therapy, targeted drugs, or immunotherapy. [Invention 53] The method according to Invention 51, wherein the prior treatment comprises a PD-1 axis-binding antagonist, a PD-L1-binding antagonist, or a PD-1-binding antagonist, wherein the PD-1 axis-binding antagonist is optionally an antibody. [Invention 54] The method according to Invention 51, wherein the prior treatment comprises an epidermal growth factor receptor antagonist, optionally cetuximab. [Invention 55] The method according to Invention 51, wherein the prior treatment comprises chemotherapy comprising a platinum compound, which is optionally selected from lipoplatin, cisplatin, carboplatin, oxaliplatin, nedaplatin, triplatin tetranitrate, phenantriplatin, satraplatin, and picoplatin. [Invention 56] The method according to Invention 51, wherein the prior treatment comprises chemotherapy comprising a chemotherapeutic agent selected from methotrexate, capecitabine, taxane, anthracycline, paclitaxel, docetaxel, paclitaxel protein-binding particles, doxorubicin, epirubicin, 5-fluorouracil, cyclophosphamide, afatinib, vincristine, etoposide, or a combination thereof. [Invention 57] The method according to Invention 51, wherein the prior treatment includes chemotherapy comprising a chemotherapeutic agent selected from FEC: 5-fluorouracil, epirubicin, cyclophosphamide; FAC: 5-fluorouracil, doxorubicin, cyclophosphamide; AC: doxorubicin, cyclophosphamide; EC: epirubicin, cyclophosphamide. [Invention 58] The method according to any one of inventions 51 to 57, wherein the subject has not received prior treatment repeatedly for 12 months or less since the last treatment or for 6 months or less since the last treatment. [Invention 59] The method according to any one of Inventions 51 to 57, wherein the subject has not received any prior adjuvant therapy (postoperative radiation and / or chemotherapy) repeatedly within 12 months or less since the last treatment, or repeatedly within 6 months or less since the last treatment. [Invention 60] The aforementioned treatment (a) Survival without progression, (b) Time until exacerbation, (c) Duration of response, (d) overall survival; (e) Objective effectiveness or objective effectiveness, (f) Total response or total response rate, (g) Partial response or partial response rate, (h) Complete response or complete response rate, (i) Disease stability rate or median disease stability, (j) Median progression-free survival, (k) median time to exacerbation, (l) Median duration of response, (m) Median overall survival, (n) Median objective response or median objective response rate, (o) Median overall response or median overall response rate, (p) median partial response or median partial response rate, (q) Median complete response or median complete response, (r) Median disease stability rate or median disease stability However, the method of any of the inventions which effectively prolongs or improves compared to treatment with a PD-1 axis-coupled antagonist alone or modified immune-responsive cells alone.

Claims

1. A pharmaceutical composition comprising a population of modified immune-responsive cells expressing or presenting a heterologous TCR that specifically binds to the MAGE-A4 antigen or its peptide antigen, for use in a method of treating, preventing, or delaying the progression of cancer in a subject, The aforementioned heterogeneous TCR includes a TCR alpha chain variable domain and a TCR beta chain variable domain, (i) The alpha-chain variable domain includes a CDR having the sequence of VSPFSN (αCDR1) of SEQ ID NO: 27 or amino acids 48-53 of SEQ ID NO: 21, LTFSEN (αCDR2) of SEQ ID NO: 28 or amino acids 71-76 of SEQ ID NO: 21, and CVVSGGTDDSWGKLQF (αCDR3) of SEQ ID NO: 29 or amino acids 111-125 of SEQ ID NO: 21, (ii) The beta-chain variable domain includes a CDR having the sequence of KGHDR (βCDR1) of SEQ ID NO: 30 or amino acids 46-50 of SEQ ID NO: 23, SFDVKD (βCDR2) of SEQ ID NO: 31 or amino acids 68-73 of SEQ ID NO: 23, and CATSGQGAYEEQFF (βCDR3) of SEQ ID NO: 32 or amino acids 110-123 of SEQ ID NO: 23, The cancer is synovial sarcoma, ovarian cancer, or melanoma, and expresses the MAGE-A4 antigen or its peptide antigen. The method includes the step of applying a therapeutic regimen comprising an effective amount of anti-PD-1 antibody and a population of modified immune-responsive cells to the subject, The aforementioned pharmaceutical composition.

2. The aforementioned anti-PD-1 antibody, (a) Pembrolizumab, Keytruda, lambrolizumab or MK-3475, (b) semiprimab, ributayo or REGN-2810, and (c) BMS / ONO, nivolumab, Opdivo, ONO-4538, BMS-936558 or MDX1106 It is selected from the group consisting of, The pharmaceutical composition according to claim 1.

3. The aforementioned anti-PD-1 antibody, (i) the heavy chain containing SEQ ID NO: 12, and the light chain containing SEQ ID NO: 13, or their variable regions or their CDRs, (ii) The heavy chain containing SEQ ID NO: 14, and the light chain containing SEQ ID NO: 15, or the variable regions thereof or their CDRs, (iii) Heavy chain containing SEQ ID NO: 16, and light chain containing SEQ ID NO: 17, or their variable regions or their CDRs including, The pharmaceutical composition according to claim 2.

4. The MAGE-A4 antigen or its peptide antigen is HLA-A2 or HLA-A ＊ 02, or HLA-A ＊ A pharmaceutical composition according to any one of claims 1 to 3, which forms a complex with 0201.

5. The pharmaceutical composition according to any one of claims 1 to 4, wherein the alpha-chain variable domain comprises an amino acid sequence having at least 90% identity with the alpha-chain variable domain of SEQ ID NO: 25, and / or the beta-chain variable domain comprises an amino acid sequence having at least 90% identity with the beta-chain variable domain of SEQ ID NO:

26.

6. A population of modified immune-responsive cells expressing or presenting heterologous TCRs further expresses or presents heterologous coreceptors, (i) The coreceptor is a CD8 coreceptor, (ii) The coreceptor is a heterodimer or homodimer, a CD8αb heterodimer or a CD8αα homodimer, a CD8 coreceptor, or (iii) The coreceptor is a CD8 coreceptor comprising CDR1, which is the amino acid sequence VLLSNPTSG of SEQ ID NO: 33; CDR2, which is the amino acid sequence YLSQNKPK of SEQ ID NO: 34; and CDR3, which is the amino acid sequence LSNSIM of SEQ ID NO:

35. Optionally, the CD8 coreceptor is selected. (a) an amino acid sequence having at least 80% sequence identity to amino acids 22-235 of SEQ ID NO: 19, or (b) Amino acid sequences having 100% sequence identity to amino acids 22-235 of the sequence of Sequence ID No. 19 including, A pharmaceutical composition according to any one of claims 1 to 5.

7. The modified immune-responsive cells, (i) Present heterogeneous costimulatory ligands, and / or (ii) B cells, T cells, natural killer (NK) cells, CD4 ＋ T cells or CD8 ＋ T cells, A pharmaceutical composition according to any one of claims 1 to 6.

8. (i) The anti-PD-1 antibody and modified immune-responsive cells are administered separately, over time, or simultaneously. (ii) The anti-PD-1 antibody is (a) administered before, simultaneously with, or after the modified immune-responsive cells, (b) administered before and after the modified immune-responsive cells, (c) administered simultaneously with and after the modified immune-responsive cells, (d) administered before and concurrently with the modified immune-responsive cells, (e) administered after the modified immune-responsive cells, or (f) administered before, simultaneously with, and after the modified immune-responsive cells, (iii) The anti-PD-1 antibody and / or the modified immune-responsive cells are administered continuously or intermittently. (iv) The modified immune-responsive cells are administered in doses of approximately 500 million to 1 billion cells, approximately 2 billion to 5 billion cells, or approximately 6 billion to 10 billion cells. (v) The anti-PD-1 antibody is (a) Approximately 1 to 9 or approximately 10 to 20 mg / kg (b) about 3 to 5 mg / kg, (c) Approximately 50-200 mg or approximately 300-500 mg, (d) Approximately 200mg It is administered in the following dose: (vi) The anti-PD-1 antibody is (a) Single dose in each of one or more administration cycles, (b) One or more doses in each of one or more administration cycles, (c) A single dose on day 1 of each of one or more administration cycles, (d) One or more doses in each of one or more administration cycles, where at least one dose is on day 1 of each cycle. It is administered as follows: (vii) The first dose of the anti-PD-1 antibody after administration of the modified immune-responsive cells is on day 17, day 21, or day 22 after administration of the immune-responsive cells, and / or (viiii) The anti-PD-1 antibody and / or modified immune-responsive cells are administered intravenously, i.e., by intravenous infusion. A pharmaceutical composition according to any one of claims 1 to 7.

9. The anti-PD-1 antibody is administered in one or more doses in each of one or more administration cycles before the administration of the modified immune-responsive cells, and in one or more doses in each of one or more administration cycles after the administration of the modified immune-responsive cells. A pharmaceutical composition according to any one of claims 1 to 8.

10. (i) The cancer is a recurrent cancer, or refractory cancer, or a recurrent cancer, or a locally recurrent cancer, or a metastatic cancer, an unresectable or locally limited cancer, a cancer for which there are no surgical or radiotherapy options, or an inoperable cancer. (ii) The subject has recurrent cancer, refractory cancer, or recurrent cancer, or locally recurrent cancer, metastatic cancer, or locally limited or inoperable cancer, (iii) The cancer expresses PD-L1 or PD-L2, and / or (iv) The cancer expresses PD-L1 or PD-L2 and has CPS ≥ 1, A pharmaceutical composition according to any one of claims 1 to 9.

11. (1) The subject has not received prior cancer treatment, (2) The subject has received pre-cancer treatment and / or is unresponsive to pre-cancer treatment, (3) The subject has received pre-cancer treatment and / or is unresponsive to pre-cancer treatment, and (a) the subject has not repeatedly received the pre-cancer treatment within 12 months or less since the last treatment or within 6 months or less since the last treatment, or (b) the subject has not repeatedly received any pre-adjuvant therapy (postoperative radiation and / or chemotherapy) within 12 months or less since the last treatment or within 6 months or less since the last treatment. The pharmaceutical composition according to any one of claims 1 to 10.

12. The aforementioned treatment (a) Survival without progression, (b) Time until exacerbation, (c) Duration of response, (d) overall survival; (e) Objective effectiveness or objective effectiveness, (f) Total response or total response rate, (g) Partial response or partial response rate, (h) Complete response or complete response rate, (i) Disease stability rate or median disease stability, (j) median progression-free survival, (k) median time to exacerbation, (l) Median duration of response, (m) Median overall survival, (n) Median objective response or median objective response rate, (o) Median overall response or median overall response rate, (p) median partial response or median partial response rate, (q) Median complete response or median complete response, (r) Median disease stability rate or median disease stability The pharmaceutical composition according to any one of claims 1 to 11, which effectively prolongs or improves the treatment compared to treatment with an anti-PD-1 antibody alone or modified immune-responsive cells alone.

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