Modified T cells for treatment of head and neck cancer

Abstract

The present disclosure relates to methods of treating head and neck cancer, and populations of modified immune responsive cells expressing heterologous TCRs for use in the methods.

Description

Technical Field

[0001] The present disclosure relates to methods of treating head and neck cancer, and populations of modified immune response cells expressing heterologous TCRs for use in the methods. Background Art

[0002] Head and neck cancers are a group of cancers that affect the mouth, nose, throat, larynx, sinuses, or salivary glands, and most are squamous cell carcinomas. Together, these cancers are the seventh most common cancer and the ninth leading cause of cancer death, affecting more than 5.5 million people worldwide. The cancers are strongly associated with smoking and drinking, although other known risk factors are of viral origin and include infection with the Epstein-Barr virus or human papillomavirus. The mutational profiles of HPV+ and HPV- head and neck cancers suggest that these are fundamentally different cancers.

[0003] Head and neck cancer usually affects individuals between the ages of 55 and 65, with men affected twice as often as women. The average 5-year survival rate after diagnosis is 42-64%. Cure rates have improved for early-stage oral cancer, however, most patients present with more advanced cancers, which are less amenable to treatment. After successful first-line therapy, a significant percentage of patients develop a second primary tumor within 20 years at a rate of 9% to 23%, which is usually due to the same carcinogenic exposure as the primary tumor.

[0004] The stage at diagnosis is based on the TNM classification system, where T is the size and configuration of the tumor, N is the presence or absence of lymph node metastases, and M is the presence or absence of distant metastases. The T, N, and M characteristics are combined to produce the "stage" of the cancer, from stage I to stage IVB.

[0005] Surgical resection and radiation therapy (including 3D conformal radiation therapy, intensity modulated radiation therapy, particle beam therapy, and brachytherapy) or concomitant chemotherapy regimens are the main treatment for most head and neck cancers as standard treatment for tumors with regional metastases (stage III or IV). For early primary cancers without regional metastases (stage I or II), surgery alone may be sufficient treatment. Typical chemotherapy agents include a combination of paclitaxel and carboplatin. Docetaxel is also approved for the treatment of advanced head and neck cancer, either alone or in combination with cisplatin and / or fluorouracil.

[0006] Immune checkpoint blockade provides additional options for treatment. Pembrolizumab is approved for first-line treatment of metastatic or unresectable recurrent HNSCC, and nivolumab is approved for the treatment of recurrent or metastatic HNSCC with disease progression during or after platinum-based chemotherapy. Some targeted antibody treatment options for squamous cell carcinoma of the head and neck include cetuximab, bevacizumab, and erlotinib, including the combination of cetuximab with the conventional chemotherapy cisplatin. Cetuximab and platinum / 5-fluorouracil (5-FU) are recognized first-line options.

[0007] For recurrent, unresectable, or metastatic (surgical resection or radiation therapy is not an option) head and neck cancer, the preferred first-line regimen includes pembrolizumab, for example in combination with a platinum-based chemotherapy agent (e.g., cisplatin or carboplatin) and 5-FU. Pembrolizumab administered as a single agent has been described for patients who express PD-L1 and have a combined positivity score (CPS) ≥1.

[0008] New therapies are needed to treat, prevent and / or delay the progression of head and neck cancer. Summary of the invention

[0009] The present inventors have determined that the use of modified T cells comprising a heterologous CD8 co-receptor and a heterologous T cell receptor capable of binding to a MAGE-A4 peptide antigen as a treatment for head and neck cancer is advantageous. Modified T cells comprising a heterologous CD8 co-receptor and a heterologous T cell receptor capable of binding to a MAGE-A4 peptide antigen can, for example, be included in the first-line treatment of head and neck cancer or recurrent head and neck cancer. The present inventors have designed exemplary treatment regimens in these regards.

[0010] Incorporating T cell therapy into first-line treatment has the potential to change the tumor microenvironment by infiltrating the tumor and further exploiting a broader immune response to enhance and maintain T cell activation. This can lead to increased frequency, depth, and persistence of responses. In addition, T cell therapy in earlier lines of treatment has the advantage of access to healthier patients, with a more favorable tumor microenvironment and better T cell harvest. In addition, healthier patients may have a greater response to treatment and have fewer adverse reactions, such as cytokine release syndrome (CRS) and cytopenias.

[0011] T cell therapy can be included in a combination therapy, such as a combination therapy containing a checkpoint inhibitor and / or an additional anticancer therapy such as chemotherapy. Including modified T cells in a combination therapy may be advantageous because it can reduce the dose of the checkpoint inhibitor or additional anticancer therapy. In turn, CRS and / or cytopenias can be reduced, particularly when the anticancer therapy is chemotherapy.

[0012] Thus, the present disclosure provides a method of treating head and neck cancer in an individual comprising administering to the individual a population of modified T cells comprising a heterologous CD8 co-receptor and a heterologous T cell receptor (TCR) capable of binding a MAGE-A4 peptide antigen.

[0013] The present disclosure also provides a population of modified T cells for use in the methods of the present disclosure, comprising a heterologous CD8 co-receptor and a heterologous T cell receptor capable of binding to a MAGE-A4 peptide antigen. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 : An exemplary treatment regimen for locally advanced head and neck cancer that has recurred following treatment with curative intent.

[0015] Figure 2 : Alternative representation of exemplary treatment regimens for locally advanced head and neck cancer that has relapsed following treatment with curative intent.

[0016] Figure 3 : Exemplary treatment regimens for newly metastatic or unresectable locally advanced head and neck cancer.

[0017] Figure 4 : Alternative representation of exemplary treatment regimens for newly metastatic or unresectable locally advanced head and neck cancer.

[0018] Figure 5: Efficacy of ADP-A2M4CD8 in patients with head and neck cancer. Figure 5A : Change in the sum of the longest diameters (SLD) of baseline target lesions in an individual patient. Figure 5B : Change in baseline target SLD within weeks from T-cell infusion. Patients with stable disease had a decrease in SLD of more than 30%, but only at one time point, and were therefore assessed as having stable disease according to RECIST v1.1. Data show the change in SLD from baseline through progression or before surgical resection. Best overall response assessed by the investigator according to RECIST v1.1. PR: partial response. RECIST: Response Evaluation Criteria in Solid Tumors. SD: stable disease.

[0019] Figure 6 : Computed tomography of a hilar mass in a patient with stage IV head and neck cancer demonstrating a partial response. Scans were performed at baseline and 4 weeks after ADP-A2M4CD8 T cell infusion. DETAILED DESCRIPTION

[0020] It should be understood that different applications of the disclosed methods and products can be customized according to specific needs in the art. It should also be understood that the terminology used herein is only for the purpose of describing specific embodiments of the present disclosure and is not intended to be limiting.

[0021] All publications, patents and patent applications cited herein, whether supra or infra, are hereby incorporated by reference in their entirety.

[0022] General Definition

[0023] Unless defined otherwise, technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs.

[0024] As used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the content clearly dictates otherwise. Thus, for example, reference to "a TCR" includes "multiple TCRs," reference to "an antibody" includes two or more such antibodies, etc.

[0025] In general, the term "comprising" is intended to mean including but not limited to. For example, the phrase "a method comprising administering a population of modified T cells" should be interpreted to mean that the method comprises the step of administering such a population, but the method may comprise additional steps, such as administering another therapeutic agent.

[0026] In some aspects of the present disclosure, the word "comprising" is replaced with the phrase "consisting of". The term "consisting of" is intended to be limiting. For example, the phrase "a method consisting of administering a population of modified T cells" should be interpreted to mean that the method comprises the step of administering such a population, and no additional steps.

[0027] The terms "protein" and "polypeptide" are used interchangeably herein and are intended to refer to a polymeric chain of amino acids of any length.

[0028] Typically, the term "about" is used to refer to a value that is within + / - 10% (eg, within + / - 5% or within + / - 2%) of the subsequent value.

[0029] For purposes of this disclosure, to determine the percent identity of two sequences (such as two polynucleotides or two polypeptide sequences), the sequences are aligned for the purpose of optimal comparison (e.g., gaps can be introduced in the first sequence for optimal alignment with the second sequence). Then, the nucleotide residues at the nucleotide positions are compared. When a position in the first sequence is occupied by the same nucleotide residue as the corresponding position in the second sequence, then the nucleotides at that position are identical. The percent identity between the two sequences is a function of the number of identical positions shared by the sequences (i.e., % identity = number of identical positions / total number of positions in the reference sequence × 100).

[0030] Typically, sequence comparisons are performed over the length of a reference sequence. For example, if a user wishes to determine whether a given ("test") sequence has a certain percentage of identity with SEQ ID NO: X, then SEQ ID NO: X would be the reference sequence. For example, to assess whether a sequence is at least 80% identical to SEQ ID NO: X (an example of a reference sequence), one skilled in the art would compare over the length of SEQ ID NO: X and identify how many positions in the test sequence are identical to those in SEQ ID NO: X. If at least 80% of the positions are identical, then the test sequence is at least 80% identical to SEQ ID NO: X. If the sequence is shorter than SEQ ID NO: X, then gap or missing positions should be considered non-identical positions.

[0031] Those skilled in the art are aware of different computer programs that can be used to determine the homology or identity between two sequences. For example, a mathematical algorithm can be used to accomplish the comparison of sequences and the determination of the percent identity between two sequences.

[0032] Methods of treating cancer

[0033] The present disclosure provides a method for treating head and neck cancer in an individual, comprising administering to the individual a population of modified T cells, the population of T cells comprising a heterologous CD8 co-receptor and a heterologous T cell receptor (TCR) capable of binding to a MAGE-A4 peptide antigen. As described above, the inventors have determined that this method is advantageous. In particular, modified T cells comprising a heterologous CD8 co-receptor and a heterologous T cell receptor capable of binding to a MAGE-A4 peptide antigen can be included in the first-line treatment of head and neck cancer.

[0034] In the context of the present disclosure, treating head and neck cancer may also include preventing and / or delaying the progression of head and neck cancer.

[0035] Head and neck cancer in individuals

[0036] The methods of the present disclosure are used to treat head and neck cancer in an individual. The individual is preferably a human. For example, the individual can be a non-human mammal, such as a mouse, rat, rabbit, cat, dog, pig, cow or horse.

[0037] Head and neck cancer can be a cancer that expresses MAGE-A4. Expression of MAGE-A4 in head and neck cancer has been reported. For example, about 20-25% of head and neck cancer solid tumors express MAGE-A4, of which 40-45% of individuals also express HLA-A*02. For example, the cancer can be a solid tumor. For example, the cancer can be head and neck squamous cell carcinoma.

[0038] At least 1% of head and neck cancer cells from an individual may express MAGE-A4, such as at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, or at least 80%. The percentage of cells expressing MAGE-A4 can be determined by any method known to the skilled person, such as immunohistochemistry (IHC), flow cytometry, or enzyme-linked immunosorbent assay (ELISA).

[0039] The expression of MAGE-A4 in head and neck cancer may have an intensity greater than or equal to (≥) 1+, such as ≥2+ or ≥3+. The intensity score can be assessed by IHC staining of the tumor, and the score is as follows: negative = no staining or staining in less than or equal to ≤10% of the stained cells; 1+ = incomplete staining in ≥10% of the stained cells; 2+ = weak to moderate staining in ≥10% of the stained cells; strong and complete staining in ≥10% of the stained cells.

[0040] The head and neck cancer may be a head and neck tumor. The head and neck cancer may be a head and neck epithelial cell carcinoma (carcinoma). For example, the head and neck cancer may be selected from any one of head and neck squamous cell carcinoma (HNSCC), oral cancer, oropharyngeal cancer, hypopharyngeal cancer, pharyngeal cancer, laryngeal cancer, tonsil cancer, tongue cancer, soft palate cancer, and pharyngeal cancer.

[0041] For example, head and neck cancer can be nasopharyngeal cancer or non-nasopharyngeal cancer. Non-nasopharyngeal cancer can include lip cancer, oral cancer, oropharyngeal cancer, hypopharyngeal cancer, glottic cancer, supraglottic cancer, ethmoid cancer, maxillary sinus cancer and occult primary cancer.

[0042] Thus, the cancer may be nasal cancer or epithelial cell cancer or squamous cell cancer, including paranasal sinus cancer and nasal cavity cancer affecting the nasal cavity and paranasal sinuses, or nasopharyngeal cancer, including cancer occurring in the nasopharynx, nasal cavity and Eustachian tube and upper throat, which may also be a lymphoepithelioma.

[0043] Cancer can be oral cancer or epithelial cancer or cancer or epithelial cancer of the mouth, including squamous cell carcinoma of the inner lip, lip, tongue, floor of mouth, gums, hard palate. Cancer can be pharyngeal cancer, such as oropharyngeal cancer, oropharyngeal squamous cell carcinoma, HPV-positive oropharyngeal cancer or HPV-positive oropharyngeal squamous cell carcinoma, optionally, wherein the squamous cell carcinoma is oropharyngeal or pharyngeal, including soft palate, tongue base and tonsils. Cancer can be hypopharyngeal cancer, including cancer of the pyriform sinus, posterior pharyngeal wall or cricoid cartilage area or its metastasis to the lymphatic network around the larynx. Cancer can be laryngeal cancer, including laryngeal cancer, glottic cancer, supraglottic laryngeal cancer or subglottic laryngeal cancer. Cancer can be tracheal cancer, cancer or squamous cell carcinoma of the salivary glands, teratoma, adenocarcinoma, adenoid cystic carcinoma and mucoepidermoid carcinoma or melanoma or lymphoma of the upper aerodigestive tract. The cancer may be metastatic head and neck cancer that has spread to the adrenal glands, skin, liver pleura, bones, lungs, or mediastinal lymph nodes.

[0044] In any case, head and neck cancer can be primary, secondary, recurrent or recurrent, metastatic or advanced. For example, head and neck cancer can be recurrent or recurrent, unresectable or metastatic head and neck cancer. For example, head and neck cancer may not be suitable for treatment by surgical resection or radiotherapy. For example, head and neck cancer can be staged as T4b, N0-3 head and neck cancer.

[0045] For example, the head and neck cancer can be a recurrent head and neck cancer. The recurrent head and neck cancer can be a locally advanced recurrent or a metastatic recurrent. For example, after a curative intent treatment for locally advanced head and neck cancer, the head and neck cancer may have recurred. Therefore, the individual can be a cancer patient who has received a curative intent treatment for locally advanced head and neck cancer. Such cancer patients form a subgroup of head and neck cancer patients known in the art. The curative intent treatment will be described in detail below. The method disclosed herein may represent a first-line treatment for recurrent head and neck cancer.

[0046] Alternatively, for example, the head and neck cancer may be a head and neck cancer that has not been treated before. In other words, the head and neck cancer may be the first occurrence of head and neck cancer in an individual. Therefore, prior to the methods of the present disclosure, the individual may not have received treatment for head and neck cancer. For example, the head and neck cancer may be a newly metastatic head and neck cancer or an unresectable locally advanced head and neck cancer. For example, the head and neck cancer may be a newly metastatic head and neck cancer or an unresectable locally advanced head and neck cancer that has not been treated before. Therefore, prior to the methods of the present disclosure, the individual may not have received treatment for newly metastatic head and neck cancer or unresectable locally advanced head and neck cancer. In any case, the methods of the present disclosure may be intended to treat previously untreated head and neck cancer. The methods of the present disclosure may represent a first-line treatment for head and neck cancer.

[0047] In the context of the present disclosure, a "line" of therapy may refer to a treatment regimen for a cancer for which curative intent treatment has failed or for which curative intent treatment may not be appropriate. Curative intent treatments are described in detail herein. For example, failure of curative intent treatment may refer to failure of curative intent treatment to eliminate cancer or induce remission. For example, failure of curative intent treatment may lead to recurrence, or spread of cancer, such as local invasion or metastasis. When the cancer is advanced, such as when the cancer is locally advanced cancer or metastatic cancer, curative intent treatment may not be appropriate. Therefore, a "line" of therapy may refer to a treatment regimen for locally advanced, metastatic, or recurrent cancer. A "line" of therapy may refer to a treatment regimen for recurrent or already spread cancer.

[0048] For example, a "line" of treatment may be a first-line therapy for cancer. In other words, the treatment regimen may be the first treatment regimen adopted for cancer after failed or curative intent treatment. For example, the treatment regimen may be the first treatment regimen adopted after cancer relapses or spreads. For example, the treatment regimen may be the first treatment regimen adopted for cancer after local progression / invasion, metastasis, or recurrence. For example, a "line" of treatment may be a second-line therapy for cancer. In other words, the treatment regimen may be the second treatment regimen adopted for cancer, such as after the failure of the first treatment regimen. For example, the failure of the first treatment regimen may lead to the relapse or further spread of cancer. For example, the failure of the first treatment regimen may lead to local progression / invasion, metastasis, or recurrence. For example, the treatment line may be a third-line therapy for cancer. In other words, the treatment regimen may be the third treatment regimen adopted for cancer, such as after the failure of the second treatment regimen. For example, the failure of the second treatment regimen may lead to the relapse or further spread of cancer. For example, the failure of the second treatment regimen may lead to local progression / invasion, metastasis, or recurrence.

[0049] Curative Intent Treatment

[0050] Treatment with curative intent may refer to any therapy that has the potential to cure an individual's cancer. For example, treatment with curative intent may refer to therapy administered to an individual in an attempt to cure head and neck cancer. For recurrent or spread cancer, treatment with curative intent may precede "lines" of treatment. Thus, cancer may have recurred or spread after treatment with curative intent. Cancer may have locally invaded, metastasized, or recurred after treatment with curative intent.

[0051] The curative intent treatment can be any known or unknown treatment for head and neck cancer. As described above, known treatments for head and neck cancer include surgical resection, radiation therapy, and systemic therapy (e.g., chemotherapy). Thus, the curative intent treatment can include (i) surgical resection, (ii) radiation therapy, and / or (iii) systemic therapy. For example, the curative intent treatment can include: (i); (ii); (iii); (i) and (ii); (i) and (iii); (ii) and (iii); or (i), (ii), and (iii). In certain aspects of the present disclosure, the curative intent treatment can include (i); (ii); or (i) and (ii).

[0052] Systemic therapy may include or consist of: (a) chemotherapy. Systemic therapy may include or consist of: (b) immunotherapy. Systemic therapy may include or consist of: (c) targeted therapy. For example, systemic therapy may include or consist of: (a); (b); (c); (a) and (b); (a) and (c); (b) and (c); or (a), (b), and (c).

[0053] Chemotherapy such as for head and neck cancer is well known in the art. For example, such chemotherapy can include anti-tumor drugs based on platinum, such as cisplatin or carboplatin. For example, such chemotherapy can include antimetabolites, such as fluorouracil (5-FU), capecitabine or methotrexate. For example, such chemotherapy can include taxane drugs, such as docetaxel or paclitaxel. For example, such chemotherapy can include podophyllotoxin derivatives, such as etoposide. For example, such chemotherapy can include alkylating agents, such as cyclophosphamide. For example, such chemotherapy can include anthracyclines, such as doxorubicin. For example, such chemotherapy can include vinca alkaloids, such as vincristine.

[0054] Immunotherapy, such as immunotherapy for head and neck cancer, is well known in the art. For example, such immunotherapy may include therapeutic immune cells, immunomodulators, checkpoint inhibitors and vaccines. Therapeutic immune cells may include T cells, such as engineered T cells, such as CAR T cells or T cells expressing engineered TCR. Immunomodulators may include, for example, interleukins, cytokines, chemokines and immunomodulatory imide drugs. For example, immunomodulators may include Multikine (interleukin, injection). For example, immunomodulators may include monalizumab, a humanized anti-NKG2A blocking antibody that prevents tumor cells expressing HLA-E from inhibiting CD8+T cells and NK cells. Checkpoint inhibitors may, for example, include CTLA-4 inhibitors or PD-1 axis binding antagonists. PD-1 axis binding antagonists may, for example, include pembrolizumab, nivolumab and penpulimab. Checkpoint inhibitors and PD-1 axis binding antagonists will be described in detail below. For example, the vaccine may include vidutolimod (CMP-001), which is a Toll-like receptor 9 (TLR9) agonist cancer vaccine.

[0055] Targeted therapies, such as targeted therapies for head and neck cancer, are well known in the art. Targeted therapy is a technical term that refers to treatments that target specific genes and proteins that help cancer cells survive and grow. For example, targeted therapies can include EGFR antagonists (such as cetuximab), tropomyosin kinase receptor antagonists (such as larotrectinib), HER2 antagonists, kinase inhibitors (such as afatinib or lenvatinib), farnesyl transferase inhibitors (such as tipifarnib), PI3 kinase inhibitors (such as buparlisib), inhibitors of apoptosis proteins (such as xevinapant (Debio 1143)).

[0056] Systemic treatment can include previously described medicines or drug combinations for the first-line therapy of head and neck cancer (such as recurrent, unresectable or metastatic head and neck cancer).Systemic treatment can include medicines or drug combinations, which have been previously described as "standard treatment" or approved treatment for head and neck cancer (such as recurrent, unresectable or metastatic head and neck cancer), such as "standard treatment" first-line treatment.For example, systemic treatment can include pembrolizumab, cetuximab, cisplatin, carboplatin, 5-FU, docetaxel, paclitaxel, etoposide, cyclophosphamide, doxorubicin, vincristine, methotrexate, capecitabine and / or afatinib.For example, systemic treatment can include monotherapy with pembrolizumab, cetuximab, cisplatin, carboplatin, 5-FU, docetaxel, paclitaxel, methotrexate or capecitabine. For example, systemic treatment may include monotherapy with: pembrolizumab, cisplatin, and 5-FU; pembrolizumab, carboplatin, and 5-FU; cetuximab, cisplatin, and 5-FU; cetuximab, carboplatin, and 5-FU; cetuximab and cisplatin; cisplatin and docetaxel; cisplatin and paclitaxel; carboplatin and docetaxel; carboplatin and paclitaxel; cisplatin and 5-FU; cisplatin, docetaxel, and cetuximab anti-drug therapy; cisplatin, paclitaxel, and cetuximab; carboplatin, docetaxel, and cetuximab; carboplatin, paclitaxel, and cetuximab; cisplatin, docetaxel, and pembrolizumab; cisplatin, paclitaxel, and pembrolizumab; carboplatin, docetaxel, and pembrolizumab; carboplatin, paclitaxel, and pembrolizumab; cetuximab and pembrolizumab; cisplatin and etoposide; carboplatin and etoposide; or cyclophosphamide, doxorubicin, and vincristine.

[0057] In one aspect of the present disclosure, the curative intent treatment does not include therapeutic T cells. Thus, prior to the methods of the present disclosure, the individual may not have received therapeutic T cells.

[0058] In any case, a cancer patient who has received curative intent treatment for locally advanced head and neck cancer may have been substantially cured of cancer by curative intent treatment. However, head and neck cancer (such as locally advanced head and neck cancer) may be prone to recurrence. For example, the cancer may be prone to recurrence or spread. The cancer may be prone to local invasion or metastasis. The methods of the present disclosure may be intended to treat recurrence, recurrence, spread, local invasion or metastasis. The methods of the present disclosure may represent a first-line treatment for recurrence, recurrence, spread, local invasion or metastasis.

[0059] Population of modified T cells

[0060] The method includes administering to an individual a population of modified T cells, the population of modified T cells comprising a heterologous CD8 co-receptor and a heterologous TCR capable of binding to a MAGE-A4 peptide antigen. The presence of the heterologous CD8 co-receptor and the heterologous TCR causes the T cells to be "modified". The heterologous CD8 co-receptor and the heterologous TCR are usually present on the surface of the modified T cells. In other words, the modified T cells can express the heterologous CD8 co-receptor and the heterologous TCR on their surface.

[0061] In the context of the present disclosure, the term "heterologous" refers to a polypeptide or nucleic acid that is foreign to a particular biological system (such as a T cell), i.e., does not naturally occur in that system. A "heterologous" polypeptide or nucleic acid can be introduced into the system artificially or recombinantly. Therefore, heterologous expression of a TCR can alter the specificity of a T cell. Heterologous expression of a CD8 co-receptor can confer functions associated with a CD8 co-receptor to a T cell. Heterologous CD8 co-receptors and heterologous TCRs will be described in detail below.

[0062] The modified T cells may include CD4+T cells. In other words, the modified T cells may include T cells expressing endogenous CD4 co-receptors. The modified T cells may include CD8+T cells. In other words, the modified T cells may include T cells expressing endogenous CD8 co-receptors. The modified T cells may include CD4+T cells and CD8+T cells. In other words, the modified T cells may include T cells expressing endogenous CD4 co-receptors and T cells expressing endogenous CD8 co-receptors. Both CD4+T cells and CD8+T cells are capable of carrying heterologous CD8 co-receptors.

[0063] The modified T cells can be allogeneic for an individual. The modified T cells can preferably be autologous for an individual. In this case, the modified T cells can be produced by modifying endogenous cells obtained from an individual. Therefore, the method can include producing a colony. The method for producing modified T cells is known in the art and is considered in the following embodiments. Generally, the modified T cells of the present disclosure are produced by cells, such as peripheral blood mononuclear cells (PBMC). T cells are usually selected from harvested cells and operated to contain the desired modifications (here, heterologous CD8 co-receptors and heterologous TCRs). Therefore, the method can include producing a colony by the following steps: (a) obtaining peripheral blood mononuclear cells (PBMCs) from the individual; (b) selecting T cells from PBMCs; and (c) modifying the selected T cells to express the heterologous CD8 co-receptors and the heterologous TCRs.

[0064] Autologous modified T cells can be produced when it is expected that an individual will need them. In other words, autologous modified T cells can be produced in advance before an individual needs to be treated with modified cells. This can help ensure that autologous modified T cells are available for administration as soon as possible after an individual is determined to need treatment. In this way, an individual does not need to wait for autologous T cells to be produced before treatment can begin. This can improve the effectiveness of treatment.

[0065] In the treatment of head and neck cancer with a high risk of recurrence, it may be particularly relevant to produce autologous T cells in advance. The risk of recurrence of an individual receiving a cure for head and neck cancer (such as locally advanced head and neck cancer) can be determined by conventional methods in the art. This method may include monitoring clinical signs or symptoms. For example, this method may include one or more magnetic resonance imaging (MRI), positron emission tomography (PET) and / or computerized tomography (CT) scans performed after the cure for the intended treatment to monitor the progression and / or recurrence of the tumor. For example, MRI, PET and / or CT scans may be performed approximately every three months (such as once every 4 to 16 weeks, or once every 8 to 12 weeks). If an individual is identified as having a high risk of recurrence, PBMCs may be obtained at this time (i.e., before recurrence) to produce autologous modified T cells prepared for administration at the time of recurrence. Thus, when the cancer is a recurrent head and neck cancer, the method of the present disclosure may include generating a population by the following steps: (a) obtaining peripheral blood mononuclear cells (PBMC) from an individual; (b) selecting T cells from the PBMC; and (c) modifying the selected T cells to express a heterologous TCR and an optional heterologous CD8 co-receptor, wherein one or more of steps (a) to (c) are performed before recurrence. Preferably, step (a) is performed before recurrence. More preferably, steps (a) and (b) are performed before recurrence. Most preferably, steps (a), (b) and (c) are performed before recurrence. Any of these options effectively allows treatment to preemptively intervene in recurrence.

[0066] In any case, the population of modified T cells can be administered, for example, in a single dose. The population of modified T cells can be administered, for example, as soon as possible after the diagnosis of head and neck cancer, for example, in a single dose. For example, after the recurrence of head and neck cancer is determined, the population of modified T cells can be administered as soon as possible. For example, the population of modified T cells can be administered as soon as possible after previously untreated head and neck cancer is determined.

[0067] In the context of the present disclosure, the term "as soon as possible" may refer to the earliest time point of a colony of modified T cells. As described above, the generation of autologous modified T cells may require some time, which may cause a delay between the diagnosis of head and neck cancer and the implementation of therapy. Therefore, administering as soon as possible after diagnosis may refer to administering as soon as possible once autologous modified T cells are produced where feasible. For example, administering as soon as possible after diagnosis may refer to administering less than about 150 days after the diagnosis of head and neck cancer, such as less than about 125 days, less than about 100 days, less than about 90 days, less than about 80 days, less than about 70 days, less than about 60 days, less than about 50 days, less than about 40 days, or less than about 30 days after the diagnosis of head and neck cancer. For example, the population can be administered to an individual about 30 to about 150 days after diagnosis of head and neck cancer, such as about 40 to about 125 days, about 50 to about 100 days, about 90 days, about 85 days, about 80 days, about 75 days, about 70 days, about 65 days, or about 60 days after diagnosis of head and neck cancer.

[0068] Heterologous TCR

[0069] The modified T cells comprise a heterologous TCR capable of binding to a MAGE-A4 peptide antigen. In other words, the modified T cells express or present a heterologous TCR capable of binding to a MAGE-A4 peptide antigen, for example on their surface. MAGE-A4 is a well-known cancer antigen whose expression in normal (i.e., non-cancerous) tissues is limited. MAGE-A4 has been shown to inhibit p53 targets such as BAX and CDKN1A and is a binding partner of the oncogene gankyrin.

[0070] The heterologous TCR is capable of binding to a MAGE-A4 peptide antigen. The heterologous TCR may, for example, bind to GVYDGREHTV (SEQ ID NO: 1), which is a peptide sequence called MAGE-A4 230-239 contained in MAGE-A4. The heterologous TCR may, for example, bind to a complex comprising a MAGE-A4 peptide antigen (e.g., GVYDGREHTV (SEQ ID NO: 1)) and an HLA molecule (e.g., an HLA-A molecule (e.g., HLA-A*02 or HLA-A*0201 molecule)). In any case, the binding may be specific. Specificity refers to the strength of binding between a heterologous TCR and its target antigen. Specificity can be described by the dissociation constant Kd, which is the ratio of the bound state to the unbound state of a receptor-ligand system. Generally, the fewer different antigens other than MAGE-A4 that a heterologous TCR is able to bind, the stronger its binding specificity.

[0071] For example, the heterologous TCR can bind to a MAGE-A4 peptide antigen (e.g., SEQ ID NO: 1), or a complex comprising a MAGE-A4 peptide antigen (e.g., SEQ ID NO: 1) and an HLA molecule, wherein the dissociation constant (Kd) is 0.01 μM to 100 μM, 0.01 μM to 50 μM, 0.01 μM to 20 μM, 10 μM to 1000 μM, 10 μM to 500 μM, or 50 μM to 500 μM. For example, in a preferred aspect of the present disclosure, the heterologous TCR binds to a MAGE-A4 peptide antigen, or a complex comprising a MAGE-A4 peptide antigen and an HLA molecule, wherein the Kd is 0.05 μM to 20.0 μM. For example, a heterologous TCR can bind to the MAGE-A4 peptide antigen, or to a complex containing the MAGE-A4 peptide antigen and an HLA molecule, with 0.01 μM, 0.02 μM, 0.03 μM, 0.04 μM, 0.05 μM, 0.06 μM, 0.07 μM, 0.08 μM, 0.0 9μM, 0.1μM, 0.15μM, 0.2μM, 0.25μM, 0.3μM, 0.35μM, 0.4μM, 0.45μM ,0.5μM, 0.55μM, 0.6μM, 0.65μM, 0.7μM, 0.75μM, 0.8μM, 0.85μM, 0.9 μM, 0.95μM, 1.0μM, 1.5μM, 2.0μM, 2.5μM, 3.0μM, 3.5μM, 4.0μM, 4.5 μM, 5.0μM, 5.5μM, 6.0μM, 6.5μM, 7.0μM, 7.5μM, 8.0μM, 8.5μM, 9.0μM , 9.5μM, 10.0μM, 20μM, 30μM, 40μM, 50μM, 60μM, 70μM, 80μM, 90μM, 1 Kd of 00μM, 150μM, 200μM, 250μM, 300μM, 350μM, 400μM, 450μM, 500μM. For example, Kd can be measured using surface plasmon resonance, optionally at 25°C, optionally between pH 6.5 and pH 6.9 or between pH 7.0 and pH 7.5. The dissociation constant Kd or koff / kon can be determined by experimentally measuring the dissociation rate constant koff and the association rate constant kon. The TCR dissociation constant can be measured using a soluble form of the TCR, wherein the TCR comprises a TCR alpha chain variable domain and a TCR beta chain variable domain.

[0072] For example, a heterologous TCR can be a recombinant or synthetic or artificial TCR. In other words, a heterologous TCR can be a TCR that does not exist in nature. For example, a heterologous TCR can be an affinity-enhanced TCR, such as a specific peptide enhanced affinity receptor (SPEAR). TM )TCR.

[0073] The heterologous TCR may, for example, comprise an alpha chain variable domain having at least 80% sequence identity to the sequence of amino acid residues 22-125 of SEQ ID NO: 2. The heterologous TCR may, for example, comprise a beta chain variable domain having at least 80% sequence identity to the sequence of amino acid residues 22-123 of SEQ ID NO: 3. The heterologous TCR may, for example, comprise an alpha chain variable domain having at least 80% sequence identity to the sequence of amino acid residues 22-125 of SEQ ID NO: 2 and a beta chain variable domain having at least 80% sequence identity to the sequence of amino acid residues 22-123 of SEQ ID NO: 3. For example, the alpha chain variable domain may have at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, or at least 99% sequence identity to the sequence of amino acid residues 22-125 of SEQ ID NO: 2. For example, the alpha chain variable domain can comprise or consist of amino acid residues 22-125 of SEQ ID NO: 2. For example, the beta chain variable domain can have at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, or at least 99% sequence identity to the sequence of amino acid residues 22-123 of SEQ ID NO: 3. For example, the beta chain amino acid sequence can comprise or consist of amino acid residues 22-123 of SEQ ID NO: 3.

[0074] The heterologous TCR may, for example, comprise an alpha chain having at least 80% sequence identity to the sequence of amino acid residues 22-282 of SEQ ID NO: 2. The heterologous TCR may, for example, comprise a beta chain having at least 80% sequence identity to the sequence of amino acid residues 22-311 of SEQ ID NO: 3. The heterologous TCR may, for example, comprise an alpha chain having at least 80% sequence identity to the sequence of amino acid residues 22-282 of SEQ ID NO: 2 and a beta chain variable domain having at least 80% sequence identity to the sequence of amino acid residues 22-311 of SEQ ID NO: 3. For example, the alpha chain variable domain may have at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, or at least 99% sequence identity to the sequence of amino acid residues 22-282 of SEQ ID NO: 2. For example, the alpha chain variable domain may comprise or consist of amino acid residues 22-282 of SEQ ID NO: 2. For example, the beta chain variable domain can have at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, or at least 99% sequence identity to the sequence of amino acid residues 22-311 of SEQ ID NO: 3. For example, the beta chain amino acid sequence can comprise or consist of amino acid residues 22-311 of SEQ ID NO: 3.

[0075] Heterologous TCRs are typically expressed at the surface of T cells with an N-terminal signal peptide that is cleaved prior to expression. In this regard, amino acids 1-21 of each of SEQ ID NO: 2 and SEQ ID NO: 3 are typically cleaved prior to expression of the TCR at the surface of the T cell. A heterologous TCR may, for example, comprise an alpha chain amino acid sequence having at least 80% sequence identity to SEQ ID NO: 2. A heterologous TCR may, for example, comprise a beta chain amino acid sequence having at least 80% sequence identity to SEQ ID NO: 3. A heterologous TCR may, for example, comprise an alpha chain amino acid sequence having at least 80% sequence identity to SEQ ID NO: 2 and a beta chain amino acid sequence having at least 80% sequence identity to SEQ ID NO: 3. For example, the alpha chain amino acid sequence may have at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 2. For example, the alpha chain amino acid sequence may comprise or consist of SEQ ID NO: 2. For example, the beta chain amino acid sequence can have at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 3. For example, the beta chain amino acid sequence can comprise or consist of SEQ ID NO: 3.

[0076] For example, a heterologous TCR may comprise: (i) an alpha chain variable domain comprising a CDR1 comprising (1) a sequence of SEQ ID NO: 4 or (2) an amino acid sequence comprising one, two or three amino acid insertions, deletions or substitutions relative to the sequence of SEQ ID NO: 4; (ii) an alpha chain variable domain comprising a CDR2 comprising (1) a sequence of SEQ ID NO: 5 or (2) an amino acid sequence comprising one, two or three amino acid insertions, deletions or substitutions relative to the sequence of SEQ ID NO: 5; (iii) an alpha chain variable domain comprising a CDR3 comprising (1) a sequence of SEQ ID NO: 6 or (2) an amino acid sequence comprising one, two or three amino acid insertions, deletions or substitutions relative to the sequence of SEQ ID NO: 6; (iv) a beta chain variable domain comprising a CDR1 comprising (1) a sequence of SEQ ID NO: 7 or (2) an amino acid sequence comprising one, two or three amino acid insertions, deletions or substitutions relative to the sequence of SEQ ID NO: 7. NO:7 comprises an amino acid sequence having one, two or three amino acids inserted, deleted or substituted; (v) a beta chain variable domain comprising CDR2, wherein CDR2 comprises (1) a sequence of SEQ ID NO:8 or (2) an amino acid sequence having one, two or three amino acids inserted, deleted or substituted relative to the sequence of SEQ ID NO:8; and / or (vi) a beta chain variable domain comprising CDR3, wherein CDR3 comprises (1) a sequence of SEQ ID NO:9 or (2) an amino acid sequence having one, two or three amino acids inserted, deleted or substituted relative to the sequence of SEQ ID NO:9.

[0077] For example, the α chain of a heterologous TCR can comprise: (i) an α chain variable domain comprising a CDR1 comprising (1) the sequence of SEQ ID NO: 4 or (2) an amino acid sequence comprising one, two or three amino acid insertions, deletions or substitutions relative to the sequence of SEQ ID NO: 4; (ii) an α chain variable domain comprising a CDR2 comprising (1) the sequence of SEQ ID NO: 5 or (2) an amino acid sequence comprising one, two or three amino acid insertions, deletions or substitutions relative to the sequence of SEQ ID NO: 5; and (iii) an α chain variable domain comprising a CDR3 comprising (1) the sequence of SEQ ID NO: 6 or (2) an amino acid sequence comprising one, two or three amino acid insertions, deletions or substitutions relative to the sequence of SEQ ID NO: 6. For example, the α chain of a heterologous TCR can comprise: (i) an α chain variable domain comprising CDR1 comprising the sequence of SEQ ID NO: 4; (ii) an α chain variable domain comprising CDR2 comprising the sequence of SEQ ID NO: 5; and (iii) an α chain variable domain comprising CDR3 comprising the sequence of SEQ ID NO: 6.

[0078] For example, the beta chain of a heterologous TCR can include: (iv) a beta chain variable domain comprising a CDR1 comprising (1) the sequence of SEQ ID NO: 7 or (2) an amino acid sequence comprising one, two or three amino acid insertions, deletions or substitutions relative to the sequence of SEQ ID NO: 7; (v) a beta chain variable domain comprising a CDR2 comprising (1) the sequence of SEQ ID NO: 8 or (2) an amino acid sequence comprising one, two or three amino acid insertions, deletions or substitutions relative to the sequence of SEQ ID NO: 8; and (vi) a beta chain variable domain comprising a CDR3 comprising (1) the sequence of SEQ ID NO: 9 or (2) an amino acid sequence comprising one, two or three amino acid insertions, deletions or substitutions relative to the sequence of SEQ ID NO: 9. For example, the β chain of a heterologous TCR can comprise: (iv) a β chain variable domain comprising CDR1 comprising the sequence of SEQ ID NO: 7; (v) a β chain variable domain comprising CDR2 comprising the sequence of SEQ ID NO: 8; and (vi) a β chain variable domain comprising CDR3 comprising the sequence of SEQ ID NO: 9.

[0079] A heterologous TCR may, for example, comprise an alpha chain comprising a CDR1 having the sequence of SEQ ID NO: 4, a CDR2 having the sequence of SEQ ID NO: 5, and a CDR3 having the sequence of SEQ ID NO: 6, and a beta chain comprising a CDR1 having the sequence of SEQ ID NO: 7, a CDR2 having the sequence of SEQ ID NO: 8, and a CDR3 having the sequence of SEQ ID NO: 9. For example, a heterologous TCR may additionally have any percent identity in the alpha and beta chains discussed herein.

[0080] Heterologous CD8 co-receptor

[0081] The modified T cell comprises a heterologous CD8 co-receptor. In other words, the modified T cell expresses a heterologous CD8 co-receptor, for example on its surface.

[0082] CD8 is a cell surface glycoprotein that is found on most cytotoxic T lymphocytes in nature and mediates potent cell-cell interactions within the immune system. CD8 serves as a co-receptor for the T cell receptor, allowing CD8 and the T cell receptor to recognize antigens displayed by antigen presenting cells in the context of class I MHC molecules. The CD8 co-receptor binds to class 1 MHC and enhances TCR signaling. Functional co-receptors can be homodimers consisting of two CD8 α chains or heterodimers consisting of one CD8 α chain and one CD8 β chain.

[0083] Therefore, the heterologous CD8 co-receptor contained in the modified T cell can be CD8α. In other words, the heterologous CD8 co-receptor can be a homodimer composed of two CD8α chains. Alternatively, the heterologous CD8 co-receptor can be a heterodimer composed of one CD8α chain and one CD8β chain. In either case, the CD8 chain can comprise or consist of an amino acid sequence having at least 80% (such as at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99% or 100%) sequence identity with SEQ ID NO: 10. Therefore, the heterologous CD8 co-receptor can comprise an amino acid sequence having at least 80% (such as at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99% or 100%) sequence identity with SEQ ID NO: 10.

[0084] Both the CD8 α chain and the CD8 β chain have significant homology with immunoglobulin variable light chains. The CD8 α chain and the CD8 β chain have CDR-like loops involved in MHC-I class binding. A heterologous CD8 co-receptor may, for example, comprise a CD8 α chain comprising: (i) an α chain CDR1 comprising (1) the sequence of SEQ ID NO: 11 or (2) an amino acid sequence comprising one, two or three amino acid insertions, deletions or substitutions relative to the sequence of SEQ ID NO: 11; (ii) an α chain CDR2 comprising (1) the sequence of SEQ ID NO: 12 or (2) an amino acid sequence comprising one, two or three amino acid insertions, deletions or substitutions relative to the sequence of SEQ ID NO: 12; and / or (iii) an α chain CDR3 comprising (1) the sequence of SEQ ID NO: 13 or (2) an amino acid sequence comprising one, two or three amino acid insertions, deletions or substitutions relative to the sequence of SEQ ID NO: 13.

[0085] The heterologous CD8 co-receptor is capable of binding to class I MHC molecules. For example, the heterologous CD8 co-receptor can bind to the α3 portion of the class I MHC molecule through the IgV-like domain of the CD8 co-receptor. The α3 portion is typically found between residues 223 and 229 of the class I MHC molecule. The ability of the heterologous CD8 co-receptor to bind to class I MHC molecules increases the ability of the modified T cell to bind to the cognate antigen through its heterologous TCR. The cognate antigen MAGE-A4 is typically present in complex with a class I MHC molecule (such as HLA-A*02). The heterologous CD8 co-receptor can improve or increase the dissociation rate (k) of the TCR / peptide-MHC I interaction in the modified cell. off). This improvement or increase may be associated with modified T cells that contain a heterologous TCR that binds to MAGE-A4 but lacks a heterologous CD8 co-receptor. For example, a heterologous CD8 co-receptor can help organize a heterologous TCR on the surface of a modified cell, thereby increasing the ability of the heterologous TCR to participate in TCR / peptide-MHCI interactions. For example, a heterologous CD8 co-receptor can bind 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. Therefore, the expression of a heterologous CD8 co-receptor can confer improved affinity and / or avidity to MAGE-A4, and / or improved activation when binding to MAGE-A4 to modified T cells. Methods for determining affinity, avidity, and T cell activation are well known in the art. Expression of a heterologous CD8 co-receptor can confer to the modified T cells an improvement or increase in CD40L expression, cytokine production, cytotoxic activity, induction of dendritic cell maturation, or induction of dendritic cell cytokine production (e.g., in response to antigen (MAGE-A4) binding). The improvement or increase may be associated with a modified T cell comprising a heterologous TCR that binds to a MAGE-A4 peptide antigen, but lacks a heterologous CD8 co-receptor.

[0086] There is a synergistic effect between CD8α and peptide antigens presented on HLA-A*0201. Therefore, in one aspect of the present disclosure, the heterologous CD8 co-receptor can be CD8α, and the heterologous TCR can bind to the MAGE-A4 peptide antigen complexed with HLA-A*0201. For example, the peptide antigen can be SEQ ID NO: 1.

[0087] Checkpoint inhibitors /

[0088] The method may include administering a checkpoint inhibitor to an individual. Therefore, the method may include administering to an individual (i) a population of modified T cells, the population of modified T cells comprising heterologous CD8 co-receptors and heterologous T cell receptors (TCRs) capable of binding to MAGE-A4 peptide antigens, and (ii) a checkpoint inhibitor. In other words, the method may include treating an individual with a population of (i) modified T cells and (ii) a checkpoint inhibitor, the population of modified T cells comprising heterologous CD8 co-receptors and heterologous T cell receptors (TCRs) capable of binding to MAGE-A4 peptide antigens. As described below, the combination may also include additional anticancer therapies, such as chemotherapy.

[0089] Check point inhibitors can be administered to individuals with the same number of treatment lines as the population of modified T cells. As described above, treatment "lines" can refer to a specific treatment regimen for a cancer for which curative intent treatment has failed or for which curative intent treatment may not be appropriate. Therefore, a population of check point inhibitors and modified T cells can be administered as part of the same treatment regimen for a cancer for which curative intent treatment has failed or for which curative intent treatment may not be appropriate. A population of check point inhibitors and modified T cells can be administered as part of a first-line treatment regimen. A population of check point inhibitors and modified T cells can be administered as part of a second-line treatment regimen. A second-line treatment regimen can be adopted, for example, after a first-line treatment regimen fails. A population of check point inhibitors and modified T cells can be administered as part of a third-line treatment regimen. A third-line treatment regimen can be adopted, for example, after a second-line treatment regimen fails.

[0090] Checkpoint inhibitor therapy is a form of cancer immunotherapy. The therapy targets immune checkpoints, key regulators of the immune system that, when stimulated, can suppress the immune response to immune stimulation. Checkpoint therapy can block inhibitory checkpoints and restore immune system function. Checkpoint inhibitors can target the molecules CTLA4, PD-1, and PD-L1.

[0091] Checkpoint inhibitors can, for example, target CTLA4. In other words, checkpoint inhibitors can comprise CTLA4 blockers. CTLA4 blockers are known in the art and include, for example, ipilimumab.

[0092] Check point inhibitors may include PD-1 axis binding antagonists. The administration of PD-1 axis binding antagonists is described in the art as a standard treatment for recurrent, unresectable or metastatic head and neck cancer, such as non-nasopharyngeal cancer, including lip cancer, oral cancer, oropharyngeal cancer, hypopharyngeal cancer, glottic laryngeal cancer, supraglottic laryngeal cancer, ethmoid sinus cancer, maxillary sinus cancer and occult primary cancer. For example, pembrolizumab may be used as a standard treatment for recurrent, unresectable or metastatic head and neck cancer with tumors expressing PD-L1 and CPS≥1. Pembrolizumab may be used together with platinum-based agents (such as cisplatin or carboplatin) and 5-fluorouracil as a standard treatment for recurrent, unresectable or metastatic head and neck cancer. In some cases, pembrolizumab may be used together with platinum-based agents (such as cisplatin or carboplatin) and docetaxel or paclitaxel as a first-line or subsequent line standard treatment for recurrent, unresectable or metastatic head and neck cancer. In some cases, pembrolizumab can be used with cetuximab as a first-line or subsequent-line standard treatment for recurrent, unresectable, or metastatic head and neck cancer. In some cases, pembrolizumab can be administered as a first-line or subsequent-line standard treatment for recurrent, unresectable, or metastatic head and neck cancer with high microsatellite instability (MSI-H), such as cancer with DNA mismatch repair (MMR). In some cases, if the cancer progresses during or after treatment with a platinum-based agent, pembrolizumab or nivolumab can be administered as a subsequent-line standard treatment for recurrent, unresectable, or metastatic head and neck cancer.

[0093] Programmed cell death protein 1 (PD-1, also known as CD279) is a protein expressed on the surface of T cells and plays a role in regulating immune responses by maintaining T cell homeostasis. The connection of PD-1 to one of its ligands (PD-L1 or PD-L2) transmits inhibitory signals within T cells. In particular, the signals generated by PD-1 prevent the phosphorylation of key TCR signaling intermediates, thereby terminating early TCR signaling and reducing T cell activation. T cell effector functions (such as proliferation, cytotoxicity, and cytokine production) are reduced, and the ability to convert into memory T cells is impaired.

[0094] PD-L1 and PD-L2 are members of the B7 family. The PD-L1 protein is upregulated on certain activated immune cells (such as macrophages, dendritic cells, T cells, and B cells), and is also expressed on certain normal tissues. PD-L1 is also highly expressed in many cancers. PD-L2 is mainly expressed by dendritic cells and some tumors. Since many cancers express the PD-1 ligand, the PD-1 axis has a defined role in cancer immune evasion and tumor resistance.

[0095] Cancer (such as solid tumors) express PD-1 ligands, making the tumor microenvironment immunosuppressive. Therefore, the function of modified T cells infiltrating tumors can be suppressed. Endogenous anti-tumor T cell responses can also be suppressed. In this way, tumors are more able to avoid the immune system. In the present disclosure, PD-1 axis binding antagonists can be administered to offset the inhibitory effects of PD-L1 and / or PD-L2 expression in the tumor microenvironment. By offsetting the inhibition, the function of modified and / or endogenous T cells can be maintained. In other words, the administration of PD-1 axis binding antagonists can maintain the function of the modified T cells and / or their offspring contained in the administration. The administration of PD-1 axis binding antagonists can maintain the function of endogenous T cells in individuals. The administration of PD-1 axis binding antagonists can maintain the function of the modified T cells (and / or their offspring) contained in the administration, as well as the function of endogenous T cells in individuals. In any case, endogenous T cells can be, for example, contained in the tumor microenvironment. For example, endogenous T cells can be tumor infiltrating lymphocytes (tumour infiltrating lymphocytes, TIL).

[0096] The function of maintaining T cells may refer to, for example, maintaining, recovering and / or enhancing the function of T cells. For example, maintaining T cell function may refer to maintaining T cell activation. In this way, the duration of effective T cell response can be extended. In other words, the activation maintained may be relevant to the improvement of the duration of effector function (such as cytokine production, cytotoxicity and / or proliferation). The activation maintained may also contribute to the ability of T cells to be converted into memory T cells. The generation of memory T cells is advantageous because it allows long-term maintenance of anti-tumor immunity, such as months or years. The method for determining the activation, cytokine production, cytotoxicity, proliferation and generation of memory T cells is well known in the art.

[0097] The administration of PD-1 axis binding antagonists can maintain the function of modified T cells and / or endogenous T cells by reducing exhaustion. Exhaustion can be reduced in the population of modified T cells and / or the offspring T cells of the population of modified T cells. Exhaustion can be reduced in endogenous T cells in an individual. The following exhaustion can be reduced: (i) in the population of modified T cells and / or the offspring T cells of the population of modified T cells, and (ii) in the endogenous T cells in an individual. Exhausted T cells usually express high levels of PD-1 and experience functional loss. For example, the ability of exhausted T cells to produce cytokines (such as IL-2 or TNFα) may have been reduced. Exhausted T cells may have reduced proliferation capacity. Exhausted T cells may have reduced cytotoxic potential. Ultimately, exhausted T cells can be targeted for destruction. Therefore, exhaustion leads to the loss of T cell function, or the loss of T cells themselves, which is unfavorable for tumor immunity. Therefore, reducing T cell exhaustion can improve the therapeutic effect.

[0098] In the context of the present disclosure, a PD-1 axis binding antagonist is a molecule that inhibits the interaction of PD-1 with a PD-1 ligand and / or the transduction of a signal generated by the interaction of PD-1 with a PD-1 ligand. The PD-1 ligand may be PD-L1 or PD-L2. The PD-1 axis binding antagonist may, for example, reduce or prevent the interaction of PD-1 with a PD-1 ligand. The PD-1 axis binding antagonist may, for example, reduce or prevent the transduction of a signal generated by the interaction of PD-1 with a PD-1 ligand. The PD-1 axis binding antagonist may block, inhibit or reduce the biological activity of PD-1 and / or a PD-1 ligand.

[0099] By inhibiting the interaction of PD-1 with PD-1 ligands and / or the transduction of signals generated by the interaction of PD-1 with PD-1 ligands, PD-1 axis binding antagonists can maintain (e.g., maintain, restore or enhance) the function of endogenous T cells. In the same manner, PD-1 axis binding antagonists can maintain (e.g., maintain, restore or enhance) the function of modified T cells administered to an individual. The maintained function can be indicated, for example, by the maintenance or improvement of T cell proliferation, cytokine production, target cell killing, activation, CD28 signaling, the ability to infiltrate tumors, the ability to recognize and bind to antigens presented by dendritic cells, and / or the ability to produce interferons. In this way, the PD-1 axis binding antagonist counteracts the immunosuppressive properties of the tumor microenvironment.

[0100] For example, the PD-1 axis binding antagonist can be a PD-1 binding antagonist. In other words, the PD-1 axis binding antagonist can inhibit (e.g., prevent or reduce) the binding of PD-1 to its binding partner. For example, the PD-1 axis binding antagonist can inhibit the binding of PD-1 to PD-L1, PD-L2, or PD-L1 and PD-L2. For example, the PD-1 binding antagonist can be an antibody that binds to PD-1, or an antigen binding variant or fragment thereof. For example, the PD-1 binding antagonist can be an antibody that binds to PD-1, or an antigen binding variant or fragment thereof. For example, the PD-1 binding antagonist can be an antibody that binds to SEQ ID NO: 14, or an antigen binding variant or fragment thereof. Antibodies that bind to PD-1 are well known in the art, and include, for example, nivolumab, pembrolizumab, cemiplizumab, dotalimumab, JTX-4014, spartalizumab, carrelizumab, sintilimab, tislelizumab, toripalimab, rivalizumab (INCMGA00012), AMP-224, MEDI0680 (AMP-514), sazanalizumab, brigalimumab, ebalizumab (BI 754091), and sepalizumab (AB122). Therefore, the PD-1 axis binding antagonist can be nivolumab, pembrolizumab, cemiprilimab, dotalimab, JTX-4014, spartalizumab, carrelizumab, sintilimab, tislelizumab, toripalimab, rivulimab (INCMGA00012), AMP-224, MEDI0680 (AMP-514), sacralizumab, buglimab, ebenlimab (BI 754091) or sepalimab (AB122), or any combination thereof. For example, the PD-1 axis binding antagonist can be pembrolizumab. Alternatively, the PD-1 binding antagonist can be any immunoadhesin, protein or oligopeptide that inhibits, reduces, prevents or interferes with signal transduction due to the interaction of PD-1 with PD-L1 and / or PD-L2.

[0101] The heavy chain sequence and light chain sequence of nivolumab are set forth in SEQ ID NOs: 17 and 18, respectively. The heavy chain sequence and light chain sequence of pembrolizumab are set forth in SEQ ID NOs: 19 and 20, respectively. The heavy chain sequence and light chain sequence of cemiplimab are set forth in SEQ ID NOs: 21 and 22, respectively. The heavy chain sequence and light chain sequence of dotalizumab are set forth in SEQ ID NOs: 31 and 32, respectively. A person skilled in the art who knows the heavy chain and light chain sequences of a given antibody can identify antigen-binding variants or fragments of the antibody using routine methods in the art. An antigen-binding variant or fragment of nivolumab may, for example, comprise the three CDRs contained in SEQ ID NO: 17 and the three CDRs contained in SEQ ID NO: 18. An antigen-binding variant or fragment of pembrolizumab may, for example, comprise the three CDRs contained in SEQ ID NO: 19 and the three CDRs contained in SEQ ID NO: 20. An antigen-binding variant or fragment of cemiplimab may, for example, comprise the three CDRs contained in SEQ ID NO: 21 and the three CDRs contained in SEQ ID NO: 22. An antigen-binding variant or fragment of dotalizumab may, for example, comprise the three CDRs contained in SEQ ID NO: 31 and the three CDRs contained in SEQ ID NO: 32. Methods for identifying CDRs in heavy or light chain sequences are routine methods in the art.

[0102] For example, the PD-1 axis binding antagonist can be a PD-L1 binding antagonist. In other words, the PD-1 axis binding antagonist can inhibit (e.g., prevent or reduce) the binding of PD-L1 to a binding partner. For example, the PD-1 axis binding antagonist can inhibit the binding of PD-L1 to PD-1. For example, the PD-L1 binding antagonist can be an antibody that binds to PD-L1, or an antigen binding variant or fragment thereof. For example, the PD-L1 binding antagonist can be an antibody that binds to SEQ ID NO: 15, or an antigen binding variant or fragment thereof. Antibodies that bind to PD-L1 are well known in the art, including, for example, durvalumab, atelizumab, avelumab, BMS 936559 (MDX-1105), enflelimab (KN035), and coxilimab (CK-301). Thus, the PD-L1 axis binding antagonist can be durvalumab, atelizumab, avelumab, BMS 936559 (MDX-1105), enfalizumab (KN035) or coxilimab (CK-301) or any combination thereof. Alternatively, the PD-L1 binding antagonist can be any immunoadhesin, protein or oligopeptide that inhibits, reduces, prevents or interferes with signal transduction due to the interaction of PD-L1 with PD-1.

[0103] The heavy chain sequence and light chain sequence of Durvalumab are set forth in SEQ ID NOs: 23 and 24, respectively. The heavy chain sequence and light chain sequence of Atelizumab are set forth in SEQ ID NOs: 25 and 26, respectively. The heavy chain sequence and light chain sequence of Avelumab are set forth in SEQ ID NOs: 27 and 28, respectively. The heavy chain sequence and light chain sequence of BMS 936559 (MDX-1105) are set forth in SEQ ID NOs: 29 and 30, respectively. A person skilled in the art who is aware of the heavy chain and light chain sequences of a given antibody can identify antigen-binding variants or fragments of the antibody using conventional methods in the art. An antigen-binding variant or fragment of Durvalumab may, for example, comprise the three CDRs contained in SEQ ID NO: 23 and the three CDRs contained in SEQ ID NO: 24. An antigen-binding variant or fragment of Atelizumab may, for example, comprise the three CDRs contained in SEQ ID NO: 25 and the three CDRs contained in SEQ ID NO: 26. An antigen-binding variant or fragment of avelumab may, for example, comprise the three CDRs contained in SEQ ID NO: 27 and the three CDRs contained in SEQ ID NO: 28. An antigen-binding variant or fragment of BMS 936559 (MDX-1105) may, for example, comprise the three CDRs contained in SEQ ID NO: 29 and the three CDRs contained in SEQ ID NO: 30. Methods for identifying CDRs in heavy or light chain sequences are routine methods in the art.

[0104] For example, a PD-1 axis binding antagonist may be a PD-L2 binding antagonist. In other words, a PD-1 axis binding antagonist may inhibit (e.g., prevent or reduce) the binding of PD-L2 to a binding partner. For example, a PD-L2 binding antagonist may be an antibody that binds to PD-L2, or an antigen binding variant or fragment thereof. For example, a PD-L2 binding antagonist may be an antibody that binds to SEQ ID NO: 16, or an antigen binding variant or fragment thereof. Alternatively, a PD-L2 binding antagonist may be any immunoadhesin, protein, or oligopeptide that inhibits, reduces, prevents, or interferes with signal transduction due to the interaction of PD-L2 with PD-1.

[0105] When the PD-1 axis binding antagonist is an antibody (such as a known antibody or an antigen-binding variant thereof), the PD-1 axis binding antagonist can be, for example, a monoclonal antibody, a human or humanized antibody, a full-length antibody, a bispecific antibody, a linear antibody, or a single-chain antibody molecule. The antibody isotype can be selected from any of the five classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, with heavy chains designated α, δ, ε, γ, and μ (M), respectively. Antibodies of the γ and α classes can be any subclass: IgG1, IgG2A, IgG2B, IgG3, IgG4, IgA1, and IgA2. When the PD-1 axis binding antagonist is an antigen-binding fragment of an antibody, the PD-1 axis binding antagonist can be, for example, Fv, Fab, Fab', Fab'-SH, F(ab')2, or scFv.

[0106] When the PD-1 axis binding antagonist is an immunoadhesin, the immunoadhesin may comprise an adhesion domain and an immunoglobulin constant domain that confer binding activity to a PD-1 axis component (e.g., PD-1, PD-L1, or PD-L2). The immunoglobulin constant domain may be from any isotype, such as IgG1, IgG2, IgG2A, IgG2B, IgG3, IgG4 subtype, IgA, IgA1, IgA2, IgE, IgD, or IgM. For example, the immunoglobulin constant domain may comprise (i) a hinge region, CH2, and CH3 of an immunoglobulin molecule, or (ii) a hinge region, CH1, CH2, and CH3. Thus, the immunoadhesin may comprise (a) an extracellular or PD-1 binding portion of PD-L1 or PD-L2, or an extracellular or PD-L1 or PD-L2 binding portion of PD-1, fused to (b) a constant domain of an immunoglobulin sequence.

[0107] At least 1% of head and neck cancer cells from an individual may express PD-L1, such as at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, or at least 80%. Head and neck cancer from an individual may express PD-L1, and its tumor proportion score (TPS) is greater than or equal to (≥) 1%, such as ≥2%, ≥10%, or ≥50%. The percentage of cells expressing PD-L1 can be determined by any method known to those skilled in the art, such as IHC, flow cytometry, or ELISA.

[0108] At least 1% of head and neck cancer cells from an individual may express PD-L2, such as at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, or at least 80%. Head and neck cancer from an individual may express PD-L2, and its tumor proportion score is greater than or equal to (≥) 1%, such as ≥2%, ≥10%, or ≥50%. The percentage of cells expressing PD-L2 can be determined by any method known to those skilled in the art, such as IHC, flow cytometry, or ELISA.

[0109] The expression of PD-1, PD-L1 and / or PD-L2 in head and neck cancer may have an intensity greater than or equal to (≥) 1+, such as ≥2+ or ≥3+. The intensity score can be assessed by IHC staining of the tumor, and the score is as follows: negative = no staining or staining in less than or equal to ≤10% of the stained cells; 1+ = incomplete staining in ≥10% of the stained cells; 2+ = weak to moderate staining in ≥10% of the stained cells; strong and complete staining in ≥10% of the stained cells.

[0110] The population of modified T cells and the checkpoint inhibitor can be administered any number of times in any order. For example, the population of modified T cells can be administered in a single dose. For example, the checkpoint inhibitor can be administered (a) before the modified T cells, (b) simultaneously with the modified T cells, and / or (c) after the modified T cells. For example, the checkpoint inhibitor can be administered, for example, as follows: (a); (b); (c); (a) and (b); (a) and (c); (b) and (c); or (a), (b) and (c). When the checkpoint inhibitor is administered simultaneously with the population of modified T cells, the checkpoint inhibitor and the population of modified T cells can be included in the same composition or in different compositions.

[0111] Administration of a checkpoint inhibitor simultaneously with modified T cells may refer to administration of a checkpoint inhibitor and modified T cells substantially simultaneously. In other words, the dose of the checkpoint inhibitor may be administered approximately simultaneously with the dose of the population of modified T cells. For example, the dose of the checkpoint inhibitor may be administered within about 2 hours, about 4 hours, about 6 hours, about 8 hours, about 12 hours, or about 24 hours with the dose of the population of modified T cells.

[0112] Applying a checkpoint inhibitor before the modified T cells may refer to applying a checkpoint inhibitor at any time before the modified T cells. In other words, the dosage of a checkpoint inhibitor can be applied at any time before the dosage of the population of modified T cells. For example, the dosage of a checkpoint inhibitor can be applied before the population of modified T cells by about 24 hours or more, about 36 hours or more, about 48 hours or more, about 72 hours or more, about 4 days or more, about 5 days or more, about 6 days or more, about 1 week or more, about 2 weeks or more, about 3 weeks or more, or about 4 weeks or more, about 5 weeks or more, about 6 weeks or more, about 8 weeks or more, or about 12 weeks or more. For example, a checkpoint inhibitor can be applied before the modified T cells to start treatment while generating autologous modified T cells.

[0113] Applying a checkpoint inhibitor after modified T cells may refer to applying a checkpoint inhibitor at any time after the modified T cells. In other words, the dosage of a checkpoint inhibitor can be applied at any time after the dosage of a population of modified T cells. For example, the dosage of a checkpoint inhibitor can be applied at about 24 hours or more, about 36 hours or more, about 48 hours or more, about 72 hours or more, about 4 days or more, about 5 days or more, about 6 days or more, about 1 week or more, about 2 weeks or more, about 3 weeks or more, or about 4 weeks or more, about 5 weeks or more, about 6 weeks or more, about 8 weeks or more, or about 12 weeks or more of the population of modified T cells.

[0114] If the check point inhibitor is applied before the population of modified T cells, the check point inhibitor can continue to be applied after the modified T cells are applied. Therefore, in one aspect of the present disclosure, the check point inhibitor is applied before the population of the modified T cells is applied, and the check point inhibitor is continued to be applied after the population of the modified T cells is applied. In other words, the dose of the check point inhibitor can be applied before the dose of the population of modified T cells, and one or more additional doses of the check point inhibitor can be subsequently applied. The dose of the check point inhibitor can be applied, for example, according to the known treatment regimen of the check point inhibitor. The purpose of the additional dose of the check point inhibitor can be to maintain the effect obtained by the initial dose application. Each of the one or more additional doses can include the same check point inhibitor as the initial dose, or a check point inhibitor different from the initial dose.

[0115] If the check point inhibitor is administered simultaneously with the population of modified T cells, the check point inhibitor can continue to be administered after the modified T cells are administered. Therefore, in one aspect of the present disclosure, the check point inhibitor is administered while the population of the modified T cells is administered, and the check point inhibitor is continued to be administered after the population of the modified T cells is administered. In other words, the dose of the check point inhibitor can be administered simultaneously with the dose of the population of modified T cells, and one or more additional doses of the check point inhibitor can be subsequently administered. The dose of the check point inhibitor can be administered, for example, according to the known treatment regimen of the check point inhibitor. The purpose of the additional dose of the check point inhibitor can be to maintain the effect obtained by the initial dose administration. Each of the one or more additional doses can include the same check point inhibitor as the initial dose, or a check point inhibitor different from the initial dose.

[0116] If the checkpoint inhibitor is administered after the population of modified T cells, the checkpoint inhibitor can continue to be administered after the initial administration. Therefore, in one aspect of the present disclosure, the administration of the checkpoint inhibitor begins after the population of modified T cells is administered, and continues after the initial administration of the checkpoint inhibitor. In other words, the dose of the checkpoint inhibitor can be administered after the dose of the population of modified T cells, and one or more additional doses of the checkpoint inhibitor can be subsequently administered. The dose of the checkpoint inhibitor can be administered, for example, according to the known treatment regimen of the checkpoint inhibitor. The purpose of the additional dose of the checkpoint inhibitor can be to maintain the effect obtained by the initial dose administration. Each of the one or more additional doses can contain the same checkpoint inhibitor as the initial dose, or a checkpoint inhibitor different from the initial dose.

[0117] In any case, one or more additional doses of the checkpoint inhibitor may be administered at any suitable interval. Suitable dose intervals for checkpoint inhibitors are known in the art and may be specific to the identity of the checkpoint inhibitor. For example, for PD-1 axis binding antagonists (such as pembrolizumab), one or more additional doses may be administered, for example, two weeks after the initial dose is administered, about every two weeks (Q2W). One or more additional doses may be administered, for example, three weeks after the initial dose is administered, about every three weeks (Q3W). One or more additional doses may be administered, for example, four weeks after the initial dose is administered, about every four weeks (Q4W). One or more additional doses may be administered, for example, five weeks after the initial dose is administered, about every five weeks (Q5W). One or more additional doses may be administered, for example, six weeks after the initial dose is administered, about every six weeks (Q6W). In a preferred aspect of the disclosure, one or more additional doses are administered approximately every three weeks (Q3W) starting three weeks after the initial dose, or approximately every six weeks (Q6W) starting six weeks after the initial dose.

[0118] Any number of additional doses of checkpoint inhibitors may be administered, such as 1 or more, 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 15 or more, 20 or more, 25 or more, 30 or more, 35 or more, 40 or more, 45 or more, or 50 or more additional doses. Additional doses may be administered until disease progression, unacceptable toxicity, withdrawal of consent, or death.

[0119] Additional anticancer therapies

[0120] The method may include administering an additional anti-cancer therapy to the individual. Thus, the method may include administering to the individual (i) a population of modified T cells, the population of modified T cells comprising a heterologous CD8 co-receptor and a heterologous T cell receptor (TCR) capable of binding to a MAGE-A4 peptide antigen, and (ii) an additional anti-cancer therapy. In other words, the method may include treating the individual with (i) a population of modified T cells and (ii) an additional anti-cancer therapy in combination, the population of modified T cells comprising a heterologous CD8 co-receptor and a heterologous T cell receptor (TCR) capable of binding to a MAGE-A4 peptide antigen. Checkpoint inhibitors, such as PD-1 axis binding antagonists, may also be included in the combination as described above.

[0121] The additional anti-cancer therapy can be administered to an individual with the same number of lines of treatment as the population of modified T cells. As described above, a "line" of treatment can refer to a specific treatment regimen for a cancer for which curative intent treatment has failed or for which curative intent treatment may not be appropriate. Therefore, the additional anti-cancer therapy and the population of modified T cells can be administered as part of the same treatment regimen for a cancer for which curative intent treatment has failed or for which curative intent treatment may not be appropriate. The additional anti-cancer therapy and the population of modified T cells can be administered as part of a first-line treatment regimen. The additional anti-cancer therapy and the population of modified T cells can be administered as part of a second-line treatment regimen. The second-line treatment regimen can be adopted, for example, after the first-line treatment regimen has failed. The additional anti-cancer therapy and the population of modified T cells can be administered as part of a third-line treatment regimen. The third-line treatment regimen can be adopted, for example, after the second-line treatment regimen has failed.

[0122] For example, the additional anticancer therapy may be an anticancer drug therapy. In other words, the additional anticancer therapy may be a systemic treatment. The additional anticancer therapy may, for example, include or consist of: (a) chemotherapy. The additional anticancer therapy may, for example, include or consist of: (b) immunotherapy. The additional anticancer therapy may, for example, include or consist of: (c) targeted therapy. For example, the additional anticancer therapy may include or consist of: (a); (b); (c); (a) and (b); (a) and (c); (b) and (c); or (a), (b) and (c).

[0123] Any chemotherapy can be included in the additional anticancer therapy. Chemotherapy such as for head and neck cancer is well known in the art. For example, such chemotherapy can include anti-tumor drugs based on platinum, such as cisplatin or carboplatin. For example, such chemotherapy can include antimetabolites, such as fluorouracil (5-FU), capecitabine or methotrexate. For example, such chemotherapy can include taxane drugs, such as docetaxel or paclitaxel. For example, such chemotherapy can include podophyllotoxin derivatives, such as etoposide. For example, such chemotherapy can include alkylating agents, such as cyclophosphamide. For example, such chemotherapy can include anthracyclines, such as doxorubicin. For example, such chemotherapy can include vinca alkaloids, such as vincristine.

[0124] Any immunotherapy may be included in additional anticancer therapies. Immunotherapy, such as immunotherapy for head and neck cancer, is well known in the art. For example, such immunotherapy may include therapeutic immune cells, immunomodulators, checkpoint inhibitors and vaccines. Therapeutic immune cells may include T cells, such as engineered T cells, such as CAR T cells or T cells expressing engineered TCR. Immunomodulators may include, for example, interleukins, cytokines, chemokines and immunomodulatory imide drugs. For example, immunomodulators may include Multikine (interleukin, injection). For example, immunomodulators may include monalizumab, a humanized anti-NKG2A blocking antibody that prevents tumor cells expressing HLA-E from inhibiting CD8+T cells and NK cells. Checkpoint inhibitors may, for example, include CTLA-4 inhibitors or PD-1 axis binding antagonists. PD-1 axis binding antagonists may, for example, include pembrolizumab, nivolumab and penampulimumab. Checkpoint inhibitors and PD-1 axis binding antagonists are described in detail herein. For example, the vaccine may include vidutolimod (CMP-001), which is a Toll-like receptor 9 (TLR9) agonist cancer vaccine.

[0125] Any targeted therapy can be included in the additional anticancer therapy. Targeted therapies, such as targeted therapies for head and neck cancer, are well known in the art. For example, targeted therapies can include EGFR antagonists (such as cetuximab), tropomyosin kinase receptor antagonists (such as larotrectinib), HER2 antagonists, kinase inhibitors (such as afatinib or lenvatinib), farnesyl transferase inhibitors (such as tipifarnib), PI3 kinase inhibitors (such as buparlisib), inhibitors of apoptosis proteins (such as xevinapant (Debio 1143)).

[0126] Additional anticancer therapy can include previously described medicines or drug combinations for the first-line therapy of head and neck cancer (such as recurrent, unresectable or metastatic head and neck cancer).Anticancer therapy can include medicines or drug combinations, which have been previously described as approved or "standard treatment" treatments (such as first-line "standard treatment" treatments) for head and neck cancer (such as recurrent, unresectable or metastatic head and neck cancer).In this case, approval can be related to approvals such as FDA, EMA or MHRA.For example, additional anticancer therapy can include pembrolizumab, cetuximab, cisplatin, carboplatin, 5-FU, docetaxel, paclitaxel, etoposide, cyclophosphamide, doxorubicin, vincristine, methotrexate, capecitabine and / or afatinib.For example, anticancer therapy can include monotherapy with pembrolizumab, cetuximab, cisplatin, carboplatin, 5-FU, docetaxel, paclitaxel, methotrexate or capecitabine. For example, anticancer therapy may include monotherapy with: pembrolizumab, cisplatin, and 5-FU; pembrolizumab, carboplatin, and 5-FU; cetuximab, cisplatin, and 5-FU; cetuximab, carboplatin, and 5-FU; cetuximab and cisplatin; cisplatin and docetaxel; cisplatin and paclitaxel; carboplatin and docetaxel; carboplatin and paclitaxel; cisplatin and 5-FU; cisplatin, docetaxel, and cetuximab anti-drug therapy; cisplatin, paclitaxel, and cetuximab; carboplatin, docetaxel, and cetuximab; carboplatin, paclitaxel, and cetuximab; cisplatin, docetaxel, and pembrolizumab; cisplatin, paclitaxel, and pembrolizumab; carboplatin, docetaxel, and pembrolizumab; carboplatin, paclitaxel, and pembrolizumab; cetuximab and pembrolizumab; cisplatin and etoposide; carboplatin and etoposide; or cyclophosphamide, doxorubicin, and vincristine.

[0127] For example, when the cancer expresses PD-L1 and the combined positive score is less than 1, additional anticancer therapy may include a combination of a PD-1 axis binding antagonist and cisplatin and 5-FU, or a combination of a PD-1 axis binding antagonist and carboplatin and 5-FU. For example, the PD-1 axis binding antagonist may include pembrolizumab. For example, when the cancer expresses PD-L1 and the combined positive score (CPS) is ≥1, anticancer therapy may include a PD-1 axis binding antagonist as a monotherapy. For example, the PD-1 axis binding antagonist may include pembrolizumab. In either case, the cancer may be recurrent, unresectable, or metastatic head and neck cancer. A person skilled in the art can determine the combined positive score, for example, by consulting FDA guidelines.

[0128] The population of modified T cells and the additional anti-cancer therapy can be administered any number of times in any order. For example, the population of modified T cells can be administered in a single dose. For example, the additional anti-cancer therapy can be administered (a) before the modified T cells, (b) simultaneously with the modified T cells, and / or (c) after the modified T cells. For example, the additional anti-cancer therapy can be administered, for example, as follows: (a); (b); (c); (a) and (b); (a) and (c); (b) and (c); or (a), (b) and (c). When the additional anti-cancer therapy is administered simultaneously with the population of modified T cells, the additional anti-cancer therapy and the population of modified T cells can be included in the same composition or in different compositions.

[0129] Administering an additional anti-cancer therapy simultaneously with the modified T cells can refer to administering the additional anti-cancer therapy and the modified T cells substantially simultaneously. In other words, the dose of the additional anti-cancer therapy can be administered at about the same time as the dose of the population of modified T cells. For example, the dose of the additional anti-cancer therapy can be administered within about 2 hours, about 4 hours, about 6 hours, about 8 hours, about 12 hours, or about 24 hours of the dose of the population of modified T cells.

[0130] Applying anticancer therapy before the modified T cell may refer to applying anticancer therapy at any time before the modified T cell. In other words, the dosage of additional anticancer therapy can be applied at any time before the dosage of the colony of the modified T cell. For example, the dosage of additional anticancer therapy can be applied before the colony of the modified T cell about 24 hours or more, about 36 hours or more, about 48 hours or more, about 72 hours or more, about 4 days or more, about 5 days or more, about 6 days or more, about 1 week or more, about 2 weeks or more, about 3 weeks or more, or about 4 weeks or more, about 5 weeks or more, about 6 weeks or more, about 8 weeks or more, or about 12 weeks or more. For example, additional anticancer therapy can be applied before the modified T cell to start treatment while producing the T cell of autologous modification.

[0131] The administration of anticancer therapy after the modified T cells may refer to the administration of anticancer therapy at any time after the modified T cells. In other words, the dosage of additional anticancer therapy can be administered at any time after the dosage of the population of modified T cells. For example, the dosage of additional anticancer therapy can be administered at about 24 hours or more, about 36 hours or more, about 48 hours or more, about 72 hours or more, about 4 days or more, about 5 days or more, about 6 days or more, about 1 week or more, about 2 weeks or more, about 3 weeks or more, or about 4 weeks or more, about 5 weeks or more, about 6 weeks or more, about 8 weeks or more, or about 12 weeks or more of the population of modified T cells. For example, additional anticancer therapy can be administered after the modified T cells to start treatment while producing autologous modified T cells.

[0132] If an additional anticancer therapy is administered before the population of modified T cells, the additional anticancer therapy can continue to be administered after the modified T cells are administered. Therefore, in one aspect of the present disclosure, the additional anticancer therapy is administered before the population of the modified T cells is administered, and the additional anticancer therapy is continued to be administered after the population of the modified T cells is administered. In other words, the dose of the additional anticancer therapy can be administered before the dose of the population of modified T cells, and one or more additional doses of the additional anticancer therapy can be subsequently administered. The dose of the additional anticancer therapy can be administered, for example, according to a known treatment regimen for the additional anticancer therapy. The purpose of the additional dose of the additional anticancer therapy can be to maintain the effect obtained by the initial dose administration. Each of the one or more additional doses can contain the same additional anticancer therapy as the initial dose, or an additional anticancer therapy different from the initial dose.

[0133] If the additional anticancer therapy is administered simultaneously with the population of modified T cells, the additional anticancer therapy can continue to be administered after the modified T cells are administered. Therefore, in one aspect of the present disclosure, the additional anticancer therapy is administered while the population of modified T cells is administered, and the additional anticancer therapy is continued to be administered after the population of modified T cells is administered. In other words, the dose of the additional anticancer therapy can be administered simultaneously with the dose of the population of modified T cells, and one or more additional doses of the additional anticancer therapy can be subsequently administered. The dose of the additional anticancer therapy can be administered, for example, according to a known treatment regimen for the additional anticancer therapy. The purpose of the additional dose of the additional anticancer therapy can be to maintain the effect obtained by the initial dose administration. Each of the one or more additional doses can contain the same additional anticancer therapy as the initial dose, or an additional anticancer therapy different from the initial dose.

[0134] If additional anticancer therapy is administered after the population of modified T cells, the additional anticancer therapy can continue to be administered after the initial administration. Therefore, in one aspect of the present disclosure, the administration of the additional anticancer therapy begins after the population of modified T cells is administered, and continues after the initial administration of the additional anticancer therapy. In other words, the dose of the additional anticancer therapy can be administered after the dose of the population of modified T cells, and one or more additional doses of the additional anticancer therapy can be subsequently administered. The dose of the additional anticancer therapy can be administered, for example, according to a known treatment regimen for the additional anticancer therapy. The purpose of the additional dose of the additional anticancer therapy can be to maintain the effect obtained by the initial dose administration. Each of the one or more additional doses can contain the same additional anticancer therapy as the initial dose, or an additional anticancer therapy different from the initial dose.

[0135] In any case, one or more additional doses of the additional anti-cancer therapy may be administered at any suitable intervals. Suitable dosage intervals for the additional anti-cancer therapy are known in the art and may be specific to the nature of the additional anti-cancer therapy.

[0136] Any number of additional doses of the additional anti-cancer therapy may be administered, such as 1 or more, 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 15 or more, 20 or more, 25 or more, 30 or more, 35 or more, 40 or more, 45 or more, or 50 or more additional doses. Additional doses may be administered until disease progression, unacceptable toxicity, withdrawal of consent, or death.

[0137] Combination therapy

[0138] Figures 1 to 4 Examples of combination treatment regimens are shown. Figure 1 and 2 It relates to the treatment of head and neck cancer that recurs after curative intent treatment for locally advanced head and neck cancer. The treatment is first-line treatment. In this case, a checkpoint inhibitor (such as a PD-1 axis binding inhibitor, such as pembrolizumab) can be administered simultaneously or substantially simultaneously with the modified T cells. In other words, the dose of the checkpoint inhibitor can be administered approximately simultaneously with the dose of the modified T cell population. For example, the dose of the checkpoint inhibitor can be administered within about 2 hours, about 4 hours, about 6 hours, about 8 hours, about 12 hours, or about 24 hours with the dose of the modified T cell population. Alternatively, the checkpoint inhibitor can be administered after the modified T cells. The administration of the checkpoint inhibitor can, for example, continue after the modified T cells are administered. In other words, the dose of the checkpoint inhibitor can be administered almost simultaneously with the dose of the modified T cell population, and one or more additional doses of the checkpoint inhibitor can be subsequently administered. The dose of the checkpoint inhibitor can be administered, for example, according to the known treatment regimen of the checkpoint inhibitor. The treatment regimen of the PD-1 axis binding inhibitor (such as pembrolizumab) is as described above. Additional anti-cancer therapies, such as chemotherapy, may be administered as an alternative to or in addition to checkpoint inhibitors.

[0139] exist Figure 1 and Figure 2In the present invention, cells (such as PBMC) used to generate a population of modified T cells are obtained from individuals before relapse. For example, when an individual is identified as having a high risk of relapse, cells can be obtained. This allows the generation of a population of modified T cells to begin (and ideally be completed) before cancer relapse. Accordingly, this can shorten the window period between relapse and administration of a population of modified T cells, i.e., allow a population of modified T cells to be administered as soon as possible after diagnosis of a relapsed cancer. This can improve the therapeutic effect.

[0140] Figure 3 and Figure 4 Relating to the treatment of previously untreated head and neck cancer (such as newly metastatic or unresectable locally advanced head and neck cancer). The treatment is a first-line treatment. In this case, a checkpoint inhibitor (such as a PD-1 axis binding inhibitor, such as pembrolizumab) can be administered before the modified T cells. In other words, the dose of the checkpoint inhibitor can be administered before the dose of the modified T cell population. For example, the dose of the checkpoint inhibitor can be administered before about 24 hours or more, about 36 hours or more, about 48 hours or more, about 72 hours or more, about 4 days or more, about 5 days or more, about 6 days or more, about 1 week or more, about 2 weeks or more, about 3 weeks or more, or about 4 weeks or more before the modified T cell population. The administration of the checkpoint inhibitor can, for example, continue after the modified T cells are administered. In other words, the dose of the checkpoint inhibitor can be administered before the dose of the modified T cell population, and one or more additional doses of the checkpoint inhibitor can be administered subsequently. The dose of the checkpoint inhibitor can be administered, for example, according to the known treatment regimen of the checkpoint inhibitor. The treatment regimen of the PD-1 axis binding inhibitor (such as pembrolizumab) is as described above. Additional anti-cancer therapies, such as chemotherapy, may be administered as an alternative to or in addition to checkpoint inhibitors.

[0141] exist Figure 3 and 4 In the present invention, once cancer is diagnosed, cells (such as PBMCs) for generating a population of modified T cells can be obtained from the individual. Then the population of modified T cells can be started. Treatment with checkpoint inhibitors can be implemented during the generation window period, and the population of modified T cells can be administered once ready. This allows treatment to be started as soon as possible after cancer is diagnosed, thereby improving the treatment effect.

[0142] Application

[0143] As mentioned above, the colony of the modified T cell can be applied to an individual as soon as possible after the diagnosis of head and neck cancer.For example, less than about 150 days after the diagnosis of head and neck cancer, such as less than about 125 days, less than about 100 days, less than about 90 days, less than about 80 days or less than about 70 days after the diagnosis of head and neck cancer, the colony is applied to the individual.For example, about 30 to about 150 days after the diagnosis of head and neck cancer, such as about 40 to about 125 days, about 50 to about 100 days, about 90 days, about 85 days, about 80 days, about 75 days, about 70 days, about 65 days or about 60 days after the diagnosis of head and neck cancer, the colony is applied to the individual.Usually, the colony of the modified T cell is applied in a single dose.However, according to the patient's factors and the judgment of the physician, one or more (such as two or more, three or more, four or more or five or more) other dosages can be applied.

[0144] The population may contain any number of therapeutically effective modified T cells. The number of modified T cells for a given individual may depend on factors such as the cancer to be treated, the severity or stage of the cancer, the age of the patient, etc. Therefore, the number of administered cells may depend on the physician's judgment and may vary for each subject. For example, the population may contain about 0.8×10 9 to about 10×10 9 Modified T cells, such as about 0.8×10 9 About 1.2×10 9 Modified T cells, about 1.2×10 9 About 6×10 9 modified T cells, or about 1.0 × 10 9 Up to 10×10 9 The population may, for example, contain 1.0×10 9 modified T cells, about 5.0×10 9 modified T cells or about 10×10 9 modified T cells.

[0145] Typically, the population of modified T cells is administered intravenously. However, any suitable route may be used, such as intramuscular, subcutaneous, intradermal, transdermal or intraperitoneal routes.

[0146] As described above, the population of modified T cells can be administered in combination with checkpoint inhibitors and / or anticancer therapies. Checkpoint inhibitors or additional anticancer therapies can be administered by any route suitable for a given therapy, such as intravenous, intramuscular, subcutaneous, intradermal, transdermal or intraperitoneal administration. As described above, one or more doses of checkpoint inhibitors or anticancer therapies can be administered.

[0147] Each dose of a checkpoint inhibitor or anticancer therapy can contain any therapeutically effective amount of a checkpoint inhibitor or anticancer therapy. The amount for a given individual can depend on factors such as the cancer to be treated, the severity or stage of the cancer, the age of the patient, etc. Therefore, the amount to be administered can depend on the physician's judgment and can vary for each subject.

[0148] For example, the checkpoint inhibitor can be a PD-1 axis binding antagonist, such as pembrolizumab. The initial dose of the PD-1 axis binding antagonist and / or any additional dose of the PD-1 axis binding antagonist can include about 200 mg to about 700 mg of the PD-1 axis binding antagonist, such as about 200 mg to about 400 mg of the PD-1 axis binding antagonist. For example, the initial dose of the PD-1 axis binding antagonist and / or any additional dose of the PD-1 axis binding antagonist can include about 200 mg, about 250 mg, about 300 mg, about 350 mg, or about 400 mg of the PD-1 axis binding antagonist. The initial dose of the PD-1 axis binding antagonist and / or any additional dose of the PD-1 axis binding antagonist can, for example, include about 200 mg to about 400 mg of the PD-1 axis binding antagonist, such as about 480 mg of the PD-1 axis binding antagonist. In a preferred aspect of the present disclosure, the initial dose of the PD-1 axis binding antagonist and any additional doses of the PD-1 axis binding antagonist comprise 200 mg Q3W of pembrolizumab or 400 mg Q6W of pembrolizumab.

[0149] In some aspects of the present disclosure, the method comprises: administering lymphodepleting chemotherapy to the individual prior to administering the population of modified T cells. In other words, lymphodepleting chemotherapy may be administered prior to step (a). The lymphodepleting chemotherapy may be administered, for example, from about 14 days before step (a) to about 1 day before step (a), such as from about 13 days before step (a) to about 2 days before step (a), from about 12 days before step (a) to about 3 days before step (a), from about 11 days before step (a) to about 4 days before step (a), from about 10 days before step (a) to about 5 days before step (a), from about 9 days before step (a) to about 6 days before step (a), from about 8 days before step (a) to about 7 days before step (a), from about 10 days before step (a) to about 1 day before step (a), from about 9 days before step (a) to about 2 days before step (a), from about 8 days before step (a) to about 3 days before step (a), from about 7 days before step (a) to about 4 days before step (a), or from about 6 days before step (a) to about 5 days before step (a). Preferably, lymphodepleting chemotherapy is administered from about 7 days prior to step (a) to about 4 days prior to step (a). The purpose of lymphodepleting chemotherapy can be to deplete the lymphocyte compartment of the individual, thereby providing space for the adoptively transferred modified T cells to expand. In this way, the effect of a given dose of modified T cells can be maximized. Lymphodepleting chemotherapy can include any suitable lymphotoxic agent. Lymphotoxic agents and suitable doses are known in the art. For example, lymphodepleting chemotherapy can include fludarabine and / or cyclophosphamide. Typically, lymphodepleting chemotherapy is administered intravenously. However, any suitable route can be used, such as intramuscular, subcutaneous, intradermal, transcutaneous or intraperitoneal routes.

[0150] Drugs and medical uses

[0151] The present disclosure provides a population of modified T cells for use in a method of treating head and neck cancer in an individual, comprising a heterologous CD8 co-receptor and a heterologous T cell receptor capable of binding to a MAGE-A4 peptide antigen.Any aspects described above in conjunction with the methods of the present disclosure may also be applied to the population used.

[0152] The present disclosure also provides a population of modified T cells for use in the preparation of a medicament for use in a method for treating head and neck cancer in an individual, wherein the modified T cells comprise a heterologous CD8 co-receptor and a heterologous T cell receptor capable of binding to a MAGE-A4 peptide antigen. Any of the aspects described above in conjunction with the methods of the present disclosure may also be applied to such use of the population.

[0153] The present disclosure also provides a population of modified T cells for use in a method of treating head and neck cancer in an individual, wherein the modified T cells comprise a heterologous CD8 co-receptor and a heterologous T cell receptor capable of binding to a MAGE-A4 peptide antigen. Any aspect described above in conjunction with the methods of the present disclosure may also be applied to such use of the population.

[0154] Example

[0155] introduction

[0156] ADP-A2M4CD8-specific peptide enhanced affinity receptor (SPEAR TM ) T cells are genetically engineered to target the tumor antigen MAGE-A4 in the context of appropriate human leukocyte antigen (HLA) expression. ADP-A2M4CD8 are autologous CD4 and CD8 positive T cells that have been transduced with a self-inactivating (SIN) lentiviral vector expressing a high affinity MAGE-A4 specific T cell receptor (TCR) and an additional CD8α co-receptor.

[0157] The affinity optimized TCR (ADP-A2M4 TCR) comprises: the alpha chain variable domain contained in SEQ ID NO: 2 and the beta chain variable domain contained in SEQ ID NO: 3. When expressed in T cells, the signal peptide is cleaved from SEQ ID NO: 2 and 3 before surface expression. The A2M4 TCR targets the tumor antigen MAGE-A4 and activates engineered T cells. When presented with the HLA-A*02-GVYDGREHTV antigen complex, it recognizes the MAGE-A 4230-239 (GVYDGREHTV; SEQ ID NO: 1) peptide sequence derived from MAGE-A4.

[0158] ADP-A2M4CD8 SPEAR TM The CD8α co-receptor included in the T cells is designed to provide additional functionality to CD4+ T cells. Because CD4+ T cells have weak effector function in response to class I antigens, the CD8α co-receptor was introduced along with the TCR to increase TCR binding avidity and enhance the polyfunctional response of the engineered CD4+ T cells to MAGE-A4 positive tumors. Co-expression of CD8α adds CD8+ killer T cell capacity to CD4+ helper T cells while also maintaining / enhancing the helper cell capacity of CD4+ T cells. The addition of the CD8α co-receptor directly affects the binding of the TCR to the HLA-peptide complex in CD4+ T cells, enhancing CD4+ T cell effector function. Thus, ADP-A2M4CD8 SPEAR TMThe T cells were engineered to be improved upon ADP-A2M4 expressing T cells.

[0159] This has been confirmed in preclinical in vitro assays, where ADP-A2M4CD8 showed a significant improvement in T cell activation (when cultured with antigen-positive cells) relative to ADP-A2M4 expressing T cells, as measured by increased CD40L surface expression, particularly in the CD4+ fraction. When dendritic cells (DC) were included in the co-culture, significant improvements were observed in ADP-A2M4CD8 T cells. Cytokine release from DC (IL-12, MIG) and T cells (IFNy, IL-2 and other Th1) was improved compared to cultures containing ADP-A2M4 cells. In addition, the transformation of CD4+T cells was observed, from being unable to kill MAGE-A4 positive 3D microspheres to having effective cytotoxic function when transduced with ADP-A2M4CD8. Therefore, CD4+T cells transduced with ADP-A2M4CD8 not only show CD4+ helper function, but also show improved T cell effector function.

[0160] Individuals with recurrent or metastatic head and neck cancer have a response rate of approximately 20% to checkpoint inhibitor monotherapy and a response rate of 36% to checkpoint inhibitor combination chemotherapy, with progression-free survival of less than 5 months. Preliminary clinical trial results related to ADP-A2M4CD8 showed that when ADP-A2M4CD8 was used to treat cancer (including head and neck cancer), a response rate of approximately 36% was observed, and previous treatment with standard treatment had failed. Among them, 3 of 4 individuals with advanced metastatic head and neck cancer who received ADP-A2M4CD8 monotherapy were confirmed to have responded.

[0161] There is a need for an improved approach to treating head and neck cancer. Examples 1 and 2 consider the use of ADP-A2M4CD8 as a first-line treatment for (1) recurrent head and neck cancer and (2) newly metastatic or unresectable locally advanced head and neck cancer, respectively.

[0162] Example 1. Recurrent head and neck cancer.

[0163] Subjects are selected for ADP-A2M4CD8 treatment. In short, eligible subjects must have been previously diagnosed with locally advanced head and neck cancer and have undergone curative intent treatment, such as surgical resection and / or radiation therapy. Eligible subjects typically receive a follow-up PET and / or CT scan every three months after curative intent treatment. This scan identifies whether the subject is at high risk of recurrence. Subjects at high risk of recurrence can be selected for treatment.

[0164] In addition, the selected subjects were positive for HLA-A*02:01, HLA-A*02:03, or HLA-A*02:06 or another HLA-A*02 allele with the same protein sequence in the peptide binding domain. Patients who were HLA-A*02:05 positive were excluded from this study because alloreactivity of ADP-A2M4 and ADP-A2M4CD8 to two HLA-A*02:05 positive cell lines has been previously observed. Patients with HLA-A*02:07 or any A*02 null allele as the only HLA-A*02 allele were also excluded because of the reduced activity of these alleles.

[0165] Selected subjects also had tumors that showed MAGE-A4 expression, defined as ≥30% of tumor cells ≥2+ by immunohistochemistry (IHC).

[0166] Autologous cells are collected from selected subjects by leukapheresis for processing and production into ADP-A2M4CD8. The heterologous TCR contained in the ADP-A2M4CD8 T cells comprises: the α chain sequence contained in SEQ ID NO: 2 and the β chain sequence contained in SEQ ID NO: 3. The heterologous CD8 co-receptor contained in the ADP-A2M4CD8 T cells comprises two CD8 α chains, each chain comprising the amino acid sequence of SEQ ID NO: 10. The surface-expressed heterologous TCR and the surface-expressed heterologous CD8 co-receptor do not contain a signal sequence. Preferably, the autologous cells are collected before the diagnosis of recurrent locally advanced or metastatic head and neck cancer.

[0167] Baseline tumor assessments were obtained prior to treatment. Subjects were then provided with lymphodepleting chemotherapy with fludarabine and cyclophosphamide (Days -7 to -4), anticipating administration of ADP-A2M4CD8. Subjects received a single intravenous infusion of ADP-A2M4CD8 on Day 1. Treatment with standard therapeutic anticancer drug therapy (typically pembrolizumab) was concurrently implemented (e.g., within approximately 14 days of ADP-A2M4CD8 administration).

[0168] The subjects were given 1×10 8 Up to 1×10 10 ADP-A2M4CD8 T cells. The initial dose selected for ADP-A2M4CD8 was 1 × 10 9 transduced cells (range: 0.8×10 9 -1.2×10 9 transduced cells).

[0169] Subjects were monitored immediately after infusion (Day 1 to Day 8). Subjects were monitored weekly until Week 4 post-infusion. Subjects were then monitored at Weeks 6, 8, 12, 16, and 24 and at least every 3 months thereafter until disease progression. Tumor response was assessed according to Response Evaluation Criteria in Solid Tumors (RECIST) v1.1. Additional data collected included:

[0170] - Core needle biopsies to directly assess the "immune landscape" within the tumor at baseline and during the study.

[0171] - Cytokine levels in serum at baseline and during the study.

[0172] - Humoral immune responses to tumor antigens at baseline and during the study using sera.

[0173] - ADP-A2M4CD8 antibodies at baseline and during the study using serum.

[0174] - Using liquid biopsies, soluble markers representing the tumor and its microenvironment. For example, markers of circulating tumor cells (CTCs), exosomes, and cell-free DNA (cfDNA) produced by dying tumor cells can be used to monitor the molecular characteristics of tumor burden (including expression of target antigens) and immune response. Analysis of such soluble markers allows for the assessment of overall tumor burden and genetic analysis, including expression and mutation analysis of MAGE-A4 mRNA. Analysis of such soluble markers also allows for the systematic assessment of immune response.

[0175] - Phenotype and activity of genetically modified T cells before and after infusion. Relevant assays can be performed using blood and tumor (if resection is performed). These assays include: (i) phenotypic analysis to determine T cell lineages in cell products and blood (and tumor, if resection is performed) after infusion; (ii) quantification of senescence and activation state of immune subsets from PBMCs; (iii) analysis of gene expression or epigenetic features to reflect the phenotypic and functional state of cells; and / or (iv) direct functional assessment of cells.

[0176] -Persistence of infused engineered cells and its correlation with therapeutic efficacy. Persistence can be determined by the number of copies of genetically modified DNA per μg DNA and / or the number of cells transduced per μL or data relative to the total number of lymphocytes. Established methods include: (i) quantification of ADP-A2M4CD8 cells by quantitative PCR of the transgene from DNA extracted from frozen PBMCs, and (ii) quantification and phenotyping of ADP-A2M4CD8 cells by flow cytometry, DNA and RNA analysis from frozen PBMCs.

[0177] Figure 1 and Figure 2 This summarizes the above process.

[0178] Up to 10×10 9 The study was conducted to determine the efficacy of ADP-A2M4CD8 in patients with recurrent head and neck cancer. ...

[0179] Example 2. Newly metastatic or unresectable locally advanced head and neck cancer.

[0180] Subjects were selected for ADP-A2M4CD8 treatment. Briefly, selected subjects had newly metastatic or unresectable locally advanced head and neck cancer. Selected subjects were also positive for HLA-A*02:01, HLA-A*02:03, or HLA-A*02:06, or another HLA-A*02 allele with the same protein sequence in the peptide binding domain. Patients who were HLA-A*02:05 positive were excluded from this study because alloreactivity of ADP-A2M4 and ADP-A2M4CD8 to two HLA-A*02:05 positive cell lines has been previously observed. Patients with HLA-A*02:07 or any A*02 null allele as the only HLA-A*02 allele were also excluded because of the reduced activity of these alleles. In addition, selected subjects had tumors showing MAGE-A4 expression, defined as ≥30% of tumor cells ≥2+ by immunohistochemistry (IHC).

[0181] Autologous cells are collected from enrolled subjects by leukapheresis for processing and production into ADP-A2M4CD8. The heterologous TCR contained in the ADP-A2M4CD8 T cells comprises: the alpha chain sequence contained in SEQ ID NO: 2 and the beta chain sequence contained in SEQ ID NO: 3. The heterologous CD8 co-receptor contained in the ADP-A2M4CD8 T cells comprises two CD8 alpha chains, each chain comprising the amino acid sequence of SEQ ID NO: 10. The surface expressed heterologous TCR and the surface expressed heterologous CD8 co-receptor do not contain a signal sequence.

[0182] Baseline tumor assessments were obtained prior to treatment. Subjects were then administered one or more cycles of standard-of-care anticancer drug therapy, typically pembrolizumab. Subjects were offered lymphodepleting chemotherapy with fludarabine and cyclophosphamide (Days -7 to -4), with anticipated administration of ADP-A2M4CD8. Subjects received a single intravenous infusion of ADP-A2M4CD8 on Day 1. Day 1 was set as soon as possible after diagnosis of newly metastatic or unresectable locally advanced head and neck cancer, i.e., as soon as autologous cells were ready. This was typically approximately 60 days after leukapheresis.

[0183] The subjects were given 1×10 8 Up to 1×10 10 ADP-A2M4CD8 T cells. The initial dose selected for ADP-A2M4CD8 was 1 × 10 9 transduced cells (range: 0.8×10 9 -1.2×10 9 transduced cells).

[0184] Subjects were monitored immediately after infusion (Day 1 to Day 8). Subjects were monitored weekly until Week 4 post-infusion. Subjects were then monitored at Weeks 6, 8, 12, 16, and 24 and at least every 3 months thereafter until disease progression. Tumor response was assessed according to Response Evaluation Criteria in Solid Tumors (RECIST) v1.1. Additional data collected included:

[0185] - Core needle biopsies to directly assess the "immune landscape" within the tumor at baseline and during the study.

[0186] - Cytokine levels in serum at baseline and during the study.

[0187] - Humoral immune responses to tumor antigens at baseline and during the study using sera.

[0188] - ADP-A2M4CD8 antibodies at baseline and during the study using serum.

[0189] - Using liquid biopsies, soluble markers representing the tumor and its microenvironment. For example, markers of circulating tumor cells (CTCs), exosomes, and cell-free DNA (cfDNA) produced by dying tumor cells can be used to monitor the molecular characteristics of tumor burden (including expression of target antigens) and immune response. Analysis of such soluble markers allows for the assessment of overall tumor burden and genetic analysis, including expression and mutation analysis of MAGE-A4 mRNA. Analysis of such soluble markers also allows for the systematic assessment of immune response.

[0190] - Phenotype and activity of genetically modified T cells before and after infusion. Relevant assays can be performed using blood and tumor (if resection is performed). These assays include: (i) phenotypic analysis to determine T cell lineages in cell products and blood (and tumor, if resection is performed) after infusion; (ii) quantification of senescence and activation state of immune subsets from PBMCs; (iii) analysis of gene expression or epigenetic features to reflect the phenotypic and functional state of cells; and / or (iv) direct functional assessment of cells.

[0191] -Persistence of infused engineered cells and its correlation with therapeutic efficacy. Persistence can be determined by the number of copies of genetically modified DNA per μg DNA and / or the number of cells transduced per μL or data relative to the total number of lymphocytes. Established methods include: (i) quantification of ADP-A2M4CD8 cells by quantitative PCR of the transgene from DNA extracted from frozen PBMCs, and (ii) quantification and phenotyping of ADP-A2M4CD8 cells by flow cytometry, DNA and RNA analysis from frozen PBMCs.

[0192] Figure 3 and Figure 4 This summarizes the above process.

[0193] Up to 10×10 9 The study showed that the first-line treatment of newly metastatic or unresectable locally advanced head and neck cancer included administration of ADP-A2M4CD8, and was well tolerated. Emerging data suggest that treatment outcomes may be improved when first-line treatment of newly metastatic or unresectable locally advanced head and neck cancer includes administration of ADP-A2M4CD8. For example, outcomes may be improved relative to "standard of care" first-line treatment of newly metastatic or unresectable locally advanced head and neck cancer, such as checkpoint inhibitor (e.g., PD-1 axis binding antagonist) monotherapy or a combination of checkpoint inhibitor and chemotherapy.

[0194] Example 3. Preliminary results from the Phase 1 SURPASS trial of ADP-A2M4CD8 (next-generation SPEAR T-cell therapy) in patients with head and neck cancer.

[0195] Autologous T cells were obtained by leukapheresis, transduced with a self-inactivating lentiviral vector expressing the MAGE-A4-specific TCR and CD8α co-receptor, and infused back into the patient as ADP-A2M4CD8 after lymphodepleting chemotherapy.

[0196] Four patients with MAGE-A4 positive head and neck cancer were treated with ADP-A2M4CD8. Prior therapy was:

[0197] -First-line: 5-fluorouracil (5FU), carboplatin, pembrolizumab;

[0198] -Third-line: cisplatin, carboplatin, paclitaxel, pembrolizumab, cetuximab, panitumumab;

[0199] -Third-line: 5FU, carboplatin, cetuximab, nivolumab, taxane, cetuximab;

[0200] -Fifth line: cisplatin, 5FU, paclitaxel, cetuximab, carboplatin, nivolumab, CX-2029.

[0201] Three of the four patients with head and neck cancer underwent human papillomavirus screening; all were negative. The following table provides the baseline characteristics of the patients:

[0202] characteristic Head and neck cancer (n=4) Gender, n (%) male 2(50.0) female 2(50.0) Age, years, median (range) 59.5(43-70) <![CDATA[H-score a , median (range)]]> 287.5(140-300) ECOG at baseline, n (%) 0 1(25) 1 3(75) Systemic bridging therapy, yes, n (%) 2(50) <![CDATA[Transduced cells, ×10 9 cells, median (range)]]> 5.46(4.6-6.5) Number of previous systemic treatment lines, median (range) 3(1-5) Baseline SLD, median (range) 69(19-164)

[0203] a H-score: 1×(percentage of 1+ cells)+2×(percentage of 2+ cells)+3×(percentage of 3+ cells). ECOG, Eastern Cooperative Oncology Group; SLD, sum of the longest diameters of target lesions /

[0204] Among patients who received systemic bridging therapy, one patient received paclitaxel and cetuximab, and one patient received tipifarnib.

[0205] According to the Response Evaluation Criteria in Solid Tumors (RECIST) version 1.1 reviewed by the investigators, the overall response rate (ORR) was 75.0% (3 partial responses; Figure 5).

[0206] The disease control rate was 100.0% (3 partial responses and 1 stable disease).

[0207] The duration of response ranged from 7.4 to 20.1 weeks, with a median of 8.7 weeks. The data presented in Figure 5 indicate the change from baseline SLD to before disease progression or surgical resection.

[0208] Figure 6 Shown is a hilar mass in a patient with stage IV head and neck cancer who had a confirmed partial response after treatment with ADP-A2M4CD8. The patient was a 69-year-old white male with stage IV squamous head and neck cancer. MAGE-A4 expression in tumor cells was: 85% 3+, 10% 2+, and 5% 1+. Baseline SLD was 111 mm (from 5 target lesions). Previous systemic treatment was platinum-based therapy, nivolumab, and taxane / cetuximab. The patient was treated with 5 billion transduced ADP-A2M4CD8 T cells. Confirmed response was initially reported at week 4 and continued through week 24.

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[0210]

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[0214]

Claims

1. A method of treating head and neck cancer in an individual, comprising administering to the individual a population of modified T cells, the population of T cells comprising a heterologous CD8 co-receptor and a heterologous T cell receptor (TCR) capable of binding a MAGE-A4 peptide antigen.

2. The method according to claim 1, wherein: The method further comprises administering a check point inhibitor to the individual, optionally wherein the population of modified T cells and the check point inhibitor are administered in the same line of treatment.

3. The method according to claim 2, wherein: (a) starting administration of the checkpoint inhibitor before administering the population of modified T cells and continuing administration of the checkpoint inhibitor after administering the population of modified T cells; or (b) administering the checkpoint inhibitor beginning simultaneously with or after administering the population of modified T cells, and continuing administering the checkpoint inhibitor after administering the population of modified T cells.

4. The method according to claim 2 or 3, wherein: The checkpoint inhibitor includes a programmed death-1 (PD-1) axis binding antagonist, optionally pembrolizumab.

5. A method according to any one of the preceding claims, wherein: The method further comprises: administering to the individual an additional anti-cancer therapy, optionally wherein: (a) the population of modified T cells and the additional anti-cancer therapy are administered in the same line of therapy; and / or (b) The additional anti-cancer therapy is chemotherapy.

6. The method according to claim 5, wherein: (a) starting administration of the additional anti-cancer therapy before administering the population of modified T cells and continuing administration of the additional anti-cancer therapy after administering the population of modified T cells; or (b) administering the additional anti-cancer therapy beginning simultaneously with or after administering the population of modified T cells, and continuing administering the additional anti-cancer therapy after administering the population of modified T cells.

7. A method according to any one of the preceding claims, wherein: The head and neck cancer is recurrent head and neck cancer.

8. The method according to claim 7, wherein: The head and neck cancer has recurred following curative intent treatment for locally advanced head and neck cancer, optionally wherein: (a) the curative intent treatment comprises surgical resection and / or radiotherapy; and / or (b) The curative intent treatment includes systemic treatment.

9. The method according to claim 7 or 8, wherein: (a) the method comprises: administering a checkpoint inhibitor to the individual, and starting administration of the checkpoint inhibitor at the same time as or after administration of the population of modified T cells, and continuing administration of the checkpoint inhibitor after administration of the population of modified T cells; and / or (b) The method comprises administering an additional anti-cancer therapy to the individual, and starting administration of the additional anti-cancer therapy at the same time as or after administration of the population of modified T cells, and continuing administration of the additional anti-cancer therapy after administration of the population of modified T cells, optionally wherein the additional anti-cancer therapy is chemotherapy.

10. The method according to any one of claims 1 to 6, wherein: The individual has not been previously treated for head and neck cancer, optionally wherein the head and neck cancer is newly metastatic or unresectable locally advanced head and neck cancer.

11. The method according to claim 10, wherein: (a) the method comprises: administering a checkpoint inhibitor to the individual, and starting administration of the checkpoint inhibitor before or simultaneously with administration of the population of modified T cells, and continuing administration of the checkpoint inhibitor after administration of the population of modified T cells; or (b) The method comprises: administering an additional anti-cancer therapy to the individual, and starting administration of the additional anti-cancer therapy before or simultaneously with administration of the population of modified T cells, and continuing administration of the additional anti-cancer therapy after administration of the population of modified T cells; optionally, wherein the additional anti-cancer therapy is chemotherapy.

12. A method according to any one of the preceding claims, wherein: (a) administering the population of modified T cells as soon as possible after diagnosis of the head and neck cancer; and / or (b) administering the population of modified T cells as a single dose.

13. A method according to any one of the preceding claims, wherein: (a) the heterologous TCR binds to GVYDGREHTV (SEQ ID NO: 1) in complex with an HLA molecule; (b) the heterologous TCR comprises an alpha chain amino acid sequence having at least 80% sequence identity to SEQ ID NO: 2 and a beta chain amino acid sequence having at least 80% sequence identity to SEQ ID NO: 3; and / or (c) The CD8 co-receptor is CD8α.

14. A method according to any one of the preceding claims, wherein: The modified T cells are autologous to the individual, optionally wherein the method comprises generating the population by the steps of: (a) obtaining peripheral blood mononuclear cells (PBMC) from the individual; (b) selecting T cells from the PBMCs; and (c) modifying the selected T cells to express the heterologous CD8 co-receptor and the heterologous TCR; Optionally, wherein the head and neck cancer is a recurrent head and neck cancer, and wherein at least one of steps (a) to (c) is performed prior to recurrence.

15. A population of modified T cells comprising a heterologous CD8 co-receptor and a heterologous T cell receptor capable of binding a MAGE-A4 peptide antigen, for use in a method as claimed in any one of the preceding claims.

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