Methods of treating prame-positive cancer

Abstract

The present disclosure provides a method of treating non-small cell lung cancer or small cell lung cancer or ovarian cancer in a subject, the method comprising administering to the subject a composition comprising (I) a heterodimeric TCR-anti-CD3 antibody fusion molecule, and (II) a second composition comprising a second active agent, e.g., wherein the second active agent is a chemotherapeutic agent (e.g., gemcitabine, decitabine, pegylated liposomal doxorubicin (PLD), or nab-paclitaxel), or a targeted therapy (e.g., bevacizumab), or a multimodal therapy (e.g., a chemotherapeutic agent and a targeted therapy), wherein the dosing regimen comprises administering: (i) a first dose comprising 2 ug to 30 ug of the TCR-anti-CD3 antibody fusion molecule, (ii) a second dose comprising 5 ug to 60 ug of the TCR-anti-CD3 antibody fusion molecule, and (iii) a maintenance dose comprising 15 ug to 200 ug of the TCR-anti-CD3 antibody fusion molecule, wherein the maintenance dose is administered in at least three phases, wherein the first phase is administered once every 6 days to 8 days, the second phase is administered once every 12 days to 16 days, and the third phase is administered once every 26 days to 30 days, wherein the second active agent is administered at least once prior to, concurrently with, and / or after the first dose comprising the TCR-anti-CD3 antibody fusion molecule.

Description

Technical Field

[0001] This disclosure provides a method for treating a subject with non-small cell lung cancer, small cell lung cancer, or ovarian cancer, the method comprising administering a composition to the subject according to a dosing regimen, the composition comprising (I) a heterodimeric TCR-anti-CD3 antibody fusion molecule, and (II) a second composition comprising a second active agent, for example, wherein the second active agent is a chemotherapeutic agent (e.g., gemcitabine, decitabine, PEGylated liposomal doxorubicin (PLD), or nab-paclitaxel), or a targeted therapy (e.g., bevacizumab), or a multimodal therapy (e.g., a chemotherapeutic agent and a targeted therapy), wherein the dosing regimen comprises administering: (a) a first dose comprising 2 μg to 30 μg of the TCR-anti-CD3 antibody fusion molecule, and (b) a second dose comprising 5 μg to 60 μg of the TCR-anti-CD3 antibody fusion molecule. (c) a maintenance dose comprising 15 μg to 200 μg of the TCR-antiCD3 antibody fusion molecule, wherein the maintenance dose is administered in at least three phases, wherein the first phase is administered every 6 to 8 days, the second phase is administered every 12 to 16 days, and the third phase is administered every 26 to 30 days, wherein the second active agent is administered at least once before, in parallel with, and / or after the first dose comprising the TCR-antiCD3 antibody fusion molecule. Background Technology

[0002] The preferred melanoma antigen (PRAME) is a nuclear receptor and transcriptional regulator, and a member of the cancer testis antigen (CTA) protein family. In normal tissues, PRAME is expressed almost exclusively in the testes. Physiologically, it acts as a blocker of the retinoic acid receptor pathway, thereby regulating cell differentiation, growth, and apoptosis. Because PRAME is recognized by tumor-reactive cytotoxic T cells, anti-PRAME immunotherapy has a theoretical basis, and PRAME immunopositivity needs to be evaluated to identify therapeutic targets. PRAME is typically highly expressed in a range of solid and hematologic malignancies, including melanoma, ovarian cancer, uterine cancer, small cell lung cancer, non-small cell lung cancer, triple-negative breast cancer, and several rare tumor types.

[0003] Currently recommended treatments for cancers such as unresectable or metastatic disease provide patients with crucial treatment options. For example, such treatments for lung cancer include anti-PD-1 based regimens, often considered the standard of care (SoC) for initial treatment in many cases. Such SoC treatments also include, for example, surgery and / or chemotherapy for ovarian cancer in first-line or induction settings. Despite these options, at least half of these subjects progress within one year. The need for alternative treatment options remains unmet for patients with advanced cancer. Cancer treatment is complex and varies significantly depending on disease stage, prior treatment, and tumor type, and the significant heterogeneity among individual patients is often underestimated. Even within a single type of cancer or tumor type, the potential complexity of patients can differ, complicating trial design and outcome interpretation. Clinical trials designed to evaluate the safety and efficacy of IMC-F106C (the first bispecific protein targeting PRAME and CD3 T-cell receptors (TCRs)) represent a novel approach to treating cancer patients. Summary of the Invention

[0004] In some aspects, this disclosure provides a method for treating a subject with non-small cell lung cancer or small cell lung cancer, the method comprising administering a composition to the subject according to a dosing regimen, the composition comprising (I) a heterodimeric TCR-antiCD3 antibody fusion molecule, and (II) a second composition comprising a second active agent (e.g., wherein the second active agent is a chemotherapeutic agent, such as gemcitabine, decitabine, PLD, or albumin-bound paclitaxel). In some aspects, the heterodimeric TCR-antiCD3 antibody fusion molecule comprises (a) an α-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 14, and (b) a β-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 16, wherein the α-chain comprises a complementarity-determining region (CDR) comprising α-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 3, 4, and 5, respectively, and the β-chain comprises β-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 9, 10, and 11, respectively. In some aspects, the dosing regimen comprises administering: (i) a first dose comprising 2 μg to 30 μg of the TCR-antiCD3 antibody fusion molecule; (ii) a second dose comprising 5 μg to 60 μg of the TCR-antiCD3 antibody fusion molecule; and (iii) a maintenance dose comprising 15 μg to 200 μg of the TCR-antiCD3 antibody fusion molecule. The maintenance dose is administered in μg of the TCR-antiCD3 antibody fusion molecule, wherein the maintenance dose is administered in at least three phases, wherein the first phase is administered every 6 to 8 days, the second phase is administered every 12 to 16 days, and the third phase is administered every 26 to 30 days; wherein the second dose contains a larger amount of the TCR-antiCD3 antibody fusion molecule than the first dose, and the maintenance dose contains a larger amount of the TCR-antiCD3 antibody fusion molecule than the second dose; and wherein the second active agent is administered at least once before, in parallel with, and / or after the first dose containing the TCR-antiCD3 antibody fusion molecule.

[0005] In some aspects, the second active agent is a chemotherapeutic agent. In some aspects, the chemotherapeutic agent is gemcitabine. In some aspects, the chemotherapeutic agent is docetaxel. In some aspects, the second active agent is a targeted therapy. In some aspects, the targeted therapy is osimertinib. In some aspects, the second active agent is a multimodal therapy. In some aspects, the multimodal therapy is carboplatin / paclitaxel / pembrolizumab. In some aspects, the multimodal therapy is carboplatin / pemetrexed / pembrolizumab.

[0006] In some aspects, this disclosure provides a method for treating a subject with ovarian cancer, the method comprising administering a composition to the subject according to a dosing regimen, the composition comprising (I) a heterodimeric TCR-antiCD3 antibody fusion molecule and (II) a second active agent (e.g., wherein the second active agent is a chemotherapeutic agent, such as gemcitabine, decitabine, PLD, or albumin-bound paclitaxel). In some aspects, the heterodimeric TCR-antiCD3 antibody fusion molecule comprises (a) an α-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 14, and (b) a β-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 16, wherein the α-chain comprises a complementarity-determining region (CDR) comprising α-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 3, 4, and 5, respectively, and the β-chain comprises β-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 9, 10, and 11, respectively. In some aspects, the dosing regimen comprises administering: (i) a first dose comprising 2 μg to 30 μg of the TCR-antiCD3 antibody fusion molecule; (ii) a second dose comprising 5 μg to 60 μg of the TCR-antiCD3 antibody fusion molecule; and (iii) a maintenance dose comprising 15 μg to 200 μg of the TCR-antiCD3 antibody fusion molecule. The maintenance dose is administered in μg of the TCR-antiCD3 antibody fusion molecule, wherein the maintenance dose is administered in at least three phases, wherein the first phase is administered every 6 to 8 days, the second phase is administered every 12 to 16 days, and the third phase is administered every 26 to 30 days; wherein the second dose contains a larger amount of the TCR-antiCD3 antibody fusion molecule than the first dose, and the maintenance dose contains a larger amount of the TCR-antiCD3 antibody fusion molecule than the second dose; and wherein the second active agent is administered at least once before, in parallel with, and / or after the first dose containing the TCR-antiCD3 antibody fusion molecule.

[0007] In some aspects, the second active agent is a chemotherapeutic agent. In some aspects, the chemotherapeutic agent is gemcitabine. In some aspects, the chemotherapeutic agent is pegylated liposomal doxorubicin (PLD). In some aspects, the chemotherapeutic agent is carboplatin / paclitaxel. In some aspects, the second active agent is a targeted therapy. In some aspects, the targeted therapy is bevacizumab. In some aspects, the second active agent is a multimodal therapy. In some aspects, the multimodal therapy is carboplatin / paclitaxel / pembrolizumab. In some aspects, the multimodal therapy is carboplatin / gemcitabine / bevacizumab.

[0008] In some aspects (e.g., the methods described herein that can provide a dosing regimen), the first dose comprises 2 μg to 4 μg of the TCR-antiCD3 antibody fusion molecule, the second dose comprises 5 μg to 14 μg of the TCR-antiCD3 antibody fusion molecule, and the maintenance dose comprises 15 μg to 55 μg of the TCR-antiCD3 antibody fusion molecule. In some aspects (e.g., the methods described herein that can provide a dosing regimen), the first dose comprises 3 μg of the TCR-antiCD3 antibody fusion molecule, the second dose comprises 10 μg of the TCR-antiCD3 antibody fusion molecule, and the maintenance dose comprises 40 μg of the TCR-antiCD3 antibody fusion molecule. In some aspects (e.g., the methods described herein that can provide a dosing regimen), the first dose comprises 15 μg to 40 μg of the TCR-antiCD3 antibody fusion molecule, the second dose comprises 30 μg to 80 μg of the TCR-antiCD3 antibody fusion molecule, and the maintenance dose comprises 140 μg to 200 μg of the TCR-antiCD3 antibody fusion molecule. In some aspects (e.g., the methods described herein that can provide a dosing regimen), the first dose comprises 20 μg of the TCR-antiCD3 antibody fusion molecule, the second dose comprises 40 μg of the TCR-antiCD3 antibody fusion molecule, and the maintenance dose comprises 160 μg of the TCR-antiCD3 antibody fusion molecule.

[0009] In some aspects, the α-chain amino acid sequence of the TCR-anti-CD3 antibody is 100% identical to the amino acid sequence of SEQ ID NO: 14. In some aspects, the β-chain amino acid sequence is 100% identical to the amino acid sequence of SEQ ID NO: 16.

[0010] In some aspects (e.g., methods described herein that can provide a dosing regimen), the first dose is administered every 7 days. In some aspects (e.g., methods described herein that can provide a dosing regimen), the first dose is administered for 1 to 3 weeks. In some aspects (e.g., methods described herein that can provide a dosing regimen), the first dose is administered for 1 week. In some aspects, the second dose is administered 6 to 8 days after the first dose. In some aspects (e.g., methods described herein that can provide a dosing regimen), the second dose is administered every 7 days. In some aspects (e.g., methods described herein that can provide a dosing regimen), the second dose is administered for 1 to 3 weeks. In some aspects (e.g., methods described herein that can provide a dosing regimen), the second dose is administered for 1 week.

[0011] In some aspects (e.g., methods that can provide a dosing regimen as described herein), the maintenance dose in the first phase is administered 6 to 8 days after the second dose. In some aspects (e.g., methods that can provide a dosing regimen as described herein), the maintenance dose in the first phase is administered once weekly. In some aspects (e.g., methods that can provide a dosing regimen as described herein), the first phase is from about 2 weeks to about 101 weeks. In some aspects (e.g., methods that can provide a dosing regimen as described herein), the first phase is from about 8 weeks to about 14 weeks. In some aspects, the first phase is 10 weeks. In some aspects (e.g., methods that can provide a dosing regimen as described herein), the maintenance dose in the second phase is administered every 2 weeks. In some aspects (e.g., techniques that can relate to the method as described herein), the second phase is from about 30 weeks to 50 weeks. In some aspects, the second phase is 38 weeks. In some aspects (e.g., methods that can provide a dosing regimen as described herein), the maintenance dose in the third phase is administered every 4 weeks. In some aspects (e.g., techniques that can relate to the method as described herein), the third phase is from about 40 weeks to about 60 weeks. In some aspects (e.g., techniques that may relate to methods described herein), the third phase is approximately 48 weeks. In some aspects (e.g., methods that can provide a dosing regimen described herein), the first phase is 10 weeks, the second phase is 38 weeks, and the third phase is 48 weeks. In some aspects (e.g., methods that can provide a dosing regimen described herein), the first dose is administered for 1 week, the second dose is administered for 1 week, and the maintenance dose is administered for at least 40 weeks. In some aspects, the dosing regimen is approximately 90 weeks to approximately 110 weeks.

[0012] In some aspects (e.g., where the present disclosure provides a method), no corticosteroids are administered within three weeks prior to and throughout the dosing regimen. In some aspects (e.g., where the present disclosure provides a method of administration to a subject), the subject is HLA-A resistant. 02 positive. In some respects, the subject tested positive for HLA-A. 02:01 Positive. In some respects, the subject has a BRAF V600 mutation. In some respects, the subject has been diagnosed with a life expectancy of more than 3 months.

[0013] In some aspects (e.g., the methods provided herein), the second active agent is co-administered with the heterodimeric TCR-antiCD3 antibody fusion molecule. In some aspects, the second active agent is first administered 1 to 6 weeks prior to the administration of the heterodimeric TCR-antiCD3 antibody fusion molecule. In some aspects, the second active agent is administered 1 to 6 weeks after the administration of the heterodimeric TCR-antiCD3 antibody fusion molecule. In some aspects, the heterodimeric TCR-antiCD3 antibody fusion molecule is administered after treatment with the second active agent has been completed. In some aspects, the second active agent is administered 1 to 6 weeks prior to the administration of the heterodimeric TCR-antiCD3 antibody fusion molecule. In some aspects (e.g., the methods described herein that provide a dosing regimen), the second active agent is administered for at least 10 weeks. In some aspects, the second active agent is administered for at least 40 weeks. In some aspects, the second active agent is administered for the duration of the dosing regimen.

[0014] In some aspects (e.g., where this disclosure provides a second active agent), the second active agent is gemcitabine. In some aspects, the gemcitabine is administered at approximately 750 mg / m². 2 Approximately 1000 mg / m 2 Administration. In some respects, gemcitabine is administered at approximately 1000 mg / m². 2 The dosage of gemcitabine is as follows. In some aspects, gemcitabine is administered from once a week to once a month. In some aspects, gemcitabine is administered generally as follows: (i) for 4 weeks and then on for 1 week, (ii) for 3 weeks and then on for 1 week, (iii) for 2 weeks and then on for 1 week, (iv) for 1 week and then on for 1 week, (v) once a week, or (vi) once every two weeks.

[0015] In some aspects (e.g., where this disclosure provides a second active agent), the second active agent is albumin-bound paclitaxel. In some aspects, the albumin-bound paclitaxel is used at approximately 75 mg / m³. 2 Approximately 100 mg / m2 Administration. In some respects, the albumin-bound paclitaxel is administered at approximately 100 mg / m². 2 Dosage administration. In some aspects, the albumin-bound paclitaxel is administered once a week to once a month. In some aspects, the albumin-bound paclitaxel is administered generally as follows: (i) for 4 weeks followed by a 1-week break, (ii) for 3 weeks followed by a 1-week break, (iii) for 2 weeks followed by a 1-week break, (iv) for 1 week followed by a 1-week break, (v) once a week, or (vi) once every two weeks.

[0016] In some aspects (e.g., where this disclosure provides a second active agent), the second active agent is decitabine, for example, low-dose decitabine. In some aspects, the decitabine is administered at a dose of about 0.07 mg / kg to about 0.6 mg / kg. In some aspects, decitabine is administered from once a week to once a month. In some aspects, the decitabine is administered generally as follows: (i) once a week, (ii) once every two weeks, (iii) once every three weeks, or (iv) once a month.

[0017] In some aspects, the second active agent is pegylated liposomal doxorubicin (PLD). In some aspects, the PLD is administered at a dose of about 40 mg / m² to 60 mg / m² every 4 to 6 weeks.

[0018] In some aspects (e.g., where the methods described herein comprise the administration of a composition), the composition is administered orally, subcutaneously, sublingually, intramuscularly, intravenously, intranasally, transdermally, or in any combination thereof. In some aspects, the second composition is administered orally, subcutaneously, sublingually, intramuscularly, intravenously, intranasally, transdermally, or in any combination thereof. In some aspects, this disclosure provides a method for treating a subject with non-small cell lung cancer or small cell lung cancer, the method comprising administering a composition to the subject according to a dosing regimen, the composition comprising (I) a heterodimeric TCR-anti-CD3 antibody fusion molecule, and (II) a second composition comprising a second active agent (e.g., wherein the second active agent is a chemotherapeutic agent, such as gemcitabine, decitabine, PLD, or albumin-bound paclitaxel), in some aspects, the heterodimeric TCR-anti-CD3 antibody fusion molecule comprising (a) an α-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 14, and (b) a β-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 16, wherein the α-chain comprises a complementarity-determining region (CDR), the CDR comprising α-chain CDRs 1, 2, and 3, the α-chain CDRs 1, 2, and 3 having SEQ ID NO: 14, respectively. The amino acid sequences of 3, 4 and 5, and the β chain contains β chain CDRs 1, 2 and 3, which have the amino acid sequences of SEQ ID NO: 9, 10 and 11, respectively. In some aspects, the dosing regimen comprises: (i) administering a first dose comprising 3 μg of the TCR-antiCD3 antibody fusion molecule; (ii) administering a second dose one week after the first dose comprising 10 μg of the TCR-antiCD3 antibody fusion molecule; and (iii) administering a maintenance dose one week after the second dose and weekly during the first phase, the maintenance dose comprising 40 μg of the TCR-antiCD3 antibody fusion molecule, wherein the first phase is approximately 9 to 10 weeks; (iv) administering the maintenance dose every 2 weeks during the second phase, wherein the second phase is approximately 38 weeks; and (v) administering the maintenance dose every 4 weeks during the third phase, wherein the third phase is approximately 48 weeks, and wherein the second active agent is administered 1 to 6 weeks prior to administration of the heterodimeric TCR-antiCD3 antibody fusion molecule, during administration of the heterodimeric TCR-antiCD3 antibody fusion molecule during the duration of the dosing regimen, and / or after administration of the heterodimeric TCR-antiCD3 antibody fusion molecule.

[0019] In some aspects, this disclosure provides a method of treating a subject with ovarian cancer, the method comprising administering a composition to the subject according to a dosing regimen, the composition comprising (I) a heterodimeric TCR-antiCD3 antibody fusion molecule, and (II) a second composition comprising a second active agent (e.g., wherein the second active agent is a chemotherapeutic agent, such as gemcitabine, decitabine, PLD, or albumin-bound paclitaxel). In some aspects, the heterodimeric TCR-antiCD3 antibody fusion molecule comprises (a) an α-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 14, and (b) a β-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 16, wherein the α-chain comprises a complementarity-determining region (CDR) comprising α-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 3, 4, and 5, respectively, and the β-chain comprises β-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 9, 10, and 11, respectively. In some aspects, the dosing regimen comprises: (i) administering a first dose comprising 3 μg of the TCR-antiCD3 antibody fusion molecule; (ii) administering a second dose one week after the first dose comprising 10 μg of the TCR-antiCD3 antibody fusion molecule; and (iii) administering a maintenance dose one week after the second dose and weekly during the first phase, the maintenance dose comprising 40 μg of the TCR-antiCD3 antibody fusion molecule, wherein the first phase is approximately 9 to 10 weeks; (iv) administering the maintenance dose every 2 weeks during the second phase, wherein the second phase is approximately 38 weeks; and (v) administering the maintenance dose every 4 weeks during the third phase, wherein the third phase is approximately 48 weeks, and wherein the second active agent is administered 1 to 6 weeks prior to administration of the heterodimeric TCR-antiCD3 antibody fusion molecule, during administration of the heterodimeric TCR-antiCD3 antibody fusion molecule during the duration of the dosing regimen, and / or after administration of the heterodimeric TCR-antiCD3 antibody fusion molecule.

[0020] In some aspects, this disclosure provides a method for treating a subject with non-small cell lung cancer or small cell lung cancer, the method comprising administering a composition to the subject according to a dosing regimen, the composition comprising (I) a heterodimeric TCR-antiCD3 antibody fusion molecule, and (II) a second composition comprising a second active agent (e.g., wherein the second active agent is a chemotherapeutic agent, such as gemcitabine, decitabine, PLD, or albumin-bound paclitaxel). In some aspects, the heterodimeric TCR-antiCD3 antibody fusion molecule comprises (a) an α-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 14, and (b) a β-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 16, wherein the α-chain comprises a complementarity-determining region (CDR) comprising α-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 3, 4, and 5, respectively, and the β-chain comprises β-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 9, 10, and 11, respectively. In some aspects, the dosing regimen comprises: (i) administering a first dose comprising 20 μg of the TCR-antiCD3 antibody fusion molecule; (ii) administering a second dose one week after the first dose comprising 40 μg of the TCR-antiCD3 antibody fusion molecule; and (iii) administering a maintenance dose once weekly during the first phase, one week after the second dose, the maintenance dose comprising 160 μg of the TCR-antiCD3 antibody fusion molecule. (iv) administering the maintenance dose every 2 weeks during the second phase, which is approximately 38 weeks, and (v) administering the maintenance dose every 4 weeks during the third phase, which is approximately 48 weeks, and wherein the second active agent is administered 1 to 6 weeks prior to administration of the heterodimeric TCR-antiCD3 antibody fusion molecule, during administration of the heterodimeric TCR-antiCD3 antibody fusion molecule for the duration of the dosing regimen, and / or after administration of the heterodimeric TCR-antiCD3 antibody fusion molecule.

[0021] In some aspects, this disclosure provides a method of treating a subject with ovarian cancer, the method comprising administering a composition to the subject according to a dosing regimen, the composition comprising (I) a heterodimeric TCR-antiCD3 antibody fusion molecule, and (II) a second composition comprising a second active agent (e.g., wherein, in the dosing regimen, the second active agent is a chemotherapeutic agent, such as gemcitabine, decitabine, PLD, or albumin-bound paclitaxel). In some aspects, the heterodimeric TCR-antiCD3 antibody fusion molecule comprises (a) an α-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 14, and (b) a β-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 16, wherein the α-chain comprises a complementarity-determining region (CDR) comprising α-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 3, 4, and 5, respectively, and the β-chain comprises β-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 9, 10, and 11, respectively. In some aspects, the dosing regimen comprises: (i) administering a first dose comprising 20 μg of the TCR-antiCD3 antibody fusion molecule; (ii) administering a second dose one week after the first dose comprising 40 μg of the TCR-antiCD3 antibody fusion molecule; and (iii) administering a maintenance dose once weekly during the first phase, one week after the second dose, the maintenance dose comprising 160 μg of the TCR-antiCD3 antibody fusion molecule. (iv) administering the maintenance dose every 2 weeks during the second phase, which is approximately 38 weeks, and (v) administering the maintenance dose every 4 weeks during the third phase, which is approximately 48 weeks, and wherein the second active agent is administered 1 to 6 weeks prior to administration of the heterodimeric TCR-antiCD3 antibody fusion molecule, during administration of the heterodimeric TCR-antiCD3 antibody fusion molecule for the duration of the dosing regimen, and / or after administration of the heterodimeric TCR-antiCD3 antibody fusion molecule.

[0022] In some aspects, this disclosure provides a kit comprising: (i) a first container containing a first dose comprising 2 μg to 4 μg of a heterodimeric TCR-antiCD3 antibody fusion molecule; (ii) a second container containing a second dose comprising 5 μg to 14 μg of the heterodimeric TCR-antiCD3 antibody fusion molecule; (iii) one or more maintenance containers, each maintenance container containing a maintenance dose comprising 15 μg to 55 μg of the heterodimeric TCR-antiCD3 antibody fusion molecule; and (iv) one or more containers containing a second active agent dose (e.g., 10-1000 mg / m²). 2 (v) a container for administering the heterodimeric TCR-antiCD3 antibody fusion molecule according to a dosing regimen, the dosing regimen specifying: (a) administering the first dose in week 1, (b) administering the second dose in week 2, (c) administering a maintenance dose once weekly for 10 weeks during the first phase, (d) administering the maintenance dose once every 2 weeks for 38 weeks during the second phase, (e) administering the maintenance dose once every 4 weeks for 48 weeks during the third phase; and (vi) a dosing regimen for administering the second active agent in combination with the heterodimeric TCR-antiCD3 antibody fusion molecule according to a dosing regimen, wherein the dosing regimen specifies: (a) administering once weekly for two weeks and stopping for one week, (b) administering once weekly for three weeks and stopping for one week, or (c) administering once weekly for two weeks and stopping for one week.

[0023] In some aspects, the second active agent is a chemotherapeutic agent. In some aspects, the chemotherapeutic agent is gemcitabine. In some aspects, the chemotherapeutic agent is docetaxel. In some aspects, the second active agent is a targeted therapy. In some aspects, the targeted therapy is osimertinib. In some aspects, the second active agent is a multimodal therapy. In some aspects, the multimodal therapy is carboplatin / paclitaxel / pembrolizumab. In some aspects, the multimodal therapy is carboplatin / pemetrexed / pembrolizumab.

[0024] In some aspects, this disclosure provides a kit comprising: (i) a first container containing a first dose comprising 20 μg of a heterodimeric TCR-antiCD3 antibody fusion molecule; (ii) a second container containing a second dose comprising 40 μg of the heterodimeric TCR-antiCD3 antibody fusion molecule; (iii) one or more maintenance containers, each maintenance container containing a maintenance dose comprising 160 μg of the heterodimeric TCR-antiCD3 antibody fusion molecule; and (iv) one or more containers containing a second active agent dose (e.g., 10-1000 mg / m²). 2 (v) a container for administering the heterodimeric TCR-antiCD3 antibody fusion molecule according to a dosing regimen, the dosing regimen specifying: (a) administering the first dose in week 1, (b) administering the second dose in week 2, (c) administering a maintenance dose once weekly for 10 weeks during the first phase, (d) administering the maintenance dose once every 2 weeks for 38 weeks during the second phase, and (e) administering the maintenance dose once every 4 weeks for 48 weeks during the third phase; and (vi) a dosing regimen for administering the second active agent in combination with the heterodimeric TCR-antiCD3 antibody fusion molecule according to a dosing regimen, wherein the dosing regimen specifies: (a) administering once weekly for two weeks and stopping for one week, (b) administering once weekly for three weeks and stopping for one week, or (c) administering once weekly for two weeks and stopping for one week.

[0025] In some aspects of the kit, the heterodimeric TCR-antiCD3 antibody fusion molecule comprises (a) an α-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 14, and (b) a β-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 16, wherein the α-chain comprises a complementarity-determining region (CDR) comprising α-chain CDR 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 3, 4, and 5, respectively, and the β-chain comprises β-chain CDR 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 9, 10, and 11, respectively.

[0026] In some aspects of the kit, the kit comprises 2 to 10 maintenance containers. In some aspects of the kit, the first dose, the second dose, and / or the maintenance dose further comprise pharmaceutically acceptable excipients to form the first pharmaceutical formulation, the second pharmaceutical formulation, and / or the maintenance pharmaceutical formulation, respectively. In some aspects of the kit, the pH of the first pharmaceutical formulation, the second pharmaceutical formulation, and / or the maintenance pharmaceutical formulation is about 6.5 to about 7.5.

[0027] In some aspects of the kit, the second active agent is a chemotherapeutic agent. In some aspects, the chemotherapeutic agent is gemcitabine. In some aspects, the dosage of gemcitabine is 1000 mg / m². 2 In some aspects, the chemotherapeutic agent is docetaxel. In some aspects, the dosage of docetaxel is 75 mg / m². 2 In some aspects, the second active agent is a targeted therapy. In some aspects, the targeted therapy is osimertinib. In some aspects, the dosage of osimertinib is 80 mg. In some aspects, the dosage of osimertinib is 40 mg. In some aspects, the second active agent is a multimodal therapy. In some aspects, the multimodal therapy is carboplatin / paclitaxel / pembrolizumab. In some aspects, the dosage of carboplatin is AUC 6. In some aspects, the dosage of paclitaxel is 200 mg / m². 2 In some aspects, the dosage of pembrolizumab is 400 mg. In some aspects, the multimodal therapy is carboplatin / pemetrexed / pembrolizumab. In some aspects, the dosage of carboplatin is AUC 5. In some aspects, the dosage of carboplatin is AUC 6. In some aspects, the dosage of pemetrexed is 500 mg / m². 2 In some cases, the dosage of pembrolizumab is 400 mg.

[0028] In some aspects, this disclosure provides a method for treating a subject with PRAME-positive cancer, the method comprising administering a composition to the subject according to a dosing regimen, the composition comprising (I) a heterodimeric TCR-antiCD3 antibody fusion molecule according to the dosing regimen, and (II) a second composition comprising a second active agent (e.g., wherein the second active agent is a chemotherapeutic agent, such as gemcitabine, decitabine, PLD, or albumin-bound paclitaxel). In some aspects, the heterodimeric TCR-antiCD3 antibody fusion molecule comprises (a) an α-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 14, and (b) a β-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 16, wherein the α-chain comprises a complementarity-determining region (CDR) comprising α-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 3, 4, and 5, respectively, and the β-chain comprises β-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 9, 10, and 11, respectively. In some aspects, the dosing regimen comprises administering: (i) a first dose comprising 2 μg to 30 μg of the TCR-antiCD3 antibody fusion molecule; (ii) a second dose comprising 5 μg to 60 μg of the TCR-antiCD3 antibody fusion molecule; and (iii) a maintenance dose comprising 15 μg to 200 μg of the TCR-antiCD3 antibody fusion molecule. The maintenance dose is administered in μg of the TCR-anti-CD3 antibody fusion molecule, wherein the maintenance dose is administered in at least three phases, wherein the first phase is administered every 6 to 8 days, the second phase every 12 to 16 days, and the third phase every 26 to 30 days; and wherein the second dose contains a larger amount of the TCR-anti-CD3 antibody fusion molecule than the first dose, and the maintenance dose contains a larger amount of the TCR-anti-CD3 antibody fusion molecule than the second dose, wherein the second active agent is administered 1 to 6 weeks before administration of the heterodimeric TCR-anti-CD3 antibody fusion molecule, during administration of the heterodimeric TCR-anti-CD3 antibody fusion molecule during the duration of the dosing regimen, and / or after administration of the heterodimeric TCR-anti-CD3 antibody fusion molecule. In some aspects, the PRAME-positive cancer is non-small cell lung cancer or ovarian cancer.

[0029] In some aspects, this disclosure provides a method for treating a subject with platinum-sensitive ovarian cancer, the method comprising administering a composition to the subject according to a dosing regimen, the composition comprising: (I) a heterodimeric TCR-antiCD3 antibody fusion molecule, and (II) a second composition comprising a second active agent. In some aspects, the heterodimeric TCR-antiCD3 antibody fusion molecule comprises (a) an α-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 14, and (b) a β-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 16, wherein the α-chain comprises a complementarity-determining region (CDR) comprising α-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 3, 4, and 5, respectively, and the β-chain comprises β-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 9, 10, and 11, respectively. In some aspects, the dosing regimen comprises administering: (i) a first dose comprising 2 μg to 30 μg of the TCR-antiCD3 antibody fusion molecule; (ii) a second dose comprising 5 μg to 60 μg of the TCR-antiCD3 antibody fusion molecule; and (iii) a maintenance dose comprising 15 μg to 200 μg of the TCR-antiCD3 antibody fusion molecule. The maintenance dose is administered in μg of the TCR-antiCD3 antibody fusion molecule, wherein the maintenance dose is administered in at least three phases, wherein the first phase is administered every 6 to 8 days, the second phase is administered every 12 to 16 days, and the third phase is administered every 26 to 30 days; and wherein the second dose contains a larger amount of the TCR-antiCD3 antibody fusion molecule than the first dose, and the maintenance dose contains a larger amount of the TCR-antiCD3 antibody fusion molecule than the second dose, wherein the second active agent is administered 1 to 6 weeks before administration of the heterodimeric TCR-antiCD3 antibody fusion molecule, during administration of the heterodimeric TCR-antiCD3 antibody fusion molecule during the duration of the dosing regimen, and / or after administration of the heterodimeric TCR-antiCD3 antibody fusion molecule.

[0030] In some aspects, the second active agent is a targeted therapy. In some aspects, the targeted therapy is bevacizumab. In some aspects, the second active agent is a chemotherapeutic agent. In some aspects, the chemotherapeutic agent is gemcitabine. In some aspects, the chemotherapeutic agent is carboplatin / paclitaxel. In some aspects, the second active agent is a multimodal therapy. In some aspects, the multimodal therapy is carboplatin / paclitaxel / pembrolizumab. In some aspects, the multimodal therapy is carboplatin / gemcitabine / bevacizumab.

[0031] In some aspects, this disclosure provides a method for treating a subject with platinum-resistant ovarian cancer, the method comprising administering a composition to the subject according to a dosing regimen, the composition comprising: (I) a heterodimeric TCR-antiCD3 antibody fusion molecule, and (II) a second composition comprising a second active agent. In some aspects, the heterodimeric TCR-antiCD3 antibody fusion molecule comprises (a) an α-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 14, and (b) a β-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 16, wherein the α-chain comprises a complementarity-determining region (CDR) comprising α-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 3, 4, and 5, respectively, and the β-chain comprises β-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 9, 10, and 11, respectively. In some aspects, the dosing regimen comprises administering: (i) a first dose comprising 2 μg to 30 μg of the TCR-antiCD3 antibody fusion molecule; (ii) a second dose comprising 5 μg to 60 μg of the TCR-antiCD3 antibody fusion molecule; and (iii) a maintenance dose comprising 15 μg to 200 μg of the TCR-antiCD3 antibody fusion molecule. The maintenance dose is administered in μg of the TCR-antiCD3 antibody fusion molecule, wherein the maintenance dose is administered in at least three phases, wherein the first phase is administered every 6 to 8 days, the second phase is administered every 12 to 16 days, and the third phase is administered every 26 to 30 days; and wherein the second dose contains a larger amount of the TCR-antiCD3 antibody fusion molecule than the first dose, and the maintenance dose contains a larger amount of the TCR-antiCD3 antibody fusion molecule than the second dose, wherein the second active agent is administered 1 to 6 weeks before administration of the heterodimeric TCR-antiCD3 antibody fusion molecule, during administration of the heterodimeric TCR-antiCD3 antibody fusion molecule during the duration of the dosing regimen, and / or after administration of the heterodimeric TCR-antiCD3 antibody fusion molecule.

[0032] In some aspects, the second active agent is a targeted therapy. In some aspects, the targeted therapy is bevacizumab. In some aspects, the second active agent is a chemotherapeutic agent. In some aspects, the chemotherapeutic agent is gemcitabine. In some aspects, the chemotherapeutic agent is carboplatin / paclitaxel. In some aspects, the second active agent is a multimodal therapy. In some aspects, the multimodal therapy is carboplatin / paclitaxel / pembrolizumab. In some aspects, the multimodal therapy is carboplatin / gemcitabine / bevacizumab.

[0033] In some aspects, this disclosure provides a method for treating a subject with non-small cell lung cancer, the method comprising administering a composition to the subject according to a dosing regimen, the composition comprising: (I) a heterodimeric TCR-antiCD3 antibody fusion molecule, and (II) a second composition comprising a second active agent. In some aspects, the heterodimeric TCR-antiCD3 antibody fusion molecule comprises (a) an α-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 14, and (b) a β-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 16, wherein the α-chain comprises a complementarity-determining region (CDR) comprising α-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 3, 4, and 5, respectively, and the β-chain comprises β-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 9, 10, and 11, respectively. In some aspects, the dosing regimen comprises administering: (i) a first dose comprising 2 μg to 30 μg of the TCR-antiCD3 antibody fusion molecule; (ii) a second dose comprising 5 μg to 60 μg of the TCR-antiCD3 antibody fusion molecule; and (iii) a maintenance dose comprising 15 μg to 200 μg of the TCR-antiCD3 antibody fusion molecule. The maintenance dose is administered in μg of the TCR-antiCD3 antibody fusion molecule, wherein the maintenance dose is administered in at least three phases, wherein the first phase is administered every 6 to 8 days, the second phase is administered every 12 to 16 days, and the third phase is administered every 26 to 30 days; and wherein the second dose contains a larger amount of the TCR-antiCD3 antibody fusion molecule than the first dose, and the maintenance dose contains a larger amount of the TCR-antiCD3 antibody fusion molecule than the second dose, wherein the second active agent is administered 1 to 6 weeks before administration of the heterodimeric TCR-antiCD3 antibody fusion molecule, during administration of the heterodimeric TCR-antiCD3 antibody fusion molecule during the duration of the dosing regimen, and / or after administration of the heterodimeric TCR-antiCD3 antibody fusion molecule.

[0034] In some aspects, the second active agent is a chemotherapeutic agent. In some aspects, the second active agent is a targeted therapy. In some aspects, the targeted therapy is osimertinib.

[0035] In some aspects, this disclosure provides a method for treating a subject with melanoma, the method comprising administering a composition to the subject according to a dosing regimen, the composition comprising: (I) a heterodimeric TCR-antiCD3 antibody fusion molecule, and (II) a second composition comprising a second active agent. In some aspects, the heterodimeric TCR-antiCD3 antibody fusion molecule comprises (a) an α-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 14, and (b) a β-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 16, wherein the α-chain comprises a complementarity-determining region (CDR), the CDR comprising α-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 3, 4, and 5, respectively, and the β-chain comprises β-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 9, 10, and 11, respectively. In some aspects, the dosing regimen comprises administration of: (i) a first dose comprising 2 μg to 30 μg. (ii) a second dose comprising 5 μg to 60 μg of the TCR-antiCD3 antibody fusion molecule, and (iii) a maintenance dose comprising 15 μg to 200 μg of the TCR-antiCD3 antibody fusion molecule. The maintenance dose is administered in μg of the TCR-antiCD3 antibody fusion molecule, wherein the maintenance dose is administered in at least three phases, wherein the first phase is administered every 6 to 8 days, the second phase is administered every 12 to 16 days, and the third phase is administered every 26 to 30 days; and wherein the second dose contains a larger amount of the TCR-antiCD3 antibody fusion molecule than the first dose, and the maintenance dose contains a larger amount of the TCR-antiCD3 antibody fusion molecule than the second dose, wherein the second active agent is administered 1 to 6 weeks before administration of the heterodimeric TCR-antiCD3 antibody fusion molecule, during administration of the heterodimeric TCR-antiCD3 antibody fusion molecule during the duration of the dosing regimen, and / or after administration of the heterodimeric TCR-antiCD3 antibody fusion molecule.

[0036] In some aspects, the second active agent is a second heterodimeric TCR-anti-CD3 antibody fusion molecule. In some aspects, the second heterodimeric TCR-anti-CD3 antibody fusion molecule is tebentafusp. In some aspects, the second active agent is a targeted therapy. In some aspects, the targeted therapy is dabrafenib / trametinib. In some aspects, the second active agent is a demethylating agent. In some aspects, the demethylating agent is decitabine. In some aspects, the demethylating agent is low-dose decitabine. In some aspects, the second active agent is a chemotherapeutic agent. In some aspects, the chemotherapeutic agent is PLD. In some aspects, the chemotherapeutic agent is carboplatin / paclitaxel. Attached Figure Description

[0037] The following figures form part of and are included in this specification to further illustrate exemplary embodiments of certain aspects of this disclosure.

[0038] Figure 1 A high-level schematic diagram of the IMC-F106C-101 clinical trial is shown, which is a study evaluating the safety and efficacy of IMC-F106C as a monotherapy or combination therapy (see also Example 1).

[0039] Figure 2 A schematic diagram depicts the stepwise dosing regimen of the IMC-F106C compound used in the IMC-F106C-101 test.

[0040] Figure 3 A schematic diagram depicts the step-through dosing regimens evaluated in the various cohorts of the IMC-F106C-101 trial.

[0041] Figure 4 depicts the response of patients treated with the combination of IMC-F106C / chemotherapy agent (by changes in tumor size relative to baseline). Figure 4A This is a graphical representation of the change in tumor size relative to baseline in patients treated with IMC-F106C / albumin-bound paclitaxel combination therapy, where each bar represents an individual patient. Figure 4B The diagram shows the differences in tumor size relative to baseline over time, with each line representing an individual patient, as represented by a spider plot of IMC-F106C / albumin-bound paclitaxel treatment. Figure 4C This is a graphical representation of the change in tumor size relative to baseline in patients treated with the IMC-F106C / gemcitabine combination therapy, where each bar represents an individual patient. Figure 4DThis is a spider diagram representation of patients treated with IMC-F106C / gemcitabine, showing the change in tumor size relative to baseline over time, with each line representing an individual patient. (Further description is provided in Example 2).

[0042] Figure 5 illustrates the effect of decitabine pretreatment on the IMC-F106C cancer cell line (also known as PRAME ImmTAC). ® Molecularly dependent T cell activation. NCI-H441 ( Figure 5A ), NCI-H2087 ( Figure 5B ) and OVCAR3 ( Figure 5C Cancer cell lines were pretreated for 5 days with tissue culture medium containing 75 nM decitabine or with untreated tissue culture medium alone, and then used as target cells in IFNγ ELISPOT assays and PRAME ImmTAC titrations performed with healthy donor PBMCs.

[0043] Figure 6 A combination treatment approach for the IMC-F106C-101 assay, based on analyses performed during the initial phase of the assay, is described. The compounds to be combined with IMC-F106C were indicated for different groups used in the assay (C-1, C-3, C-4, C-5, D-1, E-3, E-4, F-1, G-1, G-2, G-3), and also arranged according to tumor type (indicated as separate modules for lung, ovarian / uterine, melanoma, and others).

[0044] Figure 7 An exemplary low-dose administration regimen for the heterodimeric TCR-antiCD3 antibody fusion molecule IMC-F106C is provided, the low-dose administration regimen comprising a first dose phase, a second dose phase, and a maintenance dose phase.

[0045] Figure 8 An exemplary high-dose regimen for the heterodimeric TCR-antiCD3 antibody fusion molecule IMC-F106C is provided, the high-dose regimen comprising a first dose phase, a second dose phase, and a maintenance dose phase.

[0046] Figure 9 An exemplary dosing regimen is provided when the second active agent (i.e., the combination partner) is administered in combination with a heterodimeric TCR-antiCD3 antibody fusion molecule (e.g., IMC-F106C). Detailed Implementation

[0047] Unless otherwise defined herein, scientific and technical terms as used herein shall have the meanings commonly understood by one of ordinary skill in the art. Furthermore, unless the context otherwise requires, singular terms shall include plural meanings, and plural terms shall include singular meanings.

[0048] The articles “a” and “an” are used in this text to refer to one or more grammatical objects of the article (i.e., at least one). For example, “an element” means one element or more elements.

[0049] The use of the term "or" in the claims is intended to mean "and / or" unless it is explicitly indicated that it refers only to alternatives or that the alternatives are mutually exclusive, but this disclosure supports the definition of referring only to alternatives and "and / or".

[0050] As used herein, the terms “comprising” (and any variations or forms of “comprise” and “comprises”), “having” (and any variations or forms of “have” and “has”), “including” (and any variations or forms of “includes” and “include)”), or “containing” (and any variations or forms of “containing” and “contains”) are inclusive or open-ended and do not exclude additional undescribed elements or method steps.

[0051] The use of the term "for example" and its corresponding abbreviation "eg" indicates that the specific term referenced is a representative instance and embodiment of this disclosure, and unless otherwise expressly stated, the representative instance and embodiment is not intended to be limited to the specific instance referenced or cited.

[0052] As used herein, “about” may mean a value provided plus or minus 10%. Where a range is provided, the range includes boundary values. “About” may additionally or alternatively mean within 10% of the value, or within 5% of the value, or in some cases within 2.5% of the value; or “about” may mean rounded to the nearest significant figure.

[0053] As used in this article, "between" is a range that includes the endpoints of a range. For example, numbers between x and y explicitly include the numbers x and y as well as any numbers falling between x and y.

[0054] The term "administration" or "administering" refers to the route by which a compound or composition provided herein is introduced into a subject to perform its intended function. Examples of possible administration routes include, but are not limited to, intravenous fusion.

[0055] The term "subject" means any subject in need of treatment, particularly a mammalian subject. In some embodiments, the term "subject" refers to a human subject. In some embodiments, the term "subject" refers to an adult human subject. In some embodiments, the term "subject" refers to a male human subject. In some embodiments, the term "subject" refers to a female human subject. In some embodiments, the term "subject" refers to a subject for treatment in need, namely a subject with non-small cell lung cancer or small cell lung cancer, ovarian cancer, and / or PRAME-positive cancer. As used herein, "subject in need" can mean a subject who desires treatment, for example, a subject diagnosed with non-small cell lung cancer or small cell lung cancer, ovarian cancer, and / or PRAME-positive cancer as described herein. In some embodiments, the term "subject in need" may refer to a subject having one or more symptoms associated with non-small cell lung cancer or small cell lung cancer, ovarian cancer, and / or PRAME-positive cancer, such as a subject having the following symptoms: worsening or persistent cough, chest pain, shortness of breath, wheezing, hemoptysis, abnormal vaginal discharge or bleeding after menopause, bloating or a feeling of fullness in the stomach, feeling full quickly after starting to eat, new onset of urinary frequency, new onset of constipation or other changes in bowel movements, discomfort or pain in the pelvic region, abdomen, or lower back, fatigue, unexplained weight loss, etc. In some embodiments, the term "subject in need" may refer to a subject at high risk of developing non-small cell lung cancer or small cell lung cancer, ovarian cancer, and / or PRAME-positive cancer suitable for treatment with the heterodimeric TCR-anti-CD3 antibody fusion molecule and / or the second active agent as described herein, regardless of whether the subject has physical manifestations of such conditions. In some embodiments, the subject is an adult, i.e., at least 18 years of age. In some embodiments, the subject is 12-18 years of age. In some embodiments, the subject is less than 12 years of age. In the embodiments, the second active agent is a chemotherapeutic agent (e.g., gemcitabine, decitabine, pegylated liposomal doxorubicin (PLD), or albumin-bound paclitaxel) or a targeted therapy (e.g., bevacizumab) or a multimodal therapy (e.g., a chemotherapeutic agent and a targeted therapy). In some embodiments, "subject" or "subject in need" may refer to a subject whose cancer has recurred (or relapsed). For a lung cancer patient whose cancer has recurred, this may correspond to a later-line treatment setting, such as second-line, third-line, fourth-line, or more lines of treatment, where the initial therapy or induction therapy may correspond to treatment with immunotherapy and / or surgery. For an ovarian cancer patient whose cancer has recurred after first-line therapy or induction therapy with chemotherapy and / or surgery, such patients may be considered platinum-sensitive or platinum-resistant. Platinum sensitivity or platinum resistance is generally based on the time since the last treatment with a platinum-based chemotherapy agent (such as platinum-based drugs), where platinum-sensitive cancer recurs more than 6 months after the last administration of a platinum-based chemotherapy agent, and platinum-resistant cancer recurs less than 6 months after the last administration of a platinum-based chemotherapy agent. Thus, for example, when referring to a subject, the term "platinum-resistant ovarian cancer" would be a subject whose ovarian cancer recurred less than 6 months after the last treatment with a platinum-based chemotherapy agent. Similarly, for example, when referring to a subject, the term "platinum-sensitive ovarian cancer" would refer to a subject whose ovarian cancer has recurred more than 6 months after their last treatment with a platinum-based chemotherapy agent. In some embodiments, platinum-sensitive patients who relapse within 6 to 12 months are defined as partially platinum-sensitive. In some embodiments, platinum-sensitive patients who relapse more than 12 months ago are defined as fully platinum-sensitive. In some cases, recurrence can be asymptomatic, and therefore elevations in certain markers (such as elevated CA-125) are sometimes used as indicators of recurrence and can also be indicators of whether a subject can be diagnosed as platinum-sensitive or platinum-resistant. The time interval between platinum sensitivity and platinum resistance may also be affected by treatment with certain adjuvant or neoadjuvant therapies or maintenance therapies, which may be combined with chemotherapy in first-line or induction therapy or administered to the patient after chemotherapy. Thus, in some embodiments, a "subject" or "subject in need" has previously received cancer treatment (e.g., with one or more chemotherapy agents described herein). In some embodiments, in addition to prior treatment, the subject is treated with the heterodimeric TCR-anti-CD3 antibody fusion molecule and a second or more active agents described herein. In some embodiments, the subject is treated with the described heterodimeric TCR-antiCD3 antibody fusion molecule and a second or more active agents in place of the previous treatment.

[0056] In some embodiments, platinum-based antitumor agents (platinum-based agents) are chemotherapeutic agents used to treat cancer. In some embodiments, platinum-based agents are cisplatin, carboplatin, oxaliplatin, nedaplatin, triplatin, tetranitrate, phenanthriplatin, pooplatin, or satraplatin. In some embodiments, platinum-based agents are cisplatin, carboplatin, or oxaliplatin.

[0057] Current therapies for patients with recurrent cancer typically do not improve overall survival, even if progression-free survival is improved, and often involve negative treatment side effects. While platinum-based chemotherapy is currently the preferred treatment for platinum-sensitive ovarian tumors, it is the first-line drug in combination with other cytotoxic agents as second-line chemotherapy. Current treatment options for patients with recurrent ovarian cancer that is platinum-sensitive or platinum-resistant include further chemotherapy or certain targeted therapies, such as bevacizumab or PARP inhibitors, or combinations of bevacizumab and chemotherapy, or combinations of PARP inhibitors and chemotherapy.

[0058] In some embodiments, the subject's serum LDH is about 1.1 × to about 2.0 × ULN. In some embodiments, the subject is under about 65 years of age. In some embodiments, the subject is at least about 65 years of age.

[0059] In some embodiments, the subject may have at least one of the following characteristics (e.g., at least one of 2, 3, 4, 5, 6, or 7): (1) ≥ 18 years of age, including 18 years of age; (2) Eastern Cooperative Oncology Group (ECOG) status of 0 or 1 at the start of treatment; (3) HLA-A 02:01 Positive (tested by a central laboratory); (4) Meets the requirements for tumor PRAME testing (tested by a central laboratory); (5) Has a recent biopsy (e.g., performed during screening) or a archived tumor biopsy or an adequate tumor biopsy with high tumor PRAME expression; (6) Has a disease (at least one target lesion or non-target lesion) that can be assessed according to RECIST v1.1 and a disease that can be measured according to it (at least one target lesion); (7) Has a disease suitable for biopsy and consents to have a tumor biopsy during screening and treatment; (8) Has a documented EGFR exon 19 deletion and exon 21 deletion determined by a locally approved diagnostic test. L858R mutation or T790M mutation and / or (9) having one of the following advanced (metastatic or unresectable) solid tumors: lung cancer, NSCLC, SCLC, melanoma, ovarian cancer (including fallopian tube cancer and primary peritoneal cancer), uterine cancer, endometrioid carcinoma (clear cell endometrial cancer, serous endometrial cancer, and carcinosarcoma of the endometrium), triple-negative breast cancer, germ cell tumors (choriocarcinoma, seminoma, and spermatogenic seminoma), sarcomas (myxoid liposarcoma, round cell liposarcoma, and synovial sarcoma), adenoid cystic carcinoma, basal cell carcinoma of the skin, thymic carcinoma, peripheral schwannoma, Merkel cell carcinoma / neuroendocrine carcinoma of the skin, squamous cell carcinoma of the head and neck, squamous cervical carcinoma, and urothelial carcinoma. In some embodiments, the subject has no history or current evidence of brain metastases (including leptomeningeal involvement). In some embodiments, the subject does not have a pre-existing peripheral neuropathy with an NCI CTCAE grade >2.

[0060] In some embodiments, a subject with non-small cell lung cancer or small cell lung cancer, ovarian cancer, and / or PRAME-positive cancer has a mutation in BRAF. In some embodiments, the subject has a BRAF V600 mutation. In some embodiments, the subject has a BRAF V600E mutation. In some embodiments, the subject does not contain a mutation in BRAF (e.g., non-small cell lung cancer or small cell lung cancer, ovarian cancer, and / or PRAME-positive cancer contains wild-type BRAF). In some embodiments, the subject does not contain a BRAF mutant, such as a BRAF mutant with enhanced activity (e.g., enhanced kinase activity and / or activity compared to wild-type BRAF) or a BRAF gain-of-function mutant. In some embodiments, the subject does not contain a constitutively active BRAF mutant. In some embodiments, the subject does not contain a BRAF V600E mutation (e.g., non-small cell lung cancer or small cell lung cancer, ovarian cancer, and / or PRAME-positive cancer contains wild-type BRAF). In some embodiments, the subject contains wild-type BRAF (e.g., non-small cell lung cancer or small cell lung cancer, ovarian cancer, and / or PRAME-positive cancer cells have wild-type BRAF). In some embodiments, the subject includes a BRAF mutant, such as a BRAF mutant with enhanced activity (e.g., enhanced kinase activity and / or activity compared to wild-type BRAF) or a BRAF gain-of-function mutant. In some embodiments, the subject includes a constitutively active BRAF mutant.

[0061] In some embodiments, the subject's serum lactate dehydrogenase (“LDH”) level is elevated. In some embodiments, the subject's serum LDH is less than about 0.8× of the upper limit of normal (“ULN”). In some embodiments, the subject's serum LDH is from about 0.8× to about 1.1× of the ULN. In some embodiments, the subject's serum LDH is greater than about 1.1× to about 2.0× of the ULN.

[0062] In some embodiments, treatment may be administered to a subject when the subject has precancerous lesions or when the subject is first diagnosed. In some embodiments, the methods described herein may be used when a subject has been diagnosed with a life expectancy of more than 1 year, more than 6 months, more than 3 months, or more than 1 month. In some embodiments, the methods described herein may be used when a subject has been diagnosed with a life expectancy of more than 3 months. In some embodiments, the methods described herein may be used when a subject has been diagnosed with a life expectancy of less than 5 years, less than 3 years, less than 2 years, less than 1 year, or less than 6 months. Those skilled in the art will understand that determining “life expectancy” is an estimate that can be determined by those skilled in the art using methods known in the art, such as the subject’s age, cancer stage, tumor thickness and size, growth rate, malignancy, contributing factors, etc.

[0063] In some embodiments, the subject presents with HLA-A 02 positive. The term "HLA-A" is also mentioned. "02 positive" means that it has a positive result for SLLQHLIGL (SEQ ID NO: 1)-HLA-A. Subjects whose TCRs are bound to the 02 complex. In some embodiments, the subject is HLA-A. 02:01 Positive. The term "HLA-A" is mentioned. 02:01 positive means expressing HLA-A 02:01 HLA-A allele Subjects with genotype 02.

[0064] The term “treatment” means the administration of an active agent for the purpose of: curing, eliminating, alleviating, resolving, altering, remedying, improving, enhancing or influencing a symptom (e.g., disease), symptoms of a symptom, or preventing or delaying the onset of disease symptoms, complications, biochemical markers, or otherwise statistically significantly preventing or inhibiting the further development of cancer, for example, statistically significantly preventing or inhibiting the further development of non-small cell lung cancer or small cell lung cancer, ovarian cancer, and / or PRAME-positive cancer in a subject.

[0065] In some embodiments, the kits and methods described herein relate to treating a subject's cancer, such as non-small cell lung cancer or small cell lung cancer and / or ovarian cancer. However, in some embodiments, the kits and methods described herein can be used for other PRAME-positive subjects. In some embodiments, the non-small cell lung cancer or small cell lung cancer, ovarian cancer, and / or PRAME-positive cancer is metastatic. In some embodiments, the non-small cell lung cancer or small cell lung cancer, ovarian cancer, and / or PRAME-positive cancer is a metastatic malignancy. In some embodiments, the non-small cell lung cancer or small cell lung cancer, ovarian cancer, and / or PRAME-positive cancer to be treated in the subject is stage 0, stage I, stage II, stage III, or stage IV. In some embodiments, the non-small cell lung cancer or small cell lung cancer, ovarian cancer, and / or PRAME-positive cancer to be treated is stage 0, stage IA, stage IB, stage IIA, stage IIB, stage IIC, stage IIIA, stage IIIB, stage IIIC, or stage IV. In some embodiments, the non-small cell lung cancer or small cell lung cancer, ovarian cancer, and / or PRAME-positive cancer to be treated is stage III. In some embodiments, the non-small cell lung cancer or small cell lung cancer, ovarian cancer, and / or PRAME-positive cancer to be treated is stage IV. In some embodiments, the cancer stage (e.g., stage IV cancer) is histologically confirmed. Histological confirmation refers to the examination of tissue using visual means (e.g., a microscope) to determine whether the tissue shows signs of cancer, disease, or other cancer-related abnormalities. In some embodiments, the tissue used in the histological examination has been removed by biopsy or surgical sectioning. The staging of melanoma can be based on methods known to those skilled in the art. Its staging can be determined according to the standards of the TNM or AJCC staging systems created by the American Joint Committee on Cancer (AJCC) and the International Union Against Cancer (UICC) (the disclosure of which is incorporated herein by reference in its entirety). The TNM staging system is used to describe most types of cancer.

[0066] In some embodiments, this disclosure provides a method of treating a subject (e.g., a human) with non-small cell lung cancer or small cell lung cancer and / or ovarian cancer, the method comprising administering to the subject a composition comprising a heterodimeric TCR-anti-CD3 antibody fusion molecule as described herein. In some embodiments, the method of treating small cell lung cancer and / or ovarian cancer with a TCR-anti-CD3 antibody fusion molecule further comprises a second active agent. In embodiments, the second active agent is a chemotherapeutic agent (e.g., gemcitabine, decitabine, pegylated liposomal doxorubicin (PLD), or albumin-bound paclitaxel) or a targeted therapy (e.g., bevacizumab) or a multimodal therapy (e.g., a chemotherapeutic agent and a targeted therapy). In some embodiments, this disclosure relates to the treatment of PRAME-positive cancers. The term “PRAME-positive cancer” refers to melanoma cancer in which at least some cancer cells express PRAME, a preferentially expressed antigen (i.e., PRAME). PRAME was first identified as an antigen overexpressed in melanoma (Ikeda et al., Immunity, Feb. 1997;6(2):199-208); it is also known as CT130, MAPE, OIP-4, and its Uniprot accession number is P78395. The protein acts as a repressor of retinoic acid receptor signaling (Epping et al., “Human tumor antigen PRAME is a dominant repressor of retinoic acid receptor signaling”). The human tumor antigen PRAME is a dominant repressor of retinoic acid receptor signaling (Cell, 122(6): 835-47 (2005)). PRAME belongs to a family of germline-encoded antigens known as cancer testis antigens. Cancer testis antigens are attractive targets for immunotherapeutic interventions because they are typically expressed in limited or no way in normal adult tissues. PRAME is expressed in a variety of solid tumors as well as leukemia and lymphoma. The PRAME-targeted therapy of the present invention may be particularly suitable for the treatment of cancers including, but not limited to, the following: melanoma, lung cancer, breast cancer, ovarian cancer, endometrial cancer, esophageal cancer, bladder cancer, head and neck cancer, uterine cancer, acute myeloid leukemia, chronic myeloid leukemia, and Hodgkin's lymphoma.

[0067] The peptide SLLQHLIGL (SEQ ID NO: 1) corresponds to amino acid 425-433 of the full-length PRAME protein and is associated with HLA-A. 02 Complex presentation on cell surface (Kessler et al., "Efficient identification of novel HLA-A in widely expressed tumor antigen PRAME by proteasome-mediated digestion analysis"). )0201 presents cytotoxic T lymphocyte epitopes (Efficient identification of novel HLA-A( (0201-presented cytotoxic Tlymphocyte epitopes in the widely expressed tumor antigen PRAME by proteasome-mediated digestion analysis), Journal of Experimental Medicine (J Exp Med.) 193(1):73-88 (2001). This peptide-HLA complex provides a target for TCR-based immunotherapeutic interventions.

[0068] WO 2018 / 234319 and the corresponding U.S. Patent Nos. 11,427,624 and 11,718,657 describe SLLQHLIGL (SEQ ID NO: 1)-HLA-A 02. TCRs bound to the complex, each of which is incorporated herein by reference in its entirety. A TCR has been mutated relative to the α and / or β variable domains of the natural PRAME TCR to increase its binding affinity for the complex and / or binding half-life, and can associate with a therapeutic agent (covalently or otherwise). One such therapeutic agent is an anti-CD3 antibody, or a functional fragment or variant of said anti-CD3 antibody, such as a single-chain variable fragment (scFv). The anti-CD3 antibody or fragment can be covalently linked to the C-terminus or N-terminus of the α-chain or β-chain of the TCR. The resulting molecule is TCR bispecific.

[0069] TCR bispecific proteins redirect polyclonal T cells to target peptides derived from intracellular or extracellular disease-associated antigens and presented on the cell surface in complex with HLA molecules. This approach has been used in the context of different antigens to target HLA-A from gp100. A TCR bispecific protein targeting PRAME, consisting of a 02-restricted peptide and CD3 (Tebentuft), has been clinically tested. Administration of this molecule has provided overall survival (OS) benefit for uveal melanoma (Nathan et al., 2021). However, such TCR bispecific proteins targeting PRAME have not yet been clinically tested.

[0070] In other words, PRAME-positive cancer is cancer associated with PRAME expression. In some embodiments, cancer is known to be associated with PRAME expression. For example, in some embodiments, the prevalence of PRAME expression is known to be elevated in cancer, and therefore PRAME expression is not assessed or retrospectively evaluated. Alternatively, any method known in the art can be used to assess PRAME expression, including, for example, histological methods or other quantitative or qualitative measurements, including PCR, RNA expression analysis, and / or kits or sequence sets designed to measure PRAME expression levels. However, this disclosure is not intended to be limited to the treatment of cancers in which PRAME expression can be detected by histological methods. In particular, this disclosure is not intended to be limited to the treatment of subjects, for example, in whom PRAME expression can be detected by histological methods. Rather, this disclosure can be used to treat cancers and tumor types that are considered PRAME-positive. In some embodiments, a PRAME-positive subject may have cutaneous melanoma. In some embodiments, a PRAME-positive subject does not have cutaneous melanoma but has another type of PRAME-positive cancer. In some embodiments, a PRAME-positive cancer (e.g., cutaneous melanoma) may have relapsed from standard treatment regimens, be refractory to standard treatment regimens, or be intolerant to standard treatment regimens.

[0071] When detected by histological methods such as immunohistochemistry (IHC), the H-score can be used to quantify PRAME expression. First, the expression of PRAME in the subject cells or subcellular compartments within the tumor is detected, and the expression is classified as positive or negative. Positive cells can be further classified as high, intermediate, or low based on IHC signal intensity. The H-score captures both the intensity and proportion of the biomarker of interest from the IHC image and includes values ​​between 0 and 300, thereby providing a dynamic range for quantifying the abundance of a specific biomarker or gene.

[0072] The methods and kits described herein comprise a heterodimeric TCR-anti-CD3 antibody fusion molecule. The heterodimeric TCR-anti-CD3 antibody fusion molecule contains two domains: - Heterodimeric TCR; and - Anti-CD3 antibody.

[0073] The heterodimeric TCR-antiCD3 antibody fusion molecule is described, for example, in WO2023 / 099622, which is incorporated herein by reference in its entirety.

[0074] Heterodimeric TCRs can comprise both an α chain (i.e., TCR α chain) and a β chain (i.e., TCR β chain). Each chain can contain a variable region, a linker region, and a constant region, and the β chain typically also contains a short diversity region between the variable and linker regions, but this diversity region can be considered part of the linker region. Each variable region can contain three CDRs (complementarity-determining regions) embedded in the frame sequence, one of which is a hypervariable region called CDR3. There are several types of α-chain variable (Vα) regions and several types of β-chain variable (Vβ) regions, which are distinguished by their frame, CDR1 and CDR2 sequences, and by a partially defined CDR3 sequence. The Vα type is represented by a unique TRAV number in IMGT nomenclature, and the Vβ type is represented by a unique TRBV number.

[0075] In some embodiments, this disclosure provides the TCR α-chain amino acid sequence of SEQ ID NO: 14 or a TCR α-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 14. In some embodiments, this disclosure provides the TCR β-chain anti-CD3 amino acid sequence of SEQ ID NO: 16 or a TCR β-chain anti-CD3 amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 16. In some embodiments, the TCR α-chain variable domain comprises CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 3, 4, and 5, respectively, and the TCR β-chain variable domain comprises CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 9, 10, and 11, respectively.

[0076] In some embodiments, the TCR can bind to the sequence SLLQHLIGL (SEQ ID NO: 1). The peptide of SEQ ID NO: 1 corresponds to amino acids 425-433 of the full-length PRAME protein and is associated with HLA-A. 02 Complex presentation on cell surface (Kessler et al., "Efficient identification of novel HLA-A in widely expressed tumor antigen PRAME by proteasome-mediated digestion analysis"). "Cytotoxic T-lymphocyte epitopes presented by 0201", *Journal of Experimental Medicine* 193(1):73-88 (2001). This peptide-HLA complex provides a target for TCR-based immunotherapeutic interventions. WO 2018 / 234319 describes the interaction with SLLQHLIGL (SEQ ID NO: 1)-HLA-A. The TCR bound to the 02 complex is incorporated herein as a whole.

[0077] In some embodiments, the heterodimeric TCR is fused with an antibody targeting CD3 to increase the binding affinity and / or binding half-life against CD3. One such antibody is an anti-CD3 antibody, or a functional fragment or variant of said anti-CD3 antibody, such as a single-chain variable fragment (scFv). The anti-CD3 antibody or fragment may be covalently linked to the C-terminus or N-terminus of the α-chain or β-chain of the TCR. The resulting molecule is bispecific to the TCR.

[0078] TCR bispecific proteins redirect polyclonal T cells to target peptides derived from intracellular or extracellular disease-associated antigens and presented on the cell surface in complex with HLA molecules. This approach has been used in the context of different antigens to target HLA-A from gp100. The TCR bispecific protein of 02-restricted peptide and CD3 (Tebentuft) has been clinically tested. Administration of this molecule has provided OS benefit for uveal melanoma (Nathan et al., 2021).

[0079] The term "antibody" includes, but is not limited to, genetically engineered or otherwise modified forms of immunoglobulins, such as intracellular antibodies, chimeric antibodies, fully human antibodies, humanized antibodies (e.g., generated via "CDR grafting"), antibody fragments, and heteroconjugated antibodies (e.g., bispecific antibodies, biantibodies, triantibodies, tetraantibodies, etc.). The term "antibody" includes cys-biantibodies and microantibodies. Therefore, unless explicitly stated otherwise, each and every embodiment provided herein with respect to "antibody" or "antibody-like construct" is also contemplated as an embodiment of a bispecific antibody, biantibody, scFv fragment, chimeric antibody receptor (CAR) construct, biantibody, and / or microantibody. The term "antibody" includes polypeptides of the immunoglobulin family as disclosed herein or polypeptides comprising fragments of immunoglobulins capable of non-covalently, reversibly, and specifically binding to a corresponding antigen. Exemplary antibody structural units comprise tetramers. In some embodiments, a full-length antibody may consist of two pairs of identical polypeptide chains, each pair having a "light" chain and a "heavy" chain (linked by disulfide bonds). The term “antibody” also includes different classes (i.e., IgA, IgG, IgM, IgD and IgE) and different subclasses (such as IgG1, IgG2 and so on) of immunoglobulins (Ig).

[0080] As used herein, the terms "anti-CD3 antibody" and "anti-CD3 antibody fragment" refer to antibodies or antibody fragments that recognize or bind to CD3. TCR-anti-CD3 fusion molecules used in this disclosure may include one or more conserved substitutions having similar amino acid sequences and / or retaining the same function (i.e., phenotypic silence as defined herein). Those skilled in the art will recognize that individual amino acids have similar properties, and therefore substitutions between them are "conserved." One or more of these amino acids in a protein, polypeptide, or peptide may generally be substituted with one or more other such amino acids without eliminating the desired activity of said protein, polypeptide, or peptide.

[0081] In some embodiments, the TCR-anti-CD3 fusion molecule used in this invention comprises an anti-CD3 scFv covalently linked to the N-terminus of the β-chain of a TCR via a linker. This type of molecule is known as ImmTAC® (immune mobilization monoclonal TCR for cancer). ImmTAC® molecules are engineered to activate an effective T-cell response, thereby specifically killing target cancer cells. In some embodiments, the heterodimeric TCR-anti-CD3 antibody fusion molecule used in this invention is an ImmTAC molecule as described in WO 2018 / 234319 (i.e., an ImmTAC targeting PRAME), which is incorporated herein by reference in its entirety.

[0082] In some embodiments, the heterodimeric TCR-anti-CD3 antibody fusion molecule is IMC-F106C. IMC-F106C is a T-cell redirection bispecific therapeutic agent comprising the peptide-HLA-A of SLLQHLIGL (SEQ ID NO: 1). The 02 complex binds to a soluble affinity-enhanced TCR, which fuses with anti-CD3 scFv. The targeting terminus of IMC-F106C (soluble TCR) binds to the PRAME antigen via HLA-A. 02. The peptide fragments presented on the surface of cancer cells bind. HLA molecules are polymorphic; in the United States and European countries, approximately 47% of Caucasian individuals express HLA-A. 02 genotype, of which more than 95% of HLA-A HLA-A was detected in 02 positive individuals. 02:01 Allele. The effector terminus (anti-CD3 scFv) of IMC-F106C can bind to CD3 on any T cell, thereby redirecting T cells to produce effector cytokines and / or kill target-presenting cells. Additionally, IMC-F106C-mediated tumor lysis can activate an endogenous anti-tumor immune response. ImmTAC® (immune mobilization monoclonal TCRs for cancer) molecules (such as IMC-F106C) are highly potent molecules, observed to redirect T cells in tumor cell lines presenting as few as 10 to 50 target peptides:HLA complexes. IMC-F106C has been shown to be effective at concentrations as low as 1 pM to 10 pM in the presence of HLA-A. In the case of 02:01 positive / PRAME positive cell lines, T cell activity is selectively redirected, thereby inducing T cell activation and killing PRAME positive cancer cells. As mentioned above, HLA-A 02. The restricted peptide SLLQHLIGL (SEQ ID NO: 1) is derived from the germline cancer antigen PRAME. IMC-F106C has the TCR α chain amino acid sequence of SEQ ID NO: 14 and the TCR β chain-anti-CD3 amino acid sequence of SEQ ID NO: 16.

[0083] PRAME is a cancer testis antigen that is typically highly expressed in a range of solid and hematologic malignancies, including melanoma, ovarian cancer, uterine cancer, small cell lung cancer, non-small cell lung cancer, triple-negative breast cancer, and several rare tumor types.

[0084] The sequences mentioned in this article are as follows: SEQ ID NO: 1: HLA-A 02 Restricted Peptide: SLLQHLIGL SEQ ID NO: 2: Amino acid sequence of the variable domain of the TCR α chain of the ImmTAC named IMC-F106C. CDRs (CDR1, CDR2, and CDR3) are underlined and designated as SEQ ID NO: 3, 4, and 5, respectively; framework regions (FR1, FR2, FR3, and FR4) are italicized and designated as SEQ ID NO: 27, 6, 7, and 28, respectively. Mutations relative to the native α chain are in bold.

[0085] SEQ ID NO: 8: Amino acid sequence of the variable domain of the TCR β chain of the ImmTAC named IMC-F106C. CDRs (CDR1, CDR2, and CDR3) are underlined and designated as SEQ ID NO: 9, 10, and 11, respectively; framework regions (FR1, FR2, FR3, and FR4) are italicized and designated as SEQ ID NO: 29, 12, 13, and 30, respectively. Mutations relative to the native β chain are in bold.

[0086] SEQ ID NO: 14: The amino acid sequence of the TCR α chain of the ImmTAC named IMC-F106C. CDRs (CDR1, CDR2, and CDR3) are underlined and designated as SEQ ID NO: 3, 4, and 5, respectively; frame regions (FR1, FR2, FR3, and FR4) are italicized and designated as SEQ ID NO: 27, 6, 7, and 28, respectively. The constant region is shown in bold and designated as SEQ ID NO: 15. Within the constant region, non-natural cysteine ​​residues are double-underlined (at position 48 in the constant region).

[0087] SEQ ID NO: 16: The amino acid sequence of the TCR β chain of the ImmTAC named IMC-F106C, which is anti-CD3. The anti-CD3 scFv (amino acids 1-253) is shown in bold and underlined and is designated as SEQ ID NO: 17. The linker (GGGGS) immediately following the scFv is shown in light text and is designated as SEQ ID NO: 18. The CDRs (CDR1, CDR2, and CDR3) are underlined and are designated as SEQ ID NO: 9, 10, and 11, respectively; the frame regions (FR1, FR2, FR3, and FR4) are italicized and are designated as SEQ ID NO: 29, 12, 13, and 30, respectively. The constant region is shown in bold (without underline) and is designated as SEQ ID NO: 19. Within the constant region, non-native cysteine ​​residues are double-underlined (at position 57 of the constant region). The additional non-natural amino acids at positions 75 and 89 in the constant region are also underlined.

[0088] The CDR sequence of the ImmTAC named IMC-F106C is as follows: The frame region sequence of the ImmTAC named IMC-F106C: Other connector sequences mentioned in this article: GGGSG (SEQ ID NO: 20), GGSGG (SEQ ID NO: 21), GSGGG (SEQ ID NO: 22), GSGGGP (SEQ ID NO: 23), GGEPS (SEQ ID NO: 24), GGEGGGP (SEQ ID NO: 25), and GGEGGGSEGGGS (SEQ ID NO: 26) In some embodiments, the TCR-antiCD3 fusion molecule used in the methods herein comprises: The TCR α chain amino acid sequence of SEQ ID NO: 14 or a TCR α chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 14, and The TCR β-chain anti-CD3 amino acid sequence of SEQ ID NO: 16 or the TCR β-chain anti-CD3 amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 16. The TCR α-chain variable domain comprises CDR1, 2, and 3 having amino acid sequences of SEQ ID NO: 3, 4, and 5, respectively, and the TOR β-chain variable domain comprises CDR1, 2, and 3 having amino acid sequences of SEQ ID NO: 9, 10, and 11, respectively.

[0089] In other words, although the amino acid sequence of the TCR α chain may have some variation compared to the sequence of SEQ ID NO: 14 (as long as the amino acid sequence of the TCR α chain is at least 90% identical to that of SEQ ID NO: 14) and / or the amino acid sequence of the TCR β chain-anti-CD3 may have some variation compared to the sequence of SEQ ID NO: 16 (as long as the amino acid sequence of the TCR β chain-anti-CD3 is at least 90% identical to that of SEQ ID NO: 16), the CDR of the TCR α chain must have the amino acid sequences of SEQ ID NO: 3, 4 and 5, respectively, and the CDR of the TCR β chain must have the amino acid sequences of SEQ ID NO: 9, 10 and 11, respectively. Therefore, the requirement that the TCR α-chain variable domain comprises CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 3, 4, and 5, respectively, and the requirement that the TCR β-chain variable domain comprises CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 9, 10, and 11, respectively, applies to all aspects and embodiments of the invention described herein. Thus, the TCR α-chain variable domain comprises CDRs 1, 2, and 3 having 100% identity with the amino acid sequences of SEQ ID NO: 3, 4, and 5, respectively, and the TCR β-chain variable domain comprises CDRs 1, 2, and 3 having 100% identity with the amino acid sequences of SEQ ID NO: 9, 10, and 11, respectively.

[0090] In some embodiments, the heterodimeric TCR-antiCD3 antibody fusion molecule is IMC-F106C. A phenotypic silent variant of the TCR-antiCD3 fusion molecule named IMC-F106C, within the scope of this disclosure, has the TCR α-chain amino acid sequence corresponding to SEQ ID NO: 14 and the TCR β-chain antiCD3 amino acid sequence corresponding to SEQ ID NO: 16. As used herein, the term "phenotypic silent variant" should be understood to refer to a TCR-antiCD3 fusion molecule incorporating one or more additional amino acid changes (including substitutions, insertions, and deletions) compared to the sequences of SEQ ID NO: 14 and SEQ ID NO: 16, and said TCR-antiCD3 fusion molecule has a similar or identical phenotype to the TCR-antiCD3 fusion molecule named IMC-F106C. For the purposes of this application, the TCR-antiCD3 fusion molecule phenotype includes antigen binding affinity (KD and / or binding half-life) and antigen specificity. When measured under the same conditions (e.g., at 25°C and / or on the same SPR chip), the phenotypic silent variant against SLLQHLIGL (SEQ ID NO: 1) HLA-A The KD and / or binding half-life of the 02 complex can be within 50% of the measured KD and / or binding half-life of the TCR-antiCD3 fusion molecule referred to as IMC-F106C, or more preferably within 20%. Suitable conditions are further provided in Example 3 of WQ / 2018 / 234319, which is incorporated herein by reference. Antigen specificity is further defined below. As is known to those skilled in the art, variations can be incorporated into its variable domains compared to the TCR detailed above without altering its affinity for SLLQHLIGL (SEQ ID NO: 1) HLA-A. TCRs with affinity for the O2 complex interaction. In particular, such silent mutations can be incorporated into portions of the sequence that are known not to be directly involved in antigen binding. These trivial variants are included within the scope of this invention.

[0091] Phenotypic silent variants may contain one or more conserved substitutions and / or one or more tolerant substitutions. When measured under identical conditions (e.g., at 25°C and / or on the same SPR chip), tolerant and conserved substitutions may make the SLLQHLIGL (SEQ ID NO: 1) HLA-A phenotype silent variants contain one or more conserved substitutions and / or one or more tolerant substitutions. The change in KD and / or binding half-life of the 02 complex is within 50%, more preferably within 20%, or even more preferably within 10%, of the measured KD and / or binding half-life of the TCR-antiCD3 fusion molecule referred to as IMC-F106C, provided that the change in KD does not result in an affinity less than (i.e., weaker than) 200 pM. Tolerant substitution means substitution that does not fall under the conservative definition provided below but is still phenotypically silent.

[0092] The TCR-antiCD3 fusion molecule used in the methods described herein may include one or more conserved substitutions having similar amino acid sequences and / or retaining the same function (i.e., phenotypic silencing as defined above). Those skilled in the art will recognize that individual amino acids have similar properties, and therefore substitutions between them are “conserved.” One or more of these amino acids in a protein, polypeptide, or peptide can typically be substituted with one or more other such amino acids without eliminating the desired activity of the protein, polypeptide, or peptide.

[0093] Therefore, the amino acids glycine, alanine, valine, leucine, and isoleucine can generally be substituted for each other (amino acids with aliphatic side chains). Among these possible substitutions, glycine and alanine are preferred to be substituted for each other (because of their relatively short side chains), and valine, leucine, and isoleucine are preferred to be substituted for each other (due to their larger hydrophobic aliphatic side chains). Other amino acids that can often be substituted for each other include: phenylalanine, tyrosine, and tryptophan (amino acids with aromatic side chains); lysine, arginine, and histidine (amino acids with basic side chains); aspartic acid and glutamic acid (amino acids with acidic side chains); asparagine and glutamine (amino acids with amide side chains); and cysteine ​​and methionine (amino acids with sulfur-containing side chains). It should be understood that amino acid substitutions within the scope of this invention can be made using naturally occurring or non-natural amino acids. For example, it is contemplated here that the methyl group on alanine can be replaced with an ethyl group, and / or minor modifications can be made to the peptide backbone. Regardless of whether natural or synthetic amino acids are used, it is preferred that only L-amino acids are present.

[0094] This type of substitution is often referred to as "conservative" or "semi-conservative" amino acid substitution. Therefore, the present invention extends to the use of TCR-antiCD3 fusion molecules comprising the above-described amino acid sequences, but having one or more conserved substitutions and / or one or more tolerant substitutions in the sequences, such that the amino acid sequence of the TCR α chain has at least 90% identity with the amino acid sequence of SEQ ID NO: 14 (e.g., 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity), and the amino acid sequence of the TCR β chain-antiCD3 has at least 90% identity with the amino acid sequence of SEQ ID NO: 16 (e.g., 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity), provided that the variable domain of the TCR α chain comprises CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 3, 4, and 5, respectively, and the variable domain of the TCR β chain comprises CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 9, 10, and 11, respectively.

[0095] "Identity" as known in the art refers to the relationship between two or more polypeptide sequences or two or more polynucleotide sequences, as determined by sequence comparison. In the art, identity also means, as appropriate, the degree of sequence relevance between polypeptide or polynucleotide sequences, as determined by matching strings of such sequences. While many methods exist for measuring the identity between two polypeptide or polynucleotide sequences, methods commonly used to determine identity are coded in computer programs. Preferred computer programs for determining the identity between two sequences include, but are not limited to, the GCG package (Devereux et al., "A comprehensive sequence analysis program for VAX") A comprehensive set of sequence analysis programs for the VAX )”, Nucleic acids Research, 12, 387-95 (1984)), BLASTP, BLASTN and FASTA (Atschul et al., “Basic local alignment search tool”, J. Molec. Biol. 215(3), 403-10 (1990)).

[0096] Amino acid sequences can be compared using programs such as CLUSTAL. This program compares amino acid sequences and finds the optimal alignment by inserting spaces in either sequence where appropriate. Amino acid identity or similarity (identity of amino acid types plus conservation) can be calculated to achieve the optimal alignment. Programs such as BLASTx align the longest extensions of similar sequences and specify the values ​​used for fitting. Therefore, comparisons can be obtained where several similar regions are found, each with a different score. Two types of identity analysis are envisioned in this disclosure.

[0097] The percentage of identity between two amino acid sequences or two nucleic acid sequences is determined by aligning the sequences for optimal comparison purposes (e.g., introducing vacancies in the first sequence to achieve the best alignment) and comparing the amino acid residues or nucleotides at corresponding positions. The “best alignment” is the alignment of the two sequences that produces the highest percentage of identity. The percentage of identity is determined by the number of identical amino acid residues or nucleotides in the compared sequences (i.e., identity % = number of identical positions / total number of positions × 100).

[0098] The determination of the percentage of identity between two sequences can be accomplished using mathematical algorithms known to those skilled in the art. An example of a mathematical algorithm used to compare two sequences is the algorithm modified by Karlin and Altschul (1993) in Proceedings of the National Academy of Sciences (PNAS) 87:2264-2268, as described in Proceedings of the National Academy of Sciences (PNAS) 90:5873-5877. The NBLAST and XBLAST procedures of Altschul et al. (1990) Journal of Molecular Biology 215:403-410 have been incorporated into this algorithm. A BLAST nucleotide search can be performed using the NBLAST procedure (score = 100, word length = 12) to obtain nucleotide sequences homologous to the nucleic acid molecule. A BLAST protein search can be performed using the XBLAST procedure (score = 50, word length = 3) to obtain amino acid sequences homologous to the protein molecule used in this disclosure. To obtain vacancy-containing alignments for comparative purposes, vacancy-containing BLAST can be used as described below: Altschul et al. (1997) Nucleic Acid Research 25:3389-3402. Alternatively, PSI-Blast can be used for iterative searches that detect long-distance relationships between molecules (ibid.). When using BLAST, vacancy-containing BLAST, and PSI-Blast procedures, the default parameters of the respective procedures (e.g., XBLAST and NBLAST) can be used. See http: / / www.ncbi.nlm.nih.gov. Another example of a mathematical algorithm for sequence alignment is the algorithm of Myers and Miller, Computer Applications in the Biological Sciences (CABIOS) (1989). This algorithm has been incorporated into the ALIGN program (version 2.0), which is part of the CGC sequence alignment software package. Other algorithms known in the art for sequence analysis include ADVANCE and ADAM, as described below: Torellis and Robotti (1994), *Computer Applications in Biochemistry*, 10:3-5; and FASTA, as described below: Pearson and Lipman (1988), *Proceedings of the National Academy of Sciences*, 85:2444-8. Within FASTA, ktup is a control option for setting the sensitivity and speed of the search.

[0099] Mutations (including conserved and tolerant substitutions, insertions, and deletions) can be introduced into sequences provided using any suitable method, including but not limited to polymerase chain reaction (PCR) based methods, restriction enzyme-based cloning, or ligation-independent cloning (LIC) procedures. These methods are detailed in many standard molecular biology documents. For further details on polymerase chain reaction (PCR) and restriction enzyme-based cloning, see Sambrook and Russell, (2001) *Molecular Cloning - A Laboratory Manual* (3rd ed.), CSHL Press. Further information on ligation-independent cloning (LIC) procedures can be found in Rashtchian, (1995) *Curr Opin Biotechnol* 6(1):30-6. The TCR sequences provided in this disclosure can be obtained by solid-state synthesis or any other suitable method known in the art.

[0100] The TCR-antiCD3 fusion molecule used in this disclosure has HLA-A affinity for SLLQHLIGL (SEQ ID NO: 1). 02. Complex Binding Characteristics. It has been found that the TCR-anti-CD3 fusion molecule used in this disclosure strongly recognizes this epitope relative to other unrelated epitopes, and is therefore particularly suitable as a targeting vector for delivering therapeutic agents or detectable markers to cells and tissues exhibiting those epitopes. Specificity in the context of the TCR-anti-CD3 fusion molecule used in this disclosure involves its recognition of antigen-positive HLA-A. 02 The ability of target cells to simultaneously recognize antigen-negative HLA-A 02 The target cells have the least capacity.

[0101] The method disclosed herein is a dose-escalation protocol in which an increasing dose of the heterodimeric TCR-antiCD3 fusion molecule is administered to a subject sequentially. Therefore, the doses are administered in a prescribed order: a first dose, then a second dose, and then a maintenance dose. A “first dose” refers to the dose of the TCR-antiCD3 fusion molecule at a first amount within a prescribed range, i.e., the administered dose. A “second dose” refers to the dose of the TCR-antiCD3 fusion molecule at a second amount within a prescribed range, i.e., the administered dose, which is greater than the first dose. A “maintenance dose” refers to the dose of the TCR-antiCD3 fusion molecule at a maintenance amount within a prescribed range, i.e., the administered dose, which is greater than the second dose. It should be understood that, according to the method of this disclosure, the subject will receive at least three total doses (i.e., a first dose, a second dose, and a maintenance dose). However, in some embodiments, the subject may receive more than three total doses because the subject may receive one or more first doses, one or more second doses, and one or more maintenance doses of the heterodimeric TCR-antiCD3 antibody fusion molecule. As used herein, the amount in each second dose is greater than the amount in each first dose, and the amount in each maintenance dose is greater than the amount in each second dose.

[0102] In this disclosure, each dose (i.e., the first dose, the second dose, and the maintenance dose) is expressed as a specified mass, i.e., weight, of the heterodimeric TCR-antiCD3 antibody fusion molecule. Other mediators, excipients, carriers, etc., may be administered with the heterodimeric TCR-antiCD3 antibody fusion molecule but are not included in the calculated mass.

[0103] In some embodiments, the first dose is about 2 μg to about 30 μg. In some embodiments, the first dose is about 2 μg to about 20 μg, about 2 μg to about 10 μg, about 2 μg to about 5 μg, or about 2.5 μg to about 3.5 μg. In some embodiments, the first dose is about 3 μg. In some embodiments, the first dose is about 5 μg to about 25 μg, about 10 μg to about 24 μg, about 15 μg to about 23 μg, or about 18 μg to about 22 μg. In some embodiments, the first dose is about 20 μg. The dosage can be 2 μg, 3 μg, 4 μg, 5 μg, 6 μg, 7 μg, 8 μg, 9 μg, 10 μg, 11 μg, 12 μg, 13 μg, 14 μg, 15 μg, 16 μg, 17 μg, 18 μg, 19 μg, 20 μg, 21 μg, 22 μg, 23 μg, 24 μg, 25 μg, 26 μg, 27 μg, 28 μg, 29 μg, or 30 μg. In some embodiments, the first dosage is 3 μg, 10 μg, or 20 μg. In some embodiments, the first dosage is 3 μg. In some embodiments, the first dosage is 20 μg.

[0104] The first dose can be administered to the subject in various quantities. For example, in some embodiments, the first dose is administered once, i.e., once (one time). In some embodiments, the first dose is administered more than once, such as two to ten times, two to five times, or two to three times. In some embodiments, the first dose is administered twice.

[0105] The number of first doses can be determined, for example, by the occurrence or severity of adverse events (AEs) following administration of the first dose. In the presence of adverse events, the method may include administering more than one first dose before escalating to a second dose. In some embodiments, the first dose is a “range” of amount, such as 2 μg to 30 μg. Thus, in some embodiments in which multiple first doses are administered, the first dose may contain the same amount in each first dose (e.g., each first dose contains 2 μg). In some embodiments in which multiple first doses are administered, the first dose may contain different amounts in one or more doses within each first dose (e.g., first dose #1 contains 2 μg and first dose #2 contains 4 μg), as long as each dose is within the prescribed first dose.

[0106] In some embodiments, the second dose is about 5 μg to about 60 μg. In some embodiments, the second dose is about 5 μg to about 60 μg, about 6 μg to about 30 μg, about 7 μg to about 20 μg, about 8 μg to about 15 μg, or about 8 μg to about 12 μg. In some embodiments, the second dose is about 10 μg. In some embodiments, the second dose is about 10 μg to about 60 μg, about 20 μg to about 55 μg, about 30 μg to about 50 μg, about 35 μg to about 45 μg, or about 38 μg to about 42 μg. In some embodiments, the second dose is about 40 μg. The second dose can be 5 μg, 10 μg, 15 μg, 20 μg, 25 μg, 30 μg, 35 μg, 40 μg, 45 μg, 50 μg, 55 μg, or 60 μg. In some embodiments, the second dose is about 10 μg, 20 μg, 40 μg, or 60 μg. In some embodiments, the second dose is 10 μg. In some embodiments, the second dose is 40 μg.

[0107] A variety of numbers of second doses may be administered to the subject. For example, in some embodiments, the second dose is administered once, i.e., once. In some embodiments, the second dose is administered more than once, such as two to ten times, two to five times, or two to three times. In some embodiments, the second dose is administered twice.

[0108] In some embodiments, the method may include administering more than one second dose before escalating to a maintenance dose. In some embodiments, the second dose is a “range” of amount, such as 5 μg to 60 μg. Thus, in some embodiments in which multiple second doses are administered, the second dose may contain the same amount in each second dose (e.g., each second dose contains 10 μg). In some embodiments in which multiple second doses are administered, the second dose may contain different amounts in one or more doses within each second dose (e.g., second dose #1 contains 10 μg and second dose #2 contains 15 μg), as long as each dose is within the prescribed second dose range.

[0109] The term "maintenance dose" refers to the dose administered to a subject to maintain the desired therapeutic effect. In some embodiments, the maintenance dose is repeated until treatment is discontinued.

[0110] In some embodiments, the maintenance dose is about 15 μg to about 200 μg. In some embodiments, the maintenance dose is about 20 μg to about 100 μg, about 25 μg to about 50 μg, about 30 μg to about 45 μg, about 35 μg to about 45 μg, or about 38 μg to about 42 μg. In some embodiments, the maintenance dose is about 40 μg. In some embodiments, the maintenance dose is about 100 μg to about 200 μg, about 120 μg to about 180 μg, about 150 μg to about 170 μg, about 155 μg to about 165 μg, or about 158 ​​μg to about 1622 μg. In some embodiments, the maintenance dose is about 160 μg. The maintenance dose can be 25 μg, 30 μg, 35 μg, 40 μg, 45 μg, 50 μg, 55 μg, 60 μg, 120 μg, 130 μg, 140 μg, 150 μg, 160 μg, 170 μg, or 180 μg. In some embodiments, the maintenance dose is about 40 μg, 50 μg, 150 μg, or 160 μg. In some embodiments, the maintenance dose is 20 μg. At doses of 20 μg or higher, the applicant has observed durable partial responses (PR) in several solid tumors, including cutaneous melanoma (CM), uveal melanoma (UM), and ovarian cancer. In some embodiments, the maintenance dose is 40 μg. In some embodiments, the maintenance dose is 160 μg. The applicant observed that 160 μg was 1) the lowest dose at which exposure was detected throughout the dosing period, and 2) the incidence of cytokine release syndrome (CRS) was lower with 160 μg compared to the next higher dose tested (320 μg), and less preoperative dexamethasone administration was required.

[0111] Various numbers of maintenance doses may be administered to the subject. For example, in some embodiments, a maintenance dose is administered three times, for example, once in each of three prescribed phases. In some embodiments, a maintenance dose is administered more than once, for example, three to 200 times, 10 to 150 times, or 30 to 80 times. In some embodiments, a maintenance dose is administered 30 to 34 times, for example, 32 times. In some embodiments, the maintenance dose is administered until the melanoma (e.g., cutaneous melanoma) is cured, in remission, or otherwise determined by a healthcare professional. Treatment may be discontinued, for example, due to the occurrence of unacceptable toxicity or adverse events or because the subject has shown an unacceptable level of disease progression. Alternatively, treatment may be discontinued, for example, because the severity of the subject's symptoms has decreased and / or the tumor has shrunk to a level where it is considered no longer necessary to treat with TCR-antiCD3 fusion molecules.

[0112] In some embodiments, the method may include administering more than one maintenance dose. In some embodiments, the maintenance dose is a “range” of amount, such as 40 μg to 160 μg. Thus, in some embodiments where multiple maintenance doses are administered, each maintenance dose may contain the same amount (e.g., each maintenance dose contains 40 μg). In some embodiments, the maintenance dosage may contain different amounts in one or more doses within each maintenance dose (e.g., maintenance dose #1 contains 10 μg and maintenance dose #2 contains 15 μg), as long as each dose is within the prescribed maintenance dosage range.

[0113] The method disclosed herein provides a dosing regimen in which a heterodimeric TCR-antiCD3 antibody fusion molecule is administered in a prescribed amount. In some embodiments, when the first dose is administered multiple times, the first dose is administered every 4 to 14 days or every 6 to 8 days until each of the first doses has been administered. In some embodiments, when the first dose is administered multiple times, the first dose is administered every 7 days until each of the first doses has been administered.

[0114] In some embodiments, when the second dose is administered multiple times, it is administered every 4 to 14 days or every 6 to 8 days until each of the second doses has been administered. In some embodiments, when the second dose is administered multiple times, it is administered every 7 days until each of the second doses has been administered. In some embodiments, the second dose is administered 4 to 14 days or 6 to 8 days after the last administration of the first dose. In some embodiments, the second dose is administered 7 days after the last administration of the first dose.

[0115] In some embodiments, the maintenance dose is first administered 7 days after the last administration of the second dose. The maintenance dose may be administered in three phases: Phase 1, Phase 2, and Phase 3. As used herein, the term "phase" refers to a period of time in which the frequency of maintenance dose administration is substantially consistent. The term "substantially consistent" means that the frequency deviates from a given value within ±24 hours. For example, if the phase frequency is "once every 7 days," then a frequency with dosing intervals of 7 days ± 24 hours would belong to the same phase. In some embodiments, "Phase 1" may consist of maintenance dose administration every 6 to 8 days, "Phase 2" may consist of maintenance dose administration every 12 to 16 days, and / or "Phase 3" may consist of maintenance dose administration every 26 to 30 days. In some embodiments, "Phase 1" may consist of maintenance dose administration every 7 days (i.e., once a week), "Phase 2" may consist of maintenance dose administration every 14 days (i.e., every 2 weeks), and / or "Phase 3" may consist of maintenance dose administration every 28 days (i.e., every 4 weeks).

[0116] While not bound by any particular theory, in some embodiments, reducing the frequency of maintenance dose reduction from phase one to phase two and from phase two to phase three can result in a reduction in the number and severity of side effects and improved patient compliance. In some embodiments, reducing the frequency of maintenance dose reduction from phase one to phase two and from phase two to phase three does not reduce the efficacy of the heterodimeric TCR-antiCD3 antibody fusion molecule. Reducing the frequency of phase two and phase three (relative to phase one and phase two, respectively) can reduce overall costs and the time required for treatment for subjects and healthcare professionals administering the treatment.

[0117] In some embodiments, the duration of the first phase of maintenance dose is about one week to about 20 weeks, or about 4 weeks to about 15 weeks, or about 8 weeks to about 12 weeks. In some embodiments, the duration of the first phase of maintenance dose is about 10 weeks. In some embodiments, the duration of the second phase of maintenance dose is about 4 weeks to about 40 weeks, or about 10 weeks to about 30 weeks, or about 16 weeks to about 24 weeks. In some embodiments, the duration of the second phase of maintenance dose is about 20 weeks. In some embodiments, the duration of the third phase of maintenance dose is about 10 weeks to about 80 weeks, or about 40 weeks to about 60 weeks, or about 48 weeks to about 54 weeks. In some embodiments, the third phase of maintenance dose is about 40 weeks to about 60 weeks. In some embodiments, the third phase of maintenance dose is about 48 weeks.

[0118] In some embodiments, the first phase lasts from 8 to 14 weeks, the second phase lasts from about 30 to 50 weeks, and the maintenance phase lasts from about 40 to 60 weeks. In some embodiments, the first phase lasts from about 10 weeks, the second phase lasts from about 38 weeks, and / or the maintenance phase lasts from about 48 weeks.

[0119] In some embodiments, the dosing regimen comprising administering a first dose, a second dose, and a maintenance dose (all three phases) is administered for at least 50 weeks, at least 60 weeks, at least 70 weeks, at least 80 weeks, or at least 90 weeks. In some embodiments, the dosing regimen comprising administering a first dose, a second dose, and a maintenance dose (all three phases) is administered for about 90 weeks to about 110 weeks. In some embodiments, the dosing regimen comprising administering a first dose, a second dose, and a maintenance dose (all three phases) is administered for about 100 weeks to about 105 weeks. In some embodiments, the dosing regimen comprising administering a first dose, a second dose, and a maintenance dose (all three phases) is administered for about 101 weeks.

[0120] In some embodiments, a first dose is administered for 1 to 2 weeks, a second dose is administered for 1 to 2 weeks, and a maintenance dose is administered for at least 40 weeks, such as 40 to 156 weeks, 80 to 120 weeks, or 100 to 104 weeks. In some embodiments, a first dose is administered for 1 week, a second dose is administered for 1 week, and a maintenance dose is administered for at least 40 weeks.

[0121] In some embodiments, this disclosure provides low-dose regimens, wherein a first dose is about 2 μg to 4 μg, a second dose is about 5 μg to 14 μg, and a maintenance dose is about 15 μg to 55 μg. Examples of these low-dose regimens can be found in […]. Figure 7 middle.

[0122] In some embodiments, this disclosure provides high-dose regimens, wherein a first dose is about 15 μg to 40 μg, a second dose is about 30 μg to 80 μg, and a maintenance dose is about 140 μg to 200 μg. Examples of these low-dose regimens can be found in […]. Figure 8 middle.

[0123] In some embodiments, the methods described herein are first-line treatments. The term "first-line treatment" refers to the initial or first-line treatment recommended for a disease (or illness) (e.g., lung cancer or ovarian cancer). In some embodiments, this may also be referred to as primary treatment, initial treatment, or induction therapy. In some embodiments, first-line treatment may include combination therapy, such as treatment performed using another method (e.g., another active agent and / or surgery).

[0124] In some embodiments, the methods described herein are second-line treatments. In some embodiments, the methods described herein are third-line, fourth-line, fifth-line, sixth-line, etc., treatments. Second-line or more lines of therapy (third-line, fourth-line, seventh-line, etc.) may be used for several different reasons, such as first-line treatment being ineffective, first-line treatment being effective but ceasing to be effective thereafter, first-line treatment having intolerable side effects, and / or a new treatment that appears to be more effective than the current treatment becoming available.

[0125] In some embodiments, the methods described herein are for ovarian cancer patients who have relapsed after first-line therapy or induction therapy (e.g., chemotherapy) and are platinum-sensitive or platinum-resistant ovarian cancer patients. In such patients, in some embodiments, the method comprises treating platinum-sensitive or platinum-resistant ovarian cancer patients by administering a combination of a TCR-anti-CD3 antibody fusion molecule (such as a TCR-anti-CD3 antibody fusion molecule targeting PRAME, wherein such a molecule corresponds to, for example, IMC-F106C) with a chemotherapy agent or targeted therapy. In some embodiments, IMC-F106C comprises: The TCR α chain amino acid sequence of SEQ ID NO: 14 or a TCR α chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 14, and The TCR β-chain anti-CD3 amino acid sequence of SEQ ID NO: 16 or the TCR β-chain anti-CD3 amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 16. The TCR α-chain variable domain comprises CDR1, 2, and 3 having amino acid sequences of SEQ ID NO: 3, 4, and 5, respectively, and the TOR β-chain variable domain comprises CDR1, 2, and 3 having amino acid sequences of SEQ ID NO: 9, 10, and 11, respectively.

[0126] In some embodiments, the methods described herein are first-line monotherapy, i.e., the sole active agent for treating cutaneous melanoma is a heterodimeric TCR-anti-CD3 antibody fusion molecule. In some embodiments, corticosteroids are not administered prior to the administration of the heterodimeric TCR-anti-CD3 antibody fusion molecule. In some embodiments, corticosteroids are not administered 3 months, 2 months, 1 month, 3 weeks, 2 weeks, or 1 week prior to the administration of the heterodimeric TCR-anti-CD3 antibody fusion molecule. In some embodiments, corticosteroids are not administered 3 weeks prior to the administration of the heterodimeric TCR-anti-CD3 antibody fusion molecule.

[0127] In some embodiments, the methods described herein are dual therapies, such as first-line dual therapy, wherein a second active agent is co-administered during the dosing regimen, wherein the heterodimeric TCR-antiCD3 antibody fusion molecule is the first active agent. In some embodiments, the methods described herein are dual therapies, such as first-line dual therapy, wherein a second active agent, such as an anticancer drug, is co-administered 3 months, 2 months, 1 month, 3 weeks, 2 weeks, or 1 week prior to the administration of the heterodimeric TCR-antiCD3 antibody fusion molecule. In some embodiments, the methods described herein are dual therapies, such as first-line dual therapy, wherein a second active agent, such as an anticancer drug, is co-administered 3 months, 2 months, 1 month, 3 weeks, 2 weeks, or 1 week after completion of the dosing regimen. In embodiments, the second active agent is a chemotherapeutic agent (e.g., gemcitabine, decitabine, pegylated liposomal doxorubicin (PLD), or albumin-bound paclitaxel) or a targeted therapy (e.g., bevacizumab) or a multimodal therapy (e.g., a chemotherapeutic agent and a targeted therapy). In some embodiments, the methods described herein are not monotherapy, i.e., the methods comprise the administration of a heterodimeric TCR-antiCD3 antibody fusion molecule and one or more additional active agents, such as anticancer therapeutic agents.

[0128] In some embodiments, the second TCR-antiCD3 antibody fusion molecule comprises terbentufsp. In some embodiments, the second TCR-antiCD3 antibody fusion molecule is administered in a first, second, and third phase. In some embodiments, the second TCR-antiCD3 antibody fusion molecule is administered using an escalating dosing regimen identical or similar to that of the first heterodimeric TCR-antiCD3 antibody fusion molecule disclosed herein. In some embodiments, the second TCR-antiCD3 antibody fusion molecule comprises terbentufsp, which is administered at 20 mcg in the first week, 30 mcg in the second week, and 68 mcg weekly thereafter in the third week.

[0129] In some embodiments, this disclosure provides for administering the heterodimeric TCR-antiCD3 antibody fusion molecule described herein together with a second active agent (wherein the heterodimeric TCR-antiCD3 antibody fusion molecule is considered herein as a first active agent). The terms "active agent" and "therapeutic agent" are used interchangeably throughout the application. In some embodiments, this disclosure provides a dosing regimen that further comprises administering a third active agent, a fourth active agent, or more than four active agents. In some embodiments, the dosing regimen comprises administering a second active agent. In some embodiments, the second, third, fourth, or more than four active agents may be anticancer therapeutic agents. While not bound by any particular theory, this disclosure provides that, in some embodiments, administering a second active agent may enhance the activity of the heterodimeric TCR-antiCD3 antibody fusion molecule. In some embodiments, the effect of the second active agent is not additive but synergistic. In the embodiments, the second active agent is a chemotherapeutic agent (e.g., gemcitabine, decitabine, pegylated liposomal doxorubicin (PLD), or albumin-bound paclitaxel) or a targeted therapy (e.g., bevacizumab) or a multimodal therapy (e.g., a chemotherapeutic agent and a targeted therapy).

[0130] In some embodiments, a heterodimeric TCR-antiCD3 antibody fusion molecule and a second active agent are administered according to a protocol, wherein the heterodimeric TCR-antiCD3 antibody fusion molecule is administered as described herein, and the second active agent (i.e., the combination chaperone) is administered as described herein. Figure 9As outlined in [the document]. In some cases, a second active agent is administered before initiating treatment with the heterodimeric TCR-antiCD3 antibody fusion molecule. In some embodiments, the administration of the second active agent is completed before the first dose of the heterodimeric TCR-antiCD3 antibody fusion molecule. In some embodiments, the second active agent is administered before initiating treatment with the heterodimeric TCR-antiCD3 antibody fusion molecule, and then the second active agent is continued while the heterodimeric TCR-antiCD3 antibody fusion molecule is also administered. The second active agent and the heterodimeric TCR-antiCD3 antibody fusion molecule can be administered in parallel, for example, during the same regimen. In some embodiments, when administering the heterodimeric TCR-antiCD3 antibody fusion molecule, the second active agent follows [the procedure described in the document]. Figure 9 The regimen seen herein. In some embodiments, the second active agent and the heterodimeric TCR-antiCD3 antibody fusion molecule are administered on the same day (when the dosing regimen allows for the administration of both active agents on the same day). For example, the second active agent and the heterodimeric TCR-antiCD3 antibody fusion molecule may be administered sequentially on the same day.

[0131] In some cases, the application of the second active agent begins simultaneously with the application of the heterodimeric TCR-antiCD3 antibody fusion molecule. In some embodiments, the application of the second active agent is stopped before the start of the heterodimeric TCR-antiCD3 antibody fusion molecule protocol. In some embodiments, the application of the second active agent is stopped at some point during the heterodimeric TCR-antiCD3 antibody fusion molecule protocol. In some embodiments, the application of the second active agent and the application of the heterodimeric TCR-antiCD3 antibody fusion molecule are stopped simultaneously.

[0132] In some embodiments, when the heterodimeric TCR-antiCD3 antibody fusion molecule and the second active agent are administered on the same day, the heterodimeric TCR-antiCD3 antibody fusion molecule and the second active agent are not administered simultaneously, for example, through the same intravenous line or at exactly the same time.

[0133] In some embodiments, the second active agent comprises a chemotherapeutic agent, a checkpoint inhibitor, a second (i.e., different) heterodimeric TCR-antiCD3 antibody fusion molecule, a targeted therapeutic agent, a demethylating agent, and / or a multimodal therapy. In some embodiments, the second active agent comprises a checkpoint inhibitor. In some embodiments, the second active agent comprises a checkpoint inhibitor. In some embodiments, the second active agent comprises a demethylating agent. In some embodiments, the second active agent comprises a second TCR-antiCD3 antibody fusion molecule.

[0134] In some embodiments, the second active agent is a chemotherapeutic agent. In some embodiments, the second active agent is a biological agent, such as a PD-1 inhibitor, a T-cell inactivator, a BRAF inhibitor, or a MEK inhibitor. In some embodiments, the second active agent comprises a checkpoint inhibitor. In some embodiments, the second active agent is a demethylating agent. In some embodiments, the chemotherapeutic agent is a demethylating agent. In some embodiments, the second active agent is a second TCR-anti-CD3 antibody fusion molecule.

[0135] In some embodiments, the second or more active agents are chemotherapeutic agents. In some embodiments, the chemotherapeutic agent comprises gemcitabine, albumin-bound paclitaxel, pegylated liposomal doxorubicin (PLD), docetaxel, carboplatin, paclitaxel, doxorubicin, pemetrexed, or combinations thereof. In some embodiments, the chemotherapeutic agent is paclitaxel. In some embodiments, the chemotherapeutic agent is PLD. In some embodiments, the chemotherapeutic agent is carboplatin. In some embodiments, the chemotherapeutic agent is docetaxel. In some embodiments, the chemotherapeutic agent is pemetrexed. In some embodiments, the chemotherapeutic agent is doxorubicin. In some embodiments, the chemotherapeutic agent is carboplatin / paclitaxel. Gemcitabine is a nucleoside metabolism inhibitor suitable as both a monotherapy and combination therapy for various cancers. Albumin-bound paclitaxel is a second-generation semi-synthetic taxane conjugated to albumin, which inhibits microtubule depolymerization, thereby causing cell cycle arrest. PLD is an anthracycline topoisomerase inhibitor suitable for combination therapy for various cancers.

[0136] In some embodiments, the chemotherapeutic agent comprises gemcitabine or albumin-bound paclitaxel. Any of the listed chemotherapeutic agents may be administered, for example, in the amounts and regimens outlined in their respective prescribing information.

[0137] In some embodiments, a chemotherapy agent is co-administered with a heterodimeric TCR-anti-CD3 antibody fusion molecule. In some embodiments, the chemotherapy agent is administered 1 to 12 weeks, 1 to 10 weeks, 1 to 8 weeks, or 1 to 6 weeks prior to the administration of the heterodimeric TCR-anti-CD3 antibody fusion molecule. In some embodiments, the chemotherapy agent is administered 1, 2, 3, 4, 5, or 6 weeks prior to the administration of the heterodimeric TCR-anti-CD3 antibody fusion molecule. In some embodiments, the chemotherapy agent is administered 1 to 6 weeks after the administration of the heterodimeric TCR-anti-CD3 antibody fusion molecule. In some embodiments, the chemotherapy agent is administered 1, 2, 3, 4, 5, or 6 weeks after the administration of the heterodimeric TCR-anti-CD3 antibody fusion molecule. In some embodiments, the heterodimeric TCR-anti-CD3 antibody fusion molecule is administered after treatment with the chemotherapy agent has been completed.

[0138] In some embodiments, the chemotherapy agent dose comprises approximately 10 mg / m². 2 Approximately 1000 mg / m 2 In some embodiments, the chemotherapy agent dose comprises about 10 mg / m². 2 In some embodiments, the chemotherapy agent dose comprises less than about 10 mg / m². 2 In some embodiments, the chemotherapy agent dose comprises greater than about 10 mg / m². 2 In some embodiments, the chemotherapy agent dose comprises greater than about 1000 mg / m². 2 In some embodiments, the chemotherapy agent dose comprises less than about 1000 mg / m². 2 In some embodiments, the chemotherapy agent dose comprises approximately 1000 mg / m². 2 In some embodiments, the chemotherapy agent dose comprises approximately 100 mg / m². 2 In some embodiments, the chemotherapy agent dose comprises from AUC 1 to approximately AUC 10.

[0139] In some embodiments, the chemotherapy agent is administered once a week (QW) to once a month. In some embodiments, the chemotherapy agent is administered once every 2 weeks (Q2W). In some embodiments, the chemotherapy agent is administered once every 3 weeks (Q3W). In some embodiments, the chemotherapy agent is administered once every 4 weeks (Q4W). In some embodiments, the chemotherapy agent is administered once every 5 weeks (Q5W). In some embodiments, the chemotherapy agent is administered once every 6 weeks (Q6W). In some embodiments, the chemotherapy agent is administered once a week and then stopped for three weeks; administered once a week for three weeks and then stopped for one week; administered once a week for two weeks and then stopped for two weeks; administered once a week for two weeks and then stopped for one week; administered every other week; administered once a week and then stopped for two weeks; administered once a week and then stopped for four weeks; administered once a week for two weeks and then stopped for three weeks; administered once a week for three weeks and then stopped for two weeks; administered once a week for four weeks and then stopped for one week; administered once a week and then stopped for five weeks; administered once a week for two weeks and then stopped for four weeks; administered once a week for three weeks and then stopped for three weeks; administered once a week for four weeks and then stopped for two weeks; administered once a week for five weeks and then stopped for one week.

[0140] In some embodiments, the chemotherapy agent is administered for at least 10 weeks. In some embodiments, the chemotherapy agent is administered for at least 40 weeks. In some embodiments, the chemotherapy agent is administered for the duration of the dosing regimen.

[0141] In some embodiments, the second active agent comprises: atezolizumab (Tecentriq®), avelumab (Bavencio®), bevacizumab (Avastin®), binimetinib (Mektovi®), encorafenib (Braftovi®), cobimetinib fumarate (Cotellic®), dabrafenib (e.g., dabrafenib mesylate (Tafmlar®)), dacarbazine, durvalumab (Imfinzi®), galunisertib, gemcitabine (Gemzar®), interferon, talimogene laherparepvec (Imlygic®), and recombinant interferon α-2b (Intron®). A) Pembrolizumab (Keytruda®), tebentafusp-tebn (Kimmtrak®), trametinib dimethylsulfoxide (Mekinist®), merestinib, nivolumab (Opdivo®), nivolumab and relatlimab-rmbw (Opdualag®), osimertinib (Tagrisso®), aldesleukin (Proleukin®), tremelimumab (Imjudo®), ipilimumab (Yervoy®), vemurafenib (Zelboraf®), or combinations thereof. In some embodiments, the second active agent is bevacizumab. Any of the listed second active agents may be administered as outlined in their prescribing information.

[0142] In some embodiments, the second active agent comprises: atezolizumab (TECENTRIQ®), avelumumab (BAVENCIO®), bevacizumab (Avastin®), bimetinib, braftovi (encofenib), cobimethinib fumarate (Cotellic®), dabrafenib (e.g., dabrafenib mesylate (Tafmlar®)), dacarbazine, duvalumab (IMFINZI®), encofenib, ganeserti, gemcitabine, interferon, talimogene laherparepvec (Imlygic®), recombinant interferon alpha-2b (Intron®). A) Ipilimumab, pembrolizumab (Keytruda®), tebn bromide (Kimmtrak®), trametinib (Mekinist®), bimetinib (Mektovi®), mesaltinib, nivolumab (Opdivo®), nivolumab and renalalimab (Opdualag®), osimertinib, interleukin (Proleukin®), trimemumab, vemurafenib, ipilimumab (Yervoy®), vemurafenib (Zelboraf®), or combinations thereof. In some embodiments, the second active agent comprises osimertinib. In some embodiments, the checkpoint inhibitor comprises bevacizumab. Any of the listed second active agents (e.g., checkpoint inhibitors) may be administered as outlined in their prescribing information.

[0143] In some embodiments, the second active agent comprises a checkpoint inhibitor. In some embodiments, the checkpoint inhibitor comprises: atezolizumab (TECENTRIQ®), avelumab (BAVENCIO®), durvalumab (IMFINZI®), pembrolizumab (Keytruda), nivolumab (Opdivo®), nivolumab and renalalimab-rmbw (Opdualag®), trimemumab (hnjudo®), ipilimumab (Yervoy®), or combinations thereof. In some embodiments, the checkpoint inhibitor comprises atezolizumab, avelumab, durvalumab, ipilimumab, pembrolizumab, nivolumab, or combinations thereof. In some embodiments, the checkpoint inhibitor comprises pembrolizumab. In some embodiments, the checkpoint inhibitor is bevacizumab. In some embodiments, the checkpoint inhibitor is nivolumab. Any of the listed checkpoint inhibitors may be administered as outlined in their prescribing information.

[0144] In some embodiments, a checkpoint inhibitor is co-administered with a heterodimeric TCR-anti-CD3 antibody fusion molecule. In some embodiments, the checkpoint inhibitor is administered 1 to 6 weeks prior to the administration of the heterodimeric TCR-anti-CD3 antibody fusion molecule. In some embodiments, the checkpoint inhibitor is administered 1, 2, 3, 4, 5, or 6 weeks prior to the administration of the heterodimeric TCR-anti-CD3 antibody fusion molecule. In some embodiments, the checkpoint inhibitor is administered 1 to 6 weeks after the administration of the heterodimeric TCR-anti-CD3 antibody fusion molecule. In some embodiments, the checkpoint inhibitor is administered 1, 2, 3, 4, 5, or 6 weeks after the administration of the heterodimeric TCR-anti-CD3 antibody fusion molecule. In some embodiments, the heterodimeric TCR-anti-CD3 antibody fusion molecule is administered after treatment with the checkpoint inhibitor has been completed.

[0145] In some embodiments, the checkpoint inhibitor dose comprises about 10 mcg to about 500 mg. In some embodiments, the checkpoint inhibitor dose comprises about 10 mcg. In some embodiments, the checkpoint inhibitor dose comprises about 20 mcg. In some embodiments, the checkpoint inhibitor dose comprises about 30 mcg. In some embodiments, the checkpoint inhibitor dose comprises about 68 mcg. In some embodiments, the checkpoint inhibitor dose comprises about 500 mg. In some embodiments, the checkpoint inhibitor dose comprises about 400 mg. In some embodiments, the checkpoint inhibitor dose comprises about 200 mg. In some embodiments, the checkpoint inhibitor dose comprises about 800 mg. In some embodiments, the checkpoint inhibitor dose comprises about 1 mg / kg to about 100 mg / kg. In some embodiments, the checkpoint inhibitor dose comprises about 1 mg / kg. In some embodiments, the checkpoint inhibitor dose comprises about 100 mg / kg. In some embodiments, the checkpoint inhibitor dose comprises about 15 mg / kg.

[0146] In some embodiments, checkpoint inhibitors are administered once a week (QW) to once a month. In some embodiments, checkpoint inhibitors are administered every 2 weeks (Q2W). In some embodiments, checkpoint inhibitors are administered every 3 weeks (Q3W). In some embodiments, checkpoint inhibitors are administered every 4 weeks (Q4W). In some embodiments, checkpoint inhibitors are administered every 5 weeks (Q5W). In some embodiments, checkpoint inhibitors are administered every 6 weeks (Q6W). In some embodiments, the checkpoint inhibitor is administered once a week and then stopped for three weeks; once a week for three weeks and then stopped for one week; once a week for two weeks and then stopped for two weeks; once a week for two weeks and then stopped for one week; every other week; once a week and then stopped for two weeks; once a week and then stopped for four weeks; once a week for two weeks and then stopped for three weeks; once a week for three weeks and then stopped for two weeks; once a week for four weeks and then stopped for one week; once a week and then stopped for five weeks; once a week for two weeks and then stopped for four weeks; once a week for three weeks and then stopped for three weeks; once a week for four weeks and then stopped for two weeks; once a week for five weeks and then stopped for one week.

[0147] In some embodiments, checkpoint inhibitors are administered for at least 10 weeks. In some embodiments, checkpoint inhibitors are administered for at least 40 weeks. In some embodiments, checkpoint inhibitors are administered for the duration of the dosing regimen.

[0148] In some embodiments, the second active agent comprises a demethylating agent. In some embodiments, the demethylating agent comprises: azacitidine, decitabine, cytidine, cedazuridine, guadecitabine, 5-fluoro-2'-deoxycytidine, zebularine, CP-4200, RG108, nanaomycin A, or any combination or modification thereof. In some embodiments, the demethylating agent comprises decitabine. Any of the listed demethylating agents may be administered as outlined in their prescribing information.

[0149] In some embodiments, the demethylating agent is administered co-administered with the heterodimeric TCR-antiCD3 antibody fusion molecule. In some embodiments, the demethylating agent is administered 1 to 6 weeks prior to the administration of the heterodimeric TCR-antiCD3 antibody fusion molecule. In some embodiments, the demethylating agent is administered 1, 2, 3, 4, 5, or 6 weeks prior to the administration of the heterodimeric TCR-antiCD3 antibody fusion molecule. In some embodiments, the demethylating agent is administered 1 to 6 weeks after the administration of the heterodimeric TCR-antiCD3 antibody fusion molecule. In some embodiments, the demethylating agent is administered 1, 2, 3, 4, 5, or 6 weeks after the administration of the heterodimeric TCR-antiCD3 antibody fusion molecule. In some embodiments, the heterodimeric TCR-antiCD3 antibody fusion molecule is administered after treatment with the demethylating agent has been completed.

[0150] In some embodiments, the demethylating agent dosage comprises about 0.01 mg / kg to about 1 mg / kg. In some embodiments, the demethylating agent dosage comprises about 0.01 mg / kg. In some embodiments, the demethylating agent dosage comprises about 0.07 mg / kg. In some embodiments, the demethylating agent dosage comprises about 0.2 mg / kg. In some embodiments, the demethylating agent dosage comprises about 0.6 mg / kg. In some embodiments, the demethylating agent dosage comprises about 20 mg / kg. 2 / day up to 45 mg / m 2 / day. In some embodiments, the demethylating agent dosage comprises more than 20 mg / m². 2 / day up to 45 mg / m 2 / Day dose reduced by 75% to 90%.

[0151] In some embodiments, the demethylating agent is applied once a week (QW) to once a month. In some embodiments, the demethylating agent is applied every 2 weeks (Q2W). In some embodiments, the demethylating agent is applied every 3 weeks (Q3W). In some embodiments, the demethylating agent is applied every 4 weeks (Q4W). In some embodiments, the demethylating agent is applied every 5 weeks (Q5W). In some embodiments, the demethylating agent is applied every 6 weeks (Q6W). In some embodiments, the demethylating agent is applied once a week.

[0152] In some embodiments, the demethylating agent is administered for at least 10 weeks. In some embodiments, the demethylating agent is administered for at least 40 weeks. In some embodiments, the demethylating agent is administered for the duration of the dosing regimen.

[0153] In some embodiments, the second active agent comprises a “targeted therapeutic agent.” In some embodiments, the targeted therapeutic agent is an antibody, such as a monoclonal antibody. In some embodiments, the targeted therapeutic agent comprises osimertinib, bevacizumab, dabrafenib, and trametinib, or a combination thereof, such as dabrafenib / trametinib. In some embodiments, the targeted therapeutic agent comprises osimertinib. In some embodiments, the targeted therapeutic agent comprises trametinib. In some embodiments, the targeted therapeutic agent recognizes VEGF, such as VEGF-A or ANGPT2. In some embodiments, the targeted therapeutic agent directly or indirectly blocks the pro-angiogenic pathway. Any of the listed commercial targeted therapeutic agents may be administered as outlined in their prescribing information.

[0154] In some embodiments, the targeted therapy dose comprises a targeted therapy of about 0.01 mg / kg to about 100 mg / kg, about 1 mg / kg to about 50 mg / kg, about 5 mg / kg to about 25 mg / kg, about 10 mg / kg to about 20 mg / kg, or about 15 mg / kg, such as bevacizumab. In some embodiments, the targeted therapy dose comprises a targeted therapy of about 0.1 mg / day to about 500 mg / day, about 1 mg / day to about 250 mg / day, about 10 mg / day to about 200 mg / day, about 50 mg / day to about 150 mg / day, about 60 mg / day to about 100 mg / day, or about 80 mg / day, such as osimertinib. In some embodiments, the targeted therapeutic dose comprises a targeted therapeutic agent of about 50 mg / day to about 500 mg / day, about 150 mg / day to about 400 mg / day, about 200 mg / day to about 350 mg / day, about 550 mg / day to about 350 mg / day, or about 300 mg / day, such as dabrafenib.

[0155] In some embodiments, the targeted therapy is administered once a week (QW) to once a month. In some embodiments, the targeted therapy is administered once every 2 weeks (Q2W). In some embodiments, the targeted therapy is administered once every 3 weeks (Q3W). In some embodiments, the targeted therapy is administered once every 4 weeks (Q4W). In some embodiments, the targeted therapy is administered once every 5 weeks (Q5W). In some embodiments, the targeted therapy is administered once every 6 weeks (Q6W). In some embodiments, the targeted therapy is administered once a week. In some embodiments, the targeted therapy is administered once daily. In some embodiments, the targeted therapy is administered twice daily.

[0156] In some embodiments, the second active agent comprises a second heterodimeric TCR-antiCD3 antibody fusion molecule, such as terbentuft. Terbentuft is a bispecific gp100 peptide-HLA-directed CD3 T cell connector suitable for the treatment of HLA-A inflammatory tumors with unresectable or metastatic uveal melanoma. 02:01 Positive adult patient.

[0157] In some embodiments, the second heterodimeric TCR-antiCD3 antibody fusion molecule is administered at 10-40 mcg on day 1, 20-50 mcg on days 6-10, 50-75 mcg on days 13-18, and thereafter at 50-80 mcg weekly. In some embodiments, the second heterodimeric TCR-antiCD3 antibody fusion molecule is administered intravenously at 20 mcg on day 1, 30 mcg on day 8, 68 mcg on day 15, and thereafter at 68 mcg weekly. Any commercially available second heterodimeric TCR-antiCD3 antibody fusion molecule can be administered as outlined in its prescribing information.

[0158] In some embodiments, the second active agent is pegylated liposomal doxorubicin (PLD). PLD can be administered according to its prescription information. In some embodiments, PLD is administered at approximately 10 mg / m² per week. 2 For example, approximately 20 mg / m² every two weeks. 2 30 mg / m² every three weeks 2 40 mg / m² every 4 weeks 2 50 mg / m² every 5 weeks 2 Or 60 mg / m² every 6 weeks 2 The dose intensity is administered. In some embodiments, approximately 40 mg / m² is administered to the subject every 4 to 6 weeks. 2 Up to 60 mg / m 2 Or administer approximately 40 mg / m² to the subject every 4 weeks. 2 .

[0159] In some embodiments, the second active agent comprises a multimodal therapy, such as a combination of second active agents described herein. In some embodiments, the multimodal therapy may comprise a biologic and a chemotherapeutic agent as described herein. For example, in some embodiments, the multimodal therapy may comprise: a chemotherapeutic agent and a checkpoint inhibitor, a chemotherapeutic agent and a targeted therapy, a chemotherapeutic agent and a demethylating agent, a checkpoint inhibitor and a targeted therapy, a checkpoint inhibitor and a demethylating agent, or a targeted therapy and a demethylating agent. In some embodiments, the multimodal therapy comprises a targeted therapy and a chemotherapeutic agent. In some embodiments, the multimodal therapy comprises bevacizumab and a chemotherapeutic agent. In some embodiments, the multimodal therapy comprises pembrolizumab and a chemotherapeutic agent.

[0160] In some embodiments, the second active agent may comprise two, three, or four chemotherapeutic agents, two, three, or four checkpoint inhibitors, two, three, or four targeted therapies, or two, three, or four demethylating agents.

[0161] In some embodiments, the second active agent may comprise carboplatin and paclitaxel. In some embodiments, the second active agent may comprise pembrolizumab and paclitaxel. In some embodiments, the second active agent may comprise pembrolizumab, paclitaxel, and carboplatin. In some embodiments, the second active agent may comprise pembrolizumab, pemetrexed, and carboplatin. In some embodiments, the second active agent may comprise bevacizumab, gemcitabine, PLD, and carboplatin.

[0162] In some embodiments, the second active agent is administered co-administered with the heterodimeric TCR-antiCD3 antibody fusion molecule, i.e., administered on the same day as the heterodimeric TCR-antiCD3 antibody fusion molecule.

[0163] In some embodiments, the second or more active agents are administered according to known dosages and regimens for the specific active agent. In some embodiments, the second or more active agents are administered from once a week to once a month. In some embodiments, the second or more active agents are administered once a week. In some embodiments, when the second or more active agents are co-administered with the heterodimeric TCR-antiCD3 antibody fusion molecule, they may be administered at a reduced frequency or at a reduced dosage.

[0164] In some embodiments, the second active agent or more active agents or more pharmaceutical agents are administered for the duration of the dosing regimen. In some embodiments, the second active agent or more active agents are administered for a period of time known to the particular active agent. In some embodiments, the second active agent or more active agents are administered for at least 10 weeks. In some embodiments, the second active agent or more active agents are administered for at least 20 weeks, at least 30 weeks, at least 40 weeks, at least 50 weeks, at least 60 weeks, at least 70 weeks, at least 80 weeks, or at least 90 weeks. In some embodiments, the second active agent or more active agents are administered for at least 40 weeks. In some embodiments, the second active agent or more active agents are administered for a shorter period of time when co-administered with a heterodimeric TCR-antiCD3 antibody fusion molecule, relative to the second active agent or more active agents administered as a monotherapy.

[0165] In one specific embodiment of treatment with a heterodimeric TCR-antiCD3 antibody fusion molecule and a second or more active agents, this disclosure provides a method for treating a subject with non-small cell lung cancer or ovarian cancer, the method comprising administering to the subject: (I) A composition comprising a heterodimeric TCR-antiCD3 antibody fusion molecule, wherein the heterodimeric TCR-antiCD3 antibody fusion molecule comprises (a) an α-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 14, and (b) a β-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 16, wherein the α-chain comprises a complementarity-determining region (CDR) comprising α-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 3, 4, and 5, respectively, and the β-chain comprises β-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 9, 10, and 11, respectively, and (II) a second active agent. The dosing regimen described herein includes administration: (i) A first dose comprising 2 μg to 30 μg of the TCR-antiCD3 antibody fusion molecule, (ii) A second dose, the second dose comprising 5 μg to 60 μg of the TCR-antiCD3 antibody fusion molecule, and (iii) A maintenance dose comprising 15 μg to 200 μg of the TCR-antiCD3 antibody fusion molecule, wherein the maintenance dose is administered in at least three phases, wherein the first phase is administered every 6 to 8 days, the second phase every 12 to 16 days, and the third phase every 26 to 30 days; and The second dose contains a greater amount of the TCR-antiCD3 antibody fusion molecule than the first dose, and the maintenance dose contains a greater amount of the TCR-antiCD3 antibody fusion molecule than the second dose; and (B) A second active agent, wherein the second active agent is a chemotherapeutic agent, such as gemcitabine, decitabine, PLD or albumin-bound paclitaxel, and wherein the second active agent is applied once a week to once a month.

[0166] Administration of the heterodimeric TCR-anti-CD3 antibody fusion molecule and / or the second active agent can be performed by any conventional means known for administering anticancer biologics, such as intravenous (e.g., intravenous catheterization or central venous access device (CVAD)), intramuscular, subcutaneous, oral, subcutaneous, sublingual, intramuscular, intravenous, intranasal, percutaneous, or any combination thereof. In some embodiments, the heterodimeric TCR-anti-CD3 antibody fusion molecule and / or the second active agent can be administered by intravenous infusion. In some embodiments, the heterodimeric TCR-anti-CD3 antibody fusion molecule and / or the second active agent can be administered orally.

[0167] In a specific embodiment, this disclosure provides a method for treating a subject with non-small cell lung cancer, small cell lung cancer, or ovarian cancer, the method comprising administering a composition to the subject according to a dosing regimen, the composition comprising (I) a heterodimeric TCR-anti-CD3 antibody fusion molecule, and (II) a second composition comprising a second active agent, wherein the second active agent is gemcitabine, decitabine, PLD, pembrolizumab, terbentufosse, or albumin-bound paclitaxel.

[0168] In a specific embodiment, this disclosure provides a method for treating a subject with non-small cell lung cancer, small cell lung cancer, or ovarian cancer. The method comprises administering a composition to the subject according to a dosing regimen. The composition comprises (I) a heterodimeric TCR-anti-CD3 antibody fusion molecule, and (II) a second composition comprising a second active agent, wherein the second active agent is gemcitabine, decitabine, PLD, or albumin-bound paclitaxel. The heterodimeric TCR-anti-CD3 antibody fusion molecule comprises (a) an α-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 14, and (b) a β-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 16. The α-chain comprises a complementarity-determining region (CDR), the CDR comprising α-chain CDRs 1, 2, and 3, each having a sequence of SEQ ID NO: 14. The amino acid sequences NO:3, 4, and 5, and the β chain contains β chain CDRs 1, 2, and 3, which have the amino acid sequences SEQ ID NO: 9, 10, and 11, respectively. The dosing regimen includes: (i) Administering a first dose, said first dose comprising 3 μg of the TCR-antiCD3 antibody fusion molecule, (ii) One week after the first dose, a second dose is administered, said second dose comprising 10 μg of the TCR-antiCD3 antibody fusion molecule, and (iii) A maintenance dose comprising 40 μg of the TCR-antiCD3 antibody fusion molecule is administered once weekly for one week after the second dose and during the first phase, wherein the first phase is approximately 9 to 10 weeks. (iv) The maintenance dose is administered every 2 weeks during the second phase, wherein the second phase is approximately 38 weeks, and (v) The maintenance dose is administered every 4 weeks during the third phase, which is approximately 48 weeks.

[0169] In another specific embodiment, this disclosure provides a method for treating a subject with non-small cell lung cancer, small cell lung cancer, or ovarian cancer, the method comprising administering a composition to the subject according to a dosing regimen, the composition being (I) a heterodimeric TCR-anti-CD3 antibody fusion molecule, and (II) a second composition comprising a second active agent, wherein the second active agent is a chemotherapeutic agent, such as gemcitabine, decitabine, PLD, or albumin-bound paclitaxel. The heterodimeric TCR-antiCD3 antibody fusion molecule comprises (a) an α-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 14, and (b) a β-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 16, wherein the α-chain comprises a complementarity-determining region (CDR), the CDR comprising α-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 3, 4, and 5, respectively, and the β-chain comprises β-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 9, 10, and 11, respectively. The dosing regimen includes: (i) Administering a first dose, said first dose comprising 20 μg of the TCR-antiCD3 antibody fusion molecule, (ii) One week after the first dose, a second dose is administered, said second dose comprising 40 μg of the TCR-antiCD3 antibody fusion molecule, and (iii) One week after the second dose, and then weekly during the first phase, the maintenance dose comprising 160 μg of the TCR-anti-CD3 antibody fusion molecule, wherein the first phase is approximately 9 to 10 weeks. (iv) The maintenance dose is administered every 2 weeks during the second phase, wherein the second phase is approximately 38 weeks, and (v) The maintenance dose is administered every 4 weeks during the third phase, which is approximately 48 weeks.

[0170] In some embodiments, the methods provided herein can target PRAME-positive cancers, including but not limited to the following cancers: melanoma, lung cancer, breast cancer, ovarian cancer, endometrial cancer, esophageal cancer, bladder cancer, head and neck cancer, uterine cancer, acute myeloid leukemia, chronic myeloid leukemia, and Hodgkin's lymphoma. In some embodiments, this disclosure provides a method for treating a subject with PRAME-positive cancer, the method comprising administering a composition to the subject according to a dosing regimen, the composition comprising a heterodimeric TCR-anti-CD3 antibody fusion molecule. The heterodimeric TCR-antiCD3 antibody fusion molecule comprises (a) an α-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 14, and (b) a β-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 16, wherein the α-chain comprises a complementarity-determining region (CDR), the CDR comprising α-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 3, 4, and 5, respectively, and the β-chain comprises β-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 9, 10, and 11, respectively. The dosing regimen described herein includes administration: (i) A first dose comprising 2 μg to 30 μg of the TCR-antiCD3 antibody fusion molecule, (ii) A second dose, the second dose comprising 5 μg to 60 μg of the TCR-antiCD3 antibody fusion molecule, and (iii) A maintenance dose comprising 15 μg to 200 μg of the TCR-antiCD3 antibody fusion molecule, wherein the maintenance dose is administered in at least three phases, wherein the first phase is administered every 6 to 8 days, the second phase every 12 to 16 days, and the third phase every 26 to 30 days; and The second dose contains a greater amount of the heterodimeric TCR-antiCD3 antibody fusion molecule than the first dose contains, and the maintenance dose contains a greater amount of the heterodimeric TCR-antiCD3 antibody fusion molecule than the second dose contains. In some embodiments, PRAME-positive cancer is non-small cell lung cancer or small cell lung cancer. In some embodiments, PRAME-positive cancer is ovarian cancer.

[0171] In a specific embodiment, this disclosure provides a method for treating a subject with non-small cell lung cancer, small cell lung cancer, or ovarian cancer, the method comprising administering a composition to the subject according to a dosing regimen as outlined herein, the composition comprising (I) a heterodimeric TCR-anti-CD3 antibody fusion molecule, and (II) a second composition comprising a second active agent, wherein the second active agent is gemcitabine, PLD, albumin-bound paclitaxel, pembrolizumab, terbentufosse, osimertinib, docetaxel, bevacizumab, carboplatin, paclitaxel, dabrafenib, trametinib, pemetrexed, or decitabine.

[0172] In a specific embodiment, this disclosure provides a method for treating a subject with non-small cell lung cancer, small cell lung cancer, or ovarian cancer, the method comprising administering a composition to the subject, the composition comprising (I) a heterodimeric TCR-anti-CD3 antibody fusion molecule, and (II) a second composition comprising a second active agent, wherein the second active agent is gemcitabine, PLD, albumin-bound paclitaxel, pembrolizumab, or terbentuft.

[0173] In a specific embodiment, this disclosure provides a method for treating a subject with non-small cell lung cancer, small cell lung cancer, or ovarian cancer, the method comprising administering to the subject a composition comprising (I) a heterodimeric TCR-antiCD3 antibody fusion molecule, and (II) a second composition comprising osimertinib, docetaxel, or carboplatin.

[0174] In a specific embodiment, this disclosure provides a method for treating a subject with non-small cell lung cancer, small cell lung cancer, or ovarian cancer, the method comprising administering a composition to the subject, the composition comprising (I) a heterodimeric TCR-anti-CD3 antibody fusion molecule, and (II) a second composition comprising one of the following combinations: carboplatin + paclitaxel, bevacizumab + gemcitabine + carboplatin, or pembrolizumab + paclitaxel + carboplatin.

[0175] In a specific embodiment, this disclosure provides a method for treating a subject with non-small cell lung cancer, small cell lung cancer, or ovarian cancer, the method comprising administering a composition to the subject according to a dosing regimen, the composition comprising (I) a heterodimeric TCR-antiCD3 antibody fusion molecule, (II) a second composition comprising a second active agent, wherein the second active agent is carboplatin, and (III) a third composition comprising a third active agent, wherein the third active agent comprises paclitaxel.

[0176] In a specific embodiment, this disclosure provides a method for treating a subject with non-small cell lung cancer, small cell lung cancer, or ovarian cancer, the method comprising administering a composition to the subject according to a dosing regimen, the composition comprising (I) a heterodimeric TCR-antiCD3 antibody fusion molecule, (II) a second composition comprising a second active agent, wherein the second active agent is gemcitabine, and (III) a third composition comprising a third active agent, wherein the third active agent comprises osimertinib.

[0177] In a specific embodiment, this disclosure provides a method for treating a subject with non-small cell lung cancer, small cell lung cancer, or ovarian cancer, the method comprising administering a composition to the subject according to a dosing regimen, the composition comprising (I) a heterodimeric TCR-antiCD3 antibody fusion molecule, (II) a second composition comprising a second active agent, wherein the second active agent is gemcitabine, (III) a third composition comprising a third active agent, wherein the third active agent is bevacizumab, (IV) a fourth composition comprising a fourth active agent, wherein the fourth active agent is carboplatin, and (V) a fifth composition comprising a fifth active agent, wherein the fifth active agent is paclitaxel.

[0178] In a specific embodiment, this disclosure provides a method for treating a subject with non-small cell lung cancer, small cell lung cancer, or ovarian cancer, the method comprising administering a composition to the subject according to a dosing regimen, the composition comprising (I) a heterodimeric TCR-antiCD3 antibody fusion molecule, (II) a second composition comprising a second active agent, wherein the second active agent is gemcitabine, (III) a third composition comprising a third active agent, wherein the third active agent is carboplatin, and (IV) a fourth composition comprising a fourth active agent, wherein the fourth active agent is paclitaxel.

[0179] In other specific embodiments, this disclosure provides a method for treating ovarian cancer in a subject, the method comprising administering to the subject a composition comprising: (I) a heterodimeric TCR-antiCD3 antibody fusion molecule (wherein this molecule may correspond to a molecule targeting PRAME, preferably wherein this molecule is IMC-F106C), and (II) a second composition comprising a second active agent, wherein this agent is a chemotherapeutic agent (and the chemotherapeutic agent may be gemcitabine, PLD, carboplatin / paclitaxel, or any combination thereof). In some embodiments, the chemotherapeutic agent is gemcitabine. In some embodiments, the chemotherapeutic agent is PLD. In some embodiments, the chemotherapeutic agent is carboplatin / paclitaxel.

[0180] In other specific embodiments, this disclosure provides a method for treating ovarian cancer, the method comprising administering a composition to the subject, the composition comprising: (I) a heterodimeric TCR-antiCD3 antibody fusion molecule (wherein the molecule may correspond to a molecule targeting PRAME, preferably wherein the molecule is IMC-F106C), and (II) a second composition comprising a second active agent, wherein the agent is a targeted therapy (and the targeted therapy may be bevacizumab).

[0181] In other specific embodiments, this disclosure provides a method for treating ovarian cancer, the method comprising administering to the subject a composition comprising: (I) a heterodimeric TCR-antiCD3 antibody fusion molecule (wherein this molecule may correspond to a molecule targeting PRAME, preferably wherein this molecule is IMC-F106C), and (II) a second composition comprising a second active agent, wherein this agent is a multimodal therapy (and the multimodal therapy may be carboplatin / paclitaxel / pembrolizumab or carboplatin / gemcitabine / bevacizumab). In some embodiments, the multimodal therapy is carboplatin / paclitaxel / pembrolizumab. In some embodiments, the multimodal therapy is carboplatin / gemcitabine / bevacizumab.

[0182] In other specific embodiments, this disclosure provides a method for treating a subject with platinum-sensitive ovarian cancer, the method comprising administering to the subject a composition comprising: (I) a heterodimeric TCR-antiCD3 antibody fusion molecule (wherein this molecule may correspond to a molecule targeting PRAME, preferably wherein this molecule is IMC-F106C), and (II) a second composition comprising a second active agent, wherein this agent is a chemotherapeutic agent (and the chemotherapeutic agent may be gemcitabine, PLD, carboplatin / paclitaxel, or any combination thereof). In some embodiments, the chemotherapeutic agent is gemcitabine. In some embodiments, the chemotherapeutic agent is PLD. In some embodiments, the chemotherapeutic agent is carboplatin / paclitaxel.

[0183] In other specific embodiments, this disclosure provides a method for treating platinum-sensitive ovarian cancer, the method comprising administering a composition to the subject, the composition comprising: (I) a heterodimeric TCR-antiCD3 antibody fusion molecule (wherein the molecule may correspond to a molecule targeting PRAME, preferably wherein the molecule is IMC-F106C), and (II) a second composition comprising a second active agent, wherein the agent is a targeted therapy (and the targeted therapy may be bevacizumab).

[0184] In other specific embodiments, this disclosure provides a method for treating platinum-sensitive ovarian cancer, the method comprising administering to the subject a composition comprising: (I) a heterodimeric TCR-antiCD3 antibody fusion molecule (wherein this molecule may correspond to a molecule targeting PRAME, preferably wherein this molecule is IMC-F106C), and (II) a second composition comprising a second active agent, wherein this agent is a multimodal therapy (and the multimodal therapy may be carboplatin / paclitaxel / pembrolizumab or carboplatin / gemcitabine / bevacizumab). In some embodiments, the multimodal therapy is carboplatin / paclitaxel / pembrolizumab. In some embodiments, the multimodal therapy is carboplatin / gemcitabine / bevacizumab.

[0185] In other specific embodiments, this disclosure provides a method for treating a subject with platinum-resistant ovarian cancer, the method comprising administering to the subject a composition comprising: (I) a heterodimeric TCR-antiCD3 antibody fusion molecule (wherein this molecule may correspond to a molecule targeting PRAME, preferably wherein this molecule is IMC-F106C), and (II) a second composition comprising a second active agent, wherein this agent is a chemotherapeutic agent (and the chemotherapeutic agent may be gemcitabine, PLD, carboplatin / paclitaxel, or any combination thereof). In some embodiments, the chemotherapeutic agent is gemcitabine. In some embodiments, the chemotherapeutic agent is PLD. In some embodiments, the chemotherapeutic agent is carboplatin / paclitaxel.

[0186] In other specific embodiments, this disclosure provides a method for treating platinum-resistant ovarian cancer, the method comprising administering a composition to the subject, the composition comprising: (I) a heterodimeric TCR-antiCD3 antibody fusion molecule (wherein the molecule may correspond to a molecule targeting PRAME, preferably wherein the molecule is IMC-F106C), and (II) a second composition comprising a second active agent, wherein the agent is a targeted therapy (and the targeted therapy may be bevacizumab).

[0187] In other specific embodiments, this disclosure provides a method for treating platinum-resistant ovarian cancer, the method comprising administering to the subject a composition comprising: (I) a heterodimeric TCR-antiCD3 antibody fusion molecule (wherein this molecule may correspond to a molecule targeting PRAME, preferably wherein this molecule is IMC-F106C), and (II) a second composition comprising a second active agent, wherein this agent is a multimodal therapy (and the multimodal therapy may be carboplatin / paclitaxel / pembrolizumab or carboplatin / gemcitabine / bevacizumab). In some embodiments, the multimodal therapy is carboplatin / paclitaxel / pembrolizumab. In some embodiments, the multimodal therapy is carboplatin / gemcitabine / bevacizumab.

[0188] In other specific embodiments, this disclosure provides a method for treating lung cancer (preferably NSCLC), the method comprising administering a composition to the subject comprising: (I) a heterodimeric TCR antiCD3 antibody fusion molecule (wherein this molecule may correspond to a molecule targeting PRAME, preferably wherein this molecule is IMC-F106C), and (II) a second composition comprising a second active agent, wherein this agent is a chemotherapeutic agent (gemcitabine, docetaxel, or a combination thereof). In some embodiments, the chemotherapeutic agent is gemcitabine. In some embodiments, the chemotherapeutic agent is docetaxel.

[0189] In other specific embodiments, this disclosure provides a method for treating non-small cell lung cancer, the method comprising administering a composition to the subject, the composition comprising: (I) a heterodimeric TCR anti-CD3 antibody fusion molecule (wherein the molecule may correspond to a molecule targeting PRAME, preferably wherein the molecule is IMC-F106C), and (II) a second composition comprising a second active agent, wherein the agent is a targeted therapy (such as osimertinib).

[0190] In other specific embodiments, this disclosure provides a method for treating non-small cell lung cancer, the method comprising administering to the subject a composition comprising: (I) a heterodimeric TCR anti-CD3 antibody fusion molecule (wherein this molecule may correspond to a molecule targeting PRAME, preferably wherein this molecule is IMC-F106C), and (II) a second composition comprising a second active agent, wherein this agent is a multimodal therapy (and the multimodal therapy may be carboplatin / paclitaxel / pembrolizumab or carboplatin / pemetrexed / pembrolizumab). In some embodiments, the multimodal therapy is carboplatin / paclitaxel / pembrolizumab. In some embodiments, the multimodal therapy is carboplatin / pemetrexed / pembrolizumab.

[0191] In other specific embodiments, this disclosure provides a method for treating a subject with melanoma, the method comprising administering to the subject a composition comprising: (I) a heterodimeric TCR-antiCD3 antibody fusion molecule (wherein the molecule may correspond to a molecule targeting PRAME, preferably wherein the molecule is IMC-F106C), and (II) a second composition comprising a second active agent, wherein the agent is a second heterodimeric TCR-antiCD3 antibody fusion molecule (such as Tbengtavsp).

[0192] In other specific embodiments, this disclosure provides a method for treating a subject with melanoma, the method comprising administering to the subject a composition comprising: (I) a heterodimeric TCR-antiCD3 antibody fusion molecule (wherein the molecule may correspond to a molecule targeting PRAME, preferably wherein the molecule is IMC-F106C), and (II) a second composition comprising a second active agent, wherein the agent is a targeted therapy (such as dabrafenib / trametinib).

[0193] In other specific embodiments, this disclosure provides a method for treating a subject with other cancers, the method comprising administering to the subject a composition comprising: (I) a heterodimeric TCR-antiCD3 antibody fusion molecule (wherein the molecule may correspond to a molecule targeting PRAME, preferably wherein the molecule is IMC-F106C), and (II) a second composition comprising a second active agent, wherein the agent is a demethylating agent (such as decitabine, preferably low-dose decitabine).

[0194] In other specific embodiments, this disclosure provides a method for treating a subject with other cancers, the method comprising administering to the subject a composition comprising: (I) a heterodimeric TCR-antiCD3 antibody fusion molecule (wherein this molecule may correspond to a molecule targeting PRAME, preferably wherein this molecule is IMC-F106C), and (II) a second composition comprising a second active agent, wherein this agent is a chemotherapeutic agent (such as PLD or carboplatin / paclitaxel). In some embodiments, the chemotherapeutic agent is PLD. In some embodiments, the chemotherapeutic agent is carboplatin / paclitaxel.

[0195] Composition / Reagent Kit For administration to subjects, the heterodimeric TCR-antiCD3 antibody fusion molecule and / or a second active agent or more active agents of this disclosure may be provided as part of a composition together with one or more pharmaceutically acceptable carriers or excipients. This pharmaceutical composition may be in any suitable form (e.g., depending on the desired method of administration to the subject). The pharmaceutical composition may be provided in unit dosage forms and will typically be provided in sealed containers and may be provided as part of a kit. This kit will typically (but not necessarily) include instructions for use and may specifically include detailed instructions relating to the dosage and timing of administration. The kit may include multiple containers.

[0196] In some embodiments, the kit may comprise more than one container, each containing a specific amount of lyophilized heterodimeric TCR-antiCD3 antibody fusion molecules. The lyophilized heterodimeric TCR-antiCD3 antibody fusion molecules in one container may be reconstituted in a liquid medium (e.g., buffered saline solution) prior to administration to a subject, while the remaining containers remain lyophilized. In some embodiments, the kit may comprise more than one container, each containing a specific amount of heterodimeric TCR-antiCD3 antibody fusion molecules dissolved in a liquid medium, such as a buffer system, and / or one or more excipients.

[0197] Therefore, for example, the kit may comprise (i) one to five first containers, each containing a first dose comprising 2 μg to 4 μg of a heterodimeric TCR-antiCD3 antibody fusion molecule; (ii) one to five second containers, each containing a second dose comprising 5 μg to 14 μg of a heterodimeric TCR-antiCD3 antibody fusion molecule; (iii) three or more maintenance containers, each containing a maintenance dose comprising 15 μg to 55 μg of a heterodimeric TCR-antiCD3 antibody fusion molecule; and (iv) one or more containers containing 10-1000 mg / m³. 2(v) A second active agent dose; and (vi) instructions for administering the heterodimeric TCR-antiCD3 antibody fusion molecule according to the dosing regimen. In some embodiments, the instructions specify a dosing regimen comprising (a) administering a first dose once weekly, (b) administering a second dose once weekly, (c) administering a maintenance dose once weekly during a first phase, (d) administering a maintenance dose every 2 weeks during a second phase, and (e) administering a maintenance dose every 4 weeks during a third phase, and (vi) instructions for administering the combination of the second active agent and the heterodimeric TCR-antiCD3 antibody fusion molecule according to the dosing regimen, wherein the dosing regimen is specified as (a) administering once weekly for two weeks and stopping for one week, (b) administering once weekly for three weeks and stopping for one week, or (c) administering once weekly for two weeks and stopping for one week.

[0198] In some embodiments, this disclosure provides a kit comprising: (i) A first container containing a first dose, the first dose containing 2 μg to 4 μg of a heterodimeric TCR-antiCD3 antibody fusion molecule. (ii) A second container containing a second dose, the second dose containing 5 μg to 14 μg of the heterodimeric TCR-antiCD3 antibody fusion molecule. (iii) One or more maintenance containers, each containing a maintenance dose, said maintenance dose containing 15 μg to 55 μg of the heterodimeric TCR-antiCD3 antibody fusion molecule. (iv) One or more containing 10-1000 mg / m 2 The container for the second dose of the active agent; (v) Instructions for use for administering the heterodimeric TCR-antiCD3 antibody fusion molecule according to a dosing regimen, wherein the dosing regimen specifies: (a) administration of the first dose in week 1, (b) administration of the second dose in week 2, (c) administration of a maintenance dose once weekly for 10 weeks during the first phase, (d) administration of the maintenance dose once every 2 weeks for 38 weeks during the second phase, and (e) administration of the maintenance dose once every 4 weeks for 48 weeks during the third phase; and (vi) Instructions for use in combination with the heterodimeric TCR-antiCD3 antibody fusion molecule to administer the second active agent according to the dosing regimen, wherein the dosing regimen is specified as: (a) once a week for two weeks, followed by a one-week break; (b) once a week for three weeks, followed by a one-week break; or (c) once a week for two weeks, followed by a one-week break.

[0199] In some embodiments, this disclosure provides a kit comprising: (i) A first container containing a first dose, the first dose containing 20 μg of a heterodimeric TCR-antiCD3 antibody fusion molecule. (ii) A second container containing a second dose, the second dose containing 40 μg of the heterodimeric TCR-antiCD3 antibody fusion molecule. (iii) One or more maintenance containers, each containing a maintenance dose comprising 160 μg of the heterodimeric TCR-antiCD3 antibody fusion molecule. (iv) One or more containing 10-1000 mg / m 2 The container for the second dose of the active agent; (v) Instructions for use for administering the heterodimeric TCR-antiCD3 antibody fusion molecule according to a dosing regimen, wherein the dosing regimen specifies: (a) administration of the first dose in week 1, (b) administration of the second dose in week 2, (c) administration of a maintenance dose once weekly for 10 weeks during the first phase, (d) administration of the maintenance dose once every 2 weeks for 38 weeks during the second phase, and (e) administration of the maintenance dose once every 4 weeks for 48 weeks during the third phase; and (vi) Instructions for use in combination with the heterodimeric TCR-antiCD3 antibody fusion molecule to administer the second active agent according to the dosing regimen, wherein the dosing regimen is specified as: (a) once a week for two weeks, followed by a one-week break; (b) once a week for three weeks, followed by a one-week break; or (c) once a week for two weeks, followed by a one-week break.

[0200] In some embodiments, the heterodimeric TCR-antiCD3 antibody fusion molecule in any kit described herein comprises (a) an α-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 14, and (b) a β-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 16, wherein the α-chain comprises a complementarity-determining region (CDR) comprising α-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 3, 4, and 5, respectively, and the β-chain comprises β-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 9, 10, and 11, respectively.

[0201] In some embodiments, this disclosure provides a kit for an escalating dosing regimen, the kit comprising: (i) A first container containing a first dose, the first dose containing 3 μg of a heterodimeric TCR-antiCD3 antibody fusion molecule. (ii) A second container containing a second dose, the second dose containing 10 μg of the heterodimeric TCR-antiCD3 antibody fusion molecule. (iii) Three or more maintenance containers, each containing a maintenance dose, the maintenance dose containing 4 μg of the heterodimeric TCR-antiCD3 antibody fusion molecule.

[0202] In some embodiments, this disclosure provides a kit for an escalating dosing regimen, the kit comprising: (i) A first container containing a first dose, the first dose containing 20 μg of a heterodimeric TCR-antiCD3 antibody fusion molecule. (ii) A second container containing a second dose, the second dose containing 40 μg of the heterodimeric TCR-antiCD3 antibody fusion molecule. (iii) Three or more maintenance containers, each containing a maintenance dose, the maintenance dose containing 160 μg of the heterodimeric TCR-antiCD3 antibody fusion molecule.

[0203] In some embodiments, this disclosure provides a kit for a maintenance dosing regimen, the kit comprising: three or more maintenance containers, each maintenance container containing a maintenance dose comprising 40 μg of the heterodimeric TCR-antiCD3 antibody fusion molecule. In some embodiments, the kit for the maintenance dosing regimen comprises maintenance containers sufficient for 1 month, 2 months, 3 months, 4 months, 5 months, or 6 months.

[0204] In some embodiments, this disclosure provides a kit for a maintenance dosing regimen, the kit comprising: three or more maintenance containers, each maintenance container containing a maintenance dose comprising 160 μg of the heterodimeric TCR-antiCD3 antibody fusion molecule.

[0205] In some embodiments, the kit for maintenance dosing regimens includes maintenance containers sufficient for 1 month, 2 months, 3 months, 4 months, 5 months, or 6 months.

[0206] The kit provided in this disclosure may comprise one or two first containers, one or two second containers, and various numbers of maintenance containers. For example, in some embodiments, the kit comprises 2 to 60 maintenance containers, 3 to 50 maintenance containers, 4 to 40 maintenance containers, 4 to 30 maintenance containers, 4 to 20 maintenance containers, 4 to 16 maintenance containers, 4 to 12 maintenance containers, or 4 to 8 maintenance containers. For example, in some embodiments, the kit comprises 2 to 10 maintenance containers. In some embodiments, the first dose, the second dose, and / or the maintenance dose further comprise pharmaceutically acceptable excipients to form the first pharmaceutical formulation, the second pharmaceutical formulation, and / or the maintenance pharmaceutical formulation, respectively. In some embodiments, the pH of the first pharmaceutical formulation, the second pharmaceutical formulation, and / or the maintenance pharmaceutical formulation is pH-adjusted. In some embodiments, the pH of the first pharmaceutical formulation, the second pharmaceutical formulation, and / or the maintenance pharmaceutical formulation is about 6.5 to about 7.5.

[0207] In some embodiments, the kit contains only one anticancer therapeutic agent. In some embodiments, the kit contains a heterodimeric TCR-antiCD3 antibody fusion molecule and further contains a second, third, fourth, and / or fifth therapeutic agent. In some embodiments, the second, third, fourth, and / or fifth therapeutic agent in the kit contains a checkpoint inhibitor, a chemotherapy agent, another TCR-antiCD3 antibody fusion molecule, and / or a demethylating agent. In some embodiments, the second, third, fourth, and / or fifth therapeutic agents in the kit comprise gemcitabine, albumin-bound paclitaxel, pegylated liposomal doxorubicin (PLD), docetaxel, carboplatin, paclitaxel, doxorubicin, pemetrexed, aldehyde interleukin, atezolizumab (TECENTRIQ®), avelumab, bevacizumab (Avastin®), bimetinib, braftovi (encofenib), cobimethinib fumarate, Cotellic (cobimethinib fumarate), dabrafenib mesylate, dacarbazine, and duvalumab. Encofenib, ganeserte, gemcitabine, IL-2 (aldehyde interleukin), interferon, Imlygic (talimogene laherparepvec), and Intron are also included. A (recombinant interferon alpha-2b), ipilimumab, Keytruda (pembrolizumab), KIMMTRAK (terbentavspir), Mekinist (trametinib dimethyl sulfoxide), Mektovi (bimetinib), mesatinib, nivolumab, nivolumab and rennet-rmbw, Opdivo (nivolumab), Opdualag (nivolumab and rennet-rmbw), Tagrisso (osimertinib), Proleukin (aldeleukin), Tafinlar (dabrafenib mesylate), terbentavspir The kit may contain tebufenozide, trimethyl sulfoxide trametinib, tramemumab, vemurafenib, Yervoy (ipilimumab), Zelboraf (vemurafenib), azacitidine, decitabine, cytidine, chidamide, guardicotabine, 5-fluoro-2'-deoxycytidine, zabraline, CP-4200, RG108, nanamycin A, or any combination or modification thereof. In some embodiments, the second, third, fourth, and / or fifth therapeutic agents in the kit comprise chemotherapeutic agents such as gemcitabine, albumin-bound paclitaxel, decitabine, tebufenozide, or combinations thereof.

[0208] The kit described herein contains one or more containers, such as a first container containing a first dose, a second container containing a second dose, and / or a maintenance container containing a maintenance dose. Suitable containers include, for example, bottles, vials, syringes, etc. The containers can be formed from a variety of materials, such as glass or plastic. The containers contain or contain compositions comprising the heterodimeric TCR-antiCD3 antibody fusion molecule disclosed herein and may have a sterile access port (e.g., the container may be an intravenous solution bag or a vial with a stopper that can be punctured by a hypodermic needle). The instructions (e.g., a label or packaging insert) indicate that the heterodimeric TCR-antiCD3 antibody fusion molecule is intended for the treatment of eligible subjects, such as subjects who have or are predisposed to the diseases described herein, and provide specific guidance on the dosage and interval of the composition and any other medications. In some embodiments, the kit may further contain additional containers containing pharmaceutically acceptable dilution buffers, such as bacteriostatic water for injection (BWFI), phosphate-buffered saline, Ringer's solution, and / or dextran solution. The kit may further include other materials desired from a commercial and user perspective, including additional buffers, diluents, filters, needles, and syringes.

[0209] Various modifications to this disclosure, other than those shown and described herein, will become apparent to those skilled in the art from the foregoing description and fall within the scope of the appended claims. Preferred features of each aspect of this disclosure are applied only with corresponding modifications to each of the other aspects. All documents referenced herein are incorporated by reference to the fullest extent permitted by law.

[0210] Example This disclosure has been described with respect to representative examples, which are considered illustrative embodiments and do not limit the scope of this disclosure as defined solely by the claims. All references to publications, including scientific publications, monographs, textbooks, patent applications, and published patents, are hereby incorporated by reference for all purposes.

[0211] The experimental design of this invention is complex and modular (see below). The initial experimental design focused on dose safety and tolerability, as well as the determination of the maximum tolerated dose (MTD) and the recommended phase 2 dose (RP2D). During the initial phase of the trials, and as a result of analyzing the research data, the research group identified certain treatment methods and regimens that were beneficial, particularly the combination therapy of IMC-F106C with certain chemotherapy therapies for lung and ovarian cancer. Based on the overall research analysis, the research group also developed new hypotheses and then established a novel set of treatment methods (described below, including combination therapies with additional chemotherapy, targeted therapies, and demethylating agents).

[0212] Example 1: IMC-F106C Clinical Trial Patients have been treated in an ongoing Phase 1 / 2 clinical trial of IMC-F106C (IMC-F106C-101), in a study entitled "IMC-F106C as a single agent and in combination with a checkpoint inhibitor in HLA-A patients with advanced PRAME-positive cancer". 02:01 A Phase 1 / 2 First-in-Human Study of the Safety and Efficacy of IMC-F106Cas a Single Agent and in Combination with Checkpoint Inhibitors in HLA-A Positive Participants 02:01 Positive Participants with Advanced PRAME Positive Cancers. The study was designed to evaluate the safety, tolerability, pharmacokinetics (PK), immunogenicity, pharmacodynamics, and antitumor activity of IMC-F106C as a monotherapy and in combination with other therapies.

[0213] In the IMC-F106C-101 study, participants with advanced cancer included those with unresectable or metastatic melanoma, ovarian cancer, uterine cancer, NSCLC, and other PRAME-positive tumors that had relapsed from standard of care, were refractory to, or were intolerant of standard of care. The study also evaluated the role of combinations of these standard of care with different treatments in participants with advanced cancer who were eligible to receive those treatments.

[0214] Despite the development and approval of new therapies, including targeted small molecule agents and immunotherapies, many patients will be refractory or relapse after the availability of treatment options. The treatment needs of these patients remain largely unmet, and access to novel targeted therapies such as IMC-F106C has the potential to provide clinical benefit.

[0215] The trial design is complex, involving different modules depending on the tumor type, similar to a complex basket trial. Figure 1 I saw advanced research plans in it.

[0216] Among other reasons, IMC-F106C is administered using a stepwise dosing regimen to reduce tumor lysis ( Figure 2 and 3 ).

[0217] Example 2: IMC-F106C combination therapy In addition to monotherapy, IMC-F106C has been evaluated in combination with standard therapies, including checkpoint inhibitors, chemotherapy agents, or another ImmTAC molecule.

[0218] The following section further describes specific therapeutic agents used in combination therapy with IMC-F106C.

[0219] Checkpoint inhibitors PD-1 can be expressed on tumor-infiltrating lymphocytes and may help suppress anti-tumor immune responses. The binding of PD-1 to PD-L1, which may be expressed on tumor cells and antigen-presenting cells, leads to inhibition of cytotoxic T cell activity, T cell proliferation, and cytokine production. Several monoclonal antibody therapeutics targeting PD-1 or PD-L1 (PD-[L]1), including atezolizumab, avelumumab, duvalumab, nivolumab, and pembrolizumab, have been approved for the treatment of various malignancies.

[0220] T cell activation, including ImmTAC-mediated activation, induces PD-1 expression on T cells. Increased PD-L1 expression has been observed in tumors following ImmTAC treatment (Middleton et al. 2020). This induction of both ligand and receptor may impair T cell responses. In vitro studies have shown that checkpoint inhibitors can increase ImmTAC-redirected killing of PD-1-positive tumor-infiltrating T cells against PD-L1-positive cancer cells (Petrovic et al. 2021). Checkpoint inhibitors, including pembrolizumab, can enhance the initial activity of IMC-F106C and prevent T cell exhaustion, thereby maintaining the effectiveness of emerging anti-tumor immune responses. Simultaneously, the redirection of effector T cells to IMC-F106C-mediated tumors may help overcome resistance to monotherapy checkpoint inhibitors caused by poor T cell infiltration.

[0221] Pembrolizumab is administered according to the manufacturer's prescription information.

[0222] Chemotherapy Holistic analysis of the combination data and analysis of the translational data revealed that the efficacy of IMC-F106C may be related to participants' immune fitness. Based on the research data, it is understood that chemotherapy treatment may reset immune fitness. Therefore, a combination approach using other chemotherapy therapies was incorporated into the clinical trial design.

[0223] Based on these facts, gemcitabine, albumin-bound paclitaxel, and PLD were chosen as combination therapy partners for IMC-F106C chemotherapy: these agents are commonly used for various indications where PRAME expression is very widespread, relatively well-tolerated, and not lymphocyte-depleted. Gemcitabine is a nucleoside metabolism inhibitor, suitable for both monotherapy and combination therapy against multiple cancers. Albumin-bound paclitaxel is a second-generation semi-synthetic taxane conjugated to albumin that inhibits microtubule depolymerization, thereby causing cell cycle arrest. PLD is an anthracycline topoisomerase inhibitor, suitable for combination therapy against multiple cancers. In addition to direct antitumor activity, chemotherapy can also enhance immunotherapy by inducing immunogenic cell death and disrupting tumor immune evasion.

[0224] Other chemotherapy regimens have been incorporated into the study design, namely docetaxel, carboplatin / paclitaxel (alone and in combination with bevacizumab or pembrolizumab), and carboplatin / pemetrexed (in combination with pembrolizumab). These regimens offer treatment for early-stage NSCLC, ovarian cancer, and / or uterine cancer.

[0225] Immunogenic cell death involves the release of potentially immunogenic tumor cell proteins and DAMPs. For example, gemcitabine treatment of cancer cells induces classic DAMPs, cell surface calreticulin, and soluble HSP70 and HMGP (Hayashi et al. 2020). Chemotherapy, including docetaxel, carboplatin, pemetrexed, paclitaxel, and doxorubicin, can also induce immunogenic cell death (Lau 2020, Schaer 2019, Flieswasser 2020). These signals can induce dendritic cell maturation and activation, leading to increased presentation of immunogenic antigens. Because ImmTAC treatment increases the number of tumor-infiltrating T cells, the combination of ImmTAC treatment and chemotherapy may increase the likelihood of activating T cells specific to newly shed tumor antigens.

[0226] The chosen chemotherapy can also reduce immunosuppression mediated by myeloid cells, including macrophages and MDSCs. Gemcitabine treatment can deplete intratumoral MDSCs (Eriksson et al. 2016; Le et al. 2009; Suzuki et al. 2005), while paclitaxel can kill MDSCs and (at subcytotoxic concentrations) induce maturation into dendritic cells (Michels et al. 2012). In vitro, tumor cell killing by ImmTAC redirection is sensitive to the suppression of M2 macrophages (alternating activation, immunosuppressive), and in a clinical study in uveal melanoma by Tbengtavsp, a high ratio of CD163-positive macrophages to T cells was associated with poor clinical response (Hassel et al. 2021). Paclitaxel, PLD, and docetaxel have been shown to promote the conversion of macrophages from the M2 phenotype to the M1 (classically activated, pro-inflammatory) phenotype (Takayama et al. 2020; Vanderley et al. 2018; Yamaguchi et al. 2017). Drugs that convert macrophages in the tumor microenvironment from the M2 phenotype to the M1 phenotype can enhance antitumor activity and synergize with immunotherapy (Li, C. et al. 2021).

[0227] Preclinical evaluation of the ImmTAC-chemotherapy combination showed enhanced tumor cell lysis when paclitaxel was administered in combination with or sequentially with ImmTAC (Bravo-Lopes et al. 2019). Although paclitaxel impaired T cell proliferation, it enhanced tumor cell killing. Prior exposure of tumor cells or T cells to paclitaxel enhanced ImmTAC-mediated tumor cell killing without affecting T cell proliferation.

[0228] Administer gemcitabine, albumin-bound paclitaxel, PLD, docetaxel, pemetrexed, carboplatin, and paclitaxel according to the manufacturer's prescription information.

[0229] The combination of IMC-F106C and chemotherapy showed clinical activity. Figure 4A and Figure 4B Clinical activity was demonstrated in combination therapy with albumin-bound paclitaxel and IMC-F106C. Figure 4A This is a graphical representation of the change in a patient's tumor size relative to baseline, where each bar represents an individual patient. For example... Figure 4A As can be seen, participants with non-small cell lung cancer (NSCLC), ovarian cancer (OC), breast cancer, and mucosal melanoma showed a reduction in tumor size, with the largest reduction observed to date in NSCLC and OC patients. Figure 4B These are different representations of the responses of these patients (spider plot), which shows the change in tumor size relative to baseline over time, and where each line represents an individual patient.

[0230] Figure 4C and Figure 4D Clinical activity was demonstrated in combination therapy with gemcitabine and IMC-F106C. Figure 4C This is a graphical representation of the change in a patient's tumor size relative to baseline, where each bar represents an individual patient. For example... Figure 4C As can be seen in the study, participants with NSCLC and OC again showed the greatest reduction in tumor size to date. Figure 4D This is a spider diagram of the responses of these patients, showing the change in tumor size relative to baseline over time, with each line representing an individual patient.

[0231] Figure 4A The data presented in -D indicate that combination therapy with IMC-F106C and albumin-bound paclitaxel or gemcitabine produced clinical activity, with the largest response observed to date in patients with NSCLC and OC. Each of these patients had received several lines of prior therapy without response or tumor regression. Therefore, combination therapy provides a crucial option for treating cancer, particularly in patients who may not respond well to standard of care.

[0232] Other ImmTAC molecules Terbenavsp is a bispecific gp100 peptide-HLA-directed CD3 T cell connector suitable for the treatment of HLA-A inflammatory diseases in patients with unresectable or metastatic uveal melanoma. 02:01 Positive adult patient.

[0233] Malignant melanomas originate from melanocytes in any organ and are characterized by the expression of gp100 (the target protein of terbentufosse). Terbentufosse is engineered to redirect T cells to kill gp100+ tumor cells. In a randomized phase 3 study of metastatic uveal melanoma (IMCgp100-202), terbentufosse monotherapy demonstrated an overall survival (OS) benefit (HR of 0.51) compared to investigator-selected pembrolizumab, ipilimumab, or dacarbazine (Nathan et al. 2021). Terbentufosse as monotherapy in previously untreated metastatic non-uveal melanoma and in combination with anti-PD-L1 therapy has shown promising OS in previously treated metastatic non-uveal melanoma (Hamid et al. 2021; Middleton et al. 2020).

[0234] The expression and presentation of tumor antigens such as PRAME can exhibit intratumoral heterogeneity. Cells presenting reduced levels of target pHLA may be less susceptible to killing by ImmTAC-redirected T cells, providing a potential resistance mechanism. A combination of two ImmTAC molecules will simultaneously target two tumor antigens (e.g., PRAME and gp100), potentially making larger tumor cell populations more vulnerable to killing. Furthermore, in vitro data indicate increased cell killing in the presence of higher levels of target pHLA, a characteristic specific to single ImmTAC molecules. Therefore, combination therapy using two ImmTAC molecules could allow more ImmTAC molecules to bind to individual tumor cells expressing the associated pHLA complex. Thus, the combination ImmTAC approach may potentially increase tumor cell killing compared to single ImmTACs.

[0235] Tbengtaftsp, also known as KIMMTRAK, is used according to its prescription information.

[0236] Targeted therapy Leveraging the unique mechanism of IMF-F106C, certain targeted therapies can be incorporated into trial designs to provide opportunities to cover the mechanism of action of IMF-F106C as part of a base therapy, particularly for patients with relapsed disease or whose benefit from prior treatment tends to plateau and / or tends toward disease progression.

[0237] Combinations with selected targeted therapies (osimertinib, bevacizumab, and dabrafenib / trametinib) were included. Hyperactive metabolism and insufficient blood supply lead to the development of a hypoxic and acidic tumor microenvironment (TME). In response to this hypoxia, certain pro-angiogenic cytokines (such as VEGF and ANGPT2) become more active, stimulating angiogenesis. The resulting disordered angiogenesis further promotes the emergence of an immunosuppressive TME. By inhibiting VEGF, bevacizumab intervenes by blocking these pro-angiogenic pathways. This intervention normalizes blood vessels, thereby improving blood flow and oxygen levels within the tumor environment. Consequently, the hypoxic TME is reversed, and drug delivery to the tumor is enhanced. Inhibition of angiogenesis also remodels the TME by promoting T cell infiltration and DC maturation. It promotes the transformation of macrophages into an M1-like macrophage phenotype while reducing the proportion of Tregs and MDSCs (Yi et al. 2022).

[0238] Osimertinib, as a third-generation EGFR kinase inhibitor, has demonstrated the ability to trigger DAMP (Furukawa et al. 2021). Dabrafenib and trametinib are BRAF and MEK inhibitors, respectively. In laboratory settings, targeted inhibition of both EGFR and MEK (indispensable components of the MAPK pathway) has been observed to increase the expression of MHC class I molecules. Combining TCR-targeted therapy against PRAME with MAPK pathway inhibitors has enhanced cell-killing efficacy (Brea et al. 2016).

[0239] Bevacizumab, osimertinib, dabrafenib, and trametinib were administered according to their prescribing information.

[0240] Demethylating agents As another innovative approach, the use of demethylating agents that can promote PRAME expression (the target of IMC-F106C) is incorporated into clinical trial designs as part of combination therapy.

[0241] The combination of low-dose decitabine (i.e., a demethylating agent (HMA)) with IMC-F106C will be used. Currently, HMA is approved for use in hematologic malignancies (myelodysplastic syndromes and acute myeloid leukemia). Low-dose decitabine will be used to improve immune function and increase tumor PRAME expression.

[0242] To date, several clinical studies have been completed that have evaluated the safety and efficacy of HMA as a monotherapy or in combination with other systemic therapies in solid tumors (George et al. 2010; Samlowski et al. 2005; Torres et al. 2021; Taylor et al. 2020; Jang et al. 2023; Palomba et al. 2022; Short et al. 2022).

[0243] Non-cytotoxic (low) doses of decitabine have been shown to deplete the epigenetic regulator DNA methyltransferase 1 (DNMT1) (Hoff et al. 2018; Matei et al. 2018; Fu et al. 2014; Garrido-Laguna et al. 2013; Awada et al. 2020; Bauman et al. 2012; Ben-Baruch et al. 1993).

[0244] T cells within the tumor-associated membrane (TME) are antigen-exposed, and continuous antigen stimulation leads to CD8+ T cell exhaustion, manifested by the presence of inhibitory receptors, dysplastic proliferation and cytokine secretion, impaired immune memory, and metabolic alterations (Wherry and Kurachi 2015). Targeting PRAME has shown efficacy in patients with advanced cancer. The ability of PRAME-redirected T cells to recognize PRAME-positive tumor cells is based on the intensity of MHC class I expression and co-expression. To evade immune surveillance, cancer has evolved to suppress the expression of IFNγ pathway genes, proteins involved in antigen presentation (including MHC class I or β-2 microglobulin (B2M)), and cancer testis antigens through mutations or epigenetic silencing. Data suggest that epigenetic regulation can improve immune function by upregulating the expression of tumor-associated antigens (such as PRAME) and / or epigenetically reprogramming T cell intrinsic programs, as well as suppressing terminal differentiation (Pauken et al. 2016; Li, X. et al. 2023; Sen et al. 2016).

[0245] HMA can induce PRAME gene expression in multiple cancer types (Wong et al., 2021; Yao et al., 2013). This can induce immunomodulatory effects, thereby promoting higher anticancer activity, especially in tumors with low PRAME expression (Chou et al., 2012). Depletion of DNMT1 by combining non-cytotoxic doses of HMA with the PRAME-directed ImmTAC molecule IMC-F106C may improve anticancer activity.

[0246] When lung cancer cell lines and ovarian cancer cell lines were pretreated with clinically relevant concentrations of decitabine and then used as target cells in the ImmTAC-dependent T cell activation assay, increased T cell activation was observed (as measured by interferon-γ release), as shown in Figure 5.

[0247] NCI-H441, NCI-H2087, and OVCAR3 cancer cell lines were pretreated for 5 days with tissue medium containing 75 nM decitabine or tissue medium alone (untreated) and then used as target cells in IFNγ ELISPOT assays and PRAME ImmTAC titrations performed with healthy donor PBMCs.

[0248] Multimodal therapy The combination of biologics (e.g., pembrolizumab, bevacizumab) and chemotherapy is the standard of care for early-line advanced non-small cell lung cancer (NSCLC), ovarian cancer (OC), and endometrial cancer (EC). Potential benefits of using multimodal therapy in early-line therapy (e.g., first-line) for advanced disease, compared to later-line therapy, include: potentially lower tumor burden (Kim, Cassella, and Byrne 2021), reduced likelihood of treatment resistance (Long et al. 2019), and improved progression-free survival (PFS) (Bailey et al. 2012).

[0249] Clinical trial modules and combination groups Figure 6 Table A shows an overview of the different combination therapies and related tumor modules.

[0250] Table A References Awada, Hassan, Reda Z. Mahfouz, Ashwin Kishtagari, Teodora Kuzmanovic, Jibran Durrani, Cassandra M. Kerr, Bhumika J. Patel et al. 2020. “Extended Experience with a Noncytotoxic DNMT1-targeting Regimen of Decitabine to Treat Myeloid Malignancies”. British Journal of Hematology (2009-2020). British Journal of Haematology )》 188 (6):924–29.

[0251] Bailey, Christopher H, Gayle Jameson, Chao Sima, Sharon Fleck, Erica White, Daniel D Von Hoff, and Glen J Weiss. 2012. "Progression-Free Survival Decreases with Each Subsequent Therapy in Patients Presenting for Phase I Clinical Trials". *Journal of Cancer* Journal of Cancer )》 3: 7–13.

[0252] Bauman, Julie, Claire Verschraegen, Steven Belinsky, Carolyn Muller, Teresa Rutledge, M. Fekrazad, Meera Ravindranathan, Sang-Joon Lee, and Dennie Jones. 2012. “A Phase I Study of 5-Azacytidine and Erlotinib in Advanced Solid Tumor Malignancies”. *Cancer Chemotherapy and Pharmacology*. Cancer Chemotherapy and Pharmacology )》69 (2): 547–54.

[0253] Ben-Baruch, Noa, Andrea M. Denicoff, Barry R. Goldspiel, Joyce A. O'Shaughnessy, and Kenneth H. Cowan. 1993. "Phase II Study of Fazarabine (NSC 281272) in Patients with Metastatic Colon Cancer". *Investigative New Drugs* Investigational New Drugs )》 11(1): 71–74.

[0254] Bravo-Lopes, Filipa, Nora Rippaus, Kristina Petrovic, Francesca Amicarella, Adam Taylor, Laure Humbert, Rupert Kenefeck, and Adel Benlahrech. 2019. “34th Annual Meeting & Pre-Conference Programs of the Society for Immunotherapy of Cancer (SITC 2019): Part 1”. Journal of Cancer Immunotherapy ( Journal for ImmunoTherapy of Cancer )》 7 (Supplement 1): 282.

[0255] Brea, Elliott J., Claire Y. Oh, Eusebio Manchado, Sadna Budhu, Ron S. Gejman, George Mo, Patrizia Mondello et al. 2016. "Kinase Regulation of Human MHC Class I Molecule Expression on Cancer Cells". *Cancer Immunology Research* Cancer Immunology Research )》 4 (11): 936–47.

[0256] Chou, Jeffrey, Lilien N. Voong, Christie L. Mortales, Andrea MHTowlerton, Seth M. Pollack, Xiaoji Chen, Cassian Yee, Paul F. Robbins, and Edus H. Warren. 2012. “Epigenetic Modulation to Enable Antigen-Specific T-Cell Therapy of Colorectal Cancer”. Journal of Immunotherapy ( Journal of Immunotherapy )》 35 (2): 131–41.

[0257] Eriksson, Emma, ​​Jessica Wenthe, Sandra Irenaeus, Angelica Loskog, and Gustav Ullenhag. 2016. “Gemcitabine Reduces MDSCs, Tregs and TGFβ-1 While Restoring the Teff / Treg Ratio in Patients with Pancreatic Cancer.” *Journal of Translational Medicine* Journal of Translational Medicine )》 14 (1): 282.

[0258] Flieswasser, Tal, Jinthe Van Loenhout, Laurie Freire Boullosa, Astrid Van den Eynde, Jorrit De Waele, Jonas Van Audenaerde, Filip Lardon, Evelien Smits, Patrick Pauwels, and Julie Jacobs. 2020. “Clinically Relevant Chemotherapeutics Have the Ability to Induce Immunogenic Cell Death in Non-Small Cell Lung Cancer.” Cell (2009). Cells )》 9 (6): 1474.

[0259] Fu, Xiaoyu, Yan Zhang, Xiaohui Wang, Meixia Chen, Yao Wang, Jing Nie, Yuanguang Meng, and Weidong Han. 2014. “Low Dose Decitabine Combined with Taxol and Platinum Chemotherapy to Treat Refractory / Recurrent Ovarian Cancer: An Open-Label, Single-Arm, Phase I / II Study.” Current Protein and Peptide Science (2014). Current Protein&Peptide Science )》 16 (4): 329–36.

[0260] Furukawa, Rie, Hiroyuki Inoue, Yasuto Yoneshima, Hirono Tsutsumi, Eiji Iwama, Yuki Ikematsu, Nobuhisa Ando et al. 2021. "Cytotoxic Chemotherapeutic Agents and the EGFRTKI Osimertinib Induce Calreticulin Exposure in Non-Small Cell Lung Cancer". Lung Cancer (… Lung Cancer )》 155: 144–50.

[0261] Garrido-Laguna, I., KA McGregor, M. Wade, J. Weis, W. Gilcrease, L. Burr, R. Soldi et al. 2013. “A Phase I / II Study of Decitabine in Combination with Panitumumab in Patients with Wild-Type (Wt) KRAS Metastatic Colorectal Cancer”. Investigational New Drugs 31(5): 1257-64.

[0262] George, Rani E., Jill M. Lahti, Peter C. Adamson, Kejin Zhu, David Finkelstein, A. Mark Ingle, Joel M. Reid et al. 2010. "Phase I Study of Decitabine with Doxorubicin and Cyclophosphamide in Children with Neuroblastoma and Other Solid Tumors: A Children's Oncology Group Study". *Pediatric Hematology and Oncology* Pediatric Blood & Cancer )》 55 (4): 629–38.

[0263] Hamid, Omid, Jessica Hassel, Alexander Shoushtari, Friedegund Meier, Todd Bauer, April Salama, John Kirkwood, et al. 2021. “546 Results from Phase Ib Study of Tebentafusp (Tebe) in Combination with Durvalumab (Durva) and / or Tremelimumab (Treme) in Metastatic Cutaneous Melanoma (MCM)”. Journal of Cancer Immunotherapy 9 (Supplement 2): A576–A576.

[0264] Hassel, Jessica Cecile, Piotr Rutkowski, Jean-Francois Baurain, Marcus O. Butler, Max Schlaak, Ryan Sullivan, Sebastian Ochsenreither, et al. 2021. "Co-Primary Endpoint of Overall Survival for Tebentafusp (Tebe)-Induced Rash in a Phase 3 Randomized Trial Comparing Tebe versus Investigator's Choice (IC) in First-Line Metastatic Uveal Melanoma". *Journal of Clinical Oncology* Journal of Clinical Oncology )》 39 (15_Supplement): 9527–9527.

[0265] Hayashi, K., F. Nikolos, YC Lee, A. Jain, E. Tsouko, H. Gao, A. Kasabyan et al. 2020. “Tipping the Immunostimulatory and Inhibitory DAMP Balance to HarnessImmunogenic Cell Death”. Nature Communications (2020). Nature Communications )》 11 (1): 6299.

[0266] Hoff, Daniel D. Von, Drew W. Rasco, Elisabeth I. Heath, Pamela N. Munster, Jan HM Schellens, Nicolas Isambert, Christophe Le Tourneau et al. 2018. “Phase I Study of CC-486 Alone and in Combination with Carboplatin or Nab-Paclitaxel in Patients with Relapsed or Refractory Solid Tumors.” Clinical Cancer Research (Clinical Cancer Research) Clinical Cancer Research )》 24 (17): 4072–80.

[0267] Jang, H. Josh, Galen Hostetter, Alexander W. MacFarlane, Zachary Madaj, Eric A. Ross, Toshinori Hinoue, Justin R. Kulchycki et al. 2023. “A Phase II Trial of Guadecitabine plus Atezolizumab in Metastatic Urothelial Carcinoma Progressing after Initial Immune Checkpoint Inhibitor Therapy”. Clinical Cancer Research 29 (11): 2052–65.

[0268] Lau, Tat San, Loucia Kit Ying Chan, Gene Chi Wai Man, Chi Hang Wong, Jacqueline Ho Sze Lee, So Fan Yim, Tak Hong Cheung, Iain A. McNeish and Joseph Kwong. 2020. “Paclitaxel Immunogenic Cell Death in Ovarian Cancer via TLR4 / IKK2 / SNARE-Dependent Exocytosis”. Cancer Immunology Research 8 (8): 1099–1111.

[0269] Le, Hanh K., Laura Graham, Esther Cha, Johanna K. Morales, Masoud H. Manjili, and Harry D. Bear. 2009. “Gemcitabine Directly Inhibits Myeloid-Derived Suppressor Cells in BALB / c Mice Bearing 4T1 Mammary Carcinoma and Augments Expansion of T Cells from Tumor-Bearing Mice.” International Journal of Immunopharmacology (2009). International Immunopharmacology )》 9 (7–8): 900–909.

[0270] Li, Chunxiao, Xiaofei Xu, Shuhua Wei, Ping Jiang, Lixiang Xue and Junjie Wang. 2021. “Tumor-Associated Macrophages: Potential Therapeutic Strategies and Future Prospects in Cancer”. Journal of Cancer Immunotherapy 9 (1): e001341.

[0271] Li, Xiang, Yaru Li, Liang Dong, Yixin Chang, Xingying Zhang, Chunmeng Wang, Meixia Chen, et al. 2023. "Decitabine-Priming Increases Anti-PD-1 Antitumor Efficacy by Promoting CD8+ Progenitor Exhausted T-Cell Expansion in Tumor Models". *Journal of Clinical Research* (2023). Journal of Clinical Investigation )》 133 (7):e165673.

[0272] Long, Jason E., Matthew J. Wongchenko, Dorothee Nickles, Wei-JenChung, Bu-er Wang, Johannes Riegler, Ji Li et al. 2019. “Therapeutic Resistance and Susceptibility Is Shaped by Cooperative Multi-Compartment Tumor Adaptation.” Cell Death and Differentiation (… Cell Death and Differentiation )》 26 (11): 2416–29.

[0273] Matei, Daniela, Sharad Ghamande, Lynda Roman, Angeles Alvarez Secord, John Nemunaitis, Merry Jennifer Markham, Kenneth P. Nephew et al. 2018. “A Phase I Clinical Trial of Guadecitabine and Carboplatin in Platinum-Resistant, Recurrent Ovarian Cancer: Clinical, Pharmacokinetic, and Pharmacodynamic Analyses”. Clinical Cancer Research (Journal of Clinical Cancer Research). Clinical Cancer Research )》 24 (10):2285–93.

[0274] Michels, Tillmann, Galina V. Shurin, Hiam Naiditch, Alexandra Sevko, Viktor Umansky, and Michael R. Shurin. 2012. “Paclitaxel Promotes Differentiation of Myeloid-Derived Suppressor Cells into Dendritic Cells in Vitro in a TLR4-Independent Manner.” Journal of Immunotoxicology (Journal of Immunotoxicology) Journal of Immunotoxicology )》 9 (3): 292–300.

[0275] Middleton, Mark R, Cheryl McAlpine, Victoria K Woodcock, Pippa Corrie, Jeffrey R Infante, Neil M Steven, Thomas R Jeffry Evans et al. 2020. “Tebentafusp, A TCR / Anti-CD3 Bispecific Fusion Protein Targeting Gp100, Potently Activated Antitumor Immune Responses in Patients with Metastatic Melanoma”. Clinical Cancer Research: Official Journal of the American Association for Cancer Research (AAAC). Clinical Cancer Research: An Official Journal of the American Association for Cancer Research )》 26 (22): 5869–78.

[0276] Nathan, Paul, Jessica C. Hassel, Piotr Rutkowski, Jean-Francois Baurain, Marcus O. Butler, Max Schlaak, Ryan J. Sullivan, et al. 2021. “Overall Survival Benefit with Tebentafusp in Metastatic Uveal Melanoma”. The New England Journal of Medicine (NJM). New England Journal of Medicine )》 385 (13): 1196–1206.

[0277] Palomba, Maria Lia, Guillaume Cartron, Leslie Popplewell, Vincent Ribrag, Jason Westin, Ling-Yuh Huw, Shefali Agarwal et al. 2022. "The effect of the combination of atezolizumab and tazemetostat in patients with relapsed / refractory diffuse large B-cell lymphoma: Results from a Phase Ib Study." *Clinical Lymphoma, Myeloma and Leukemia* Clinical Lymphoma Myeloma and Leukemia )》22 (7): 504–12.

[0278] Pauken, Kristen E., Morgan A. Sammons, Pamela M. Odorizzi, SasikanthManne, Jernej Godec, Omar Khan, Adam M. Drake et al. 2016. "Epigenetic Stability of Exhausted T Cells Limits Durability of Reinvigoration by PD-1 Blockade". Science (2016). Science )》354 (6316): 1160–65.

[0279] Petrovic, K., D. Depoil, DM Gascoyne, K. Page, A. Curnock, and A. Benlahrech. 2021. “1016P ImmTAC Redirect Exhausted Tumor-Infiltrating T-Cells: An Effect Enhanced by Pembrolizumab against PD-L1+ Tumors”. Annals of Oncology ( Annals of Oncology )》 32: S856.

[0280] Samlowski, Wolfram E., Sancy A. Leachman, Mark Wade, Pamela Cassidy, Patricia Porter-Gill, Leslie Busby, Richard Wheeler et al. 2005. “Evaluation of a 7-Day Continuous Intravenous Infusion of Decitabine: Inhibition of Promoter-Specific and Global Genomic DNA Methylation”. Journal of Clinical Oncology 23(17): 3897–3905.

[0281] Schaer, David A., Sandaruwan Geeganage, Nelusha Amaladas, Zhao HaiLu, Erik R. Rasmussen, Andreas Sonyi, Darin Chin et al. 2019. “The Folate Pathway Inhibitor Pemetrexed Pleiotropically Enhances Effects of Cancer Immunotherapy”. Clinical Cancer Research 25(23): 7175–88.

[0282] Sen, Debattama R., James Kaminski, R. Anthony Barnitz, Makoto Kurachi, Ulrike Gerdemann, Kathleen B. Yates, Hsiao-Wei Tsao et al. 2016. “The Epigenetic Landscape of T Cell Exhaustion”. Science ( Science )》 354 (6316): 1165–69.

[0283] Short, Nicholas J., Gautam Borthakur, Naveen Pemmaraju, Courtney D. Dinardo, Tapan M. Kadia, Elias Jabbour, Marina Konopleva et al. 2022. “A Multi-Arm Phase Ib / II Study Designed for Rapid, Parallel Evaluation of Novel Immunotherapy Combinations in Relapsed / Refractory Acute Myeloid Leukemia.” (Leukemia & Lymphoma) Leukemia & Lymphoma )》 63 (9): 2161–70.

[0284] Suzuki, Eiji, Veena Kapoor, Arminder Singh Jassar, Larry R. Kaiser, and Steven M. Albelda. 2005. “Gemcitabine selectively eliminates splenic Gr-1+ / CD11b+ myeloid suppressor cells in tumor-bearing animals and enhances antitumor immune activity.” Clinical Cancer Research 11 (18): 6713–21.

[0285] Takayama, Takuma, Taro Shimizu, Amr S. Abu Lila, Yuki Kanazawa, Hidenori Ando, ​​Yu Ishima, and Tatsuhiro Ishida. 2020. “Adjuvant Antitumor Immunity Contributes to the Overall Antitumor Effect of Pegylated Liposomal Doxorubicin (Doxil®) in C26 Tumor-Bearing Immunocompetent Mice.” Pharmaceutics ( Pharmaceutics )》 12 (10): 990.

[0286] Taylor, Kirsty, Helen Loo Yau, Ankur Chakravarthy, Ben Wang, Shu YiShen, Ilias Ettayebi, Charles A Ishak et al. 2020. “An Open-Label, Phase II Multicohort Study of an Oral Hypomethylating Agent CC-486 and Durvalumab in Advanced Solid Tumors”. Journal of Cancer Immunotherapy 8(2): e000883.

[0287] Torres, Evanthia T Roussos, Christine Rafie, Chenguang Wang, David Lim, Adam Brufsky, Patricia LoRusso, Joseph Paul Eder et al. 2021. “Phase I Study of Entinostat and Nivolumab with or without Ipilimumab in Advanced Solid Tumors (ETCTN-9844)”. Clinical Cancer Research: The Official Journal of the American Association for Cancer Research 27(21):5828–37.

[0288] Wanderley, Carlos W, David F Colon, Joao Paulo Mesquita Luiz, Francisco F Oliveira, Paula R. Viacava, Caio A Leite, Janaina A Pereira et al. 2018. "Paclitaxel Reduces Tumor Growth by Reprogramming Tumor-Associated Macrophages to an M1 Profile in a TLR4-Dependent Manner." Cancer Research (Journal of Cancer Research). Cancer Research )》 78 (20): canres.3480.2017.

[0289] Wherry, E. John, and Makoto Kurachi. 2015. “Molecular and Cellular Insights into T Cell Exhaustion”. *Nature Reviews Immunology*. Nature Reviews Immunology )》 15 (8): 486–99.

[0290] Wong, Kah Keng, Rosline Hassan, and Nik Soriani Yaacob. 2021. “Hypomethylating Agents and Immunotherapy: Therapeutic Synergism in Acute Myeloid Leukemia and Myelodysplastic Syndromes”. Frontiers in Oncology (… Frontiers in Oncology )》 11: 624742.

[0291] Yamaguchi, Takahisa, Sachio Fushida, Yasuhiko Yamamoto, Tomoya Tsukada, Jun Kinoshita, Katsunobu Oyama, Tomoharu Miyashita, et al. 2017. "Low-Dose Paclitaxel Suppresses the Induction of M2 Macrophages in Gastric Cancer". *Oncology Reports* Oncology Reports )》 37 (6): 3341–50.

[0292] Yao, Yushi, Jihao Zhou, Lixin Wang, Xiaoning Gao, Qiaoyang Ning, Mengmeng Jiang, Jia Wang, Lili Wang, and Li Yu. 2013. "Increased PRAME-Specific CTL Killing of Acute Myeloid Leukemia Cells by Either a Novel Histone Deacetylase Inhibitor Chidamide Alone or Combined Treatment with Decitabine". PLOS ONE (Public Library of Science, General) PLoS ONE )》 8 (8): e70522.

[0293] Yi, Ming, Xiaoli Zheng, Mengke Niu, Shuangli Zhu, Hong Ge, and Kongming Wu. 2022. “Combination Strategies with PD-1 / PD-L1 Blockade: Current Advances and Future Directions”. Molecular Cancer (Molecular Cancer) Molecular Cancer )》 21 (1): 28.

Claims

1. A method of treating a subject with non-small cell lung cancer or small cell lung cancer, the method comprising administering a composition to the subject according to a dosing regimen, the composition comprising (I) a heterodimeric TCR-anti-CD3 antibody fusion molecule, and (II) a second composition comprising a second active agent; The heterodimeric TCR-antiCD3 antibody fusion molecule comprises (a) an α-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 14, and (b) a β-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 16, wherein the α-chain comprises a complementarity-determining region (CDR), the CDR comprising α-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 3, 4, and 5, respectively, and the β-chain comprises β-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 9, 10, and 11, respectively. The administration regimen mentioned above includes the following: (i) A first dose comprising 2 μg to 30 μg of the TCR-antiCD3 antibody fusion molecule, (ii) A second dose, the second dose comprising 5 μg to 60 μg of the TCR-antiCD3 antibody fusion molecule, and (iii) A maintenance dose comprising 15 μg to 200 μg of the TCR-antiCD3 antibody fusion molecule, wherein the maintenance dose is administered in at least three phases, wherein the first phase is administered every 6 to 8 days, the second phase every 12 to 16 days, and the third phase every 26 to 30 days; and The second dose contains a greater amount of the TCR-antiCD3 antibody fusion molecule than the first dose, and the maintenance dose contains a greater amount of the TCR-antiCD3 antibody fusion molecule than the second dose; and The second active agent is administered at least once before, in parallel with, and / or after the first dose containing the TCR-antiCD3 antibody fusion molecule.

2. The method according to claim 1, wherein the second active agent is a chemotherapeutic agent.

3. The method according to claim 2, wherein the chemotherapeutic agent is gemcitabine.

4. The method of claim 2, wherein the chemotherapeutic agent is docetaxel.

5. The method of claim 1, wherein the second active agent is a targeted therapy.

6. The method of claim 5, wherein the targeted therapy is osimertinib.

7. The method of claim 1, wherein the second active agent is a multimodal therapy.

8. The method of claim 7, wherein the multimodal therapy is carboplatin / paclitaxel / pembrolizumab.

9. The method of claim 7, wherein the multimodal therapy is carboplatin / pemetrexed / pembrolizumab.

10. A method of treating a subject with ovarian cancer, the method comprising administering a composition to the subject according to a dosing regimen, the composition comprising (I) a heterodimeric TCR-antiCD3 antibody fusion molecule, and (II) a second active agent. The heterodimeric TCR-antiCD3 antibody fusion molecule comprises (a) an α-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 14, and (b) a β-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 16, wherein the α-chain comprises a complementarity-determining region (CDR), the CDR comprising α-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 3, 4, and 5, respectively, and the β-chain comprises β-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 9, 10, and 11, respectively. The administration regimen mentioned above includes the following: (i) A first dose comprising 2 μg to 30 μg of the TCR-antiCD3 antibody fusion molecule, (ii) A second dose, the second dose comprising 5 μg to 60 μg of the TCR-antiCD3 antibody fusion molecule, and (iii) A maintenance dose comprising 15 μg to 200 μg of the TCR-antiCD3 antibody fusion molecule, wherein the maintenance dose is administered in at least three phases, wherein the first phase is administered every 6 to 8 days, the second phase every 12 to 16 days, and the third phase every 26 to 30 days; and The second dose contains a greater amount of the TCR-antiCD3 antibody fusion molecule than the first dose, and the maintenance dose contains a greater amount of the TCR-antiCD3 antibody fusion molecule than the second dose; and The second active agent is administered at least once before, in parallel with, and / or after the first dose containing the TCR-antiCD3 antibody fusion molecule.

11. The method of claim 10, wherein the second active agent is a chemotherapeutic agent.

12. The method of claim 11, wherein the chemotherapeutic agent is gemcitabine.

13. The method of claim 11, wherein the chemotherapeutic agent is pegylated liposomal doxorubicin (PLD).

14. The method of claim 11, wherein the chemotherapeutic agent is carboplatin / paclitaxel.

15. The method of claim 10, wherein the second active agent is a targeted therapy.

16. The method of claim 15, wherein the targeted therapy is bevacizumab.

17. The method of claim 10, wherein the second active agent is a multimodal therapy.

18. The method of claim 17, wherein the multimodal therapy is carboplatin / paclitaxel / pembrolizumab.

19. The method of claim 17, wherein the multimodal therapy is carboplatin / gemcitabine / bevacizumab.

20. The method according to any one of claims 1 to 19, wherein the first dose comprises 2 μg to 4 μg of the TCR-antiCD3 antibody fusion molecule, the second dose comprises 5 μg to 14 μg of the TCR-antiCD3 antibody fusion molecule, and the maintenance dose comprises 15 μg to 55 μg of the TCR-antiCD3 antibody fusion molecule.

21. The method of claim 20, wherein the first dose comprises 3 μg of the TCR-antiCD3 antibody fusion molecule, the second dose comprises 10 μg of the TCR-antiCD3 antibody fusion molecule, and the maintenance dose comprises 40 μg of the TCR-antiCD3 antibody fusion molecule.

22. The method of claim 1, wherein the first dose comprises 15 μg to 40 μg of the TCR-antiCD3 antibody fusion molecule, the second dose comprises 30 μg to 80 μg of the TCR-antiCD3 antibody fusion molecule, and the maintenance dose comprises 140 μg to 200 μg of the TCR-antiCD3 antibody fusion molecule.

23. The method of claim 22, wherein the first dose comprises 20 μg of the TCR-antiCD3 antibody fusion molecule, the second dose comprises 40 μg of the TCR-antiCD3 antibody fusion molecule, and the maintenance dose comprises 160 μg of the TCR-antiCD3 antibody fusion molecule.

24. The method according to any one of claims 1 to 23, wherein the α-chain amino acid sequence is 100% identical to the amino acid sequence of SEQ ID NO:

14.

25. The method according to any one of claims 1 to 24, wherein the β-chain amino acid sequence is 100% identical to the amino acid sequence of SEQ ID NO:

16.

26. The method according to any one of claims 1 to 25, wherein the first dose is administered once every 7 days.

27. The method according to any one of claims 1 to 25, wherein the first dose is administered for 1 to 3 weeks.

28. The method of claim 27, wherein the first dose is administered for one week.

29. The method according to any one of claims 1 to 28, wherein the second dose is administered 6 to 8 days after the first dose.

30. The method according to any one of claims 1 to 29, wherein the second dose is administered once every 7 days.

31. The method according to any one of claims 1 to 30, wherein the second dose is administered for 1 to 3 weeks.

32. The method of claim 31, wherein the second dose is administered for one week.

33. The method according to any one of claims 1 to 32, wherein the maintenance dose in the first phase is administered 6 to 8 days after the second dose.

34. The method according to any one of claims 1 to 33, wherein the maintenance dose in the first stage is administered once a week.

35. The method according to any one of claims 1 to 34, wherein the first stage is from about 2 weeks to about 101 weeks.

36. The method of claim 35, wherein the first stage is from about 8 weeks to about 14 weeks.

37. The method of claim 36, wherein the first stage is 10 weeks.

38. The method according to any one of claims 1 to 37, wherein the maintenance dose in the second stage is administered once every 2 weeks.

39. The method according to any one of claims 1 to 38, wherein the second stage is about 30 to 50 weeks.

40. The method of claim 39, wherein the second phase is 38 weeks.

41. The method according to any one of claims 1 to 40, wherein the maintenance dose in the third stage is administered once every 4 weeks.

42. The method according to any one of claims 1 to 41, wherein the third stage is about 40 weeks to about 60 weeks.

43. The method of claim 42, wherein the third stage lasts approximately 48 weeks.

44. The method according to any one of claims 1 to 43, wherein the first stage is 10 weeks, the second stage is 38 weeks, and the third stage is 48 weeks.

45. The method according to any one of claims 1 to 44, wherein the first dose is administered for 1 week, the second dose is administered for 1 week, and the maintenance dose is administered for at least 40 weeks.

46. ​​The method according to any one of claims 1 to 45, wherein the dosing regimen is from about 90 weeks to about 110 weeks.

47. The method according to any one of claims 1 to 46, wherein no corticosteroids are administered within three weeks prior to and throughout the dosing regimen.

48. The method according to any one of claims 1 to 47, wherein the subject is HLA-A 02 positive.

49. The method according to any one of claims 1 to 48, wherein the subject is HLA-A 02:01 Positive.

50. The method according to any one of claims 1 to 49, wherein the subject has a BRAF V600 mutation.

51. The method according to any one of claims 1 to 50, wherein the subject has been diagnosed with a life expectancy of more than 3 months.

52. The method according to any one of claims 1 to 51, wherein the second active agent is co-administered with the heterodimeric TCR-antiCD3 antibody fusion molecule.

53. The method according to any one of claims 1 to 51, wherein the second active agent is first applied 1 to 6 weeks prior to the application of the heterodimeric TCR-antiCD3 antibody fusion molecule.

54. The method according to any one of claims 1 to 51, wherein the second active agent is applied 1 to 6 weeks after the application of the heterodimeric TCR-antiCD3 antibody fusion molecule.

55. The method according to any one of claims 1 to 51, wherein the heterodimeric TCR-antiCD3 antibody fusion molecule is applied after treatment with the second active agent has been completed.

56. The method according to any one of claims 1 to 55, wherein the second active agent is applied 1 to 6 weeks prior to the application of the heterodimeric TCR-antiCD3 antibody fusion molecule.

57. The method according to any one of claims 1 to 56, wherein the second activator is applied for at least 10 weeks.

58. The method of claim 56, wherein the second activator is applied for at least 40 weeks.

59. The method of claim 56, wherein the second active agent is administered during the duration of the administration regimen.

60. The method according to any one of claims 1 or 59, wherein the second active agent is gemcitabine.

61. The method of claim 60, wherein the gemcitabine is administered at approximately 750 mg / m². 2 Approximately 1000 mg / m 2 Apply.

62. The method of claim 60, wherein the gemcitabine is administered at approximately 1000 mg / m². 2 Dosage administration.

63. The method according to any one of claims 60 to 62, wherein the gemcitabine is applied once a week to once a month.

64. The method according to any one of claims 60 to 63, wherein the gemcitabine is administered generally as follows: (i) for 4 weeks followed by a 1-week break, (ii) for 3 weeks followed by a 1-week break, (iii) for 2 weeks followed by a 1-week break, (iv) for 1 week followed by a 1-week break, (v) once a week, or (vi) once every two weeks.

65. The method according to any one of claims 1 to 59, wherein the second active agent is albumin-bound paclitaxel.

66. The method of claim 65, wherein the albumin-bound paclitaxel is administered at approximately 75 mg / m³. 2 Approximately 100 mg / m 2 Apply.

67. The method of claim 65, wherein the albumin-bound paclitaxel is administered at approximately 100 mg / m³. 2 Dosage administration.

68. The method according to any one of claims 65 to 67, wherein the albumin-bound paclitaxel is administered once a week to once a month.

69. The method according to any one of claims 65 to 68, wherein the albumin-bound paclitaxel is administered generally as follows: (i) for 4 weeks followed by a 1-week break, (ii) for 3 weeks followed by a 1-week break, (iii) for 2 weeks followed by a 1-week break, (iv) for 1 week followed by a 1-week break, (v) once a week, or (vi) once every two weeks.

70. The method according to any one of claims 1 to 59, wherein the second active agent is decitabine.

71. The method of claim 70, wherein the decitabine is administered at about 0.07 mg / kg to about 0.6 mg / kg.

72. The method according to claim 70 or 71, wherein the decitabine is applied once a week to once a month.

73. The method according to any one of claims 70 to 72, wherein the decitabine is administered generally as follows: (i) once a week, (ii) once every two weeks, (iii) once every three weeks, or (iv) once a month.

74. The method according to any one of claims 1 to 59, wherein the second activator is polyethylene glycol-modified liposome doxorubicin (PLD).

75. The method of claim 74, wherein approximately 40 mg / m² is administered every 4 to 6 weeks. 2 Up to 60 mg / m 2 Apply the PLD.

76. The method according to any one of claims 1 to 75, wherein the composition is administered orally, subcutaneously, sublingually, intramuscularly, intravenously, intranasally, transdermally, or in any combination thereof.

77. The method according to any one of claims 1 to 76, wherein the second composition is administered orally, subcutaneously, sublingually, intramuscularly, intravenously, intranasally, transdermally, or in any combination thereof.

78. A method of treating a subject with non-small cell lung cancer or small cell lung cancer, the method comprising administering a composition to the subject according to a dosing regimen, the composition comprising (I) a heterodimeric TCR-anti-CD3 antibody fusion molecule, and (II) a second composition comprising a second active agent. The heterodimeric TCR-antiCD3 antibody fusion molecule comprises (a) an α-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 14, and (b) a β-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 16, wherein the α-chain comprises a complementarity-determining region (CDR), the CDR comprising α-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 3, 4, and 5, respectively, and the β-chain comprises β-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 9, 10, and 11, respectively. The dosing regimen includes: (i) Administering a first dose, said first dose comprising 3 μg of the TCR-antiCD3 antibody fusion molecule, (ii) One week after the first dose, a second dose is administered, said second dose comprising 10 μg of the TCR-antiCD3 antibody fusion molecule, and (iii) A maintenance dose comprising 40 μg of the TCR-antiCD3 antibody fusion molecule is administered once weekly for one week after the second dose and during the first phase, wherein the first phase is approximately 9 to 10 weeks. (iv) The maintenance dose shall be administered every 2 weeks during the second phase, wherein the second phase is approximately 38 weeks, and (v) The maintenance dose is administered every 4 weeks during the third phase, which is approximately 48 weeks. The second active agent is administered 1 to 6 weeks prior to the administration of the heterodimeric TCR-antiCD3 antibody fusion molecule, during the duration of administration of the heterodimeric TCR-antiCD3 antibody fusion molecule, and / or after the administration of the heterodimeric TCR-antiCD3 antibody fusion molecule.

79. A method of treating a subject with ovarian cancer, the method comprising administering a composition to the subject according to a dosing regimen, the composition comprising (I) a heterodimeric TCR-antiCD3 antibody fusion molecule, and (II) a second composition comprising a second active agent. The heterodimeric TCR-antiCD3 antibody fusion molecule comprises (a) an α-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 14, and (b) a β-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 16, wherein the α-chain comprises a complementarity-determining region (CDR), the CDR comprising α-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 3, 4, and 5, respectively, and the β-chain comprises β-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 9, 10, and 11, respectively. The dosing regimen includes: (i) Administering a first dose, said first dose comprising 3 μg of the TCR-antiCD3 antibody fusion molecule, (ii) One week after the first dose, a second dose is administered, said second dose comprising 10 μg of the TCR-antiCD3 antibody fusion molecule, and (iii) A maintenance dose comprising 40 μg of the TCR-antiCD3 antibody fusion molecule is administered once weekly for one week after the second dose and during the first phase, wherein the first phase is approximately 9 to 10 weeks. (iv) The maintenance dose shall be administered every 2 weeks during the second phase, wherein the second phase is approximately 38 weeks, and (v) The maintenance dose is administered every 4 weeks during the third phase, which is approximately 48 weeks. The second active agent is administered 1 to 6 weeks prior to the administration of the heterodimeric TCR-antiCD3 antibody fusion molecule, during the duration of administration of the heterodimeric TCR-antiCD3 antibody fusion molecule, and / or after the administration of the heterodimeric TCR-antiCD3 antibody fusion molecule.

80. A method of treating a subject with non-small cell lung cancer or small cell lung cancer, the method comprising administering a composition to the subject according to a dosing regimen, the composition comprising (I) a heterodimeric TCR-anti-CD3 antibody fusion molecule, and (II) a second composition comprising a second active agent. The heterodimeric TCR-antiCD3 antibody fusion molecule comprises (a) an α-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 14, and (b) a β-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 16, wherein the α-chain comprises a complementarity-determining region (CDR), the CDR comprising α-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 3, 4, and 5, respectively, and the β-chain comprises β-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 9, 10, and 11, respectively. The dosing regimen includes: (i) Administering a first dose, said first dose comprising 20 μg of the TCR-antiCD3 antibody fusion molecule, (ii) One week after the first dose, a second dose is administered, said second dose comprising 40 μg of the TCR-antiCD3 antibody fusion molecule, and (iii) One week after the second dose, and then weekly during the first phase, the maintenance dose comprising 160 μg of the TCR-anti-CD3 antibody fusion molecule, wherein the first phase is approximately 9 to 10 weeks. (iv) The maintenance dose shall be administered every 2 weeks during the second phase, wherein the second phase is approximately 38 weeks, and (v) The maintenance dose is administered every 4 weeks during the third phase, which is approximately 48 weeks. The second active agent is administered 1 to 6 weeks prior to the administration of the heterodimeric TCR-antiCD3 antibody fusion molecule, during the duration of administration of the heterodimeric TCR-antiCD3 antibody fusion molecule, and / or after the administration of the heterodimeric TCR-antiCD3 antibody fusion molecule.

81. A method of treating a subject with ovarian cancer, the method comprising administering a composition to the subject according to a dosing regimen, the composition comprising (I) a heterodimeric TCR-antiCD3 antibody fusion molecule, and (II) a second composition comprising a second active agent. The heterodimeric TCR-antiCD3 antibody fusion molecule comprises (a) an α-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 14, and (b) a β-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 16, wherein the α-chain comprises a complementarity-determining region (CDR), the CDR comprising α-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 3, 4, and 5, respectively, and the β-chain comprises β-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 9, 10, and 11, respectively. The dosing regimen includes: (i) Administering a first dose, said first dose comprising 20 μg of the TCR-antiCD3 antibody fusion molecule, (ii) One week after the first dose, a second dose is administered, said second dose comprising 40 μg of the TCR-antiCD3 antibody fusion molecule, and (iii) One week after the second dose, and then weekly during the first phase, the maintenance dose comprising 160 μg of the TCR-anti-CD3 antibody fusion molecule, wherein the first phase is approximately 9 to 10 weeks. (iv) The maintenance dose shall be administered every 2 weeks during the second phase, wherein the second phase is approximately 38 weeks, and (v) The maintenance dose is administered every 4 weeks during the third phase, which is approximately 48 weeks. The second active agent is administered 1 to 6 weeks prior to the administration of the heterodimeric TCR-antiCD3 antibody fusion molecule, during the duration of administration of the heterodimeric TCR-antiCD3 antibody fusion molecule, and / or after the administration of the heterodimeric TCR-antiCD3 antibody fusion molecule.

82. The method according to any one of claims 78 to 81, wherein the second active agent is a chemotherapeutic agent.

83. The method of claim 82, wherein the chemotherapeutic agent is gemcitabine.

84. The method of claim 82, wherein the chemotherapeutic agent is docetaxel.

85. The method according to any one of claims 78 to 81, wherein the second active agent is a targeted therapy.

86. The method of claim 85, wherein the targeted therapy is osimertinib.

87. The method according to any one of claims 78 to 81, wherein the second active agent is a multimodal therapy.

88. The method of claim 87, wherein the multimodal therapy is carboplatin / paclitaxel / pembrolizumab.

89. The method of claim 87, wherein the multimodal therapy is carboplatin / pemetrexed / pembrolizumab.

90. A reagent kit comprising: (i) A first container containing a first dose, the first dose containing 2 μg to 4 μg of a heterodimeric TCR-antiCD3 antibody fusion molecule. (ii) A second container containing a second dose, the second dose containing 5 μg to 14 μg of the heterodimeric TCR-antiCD3 antibody fusion molecule. (iii) One or more maintenance containers, each containing a maintenance dose, said maintenance dose containing 15 μg to 55 μg of the heterodimeric TCR-antiCD3 antibody fusion molecule. (iv) One or more containers containing a second dose of the active agent; (v) Instructions for use for administering the heterodimeric TCR-antiCD3 antibody fusion molecule according to a dosing regimen, wherein the dosing regimen specifies: (a) administration of the first dose in week 1, (b) administration of the second dose in week 2, (c) administration of a maintenance dose once weekly for 10 weeks during the first phase, (d) administration of the maintenance dose once every 2 weeks for 38 weeks during the second phase, and (e) administration of the maintenance dose once every 4 weeks for 48 weeks during the third phase; and (vi) Instructions for use in combination with the heterodimeric TCR-antiCD3 antibody fusion molecule to administer the second active agent according to the dosing regimen, wherein the dosing regimen is specified as: (a) once a week for two weeks, followed by a one-week break; (b) once a week for three weeks, followed by a one-week break; or (c) once a week for two weeks, followed by a one-week break.

91. A reagent kit comprising: (i) A first container containing a first dose, the first dose containing 20 μg of a heterodimeric TCR-antiCD3 antibody fusion molecule. (ii) A second container containing a second dose, the second dose containing 40 μg of the heterodimeric TCR-antiCD3 antibody fusion molecule. (iii) One or more maintenance containers, each containing a maintenance dose comprising 160 μg of the heterodimeric TCR-antiCD3 antibody fusion molecule. (iv) One or more containers containing a second dose of the active agent; (v) Instructions for use for administering the heterodimeric TCR-antiCD3 antibody fusion molecule according to a dosing regimen, wherein the dosing regimen specifies: (a) administration of the first dose in week 1, (b) administration of the second dose in week 2, (c) administration of a maintenance dose once weekly for 10 weeks during the first phase, (d) administration of the maintenance dose once every 2 weeks for 38 weeks during the second phase, and (e) administration of the maintenance dose once every 4 weeks for 48 weeks during the third phase; and (vi) Instructions for use in combination with the heterodimeric TCR-antiCD3 antibody fusion molecule to administer the second active agent according to the dosing regimen, wherein the dosing regimen is specified as: (a) once a week for two weeks, followed by a one-week break; (b) once a week for three weeks, followed by a one-week break; or (c) once a week for two weeks, followed by a one-week break.

92. The kit according to claim 90 or 91, wherein the heterodimeric TCR-antiCD3 antibody fusion molecule comprises (a) an α-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 14, and (b) a β-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 16, wherein the α-chain comprises a complementarity-determining region (CDR), the CDR comprising α-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 3, 4, and 5, respectively, and the β-chain comprises β-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 9, 10, and 11, respectively.

93. The kit according to any one of claims 90 to 92, wherein the kit comprises 2 to 10 maintenance containers.

94. The kit according to any one of claims 90 to 93, wherein the first dose, the second dose, and / or the maintenance dose further comprise pharmaceutically acceptable excipients to form the first pharmaceutical formulation, the second pharmaceutical formulation, and / or the maintenance pharmaceutical formulation, respectively.

95. The kit of claim 94, wherein the pH of the first pharmaceutical preparation, the second pharmaceutical preparation and / or the maintenance pharmaceutical preparation is about 6.5 to about 7.

5.

96. The kit according to any one of claims 90 to 95, wherein the second active agent is a chemotherapeutic agent.

97. The kit according to claim 96, wherein the chemotherapeutic agent is gemcitabine.

98. The kit according to claim 97, wherein the dosage of gemcitabine is 1000 mg / m². 2 .

99. The kit according to claim 96, wherein the chemotherapeutic agent is docetaxel.

100. The kit according to claim 99, wherein the dosage of docetaxel is 75 mg / m². 2 .

101. The kit according to claim 96, wherein the chemotherapeutic agent is pegylated liposomal doxorubicin (PLD).

102. The kit according to claim 96, wherein the chemotherapeutic agent is carboplatin / paclitaxel.

103. The kit according to any one of claims 90 to 95, wherein the second active agent is a targeted therapy.

104. The kit according to claim 103, wherein the targeted therapy is osimertinib.

105. The kit according to claim 104, wherein the dosage of osimertinib is 80 mg.

106. The kit according to claim 104, wherein the dosage of osimertinib is 40 mg.

107. The kit according to claim 103, wherein the targeted therapy is bevacizumab.

108. The kit according to any one of claims 90 to 95, wherein the second active agent is a multimodal therapy.

109. The kit according to claim 108, wherein the multimodal therapy is carboplatin / paclitaxel / pembrolizumab.

110. The kit according to claim 109, wherein the dosage of carboplatin is AUC 6.

111. The kit according to claim 109 or 110, wherein the dosage of paclitaxel is 200 mg / m². 2 .

112. The kit according to any one of claims 109 to 111, wherein the dosage of pembrolizumab is 400 mg.

113. The kit according to claim 108, wherein the multimodal therapy is carboplatin / pemetrexed / pembrolizumab.

114. The kit according to claim 113, wherein the dosage of carboplatin is AUC 5.

115. The kit according to claim 113, wherein the dosage of carboplatin is AUC 6.

116. The kit according to any one of claims 113 to 115, wherein the dosage of pemetrexed is 500 mg / m². 2 .

117. The kit according to any one of claims 113 to 116, wherein the dosage of pembrolizumab is 400 mg.

118. A method of treating a subject with PRAME-positive cancer, the method comprising administering a composition to the subject according to a dosing regimen, the composition comprising (I) a heterodimeric TCR-antiCD3 antibody fusion molecule according to the dosing regimen, and (II) a second composition comprising a second active agent. The heterodimeric TCR-antiCD3 antibody fusion molecule comprises (a) an α-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 14, and (b) a β-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 16, wherein the α-chain comprises a complementarity-determining region (CDR), the CDR comprising α-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 3, 4, and 5, respectively, and the β-chain comprises β-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 9, 10, and 11, respectively. The administration regimen mentioned above includes the following: (i) A first dose comprising 2 μg to 30 μg of the TCR-antiCD3 antibody fusion molecule, (ii) A second dose, the second dose comprising 5 μg to 60 μg of the TCR-antiCD3 antibody fusion molecule, and (iii) A maintenance dose comprising 15 μg to 200 μg of the TCR-antiCD3 antibody fusion molecule, wherein the maintenance dose is administered in at least three phases, wherein the first phase is administered every 6 to 8 days, the second phase every 12 to 16 days, and the third phase every 26 to 30 days; and The second dose contains a greater amount of the TCR-antiCD3 antibody fusion molecule than the first dose, and the maintenance dose contains a greater amount of the TCR-antiCD3 antibody fusion molecule than the second dose. The second active agent is administered 1 to 6 weeks prior to the administration of the heterodimeric TCR-antiCD3 antibody fusion molecule, during the duration of administration of the heterodimeric TCR-antiCD3 antibody fusion molecule, and / or after the administration of the heterodimeric TCR-antiCD3 antibody fusion molecule.

119. The method of claim 118, wherein the PRAME-positive cancer is non-small cell lung cancer or ovarian cancer.

120. A method of treating a subject with platinum-sensitive ovarian cancer, the method comprising administering a composition to the subject according to a dosing regimen, the composition comprising: (I) a heterodimeric TCR-antiCD3 antibody fusion molecule, and (II) a second composition comprising a second active agent. The heterodimeric TCR-antiCD3 antibody fusion molecule comprises (a) an α-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 14, and (b) a β-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 16, wherein the α-chain comprises a complementarity-determining region (CDR), the CDR comprising α-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 3, 4, and 5, respectively, and the β-chain comprises β-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 9, 10, and 11, respectively. The administration regimen mentioned above includes the following: (i) A first dose comprising 2 μg to 30 μg of the TCR-antiCD3 antibody fusion molecule, (ii) A second dose, the second dose comprising 5 μg to 60 μg of the TCR-antiCD3 antibody fusion molecule, and (iii) A maintenance dose comprising 15 μg to 200 μg of the TCR-antiCD3 antibody fusion molecule, wherein the maintenance dose is administered in at least three phases, wherein the first phase is administered every 6 to 8 days, the second phase every 12 to 16 days, and the third phase every 26 to 30 days; and The second dose contains a greater amount of the TCR-antiCD3 antibody fusion molecule than the first dose, and the maintenance dose contains a greater amount of the TCR-antiCD3 antibody fusion molecule than the second dose. The second active agent is administered 1 to 6 weeks prior to the administration of the heterodimeric TCR-antiCD3 antibody fusion molecule, during the duration of administration of the heterodimeric TCR-antiCD3 antibody fusion molecule, and / or after the administration of the heterodimeric TCR-antiCD3 antibody fusion molecule.

121. The method of claim 120, wherein the second active agent is a targeted therapy.

122. The method of claim 121, wherein the targeted therapy is bevacizumab.

123. The method of claim 120, wherein the second active agent is a chemotherapeutic agent.

124. The method of claim 123, wherein the chemotherapeutic agent is gemcitabine.

125. The method of claim 123, wherein the chemotherapeutic agent is carboplatin / paclitaxel.

126. The method of claim 123, wherein the chemotherapeutic agent is pegylated liposomal doxorubicin (PLD).

127. The method of claim 120, wherein the second active agent is a multimodal therapy.

128. The method of claim 127, wherein the multimodal therapy is carboplatin / paclitaxel / pembrolizumab.

129. The method of claim 127, wherein the multimodal therapy is carboplatin / gemcitabine / bevacizumab.

130. A method of treating a subject with platinum-resistant ovarian cancer, the method comprising administering a composition to the subject according to a dosing regimen, the composition comprising: (I) a heterodimeric TCR-antiCD3 antibody fusion molecule, and (II) a second composition comprising a second active agent. The heterodimeric TCR-antiCD3 antibody fusion molecule comprises (a) an α-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 14, and (b) a β-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 16, wherein the α-chain comprises a complementarity-determining region (CDR), the CDR comprising α-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 3, 4, and 5, respectively, and the β-chain comprises β-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 9, 10, and 11, respectively. The administration regimen mentioned above includes the following: (i) A first dose comprising 2 μg to 30 μg of the TCR-antiCD3 antibody fusion molecule, (ii) A second dose, the second dose comprising 5 μg to 60 μg of the TCR-antiCD3 antibody fusion molecule, and (iii) A maintenance dose comprising 15 μg to 200 μg of the TCR-antiCD3 antibody fusion molecule, wherein the maintenance dose is administered in at least three phases, wherein the first phase is administered every 6 to 8 days, the second phase every 12 to 16 days, and the third phase every 26 to 30 days; and The second dose contains a greater amount of the TCR-antiCD3 antibody fusion molecule than the first dose, and the maintenance dose contains a greater amount of the TCR-antiCD3 antibody fusion molecule than the second dose. The second active agent is administered 1 to 6 weeks prior to the administration of the heterodimeric TCR-antiCD3 antibody fusion molecule, during the duration of administration of the heterodimeric TCR-antiCD3 antibody fusion molecule, and / or after the administration of the heterodimeric TCR-antiCD3 antibody fusion molecule.

131. The method of claim 130, wherein the second active agent is a targeted therapy.

132. The method of claim 131, wherein the targeted therapy is bevacizumab.

133. The method of claim 131, wherein the second active agent is a chemotherapeutic agent.

134. The method of claim 133, wherein the chemotherapeutic agent is gemcitabine.

135. The method of claim 133, wherein the chemotherapeutic agent is carboplatin / paclitaxel.

136. The method of claim 133, wherein the chemotherapeutic agent is pegylated liposomal doxorubicin (PLD).

137. The method of claim 131, wherein the second active agent is a multimodal therapy.

138. The method of claim 137, wherein the multimodal therapy is carboplatin / paclitaxel / pembrolizumab.

139. The method of claim 133, wherein the multimodal therapy is carboplatin / gemcitabine / bevacizumab.

140. A method of treating a subject with non-small cell lung cancer, the method comprising administering a composition to the subject according to a dosing regimen, the composition comprising: (I) a heterodimeric TCR-anti-CD3 antibody fusion molecule, and (II) a second composition comprising a second active agent. The heterodimeric TCR-antiCD3 antibody fusion molecule comprises (a) an α-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 14, and (b) a β-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 16, wherein the α-chain comprises a complementarity-determining region (CDR), the CDR comprising α-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 3, 4, and 5, respectively, and the β-chain comprises β-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 9, 10, and 11, respectively. The administration regimen mentioned above includes the following: (i) A first dose comprising 2 μg to 30 μg of the TCR-antiCD3 antibody fusion molecule, (ii) A second dose, the second dose comprising 5 μg to 60 μg of the TCR-antiCD3 antibody fusion molecule, and (iii) A maintenance dose comprising 15 μg to 200 μg of the TCR-antiCD3 antibody fusion molecule, wherein the maintenance dose is administered in at least three phases, wherein the first phase is administered every 6 to 8 days, the second phase every 12 to 16 days, and the third phase every 26 to 30 days; and The second dose contains a greater amount of the TCR-antiCD3 antibody fusion molecule than the first dose, and the maintenance dose contains a greater amount of the TCR-antiCD3 antibody fusion molecule than the second dose. The second active agent is administered 1 to 6 weeks prior to the administration of the heterodimeric TCR-antiCD3 antibody fusion molecule, during the duration of administration of the heterodimeric TCR-antiCD3 antibody fusion molecule, and / or after the administration of the heterodimeric TCR-antiCD3 antibody fusion molecule.

141. The method of claim 140, wherein the second active agent is a chemotherapeutic agent.

142. The method of claim 141, wherein the chemotherapeutic agent is gemcitabine.

143. The method of claim 141, wherein the chemotherapeutic agent is docetaxel.

144. The method of claim 141, wherein the chemotherapeutic agent is pegylated liposomal doxorubicin (PLD).

145. The method of claim 141, wherein the chemotherapeutic agent is carboplatin / paclitaxel.

146. The method of claim 140, wherein the second active agent is a targeted therapy.

147. The method of claim 146, wherein the targeted therapy is osimertinib.

148. The method of claim 140, wherein the second active agent is a targeted therapy.

149. The method of claim 148, wherein the targeted therapy is osimertinib.

150. The method of claim 148, wherein the targeted therapy is bevacizumab.

151. The method of claim 140, wherein the second active agent is a multimodal therapy.

152. The method of claim 151, wherein the multimodal therapy is carboplatin / paclitaxel / pembrolizumab.

153. A method of treating a subject with melanoma, the method comprising administering a composition to the subject according to a dosing regimen, the composition comprising: (I) a heterodimeric TCR-antiCD3 antibody fusion molecule, and (II) a second composition comprising a second active agent. The heterodimeric TCR-antiCD3 antibody fusion molecule comprises (a) an α-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 14, and (b) a β-chain amino acid sequence having at least 90%, at least 95%, or 100% identity with the amino acid sequence of SEQ ID NO: 16, wherein the α-chain comprises a complementarity-determining region (CDR), the CDR comprising α-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 3, 4, and 5, respectively, and the β-chain comprises β-chain CDRs 1, 2, and 3 having the amino acid sequences of SEQ ID NO: 9, 10, and 11, respectively. The administration regimen mentioned above includes the following: (i) A first dose comprising 2 μg to 30 μg of the TCR-antiCD3 antibody fusion molecule, (ii) A second dose, the second dose comprising 5 μg to 60 μg of the TCR-antiCD3 antibody fusion molecule, and (iii) A maintenance dose comprising 15 μg to 200 μg of the TCR-antiCD3 antibody fusion molecule, wherein the maintenance dose is administered in at least three phases, wherein the first phase is administered every 6 to 8 days, the second phase every 12 to 16 days, and the third phase every 26 to 30 days; and The second dose contains a greater amount of the TCR-antiCD3 antibody fusion molecule than the first dose, and the maintenance dose contains a greater amount of the TCR-antiCD3 antibody fusion molecule than the second dose. The second active agent is administered 1 to 6 weeks prior to the administration of the heterodimeric TCR-antiCD3 antibody fusion molecule, during the duration of administration of the heterodimeric TCR-antiCD3 antibody fusion molecule, and / or after the administration of the heterodimeric TCR-antiCD3 antibody fusion molecule.

154. The method of claim 153, wherein the second active agent is a second heterodimeric TCR-antiCD3 antibody fusion molecule.

155. The method of claim 154, wherein the second heterodimer TCR-antiCD3 antibody fusion molecule is tebentafusp.

156. The method of claim 153, wherein the second active agent is a targeted therapy.

157. The method of claim 156, wherein the targeted therapy is dabrafenib / trametinib.

158. The method of claim 153, wherein the second activator is a demethylating agent.

159. The method of claim 158, wherein the demethylating agent is decitabine.

160. The method according to claim 158 or 159, wherein the demethylating agent is a low dose of decitabine.

161. The method of claim 153, wherein the second active agent is a chemotherapeutic agent.

162. The method of claim 161, wherein the chemotherapeutic agent is a PLD.

163. The method of claim 161, wherein the chemotherapeutic agent is carboplatin / paclitaxel.

164. The method according to any one of claims 1 to 9, 78, 80 or 82 to 89, wherein the lung cancer is non-small cell lung cancer (NSCLC).

165. The method according to any one of claims 10 to 77, 79, 81, 82 to 89 or 119, wherein the ovarian cancer is platinum-sensitive ovarian cancer.

166. The method according to any one of claims 10 to 77, 79, 81, 82 to 89 or 119, wherein the ovarian cancer is platinum-resistant ovarian cancer.

167. The kit according to claim 90 or 91, wherein the dose of the second active agent is about 10-1000 mg / m³. 2 .

Images