Method of treating cutaneous melanoma

Abstract

The present disclosure provides a method of treating cutaneous melanoma in a subject comprising administering to the subject a composition comprising a heterodimeric TCR-anti- CD3 antibody fusion molecule in a dosing regimen, wherein the dosing regimen comprises administering: (i) a first dose comprising 2 to 30 ug of the TCR-anti-CD3 antibody fusion molecule, (ii) a second dose comprising 5 to 60 µg of the TCR-anti-CD3 antibody fusion molecule, and (iii) a maintenance dose comprising 15 to 200 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 once every 6 to 8 days, the second phase is administered once every 12 to 16 days, and the third phase is administered once every 26 to 30 days, wherein the method is a first line treatment.

Description

Atty. Docket No.0282-0001PR1 METHOD OF TREATING CUTANEOUS MELANOMA FIELD OF THE INVENTION

[0001] The present disclosure provides a method of treating cutaneous melanoma in a subject comprising administering to the subject a composition comprising a heterodimeric TCR-anti- CD3 antibody fusion molecule in a dosing regimen, wherein the dosing regimen comprises administering: (i) a first dose comprising 2 to 30 ug of the TCR-anti-CD3 antibody fusion molecule, (ii) a second dose comprising 5 to 60 µg of the TCR-anti-CD3 antibody fusion molecule, and (iii) a maintenance dose comprising 15 to 200 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 once every 6 to 8 days, the second phase is administered once every 12 to 16 days, and the third phase is administered once every 26 to 30 days, wherein the method is a first line treatment. BACKGROUND

[0002] Melanoma is a malignant transformation of the melanocytes. It accounts for approximately 1% of all skin cancers; however, it carries the highest mortality rate among all skin cancers. Historically, melanoma treatment was limited to chemotherapy and nonspecific immunotherapy agents that display poor curative potential and high toxicity. However, the last decade has led to the development of targeted therapy and specific immunotherapy agents that have transformed the landscape of advanced melanoma treatment. Despite the considerable strides in understanding the clinical implications of these agents, there is a scarcity of randomized clinical trials that directly compare the efficacy of agents, and often, the introduction of these agents is associated with a variety of dermatologic adverse events.

[0003] The current recommended first-line treatment of unresectable or metastatic disease includes anti-PD-1-based regimens for subjects and are considered as standard of care (SoC) for initial treatment. Despite these options, at least half of these subjects will progress within 1 year. Preferentially expressed antigen in melanoma (PRAME) is an attractive target in cancer immunotherapy because it is the most broadly expressed cancer testis antigen (CTA), which is homogenously expressed in multiple tumors. PRAME is a CTA initially identified in metastatic cutaneous melanoma. IMC-F106C is the first T-cell receptor (TCR) bispecific protein targeting CD3 and PRAME.Atty. Docket No.0282-0001PR1 SUMMARY OF THE INVENTION

[0004] In some aspects, the techniques described herein relate to a method of treating a cutaneous melanoma in a subject including administering to the subject a composition including a heterodimeric TCR-anti-CD3 antibody fusion molecule in a dosing regimen, wherein the heterodimeric TCR-anti-CD3 antibody fusion molecule includes (a) an alpha chain amino acid sequence that has at least 90%, at least 95% or 100% identity to the amino acid sequence of SEQ ID NO: 14, and (b) a beta chain amino acid sequence that has at least 90%, at least 95% or 100% identity to the amino acid sequence of SEQ ID NO: 16, wherein the alpha chain includes complementarity-determining regions (CDRs) including alpha chain CDRs 1, 2 and 3 having the amino acid sequences of SEQ ID NOs: 3, 4 and 5 respectively and the beta chain includes beta chain CDRs 1, 2 and 3 having the amino acid sequences of SEQ ID NOs: 9, 10 and 11 respectively, wherein the dosing regimen includes administering: (i) a first dose including 2 to 30 ug of the TCR-anti-CD3 antibody fusion molecule, (ii) a second dose including 5 to 60 µg of the TCR-anti-CD3 antibody fusion molecule, and (iii) a maintenance dose including 15 to 200 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 once every 6 to 8 days, the second phase is administered once every 12 to 16 days, and the third phase is administered once every 26 to 30 days; and wherein the second dose includes a larger amount of the TCR-anti-CD3 antibody fusion molecule than the first dose, and the maintenance dose includes a larger amount of the TCR-anti-CD3 antibody fusion molecule than the second dose, wherein the method is a first line treatment.

[0005] In some aspects, the techniques described herein relate to a method, wherein the first dose includes 2 to 4 ug of the TCR-anti-CD3 antibody fusion molecule, the second dose includes 5 to 14 µg of the TCR-anti-CD3 antibody fusion molecule, and the maintenance dose includes 15 to 55 µg of the TCR-anti-CD3 antibody fusion molecule. In some aspects, the techniques described herein relate to a method, wherein the first dose includes 3 ug of the TCR-anti-CD3 antibody fusion molecule, the second dose includes 10 µg of the TCR-anti- CD3 antibody fusion molecule, and the maintenance dose includes 40 µg of the TCR-anti- CD3 antibody fusion molecule.

[0006] In some aspects, the techniques described herein relate to a method, wherein the first dose includes 15 to 40 µg of the TCR-anti-CD3 antibody fusion molecule, the second dose includes 30 to 80 µg of the TCR-anti-CD3 antibody fusion molecule, and the maintenance dose includes 140 to 200 µg of the TCR-anti-CD3 antibody fusion molecule. In someAtty. Docket No.0282-0001PR1 aspects, the techniques described herein relate to a method, wherein the first dose includes 20 µg of the TCR-anti-CD3 antibody fusion molecule, the second dose includes 40 µg of the TCR-anti-CD3 antibody fusion molecule, and the maintenance dose includes 160 µg of the TCR-anti-CD3 antibody fusion molecule.

[0007] In some aspects, the techniques described herein relate to a method as described herein, wherein the alpha chain amino acid sequence has 100% identity to the amino acid sequence of SEQ ID NO: 14. In some aspects, the techniques described herein relate to a method as described herein, wherein the beta chain amino acid sequence has 100% identity to the amino acid sequence of SEQ ID NO: 16.

[0008] In some aspects, the techniques described herein relate to a method as described herein, wherein the first dose is administered once every 7 days.

[0009] In some aspects, the techniques described herein relate to a method as described herein, wherein the first dose is administered for 1 week to about 3 weeks. In some aspects, the techniques described herein relate to a method as described herein, wherein the first dose is administered for 1 week.

[0010] In some aspects, the techniques described herein relate to a method as described herein, wherein the second dose is administered 6 to 8 days following the first dose.

[0011] In some aspects, the techniques described herein relate to a method as described herein, wherein the second dose is administered once every 7 days.

[0012] In some aspects, the techniques described herein relate to a method as described herein, wherein the second dose is administered for 1 week to about 3 weeks. In some aspects, the techniques described herein relate to a method as described herein, wherein the second dose is administered for 1 week.

[0013] In some aspects, the techniques described herein relate to a method as described herein, wherein the maintenance dose in the first phase is administered 6 to 8 days following the second dose.

[0014] In some aspects, the techniques described herein relate to a method as described herein, wherein the maintenance dose in the first phase is administered once a week.

[0015] In some aspects, the techniques described herein relate to a method as described herein, wherein the first phase is about 2 weeks to about 101 weeks. In some aspects, the techniques described herein relate to a method as described herein, wherein the first phase isAtty. Docket No.0282-0001PR1 about 8 weeks to about 14 weeks. In some aspects, the techniques described herein relate to a method as described herein, wherein the first phase is 10 weeks.

[0016] In some aspects, the techniques described herein relate to a method as described herein, wherein the maintenance dose in the second phase is administered once every 2 weeks.

[0017] In some aspects, the techniques described herein relate to a method as described herein, wherein the second phase is about 30 weeks to 50 weeks. In some aspects, the techniques described herein relate to a method as described herein, wherein the second phase is 38 weeks.

[0018] In some aspects, the techniques described herein relate to a method as described herein, wherein the maintenance dose in the third phase is administered every 4 weeks.

[0019] In some aspects, the techniques described herein relate to a method as described herein, wherein the third phase is about 40 weeks to about 60 weeks. In some aspects, the techniques described herein relate to a method as described herein, wherein the third phase is about 48 weeks.

[0020] In some aspects, the techniques described herein relate to a method as described herein, wherein the first phase is 10 weeks, the second phase is 38 weeks, and the third phase is 48 weeks.

[0021] In some aspects, the techniques described herein relate to a method as described herein, 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. In some aspects, the techniques described herein relate to a method as described herein, wherein the dosing regimen is about 90 weeks to about 110 weeks.

[0022] In some aspects, the techniques described herein relate to a method as described herein, wherein no corticosteroids are administered within three weeks prior to the dosing regimen and throughout the dosing regimen. In some aspects, the techniques described herein relate to a method as described herein, wherein no other anti-cancer drugs are co- administered during the dosage regimen.

[0023] In some aspects, the techniques described herein relate to a method as described herein, wherein the subject is HLA-A*02 positive. In some aspects, the techniques described herein relate to a method as described herein, wherein the subject is HLA-A*02:01 positive.Atty. Docket No.0282-0001PR1 In some aspects, the techniques described herein relate to a method as described herein, wherein the dosing regimen further includes administering a second therapeutic agent. In some aspects, the techniques described herein relate to a method as described herein, wherein the second therapeutic agent includes a checkpoint inhibitor. In some aspects, the techniques described herein relate to a method as described herein, wherein the checkpoint inhibitor includes of: Aldesleukin, Atezolizumab (TECENTRIQ®), Avelumab, Bevacizumab (Avastin®) Binimetinib, Braftovi (Encorafenib), Cobimetinib Fumarate, Cotellic (Cobimetinib Fumarate), Dabrafenib Mesylate, Dacarbazine, Durvalumab. Encorafenib, Galunisertib, Gemcitabine, IL-2 (Aldesleukin), Interferon, Imlygic (Talimogene Laherparepvec), Intron A (Recombinant Interferon Alfa-2b), Ipilimumab, Keytruda (Pembrolizumab), Kimmtrak (Tebentafusp-tebn), Mekinist (Trametinib Dimethyl Sulfoxide), Mektovi (Binimetinib), Merestinib, Nivolumab, Nivolumab and Relatlimab-rmbw, Opdivo (Nivolumab), Opdualag (Nivolumab and Relatlimab-rmbw), Pembrolizumab, Proleukin (Aldesleukin), Recombinant Interferon Alfa-2b, Tafinlar (Dabrafenib Mesylate), Talimogene Laherparepvec, Tebentafusp-tebn, Trametinib Dimethyl Sulfoxide, Tremelimumab, Vemurafenib, Yervoy (Ipilimumab), Zelboraf (Vemurafenib) or a combination thereof. In some aspects, the techniques described herein relate to a method as described herein, wherein the checkpoint inhibitor includes Nivolumab, Relatimab, or a combination thereof. In some aspects, the techniques described herein relate to a method as described herein, wherein the checkpoint inhibitor includes Nivolumab.

[0024] In some aspects, the techniques described herein relate to a method as described herein, wherein the second therapeutic agent is administered once a week to once a month. In some aspects, the techniques described herein relate to a method as described herein, wherein the second therapeutic agent is administered once a week. In some aspects, the techniques described herein relate to a method as described herein, wherein the second therapeutic agent is administered for at least 10 weeks. In some aspects, the techniques described herein relate to a method as described herein, wherein the second therapeutic agent is administered for at least 40 weeks. In some aspects, the techniques described herein relate to a method as described herein, wherein the second therapeutic agent is administered for the duration of the dosing regimen.

[0025] In some aspects, the techniques described herein relate to a method as described herein, wherein the cutaneous melanoma is a Stage IV melanoma. In some aspects, the techniques described herein relate to a method as described herein, wherein Stage IV melanoma is histologically confirmed.Atty. Docket No.0282-0001PR1

[0026] In some aspects, the techniques described herein relate to a method as described herein, wherein the cutaneous melanoma is an unresectable Stage III melanoma .

[0027] In some aspects, the techniques described herein relate to a method as described herein, wherein the subject has BRAF V600 mutation.

[0028] In some aspects, the techniques described herein relate to a method as described herein, wherein the subject has been diagnosed with a life expectancy greater than 3 months.

[0029] In some aspects, the techniques described herein relate to a method as described herein, wherein the composition is administered by intravenous (IV) infusion.

[0030] In some aspects, the techniques described herein relate to a method of treating a cutaneous melanoma in a subject including administering to the subject a composition including a heterodimeric TCR-anti-CD3 antibody fusion molecule in a dosing regimen, wherein the heterodimeric TCR-anti-CD3 antibody fusion molecule includes (a) an alpha chain amino acid sequence that has at least 90%, at least 95% or 100% identity to the amino acid sequence of SEQ ID NO: 14, and (b) a beta chain amino acid sequence that has at least 90%, at least 95% or 100% identity to the amino acid sequence of SEQ ID NO: 16, wherein the alpha chain includes complementarity-determining regions (CDRs) including alpha chain CDRs 1, 2 and 3 having the amino acid sequences of SEQ ID NOs: 3, 4 and 5 respectively and the beta chain includes beta chain CDRs 1, 2 and 3 having the amino acid sequences of SEQ ID NOs: 9, 10 and 11 respectively, wherein the dosing regimen includes: (i) administering one first dose including 3 µg of the TCR-anti-CD3 antibody fusion molecule, (ii) administering one second dose including 10 µg of the TCR-anti-CD3 antibody fusion molecule one week after the first dose, and (iii) administering a maintenance dose including 40 µg of the TCR-anti-CD3 antibody fusion molecule one week after the second dose, and every week during the first phase, wherein the first phase is about 9 to 10 weeks, (iv) administering the maintenance dose every 2 weeks during the second phase, wherein the second phase is about 38 weeks, and (v) administering the maintenance dose every 4 weeks during the third phase, wherein the third phase is about 48 weeks, wherein the method is a first line treatment.

[0031] In some aspects, the techniques described herein relate to a method of treating a cutaneous melanoma in a subject including administering to the subject a composition including a heterodimeric TCR-anti-CD3 antibody fusion molecule in a dosing regimen, wherein the heterodimeric TCR-anti-CD3 antibody fusion molecule includes (a) an alpha chain amino acid sequence that has at least 90%, at least 95% or 100% identity to the aminoAtty. Docket No.0282-0001PR1 acid sequence of SEQ ID NO: 14, and (b) a beta chain amino acid sequence that has at least 90%, at least 95% or 100% identity to the amino acid sequence of SEQ ID NO: 16, wherein the alpha chain includes complementarity-determining regions (CDRs) including alpha chain CDRs 1, 2 and 3 having the amino acid sequences of SEQ ID NOs: 3, 4 and 5 respectively and the beta chain includes beta chain CDRs 1, 2 and 3 having the amino acid sequences of SEQ ID NOs: 9, 10 and 11 respectively, wherein the dosing regimen includes: (i) administering one first dose including 20 µg of the TCR-anti-CD3 antibody fusion molecule, (ii) administering one second dose including 40 µg of the TCR-anti-CD3 antibody fusion molecule one week after the first dose, and (iii) administering a maintenance dose including 160 µg of the TCR-anti-CD3 antibody fusion molecule one week after the second dose, and then every week during the first phase, wherein the first phase is about 9 to 10 weeks, (iv) administering the maintenance dose every 2 weeks during the second phase, wherein the second phase is about 38 weeks, and (v) administering the maintenance dose every 4 weeks during the third phase, wherein the third phase is about 48 weeks, wherein the method is a first line treatment.

[0032] In some aspects, the disclosure provides a kit including: (i) one first container including a first dose including 2 to 4 µg of a heterodimeric TCR-anti-CD3 antibody fusion molecule, (ii) one second container including a second dose including 5 to 14 µg of the heterodimeric TCR-anti-CD3 antibody fusion molecule, (iii) one or more maintenance containers, each maintenance container including a maintenance dose including 15 to 55 µg of the heterodimeric TCR-anti-CD3 antibody fusion molecule, and (iv) instructions for administering the heterodimeric TCR-anti-CD3 antibody fusion molecule in a dosing regimen, wherein the dosage regimen is specified as (a) administering the first dose on week 1, (b) administering the second dose on week 2, (c) administering a maintenance dose weekly during the first phase for 10 weeks, (d) administering the maintenance dose every 2 weeks during the second phase for 38 weeks, and (e) administering the maintenance dose every 4 weeks during the third phase for 48 weeks.

[0033] In some aspects, the disclosure provides a kit including: (i) one first container including a first dose including 20 µg of a heterodimeric TCR-anti-CD3 antibody fusion molecule, (ii) one second container including a second dose including 40 µg of the heterodimeric TCR-anti-CD3 antibody fusion molecule, (iii) one or more maintenance containers, each maintenance container including a maintenance dose including 160 µg of the heterodimeric TCR-anti-CD3 antibody fusion molecule, and (iv) instructions for administering the heterodimeric TCR-anti-CD3 antibody fusion molecule in a dosingAtty. Docket No.0282-0001PR1 regimen, wherein the dosage regimen is specified as (a) administering the first dose on week 1, (b) administering the second dose on week 2, (c) administering a maintenance dose weekly during the first phase for 10 weeks, (d) administering the maintenance dose every 2 weeks during the second phase for 38 weeks, and (e) administering the maintenance dose every 4 weeks during the third phase for 48 weeks.

[0034] In some aspects, the heterodimeric TCR-anti-CD3 antibody fusion molecule in the kits described herein includes (a) an alpha chain amino acid sequence that has at least 90%, at least 95% or 100% identity to the amino acid sequence of SEQ ID NO: 14, and (b) a beta chain amino acid sequence that has at least 90%, at least 95% or 100% identity to the amino acid sequence of SEQ ID NO: 16, wherein the alpha chain includes complementarity- determining regions (CDRs) including alpha chain CDRs 1, 2 and 3 having the amino acid sequences of SEQ ID NOs: 3, 4 and 5 respectively and the beta chain includes beta chain CDRs 1, 2 and 3 having the amino acid sequences of SEQ ID NOs: 9, 10 and 11 respectively.

[0035] In some aspects, the kit includes 2 to 10 maintenance containers. In some aspects, the first dose, the second dose and / or the maintenance dose of the kit or methods described herein further include(s) a pharmaceutically acceptable excipient to form a first pharmaceutical formulation, second pharmaceutical formulation, and / or maintenance pharmaceutical formulation, respectively.

[0036] In some aspects, the first, second and / or maintenance pharmaceutical formulation of the methods or kits described herein has a pH of about 6.5 to about 7.5.

[0037] In some aspects, the kit further includes a second therapeutic agent.

[0038] In some aspects, the second therapeutic agent in the kits described herein includes a checkpoint inhibitor. In some aspects, the checkpoint inhibitor of the kit includes of: Aldesleukin, Atezolizumab (TECENTRIQ®), Avelumab, Bevacizumab (Avastin®) Binimetinib, Braftovi (Encorafenib), Cobimetinib Fumarate, Cotellic (Cobimetinib Fumarate), Dabrafenib Mesylate, Dacarbazine, Durvalumab. Encorafenib, Galunisertib, Gemcitabine, IL-2 (Aldesleukin), Interferon, Imlygic (Talimogene Laherparepvec), Intron A (Recombinant Interferon Alfa-2b), Ipilimumab, Keytruda (Pembrolizumab), Kimmtrak (Tebentafusp-tebn), Mekinist (Trametinib Dimethyl Sulfoxide), Mektovi (Binimetinib), Merestinib, Nivolumab, Nivolumab and Relatlimab-rmbw, Opdivo (Nivolumab), Opdualag (Nivolumab and Relatlimab-rmbw), Pembrolizumab, Proleukin (Aldesleukin), Recombinant Interferon Alfa-2b, Tafinlar (Dabrafenib Mesylate), Talimogene Laherparepvec,Atty. Docket No.0282-0001PR1 Tebentafusp-tebn, Trametinib Dimethyl Sulfoxide, Tremelimumab, Vemurafenib, Yervoy (Ipilimumab), Zelboraf (Vemurafenib) or a combination thereof.

[0039] In some aspects, the checkpoint inhibitor of the kit includes Nivolumab, Relatimab, or a combination thereof. In some aspects, the checkpoint inhibitor of the kit includes Nivolumab.

[0040] In some aspects, the techniques described herein relate to a method of treating a PRAME-positive cancer in a subject including administering to the subject a composition including a heterodimeric TCR-anti-CD3 antibody fusion molecule in a dosing regimen, wherein the heterodimeric TCR-anti-CD3 antibody fusion molecule includes (a) an alpha chain amino acid sequence that has at least 90%, at least 95% or 100% identity to the amino acid sequence of SEQ ID NO: 14, and (b) a beta chain amino acid sequence that has at least 90%, at least 95% or 100% identity to the amino acid sequence of SEQ ID NO: 16, wherein the alpha chain includes complementarity-determining regions (CDRs) including alpha chain CDRs 1, 2 and 3 having the amino acid sequences of SEQ ID NOs: 3, 4 and 5 respectively and the beta chain includes beta chain CDRs 1, 2 and 3 having the amino acid sequences of SEQ ID NOs: 9, 10 and 11 respectively, wherein the dosing regimen includes administering: (i) a first dose including 2 to 30 ug of the TCR-anti-CD3 antibody fusion molecule, (ii) a second dose including 5 to 60 µg of the TCR-anti-CD3 antibody fusion molecule, and (iii) a maintenance dose including 15 to 200 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 once every 6 to 8 days, the second phase is administered once every 12 to 16 days, and the third phase is administered once every 26 to 30 days; and wherein the second dose includes a larger amount of the TCR-anti-CD3 antibody fusion molecule than the first dose, and the maintenance dose includes a larger amount of the TCR-anti-CD3 antibody fusion molecule than the second dose, wherein the method is a first line treatment.

[0041] In some aspects, the PRAME-positive cancer is an unresectable Stage III melanoma.

[0042] In some aspects, the techniques described herein relate to a method of treating a cutaneous melanoma in a subject including administering to the subject (A). a composition including a heterodimeric TCR-anti-CD3 antibody fusion molecule in a dosing regimen, wherein the heterodimeric TCR-anti-CD3 antibody fusion molecule includes (a) an alpha chain amino acid sequence that has at least 90%, at least 95% or 100% identity to the amino acid sequence of SEQ ID NO: 14, and (b) a beta chain amino acid sequence that has at least 90%, at least 95% or 100% identity to the amino acid sequence of SEQ ID NO: 16, wherein the alpha chain includes complementarity-determining regions (CDRs) including alpha chainAtty. Docket No.0282-0001PR1 CDRs 1, 2 and 3 having the amino acid sequences of SEQ ID NOs: 3, 4 and 5 respectively and the beta chain includes beta chain CDRs 1, 2 and 3 having the amino acid sequences of SEQ ID NOs: 9, 10 and 11 respectively, wherein the dosing regimen includes administering: (i) a first dose including 2 to 30 ug of the TCR-anti-CD3 antibody fusion molecule, (ii) a second dose including 5 to 60 µg of the TCR-anti-CD3 antibody fusion molecule, and (iii) a maintenance dose including 15 to 200 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 once every 6 to 8 days, the second phase is administered once every 12 to 16 days, and the third phase is administered once every 26 to 30 days; and wherein the second dose includes a larger amount of the TCR-anti-CD3 antibody fusion molecule than the first dose, and the maintenance dose includes a larger amount of the TCR-anti-CD3 antibody fusion molecule than the second dose; and (B) a second therapeutic agent, wherein the second therapeutic agent is administered once every week to once every month; wherein the method is a first line treatment.

[0043] In some aspects, the second therapeutic agent of the kits described herein includes a checkpoint inhibitor. In some aspects, the checkpoint inhibitor of the kits described herein includes of: Aldesleukin, Atezolizumab (TECENTRIQ®), Avelumab, Bevacizumab (Avastin®) Binimetinib, Braftovi (Encorafenib), Cobimetinib Fumarate, Cotellic (Cobimetinib Fumarate), Dabrafenib Mesylate, Dacarbazine, Durvalumab. Encorafenib, Galunisertib, Gemcitabine, IL-2 (Aldesleukin), Interferon, Imlygic (Talimogene Laherparepvec), Intron A (Recombinant Interferon Alfa-2b), Ipilimumab, Keytruda (Pembrolizumab), Kimmtrak (Tebentafusp-tebn), Mekinist (Trametinib Dimethyl Sulfoxide), Mektovi (Binimetinib), Merestinib, Nivolumab, Nivolumab and Relatlimab-rmbw, Opdivo (Nivolumab), Opdualag (Nivolumab and Relatlimab-rmbw), Pembrolizumab, Proleukin (Aldesleukin), Recombinant Interferon Alfa-2b, Tafinlar (Dabrafenib Mesylate), Talimogene Laherparepvec, Tebentafusp-tebn, Trametinib Dimethyl Sulfoxide, Tremelimumab, Vemurafenib, Yervoy (Ipilimumab), Zelboraf (Vemurafenib) or a combination thereof. In some aspects, the checkpoint inhibitor of the kits described herein includes Nivolumab, Relatimab, or a combination thereof. In some aspects, the checkpoint inhibitor of the kits described herein includes Nivolumab. BRIEF DESCRIPTION OF THE DRAWINGS

[0044] The following drawings form part of the present specification and are included to further demonstrate exemplary embodiments of certain aspects of the present disclosure.Atty. Docket No.0282-0001PR1

[0045] FIG.1 shows a clinical study design as described in Example 1.

[0046] FIG.2 shows a spider plot for percent change from baseline in tumor size in all melanoma participants – Study IMC-F106C-101 (IMC-F106C Monotherapy IV). DETAILED DESCRIPTION

[0047] Unless otherwise defined herein, scientific and technical terms used in the present disclosure shall have meanings that are commonly understood by one of ordinary skill in the art. Further, unless otherwise required by context, singular terms shall include pluralities and plural terms shall include the singular.

[0048] The articles “a” and “an” are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, “an element” means one element or more than one element.

[0049] The use of the term “or” in the claims is used to mean “and / or,” unless explicitly indicated to refer only to alternatives or the alternatives are mutually exclusive, although the disclosure supports a definition that refers to only alternatives and “and / or.”

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

[0051] The use of the term “for example” and its corresponding abbreviation “e.g.” means that the specific terms recited are representative examples and embodiments of the disclosure that are not intended to be limited to the specific examples referenced or cited unless explicitly stated otherwise.

[0052] As used herein, “about” can mean plus or minus 10% of the provided value. Where ranges are provided, they are inclusive of the boundary values. “About” can additionally or alternately mean either within 10% of the stated value, or within 5% of the stated value, or in some cases within 2.5% of the stated value; or “about” can mean rounded to the nearest significant digit.

[0053] As used herein, “between” is a range inclusive of the ends of the range. For example, a number between x and y explicitly includes the numbers x and y and any numbers that fall within x and y.Atty. Docket No.0282-0001PR1

[0054] The term “administration” or “administering” refers to routes of introducing a compound or composition provided herein to a subject to perform its intended function. An example of a route of administration that can be used includes, but is not limited to, intravenous fusion.

[0055] The term “subject” means any subject, particularly a mammalian subject, in need of treatment, e.g., with a composition comprising a compound of formula (I). 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 administration to a subject in need thereof, i.e., a subject having cutaneous melanoma and / or a subject having a PRAME- positive cancer. As used herein, a “subject in need thereof” can refer to the subject for whom it is desirable to treat, e.g., a subject being diagnosed with cutaneous melanoma and / or a PRAME-positive cancer as described herein. In some embodiments, the term “subject in need thereof” can refer to a subject having one or more symptoms associated with cutaneous melanoma and / or a PRAME-positive cancer, e.g., a subject having a significant change in an existing mole, a subjecting having developed a new pigmented or unusual-looking growth on subject’s skin, etc. In some embodiments, the term “subject in need thereof” can refer to a subject at high risk for suffering from a cutaneous melanoma and / or a PRAME-positive cancer suitable to treatment with the heterodimeric TCR-anti-CD3 antibody fusion molecule as described herein, independently of whether the subject has physical manifestations of such condition. In some embodiments, the subject is an adult, i.e., at least 18 years old. In some embodiments, the subject is 12-18 years old. In some embodiments, the subject is less than 12 years old.

[0056] In some embodiments, the subject has serum LDH of between about 1.1 x to about 2.0 x ULN. In some embodiments, the subject is under about 65 years old. In some embodiments, the subject is at least about 65 years old (for example at least about).

[0057] In some embodiments, the subject has one or more of the characteristics described in Examples 1 of the present disclosure. For example, the subject can have at least one (e.g., at least any of 2, 3, 4, 5, 6, or 7) of the following characteristics: (1) Histologically or cytologically confirmed cutaneous malignant melanoma with evidence of metastasis (Stage IV); (2) No prior cytotoxic chemotherapy for metastatic malignant cutaneous melanoma; (3) No prior adjuvant cytotoxic chemotherapy; (4) Male or non-pregnant and non-lactating female >18 years of age; (5) No other current active malignancy within the past 3 years; (6)Atty. Docket No.0282-0001PR1 Radiographically documented measurable disease (for example, the presence of at least 1 radiographically documented measurable lesion); and (7) ECOG performance status 0-1. In some embodiments, the subject does not have history or current evidence of brain metastases, including leptomeningeal involvement. In some embodiments, the subject does not have pre- existing peripheral neuropathy of NCI CTCAE Scale of Grade > 2.

[0058] In some embodiments, the subject having cutaneous melanoma 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 comprise a mutation in BRAF (e.g., the melanoma comprises wild-type BRAF). In some embodiments, the subject does not comprise BRAF mutant such as a BRAF mutant with increased activity (for example, increased kinase activity, and / or increased activity as compared to wild-type BRAF) or a BRAF gain-of-function mutant. In some embodiments, the subject does not comprise a constitutive active BRAF mutant. In some embodiments, the subject does not comprise BRAF V600E mutation (e.g., the melanoma comprises wild-type BRAF). In some embodiments, the subject comprises wild-type BRAF (e.g., the cutaneous melanoma cells have wild-type BRAF). In some embodiments, the subject comprises a BRAF mutant such as a BRAF mutant with increased activity (for example, increased kinase activity, and / or increased activity as compared to wild-type BRAF) or a BRAF gain-of- function mutant. In some embodiments, the subject comprises a constitutive active BRAF mutant.

[0059] In some embodiments, the subject has elevated serum lactate dehydrogenase (“LDH”) level. In some embodiments, the subject has serum LDH of less than about 0.8 χ upper limit of normal (“ULN”). In some embodiments, the subject has serum LDH at about 0.8 χ to about 1.1 x ULN. In some embodiments, the subject has serum LDH of between greater than about 1.1 x to about 2.0 x ULN.

[0060] In some embodiments, the subject can be treated when the subject has pre-cancerous lesions, or when the subject is first diagnosed. In some embodiments, the methods described herein can be used when the subject has been diagnosed with a life expectancy of greater than 1 year, greater than 6 months, greater than 3 months, or greater than 1 month. In some embodiments, the methods described herein can be used when the subject has been diagnosed with a life expectancy of greater than 3 months. The skilled artisan can appreciate that determining “life expectancy” is an estimate that can be determined by skilled artisans using known methods in the art, e.g., age of subject, stage of melanoma, thickness and size of tumor, speed of growth, malignancy, contributory factors, etc.Atty. Docket No.0282-0001PR1

[0061] In some embodiments, the subject is HLA-A*02 positive. The term “HLA-A*02 positive” refers to a subject having TCRs that bind to the SLLQHLIGL(SEQ ID NO: 1)- HLA-A*02 complex. In some embodiments, the subject is HLA-A*02:01 positive. The term “HLA-A*02:01 positive refers to a subject that expresses the HLA-A*02 genotype with the HLA-A*02:01 allele.

[0062] The term “treating” refers to administering an active agent with the purpose to cure, heal, alleviate, relieve, alter, remedy, ameliorate, improve, or affect a condition (e.g., a disease), the symptoms of the condition, or to prevent or delay the onset of the symptoms, complications, biochemical indicia of a disease, or otherwise arrest or inhibit further development of the cutaneous melanoma and / or a subject having a PRAME-positive cancer in a statistically significant manner.

[0063] In some embodiments, the kits and methods described herein are directed to treatment of a cutaneous melanoma, e.g., a cancer that starts in the pigment-making cells of the skin. However, in some embodiments, the kits and methods described herein can be directed to other melanomas and / or other PRIME positive subjects. In some embodiments, the melanoma is metastatic melanoma. In some embodiments, the melanoma is metastatic malignant melanoma. In some embodiments, the melanoma to be treated in the subject is stage 0, stage I, stage II, stage III, or stage IV. In some embodiments, the melanoma 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 melanoma to be treated is stage III. In some embodiments, the melanoma to be treated is stage IV. In some embodiments, the stage of cancer, e.g., stage IV cancer, is histologically confirmed. Histological confirmation refers to the use of visual means, e.g., microscope, to look at a tissue to determine whether it has signs of the cancer, disease or other abnormalities associated with melanoma. In some embodiments, the tissue used in the histological examination has been excised by biopsy or surgical section. In some embodiments, the melanoma to be treated is stage IV and is histologically confirmed. In some embodiments, the melanoma to be treated is stage III and is histologically confirmed.

[0064] In some embodiments, the metastatic melanoma is at stage M1a. In some embodiments, the metastatic melanoma is at stage Mlb. In some embodiments, the metastatic melanoma is at stage Mlc. Staging of melanoma can be based on a method known to one skilled in the art. Staging of melanoma can be according to the criteria included in 2009 AJCC Melanoma Staging and Classification. See Balch CM et al., J Clin Oncol.2009, 27(36):6199-206 (the contents disclosed therein is incorporated by reference in its entirety).Atty. Docket No.0282-0001PR1

[0065] In some embodiments, the melanoma is stage IV melanoma (e.g., stage IV cutaneous melanoma). In some embodiments, the metastatic melanoma is at stage M1a. In some embodiments, the metastatic melanoma is at stage Mlb. In some embodiments, the metastatic melanoma is at stage Mlc. In some embodiments, the subject has not received prior therapy (e.g., prior cytotoxic chemotherapy) for the melanoma (e.g., cutaneous melanoma). In some embodiments, the cutaneous melanoma is unresectable, i.e., cannot be treated with surgery. In some embodiments, the cutaneous melanoma in an unresectable be stage III cutaneous melanoma. In some embodiments, the cutaneous melanoma is an unresectable be stage IV cutaneous melanoma.

[0066] In some embodiments, the melanoma is superficial spreading melanoma. In some embodiments, the melanoma is nodular melanoma. In some embodiments, the melanoma is acral lentiginous melanoma. In some embodiments, the melanoma is lentigo maligna melanoma.

[0067] In some embodiments, the kits and methods described herein are directed to various other melanomas. For example, in some embodiments, the melanoma is mucosal melanoma (e.g., mucosal melanoma in nose, mouth, throat, or genital area). In some embodiments, the melanoma is ocular melanoma. In some embodiments, the melanoma is uveal melanoma. In some embodiments, the melanoma is choroidal melanoma.

[0068] In some embodiments, the disclosure provides a method of treating cutaneous melanoma (e.g., metastatic or stage IV cutaneous melanoma) in a subject (e.g., human) comprising administering to the subject a composition comprising the heterodimeric TCR- anti-CD3 antibody fusion molecule as described herein. In some embodiments, the methods comprising administration of heterodimeric TCR-anti-CD3 antibody fusion molecule for treating melanoma is used as a monotherapy. In some embodiments, the method of treating melanoma using TCR-anti-CD3 antibody fusion molecule further comprises one other therapeutic agent.

[0069] In some embodiments, the present disclosure relates to the treatment of PRAME positive cancers. The term "PRAME positive cancer” refers to a PReferentially expressed Antigen in MElanoma (i.e., PRAME) cancer in which at least some of the cancer cells express PRAME. PRAME was first identified as an antigen that is over expressed in melanoma (Ikeda et al Immunity.1997 Feb;6(2):199-208); it is also known as CT130, MAPE, OIP-4 and has Uniprot accession number P78395. The protein functions as a repressor of retinoic acid receptor signaling (Epping et al., Cell.2005 Sep 23; 122(6): 835- 47). PRAME belongs to the family of germline-encoded antigens known as cancer testisAtty. Docket No.0282-0001PR1 antigens. Cancer testis antigens are attractive targets for immunotherapeutic intervention since they typically have limited or no expression in normal adult tissues. PRAME is expressed in a number of solid tumors as well as in leukemias and lymphomas. PRAME targeting therapies of the invention may be particularly suitable for treatment of cancers including, but not limited to, 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.

[0070] The peptide SLLQHLIGL (SEQ ID NO: 1) corresponds to amino acids 425-433 of the full length PRAME protein and is presented on the cell surface in complex with HLA- A*02 (Kessler et al., J Exp Med.2001 Jan 1 ;193(1):73-88). This peptide-HLA complex provides a useful target for TCR-based immunotherapeutic intervention.

[0071] WO 2018 / 234319, and corresponding US Pat. Nos.11,427,624 and 11,718,657, describes TCRs that bind to the SLLQHLIGL(SEQ ID NO: 1)-HLA-A*02 complex, each of which is incorporated by reference herein in their entirety. The TCRs are mutated relative to a native PRAME TCR alpha and / or beta variable domains to have improved binding affinities for, and / or binding half-lives, for the complex, and can be associated (covalently or otherwise) with a therapeutic agent. 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 may be covalently linked to the C- or N- terminus of the alpha or beta chain of the TCR. The resulting molecule is a TCR bispecific.

[0072] TCR bispecific proteins redirect polyclonal T cells to target peptides derived from intra- or extra-cellular disease associated antigens and presented on the cell surface in complex with an HLA molecule. This approach has been tested clinically in the context of a different antigen with a TCR bispecific protein targeting an HLA-A*02 restricted peptide from gp100 and CD3 (tebentafusp). Administration of this molecule provided an OS benefit in uveal melanoma (Nathan P, et al. Overall Survival Benefit with Tebentafusp in Metastatic Uveal Melanoma. N Engl J Med 2021; 385:1196-1206). However, no such TCR bispecific proteins targeting PRAME have been tested clinically.

[0073] In other words, a PRAME positive cancer is a cancer associated with PRAME expression. In some embodiments, the cancer is known to be associated with expression of PRAME. For example, in some embodiments, the prevalence of PRAME expression is known to be elevated in a cancer and thus PRAME expression is not assessed or is assessed retrospectively. Alternatively, PRAME expression can be assessed using any method known in the art, including, for example, histological methods or other quantitative or qualitativeAtty. Docket No.0282-0001PR1 measurements, including PCR, RNA expression analysis, and / or kits or sequence panels designed to measure the expression level of PRAME. However, the disclosure is not intended to be limited to the treatment of cancers for which PRAME expression can be detected by histological methods. In particular, the disclosure is not intended to be limited to the treatment of subject in whom PRAME expression can be detected, for example by histological methods. Rather, the disclosure is useful for the treatment of cancers and tumor types which are considered to be PRAME positive. In some embodiments, a PRAME positive subject can have a cutaneous melanoma. In some embodiments, a PRAME positive subject does not have cutaneous melanoma, but has another typey of PRAME positive cancer. In some embodiments, the PRAME positive cancer, e.g., cutaneous melanoma, can have relapsed from, be refractory to, or be intolerant of standard treatment regimens.

[0074] PRAME expression, when detected by histological methods like immunohistochemistry (IHC), can be quantified using an H-score. Expression of PRAME in subject cells or their sub-cellular compartments within a tumor is first detected and classified as either positive or negative. The positive cells can be further classified into high, medium, or low based on the IHC signal intensity. The H-score captures both the intensity and the proportion of the biomarker of interest from the IHC image and comprises values between 0 and 300, thereby offering a dynamic range to quantify abundance or a particular marker or gene.

[0075] The methods and kits described herein comprise heterodimeric TCR-anti-CD3 antibody fusion molecules. The heterodimeric TCR-anti-CD3 antibody fusion molecules comprise two domains: – a heterodimeric TCR; and – an anti-CD3 antibody.

[0076] Heterodimeric TCR-anti-CD3 antibody fusion molecules are described in, e.g., WO 2023 / 099622, incorporated by reference in its entirety.

[0077] Heterodimeric TCR can comprise a TCR comprising an alpha chain (i.e., TCR alpha chain) and a beta chain (i.e., TCR beta chain). Each chain can comprise variable, joining and constant regions, and the beta chain also usually contains a short diversity region between the variable and joining regions, but this diversity region can be considered as part of the joining region. Each variable region can comprise three CDRs (Complementarity Determining Regions) embedded in a framework sequence, one being the hypervariable region named CDR3. There are several types of alpha chain variable (Vα) regions and several types of betaAtty. Docket No.0282-0001PR1 chain variable (Vβ) regions distinguished by their framework, CDR1 and CDR2 sequences, and by a partly defined CDR3 sequence. The Vα types are referred to in IMGT nomenclature by a unique TRAV number, Vβ types are referred to by a unique TRBV number.

[0078] In some embodiments, the disclosure provides a TCR alpha chain amino acid sequence of SEQ ID NO: 14 or a TCR alpha chain amino acid sequence that has at least 90%, at least 95% or 100% identity to the amino acid sequence of SEQ ID NO: 14. In some embodiments, the disclosure provides a TCR beta chain-anti-CD3 amino acid sequence of SEQ ID NO: 16 or a TCR beta chain-anti-CD3 amino acid sequence that has at least 90%, at least 95% or 100% identity to the amino acid sequence of SEQ ID NO: 16. In some embodiments, the TCR alpha chain variable domain comprises CDRs 1, 2 and 3 having the amino acid sequences of SEQ ID NOs: 3, 4 and 5 respectively and the TCR beta chain variable domain comprises CDRs 1 , 2 and 3 having the amino acid sequences of SEQ ID NOs: 9, 10 and 11 respectively.

[0079] In some embodiments, the TCR can bind the sequence SLLQHLIGL (SEQ ID NO: 1). The peptide of SEC ID NO: 1 corresponds to amino acids 425-433 of the full length PRAME protein and is presented on the cell surface in complex with HLA-A*02 (Kessler et al., J Exp Med.2001 Jan 1 ;193(1):73-88). This peptide-HLA complex provides a useful target for TCR-based immunotherapeutic intervention. WO 2018 / 234319 describes TCRs that bind to the SLLQHLIGL(SEQ ID NO: 1)-HLA-A*02 complex and is incorporated herein in its entirety.

[0080] In some embodiments, the heterodimeric TCR is fused with an antibody to CD3 to have improved binding affinities for, and / or binding half-lives. 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- or N- terminus of the alpha or beta chain of the TCR. The resulting molecule is a TCR bispecific.

[0081] TCR bispecific proteins redirect polyclonal T cells to target peptides derived from intra- or extra-cellular disease associated antigens and presented on the cell surface in complex with an HLA molecule. This approach has been tested clinically in the context of a different antigen with a TCR bispecific protein targeting an HLA-A*02 restricted peptide from gp100 and CD3 (tebentafusp). Administration of this molecule provided an OS benefit in uveal melanoma (Nathan P, et al. Overall Survival Benefit with Tebentafusp in Metastatic Uveal Melanoma. N Engl J Med 2021; 385:1196-1206).Atty. Docket No.0282-0001PR1

[0082] The term “antibody” includes, but is not limited to, genetically engineered or otherwise modified forms of immunoglobulins, such as intrabodies, chimeric antibodies, fully human antibodies, humanized antibodies (e.g. generated by “CDR-grafting”), antibody fragments, and heteroconjugate antibodies (e.g., bispecific antibodies, diabodies, triabodies, tetra-bodies, etc.). The term “antibody” includes cys-diabodies and minibodies. Thus, each and every embodiment provided herein in regard to “antibodies”, or “antibody like constructs” is also envisioned as, bi-specific antibodies, diabodies, scFv fragments, chimeric antibody receptor (CAR) constructs, diabody and / or minibody embodiments, unless explicitly denoted otherwise. The term “antibody” includes a polypeptide of the immunoglobulin family or a polypeptide comprising fragments of an immunoglobulin that is capable of non- covalently, reversibly, and in a specific manner binding a corresponding antigen, as disclosed herein. An exemplary antibody structural unit comprises a tetramer. In some embodiments, a full-length antibody can be composed of two identical pairs of polypeptide chains, each pair having one “light” and one “heavy” chain (connected through a disulfide bond). The term “antibody” also comprises immunoglobulins (Ig's) of different classes (i.e., IgA, IgG, IgM, IgD and IgE) and subclasses (such as IgG1, IgG2 etc.). The terms "anti-CD3 antibody" and "anti-CD3 antibody fragment," as used herein, mean antibodies or antibody fragments which recognize or bind to CD3.The TCR-anti-CD3 fusion molecule for use in the present disclosure can include one or more conservative substitutions which have a similar amino acid sequence and / or which retain the same function (i.e. are phenotypically silent as defined above). The skilled person is aware that various amino acids have similar properties and thus substitutions between them are “conservative”. One or more such amino acids of a protein, polypeptide or peptide can often be substituted by one or more other such amino acids without eliminating a desired activity of that protein, polypeptide, or peptide.

[0083] In some embodiments, the TCR-anti-CD3 fusion molecule for use in the invention comprises an anti-CD3 scFv covalently linked to the N-terminus of the beta chain of a TCR via a linker. This type of molecule is known as an ImmTAC® (Immune Mobilizing Monoclonal TCRs Against Cancer). ImmTAC® molecules are engineered to activate a potent T cell response to specifically kill target cancer cells. In some embodiments, the heterodimeric TCR-anti-CD3 antibody fusion molecules for use in the invention are ImmTAC molecules (i.e., ImmTACs targeting PRAME) as described in WO 2018 / 234319, which is incorporated by reference herein in its entirety.Atty. Docket No.0282-0001PR1

[0084] In some embodiments, the heterodimeric TCR-anti-CD3 antibody fusion molecules is IMC-F106C. IMC-F106C is a T cell redirecting bispecific therapeutic agent comprising a soluble affinity enhanced TCR that binds to the SLLQHLIGL(SEQ ID NO: 1) peptide-HLA- A*02 complex, fused to an anti-CD3 scFv. The targeting end of IMC-F106C (the soluble TCR) binds to a peptide fragment of the PRAME antigen presented by HLA-A*02 on the surface of cancer cells. HLA molecules are polymorphic; approximately 47% of Caucasian individuals in the US and European countries express the HLA-A*02 genotype with the HLA-A*02:01 allele detected in more than 95% of HLA-A*02-positive individuals. The effector end of IMC-F106C (anti-CD3 scFv) can bind to CD3 on any T cell, redirecting the T cell to produce effector cytokines and / or kill the cell presenting the target. In addition, IMC- F106C-mediated tumor lysis may prime an endogenous anti-tumor immune response. ImmTAC® (Immune Mobilizing Monoclonal TCRs Against Cancer) molecules such as IMC-F106C are highly potent molecules, with redirection of T-cell activity observed against tumor cell lines presenting as few as 10 to 50 target peptide:HLA complexes. IMC-F106C has been shown to selectively redirect T cell activity in the presence of HLA-A*02:01- positive / PRAME-positive cell lines, leading to T cell activation and killing of PRAME- positive cancer cells, at concentrations as low as 1 pM to 10 pM. As described above, the HLA-A*02 restricted peptide SLLQHLIGL (SEQ ID NO: 1) is derived from the germline cancer antigen PRAME. IMC-F106C has a TCR alpha chain amino acid sequence of SEQ ID NO: 14 and a TCR beta chain-anti-CD3 amino acid sequence of SEQ ID NO: 16.

[0085] The sequences referred to herein are as follows:

[0086] SEQ ID NO: 1 HLA-A*02 restricted peptide: SLLQHLIGL

[0087] SEQ ID NO: 2 Amino acid sequence of the TCR alpha chain variable domain of the ImmTAC designated as IMC-F106C. CDRs (CDR1 , CDR2 and CDR3) are underlined and are designated SEQ ID NO: 3, 4 and 5 respectively, framework regions (FR1 , FR2, FR3 and FR4) are in italics and are designated SEQ ID NO: 27, 6, 7 and 28 respectively. Mutations with respect to native alpha chain are in bold. GDAKTTQPNSMESNEEEPVHLPCNHSTISGTDYIHWYRQLPSQGPEYVIHGLTSNVNNR MASLAIAEDRKSSTLILHRATLRDAAVYYCILILGHSRLGNYIATFGKGTKLSVIP

[0088] SEQ ID NO: 8 Amino acid sequence of the TCR beta chain variable domain of the ImmTAC designated as IMC-F106C. CDRs (CDR1 , CDR2 and CDR3) are underlined and are designated SEQ ID NO: 9, 10 and 11 respectively, framework regions (FR1, FR2, FR3Atty. Docket No.0282-0001PR1 and FR4) are in italics and are designated SEQ ID NO: 29, 12, 13 and 30 respectively. Mutations with respect to native beta chain are in bold. DGGITQSPKYLFRKEGQNVTLSCEQNLNHDAMYWYRQDPGQGLRLIYYSQIMGDEQKG DIAEGYSVSREKKESFPLTVTSAQKNPTAFYLCASSWWTGGASPIRFGPGTRLTVT

[0089] SEQ ID NO: 14 Amino acid sequence of the TCR alpha chain of the ImmTAC designated as IMC-F106C. CDRs (CDR1, CDR2 and CDR3) are underlined and are designated SEQ ID NO: 3, 4 and 5 respectively, framework regions (FR1 , FR2, FR3 and FR4) are in italics and are designated SEQ ID NO: 27, 6, 7 and 28 respectively. The constant region is shown in bold and is designated SEQ ID NO: 15. Within the constant region, the nonnative cysteine residue is double underlined (at position 48 of constant region). GDAKTTQPNSMESNEEEPVHLPCNHSTISGTDYIHWYRQLPSQGPEYVIHGLTSNVNNR MASLAIAEDRKSSTLILHRATLRDAAVYYCILILGHSRLGNYIATFGKGTKLSVIPNIQNPD PAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKCVLDMRSMDFKSN SAVAWSNKSDFACANAFNNSIIPEDT

[0090] SEQ ID NO: 16 Amino acid sequence of the TCR beta chain-anti-CD3 of the ImmTAC designated as IMC-F106C. Anti-CD3 scFv (amino acids 1-253) is shown in bold and underline and is designated SEQ ID NO: 17. The linker (GGGGS) appears immediately after the scFv, is shown in paler text and is designated SEQ ID NO: 18. CDRs (CDR1 , CDR2 and CDR3) are underlined and are designated SEQ ID NO: 9, 10 and 11 respectively, framework regions (FR1, FR2, FR3 and FR4) are in italics and are designated SEQ ID NO: 29, 12, 13 and 30 respectively. Constant region is shown in bold (no underline) and is designated SEQ ID NO: 19. Within the constant region, the nonnative cysteine residue is double underlined (at position 57 of constant region). Additional non-native amino acids at position 75 and position 89 of the constant region are also double underlined. AIQMTQSPSSLSASVGDRVTITCRASQDIRNYLNWYQQKPGKAPKLLIYYTSRLE SGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCQQGNTLPWTFGQGTKVEIKGG GGSGGGGSGGGGSGGGGSGGGSEVQLVESGGGLVQPGGSLRLSCAASGYSFTG YTMNWVRQAPGKGLEWVALINPYKGVSTYNQKFKDRFTISVDKSKNTAYLQM NSLRAEDTAVYYCARSGYYGDSDWYFDVWGQGTLVTVSSGGGGSDGGITQSPKY LFRKEGQNVTLSCEQNLNHDAMYWYRQDPGQGLRLIYYSQIMGDEQKGDIAEGYSVSRE KKESFPLTVTSAQKNPTAFYLCASSWWTGGASPIRFGPGTRLTVTEDLKNVFPPEVAVF EPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKEVHSGVCTDPQPLKEQPAtty. Docket No.0282-0001PR1 ALNDSRYALSSRLRVSATFWQDPRNHFRCQVQFYGLSENDEWTQDRAKPVTQI VSAEAWGRAD

[0091] CDR sequences of the ImmTAC designated as IMC-F106C as follows: Alpha chain Alpha chain Alpha chain Beta chain Beta chain Beta chain CDR1 CDR2 CDR3 CDR1 CDR2 CDR3 TISGTDY GLTSN CILILGHSR LNHDA SQIMGDE CASSWWT (SEQ ID (SEQ ID LGNYIATF (SEQ ID (SEQ ID GGASPIRF NO:3) NO: 4) (SEQ ID NO: 9) NO: 10) (SEQ ID NO: 5) NO: 11)

[0092] Framework region sequences of the ImmTAC designated as IMC-F106C: Alpha chain FR1 Alpha chain FR2 Alpha chain FR3 Alpha chain FR4 GDAKTTQPNSMES IHWYRQLP VNNRMASLAIA GKGTKLSVIP NEEEPVHLPCNHS SQGPEYVIH EDRKSSTLILHR (SEQ ID NO: 28) (SEQ ID NO:27) (SEQ ID NO: 6) ATLRDAAVYY (SEQ ID NO: 7) Beta chain FR1 Beta chain FR2 Beta chain FR3 Beta chain FR4 DGGITQSPKYLFRK MYWYRQDP QKGDIAEGYSV GPGTRLTVT EGQNVTLSCEQN GQGLRLIYY SREKKESFPLTV (SEQ ID NO: 30) (SEQ ID NO:29) (SEQ ID NO: 12) TSAQKNPTAFYL (SEQ ID NO: 13)

[0093] Additional linker sequences referred to herein: 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)

[0094] In some embodiments, the TCR-anti-CD3 fusion molecule for use in the methods herein comprises: - a TCR alpha chain amino acid sequence of SEQ ID NO: 14 or a TOR alpha chain amino acid sequence that has at least 90%, at least 95% or 100% identity to the amino acid sequence of SEQ ID NO: 14, and - a TCR beta chain-anti-CD3 amino acid sequence of SEQ ID NO: 16 or a TOR beta chain-anti-CD3 amino acid sequence that has at least 90%, at least 95% or 100% identity to the amino acid sequence of SEQ ID NO: 16, wherein the TOR alpha chain variable domain comprises CDRs 1, 2 and 3 having the amino acid sequences of SEQ ID NOs: 3, 4 and 5 respectively and the TOR beta chainAtty. Docket No.0282-0001PR1 variable domain comprises CDRs 1 , 2 and 3 having the amino acid sequences of SEQ ID NOs: 9, 10 and 11, respectively.

[0095] In other words, although the molecule may have some variation in the TCR alpha chain amino acid sequence compared to the sequence of SEQ ID NO: 14 (as long as the TCR alpha chain amino acid sequence has at least 90% identity to SEQ ID NO: 14) and / or some variation in the TCR beta chain-anti-CD3 amino acid sequence compared to the sequence of SEQ ID NO: 16 (as long as the TCR beta chain-anti-CD3 amino acid sequence has at least 90% identity to the amino acid sequence of SEQ ID NO: 16), the CDRs of the TCR alpha chain must have the amino acid sequences of SEQ ID NOs: 3, 4 and 5 respectively and the CDRs of TCR beta chain must have the amino acid sequences of SEQ ID NOs: 9, 10 and 11 respectively. The requirement for the TCR alpha chain variable domain to comprise CDRs 1 , 2 and 3 having the amino acid sequences of SEQ ID NOs: 3, 4 and 5 respectively and the requirement for the TCR beta chain variable domain to comprise CDRs 1, 2 and 3 having the amino acid sequences of SEQ ID NOs: 9, 10 and 11 respectively thus applies to all aspects and embodiments of the invention described herein. The TCR alpha chain variable domain thus comprises CDRs 1 , 2 and 3 having 100% identity to the amino acid sequences of SEQ ID NOs: 3, 4 and 5 respectively and the TCR beta chain variable domain comprises CDRs 1 , 2 and 3 having 100% identity to the amino acid sequences of SEQ ID NOs: 9, 10 and 11 respectively.

[0096] In some embodiments, the heterodimeric TCR-anti-CD3 antibody fusion molecule is IMC-F106C. Within the scope of the present disclosure are phenotypically silent variants of the TCR-anti-CD3 fusion molecule designated as IMC-F106C, which has a TCR alpha chain amino acid sequence corresponding to SEQ ID NO: 14 and a TCR beta chain-anti-CD3 amino acid sequence corresponding to SEQ ID NO: 16. As used herein the term “phenotypically silent variant” is understood to refer to a TCR-anti-CD3 fusion molecule which incorporates one or more further amino acid changes, including substitutions, insertions and deletions, compared to the sequences of SEQ ID NO: 14 and SEQ ID NO: 16 and which TCR-anti-CD3 fusion molecule has a similar phenotype to or the same phenotype as the TCR-anti-CD3 fusion molecule designated as IMC-F106C. For the purposes of this application, TCR-anti-CD3 fusion molecule phenotype comprises antigen binding affinity (KD and / or binding half-life) and antigen specificity. A phenotypically silent variant may have a KD and / or binding half-life for the SLLQHLIGL (SEQ ID NO: 1) HLA-A*02 complex within 50%, or more preferably within 20%, of the measured KD and / or binding half-life of the TCR-anti-CD3 fusion molecule designated as IMC-F106C, when measuredAtty. Docket No.0282-0001PR1 under identical conditions (for example at 25°C and / or on the same SPR chip). 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, it may be possible to produce TCRs that incorporate changes in the variable domains thereof compared to those detailed above without altering the affinity of the interaction with the SLLQHLIGL (SEQ ID NO: 1) HLA-A*02 complex. In particular, such silent mutations may be incorporated within parts of the sequence that are known not to be directly involved in antigen binding. Such trivial variants are included in the scope of this invention.

[0097] Phenotypically silent variants may contain one or more conservative substitutions and / or one or more tolerated substitutions. Tolerated and conservative substitutions may result in a change in the KD and / or binding half-life for the SLLQHLIGL (SEQ ID NO: 1) HLA-A*02 complex within 50%, or more preferably within 20%, even more preferably within 10%, of the measured KD and / or binding half-life of the TCR-anti-CD3 fusion molecule designated as IMC-F106C, when measured under identical conditions (for example at 25°C and / or the same SPR chip), provided that the change in KD does not result in the affinity being less than (i.e. weaker than) 200 pm. By tolerated substitutions it is meant those substitutions which do not fall under the definition of conservative as provided below but are nonetheless phenotypically silent.

[0098] The TCR-anti-CD3 fusion molecule for use in the methods described herein can include one or more conservative substitutions which have a similar amino acid sequence and / or which retain the same function (i.e., are phenotypically silent as defined above). The skilled person is aware that various amino acids have similar properties and thus substitutions between them are “conservative”. One or more such amino acids of a protein, polypeptide or peptide can often be substituted by one or more other such amino acids without eliminating a desired activity of that protein, polypeptide, or peptide.

[0099] Thus, the amino acids glycine, alanine, valine, leucine, and isoleucine can often be substituted for one another (amino acids having aliphatic side chains). Of these possible substitutions it is preferred that glycine and alanine are used to substitute for one another (since they have relatively short side chains) and that valine, leucine and isoleucine are used to substitute for one another (since they have larger aliphatic side chains which are hydrophobic). Other amino acids which can often be substituted for one another include: phenylalanine, tyrosine and tryptophan (amino acids having aromatic side chains); lysine, arginine and histidine (amino acids having basic side chains); aspartate and glutamate (aminoAtty. Docket No.0282-0001PR1 acids having acidic side chains); asparagine and glutamine (amino acids having amide side chains); and cysteine and methionine (amino acids having Sulphur containing side chains). It should be appreciated that amino acid substitutions within the scope of the present invention can be made using naturally occurring or non-naturally occurring amino acids. For example, it is contemplated herein that the methyl group on an alanine may be replaced with an ethyl group, and / or that minor changes may be made to the peptide

[0100] backbone. Whether or not natural or synthetic amino acids are used, it is preferred that only L- amino acids are present.

[0101] Substitutions of this nature are often referred to as “conservative” or “semi- conservative” amino acid substitutions. The present invention therefore extends to use of a TCR-anti-CD3 fusion molecule comprising an amino acid sequence described above but with one or more conservative substitutions and / or one or more tolerated substitutions in the sequence, such that the TCR alpha chain amino acid sequence has at least 90% identity (such as 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity) to the amino acid sequence of SEQ ID NO: 14, and the TCR beta chain-anti-CD3 amino acid sequence has at least 90% identity (such as 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity) to the amino acid sequence of SEQ ID NO: 16, and provided that the TCR alpha chain variable domain comprises CDRs 1, 2 and 3 having the amino acid sequences of SEQ ID NOs: 3, 4 and 5 respectively and the TCR beta chain variable domain comprises CDRs 1, 2 and 3 having the amino acid sequences of SEQ ID NOs: 9, 10 and 11 respectively.

[0102] “Identity” as known in the art is the relationship between two or more polypeptide sequences or two or more polynucleotide sequences, as determined by comparing the sequences. In the art, identity also means the degree of sequence relatedness between polypeptide or polynucleotide sequences, as the case can be, as determined by the match between strings of such sequences. While there exist a number of methods to measure identity between two polypeptide or two polynucleotide sequences, methods commonly employed to determine identity are codified in computer programs. Preferred computer programs to determine identity between two sequences include, but are not limited to, GCG program package (Devereux, et al., Nucleic Acids Research, 12, 387 (1984), BLASTP, BLASTN, and FASTA (Atschul et al., J. Molec. Biol.215, 403 (1990)).

[0103] One can use a program such as the CLUSTAL program to compare amino acid sequences. This program compares amino acid sequences and finds the optimal alignment by inserting spaces in either sequence as appropriate. It is possible to calculate amino acid identity or similarity (identity plus conservation of amino acid type) for an optimal alignment.Atty. Docket No.0282-0001PR1 A program like BLASTx will align the longest stretch of similar sequences and assign a value to the fit. It is thus possible to obtain a comparison where several regions of similarity are found, each having a different score. Both types of identity analysis are contemplated in the present disclosure.

[0104] The percent identity of two amino acid sequences or of two nucleic acid sequences is determined by aligning the sequences for optimal comparison purposes (e.g., gaps can be introduced in the first sequence for best alignment with the sequence) and comparing the amino acid residues or nucleotides at corresponding positions. The “best alignment” is an alignment of two sequences which results in the highest percent identity. The percent identity is determined by the number of identical amino acid residues or nucleotides in the sequences being compared (i.e., % identity = number of identical positions / total number of positions x 100).

[0105] The determination of percent identity between two sequences can be accomplished using a mathematical algorithm known to those of skill in the art. An example of a mathematical algorithm for comparing two sequences is the algorithm of Karlin and Altschul (1990) Proc. Natl. Acad. Sci. USA 87:2264-2268, modified as in Karlin and Altschul (1993) Proc. Natl. Acad. Sci. USA 90:5873-5877. The NBLAST and XBLAST programs of Altschul, et al. (1990) J. Mol. Biol.215:403-410 have incorporated such an algorithm. BLAST nucleotide searches can be performed with the NBLAST program, score = 100, wordlength = 12 to obtain nucleotide sequences homologous to nucleic acid molecules. BLAST protein searches can be performed with the XBLAST program, score = 50, wordlength = 3 to obtain amino acid sequences homologous to protein molecules for use in the disclosure. To obtain gapped alignments for comparison purposes, Gapped BLAST can be utilized as described in Altschul et al. (1997) Nucleic Acids Res.25:3389-3402. Alternatively, PSI-Blast can be used to perform an iterated search which detects distant relationships between molecules (Id.). When utilizing BLAST, Gapped BLAST, and PSI- Blast programs, the default parameters of the respective programs (e.g., XBLAST and NBLAST) can be used. See http: / / www.ncbi.nlm.nih.gov. Another example of a mathematical algorithm utilized for the comparison of sequences is the algorithm of Myers and Miller, CABIOS (1989). The ALIGN program (version 2.0) which is part of the CGC sequence alignment software package has incorporated such an algorithm. Other algorithms for sequence analysis known in the art include ADVANCE and ADAM as described in Torellis and Robotti (1994) Comput. Appl. Biosci., 10 :3-5; and FASTA described in PearsonAtty. Docket No.0282-0001PR1 and Lipman (1988) Proc. Natl. Acad. Sci.85:2444-8. Within FASTA, ktup is a control option that sets the sensitivity and speed of the search.

[0106] Mutations, including conservation and tolerated substitutions, insertions, and deletions, can be introduced into the sequences provided using any appropriate method including, but not limited to, those based on polymerase chain reaction (PCR), restriction enzyme-based cloning, or ligation independent cloning (LIC) procedures. These methods are detailed in many of the standard molecular biology texts. For further details regarding polymerase chain reaction (PCR) and restriction enzyme-based cloning, see Sambrook & Russell, (2001) Molecular Cloning - A Laboratory Manual (3rdEd.) 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 by the disclosure can be obtained from solid state synthesis, or any other appropriate method known in the art.

[0107] The TCR-anti-CD3 fusion molecules for use in the present disclosure have the property of binding the SLLQHLIGL (SEQ ID NO: 1) HLA-A*02 complex. TCR-anti-CD3 fusion molecules for use in the present disclosure have been found to strongly recognize this epitope relative to other, irrelevant epitopes, and are thus particularly suitable as targeting vectors for delivery of therapeutic agents or detectable labels to cells and tissues displaying those epitopes. Specificity in the context of TCR-anti-CD3 fusion molecule for use in the present disclosure relates to their ability to recognize HLA-A*02 target cells that are antigen positive, whilst having minimal ability to recognize HLA-A*02 target cells that are antigen negative.

[0108] The method of the present disclosure is a dose escalation regimen, in which increasing doses of the heterodimeric TCR-anti-CD3 fusion molecule are sequentially administered to the subject. Doses are thus administered in the specified order: first dose, then second dose, then maintenance dose. "First dose" refers to a dose, i.e., dosage amount, of the TCR-anti- CD3 fusion molecule at a first amount within the specified range. "Second dose" refers to a dose, i.e., dosage amount, of the TCR-anti-CD3 fusion molecule at a second amount within the specified range, which is greater than the first dose. "Maintenance dose" refers to a dose, i.e., dosage amount, of the TCR-anti-CD3 fusion molecule at a maintenance amount within the specified range, which is greater than the second dose. It will be appreciated that according to the method of the present disclosure, a 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 can receive more than three total doses, as the subject can receive one or more first doses, one or more second doses, and one or more maintenance doses of theAtty. Docket No.0282-0001PR1 heterodimeric TCR-anti-CD3 antibody fusion molecule. As used herein, the amount in each second dose is larger than the amount in each first dose, and the amount in each maintenance dose is larger than the amount in each second dose.

[0109] In the present disclosure, each dose (i.e., first dose, second dose, maintenance) are expressed as a specified mass, i.e., weight, of the heterodimeric TCR-anti-CD3 antibody fusion molecule. Other vehicles, excipients, carriers, etc., may be administered with the heterodimeric TCR-anti-CD3 antibody fusion molecule but are not included in the calculated mass.

[0110] 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 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 to about 22 µg. In some embodiments, the first dose is about 20 µg. The dose 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 µg, 26 µg, 27 µg, 28 µg, 29 µg or 30 µg. In some embodiments, the first dose is 3 µg, 10 µg or 20 µg. In some embodiments, the first dose is 3 µg. In some embodiments, the first dose is 20 µg.

[0111] Various numbers of first doses can be administered to the subject. For example, in some embodiments, the first dose is administered once, i.e., one time. In some embodiments, the first dose is administered more than one time, e.g., two to 10 times, two to five times, or two to three times. In some embodiments, the first dose is administered two times.

[0112] The number of first doses can be determined, for example, by the occurrence or severity of an adverse event (AE) following administration of the first dose. In cases with adverse events, the method can include administration of more than one first doses before escalating to the second dose. In some embodiments, the first dose is a “range” of amounts, e.g., 2 to 30 µg. Thus, in some embodiments wherein multiple first doses are administered, the first dose can comprise the same amount in each first dose (e.g., each first dose comprises 2 µg). In some embodiments wherein multiple first doses are administered, the first dose amount can comprise different amounts in one or more of each first dose (e.g., first dose #1 comprises 2 µg and first dose #2 comprises 4 µg), as long as each dose is within the first dose amount specified.Atty. Docket No.0282-0001PR1

[0113] 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 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 about55 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.

[0114] Various numbers of second doses can be administered to the subject. For example, in some embodiments, the second dose is administered once, i.e., one time. In some embodiments, the second dose is administered more than one time, e.g., two to 10 times, two to five times, or two to three times. In some embodiments, the second dose is administered two times.

[0115] In some embodiments the method can include administration of more than one second dose before escalating to the maintenance dose. In some embodiments, the second dose is a “range” of amounts, e.g., 5 to 60 µg. Thus, in some embodiments wherein multiple second doses are administered, the second dose can comprise the same amount in each second dose (e.g., each second dose comprises 10 µg). In some embodiments wherein multiple second doses are administered, the second dose amount can comprise different amounts in one or more of each second dose (e.g., second dose #1 comprises 10 µg and second dose #2 comprises 15 µg), as long as each dose is within the second dose amount specified.

[0116] The term “maintenance dose(s)” a dosage administered to the subject to maintain the desired therapeutic effect. In some embodiments, the maintenance dose is administered repeatedly until treatment is stopped.

[0117] 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 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, 160Atty. Docket No.0282-0001PR1 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 20 µg or greater, Applicant observed durable partial responses (PRs) across several solid tumors including cutaneous melanoma (CM), uveal melanoma (UM), and ovarian carcinoma. In some embodiments, the maintenance dose is 40 µg. In some embodiments, the maintenance dose is 160 µg. Applicant observed that 160 µg was 1) the lowest dose where exposure was detected throughout the dosing period, and 2) compared to the next higher dose tested (320 µg), 160 µg has a lower incidence of cytokine release syndrome (CRS) and requires less dexamethasone premedication.

[0118] Various numbers of maintenance doses can be administered to the subject. For example, in some embodiments, the maintenance dose is administered three times, e.g., one time in each of the three specified phases. In some embodiments, maintenance dose is administered more than one time, e.g., three to 200 times, 10 to 150 times, or 30 to 80 times. In some embodiments, the maintenance dose is administered 30 to 34 times, e.g., 32 times. In some embodiments, the maintenance dose is administered until the melanoma, e.g., cutaneous melanoma, is cured, in remission, or is as otherwise determined by a healthcare professional. Treatment can be stopped, for example due to unacceptable toxicity or the occurrence of adverse events, or because the subject has shown an unacceptable level of disease progression. Alternatively, treatment can be stopped, for example, because the subject's symptoms have reduced in severity and / or tumor has shrunk to a level at which treatment with the TCR-anti-CD3 fusion molecule is deemed no longer necessary.

[0119] In some embodiments the method can include administration of more than one maintenance dose. In some embodiments, the maintenance dose is a “range” of amounts, e.g., 40 to 160 µg. Thus, in some embodiments wherein multiple maintenance doses are administered, the maintenance dose can comprise the same amount in each maintenance dose (e.g., each maintenance dose comprises 40 µg). In some embodiments, the maintenance dose amount can comprise different amounts in one or more of each maintenance dose (e.g., maintenance dose #1 comprises 10 µg and maintenance dose #2 comprises 15 µg), as long as each dose is within the maintenance dose amount specified.

[0120] The methods of the present disclosure provide a dosing regimen, wherein the heterodimeric TCR-anti-CD3 antibody fusion molecule is administered in specified amount. In some embodiments, when the first dose is administered multiple times, the first dose is administered every 4 days to every 14 days, or every 6 days to every 8 days, until each of the first doses is administered. In some embodiments, when the first dose is administeredAtty. Docket No.0282-0001PR1 multiple times, the first dose is administered every 7 days until each of the first doses is administered.

[0121] In some embodiments, when the second dose is administered multiple times, the second dose is administered every 4 days to every 14 days, or every 6 days to every 8 days, until each of the second doses is administered. In some embodiments, when the second dose is administered multiple times, the second dose is administered every 7 days until each of the second doses is administered. In some embodiments, the second dose is administered 4 to 14, 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.

[0122] In some embodiments, the maintenance dose is first administered 7 days after the last administration of the second dose. The maintenance dose can be administered in three phases, a first phase, a second phase and a third phase. As used herein, the term “phase” refers to a period of time in which the frequency of administration of the maintenance dose is substantially consistent. The term “substantially consistent” refers to a frequency that is ±24 hours of a given value. For example, if the phase frequency was “every 7 days,” then a frequency of 7 days ±24 hours would be in the same phase. In some embodiments, the “first phase” can comprise administering the maintenance dose every 6 to 8 days, the “second phase” can comprise administering the maintenance dose every 12 to 16 days, and / or the “third phase” can comprise administering the maintenance dose every 26 to 30 days. In some embodiments, the “first phase” can comprise administering the maintenance dose every 7 days, i.e., once a week, the “second phase” can comprise administering the maintenance dose every 14 days, i.e., every 2 weeks, and / or the “third phase” can comprise administering the maintenance dose every 28 days, i.e., every 4 weeks.

[0123] While not being bound by any particular theory, in some embodiments reducing the frequency from first phase to the second phase, and from second phase to the third phase of the maintenance dose can result in reduced number of side effects, reduced severity of side effects, and high patient compliance. In some embodiments, reducing the frequency from first phase to the second phase, and from second phase to the third phase of the maintenance dose does not reduce the efficacy of the heterodimeric TCR-anti-CD3 antibody fusions molecule. Reducing the frequency of the second and third phases (relative to the first and second phase, respectfully) can reduce overall costs and reduce the time needed for treatment of both the subject and the medical professional administering the treatment.

[0124] In some embodiments, the duration of the first phase of the maintenance dose is about one week to about 20 weeks, or about 4 weeks to about 15 weeks, or about 8 weeks to aboutAtty. Docket No.0282-0001PR1 12 weeks. In some embodiments, the duration of the first phase of the maintenance dose is about 10 weeks. In some embodiments, the duration of the second phase of the 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 the maintenance dose is about 20 weeks. In some embodiments, the duration of the third phase of the 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 the maintenance dose is about 40 weeks to about 60 weeks. In some embodiments, the third phase of the maintenance dose is about 48 weeks.

[0125] In some embodiments, the first phase is 8 weeks to about 14 weeks, the second phase is about 30 weeks to about 50 weeks, and the maintenance phase is about 40 weeks to about 60 weeks. In some embodiments, the first phase is about 10 weeks, the second phase is about 38 weeks, and / or the maintenance phase is about 48 weeks.

[0126] In some embodiments, the dosage regimen, including administration of the first dose, the second dose and the maintenance dose (all three phases), is 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 dosage regimen, including administration of the first dose, the second dose and the maintenance dose (all three phases), is about 90 weeks to about 110 weeks. In some embodiments, the dosage regimen, including administration of the first dose, the second dose and the maintenance dose (all three phases), is about 100 weeks to about 105 weeks. In some embodiments, the dosage regimen, including administration of the first dose, the second dose and the maintenance dose (all three phases), is about 101 weeks.

[0127] In some embodiments, the first dose is administered for 1 to 2 weeks, the second dose is administered for 1 to 2 weeks, and the maintenance dose is administered for at least 40 weeks, e.g., 40 weeks to 156 weeks, 80 weeks to 120 weeks, or 100 weeks to 104 weeks. In some embodiments, 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.

[0128] In some embodiments, the present disclosure provides a low dose regimen, wherein the first dose is about 2 to 4 µg, the second dose is about 5 to14 µg, and the maintenance dose is about 15 to 55 µg. Examples of these low dose regimens can be found below in Table A.Atty. Docket No.0282-0001PR1 Table AFirst dose Second dose Maintenance dose First phase Second phase Third phase 1 2-4 1-2 6-8 5-14 1-2 6-8 15- 10-14 6-8 15- 15- 12- 15- 10- 26-30 55 55 25 16 55 14 2 2-4 1 6-8 5-14 1 6-8 35- 10-14 6-8 35- 15- 12- 35- 10- 26-30 45 45 25 16 45 14 3 2-4 1 7 5-14 1 7 35- 10-14 7 35- 15- 14 35- 10- 28 45 45 25 45 14 4 3 1-2 6-8 10 1-2 6-8 40 10-14 6-8 40 15- 12- 40 10- 26-30 25 16 14 5 3 1 6-8 10 1 6-8 40 10-14 6-8 40 15- 12- 40 >10 26-30 25 16 6 3 1 NA 10 1 NA 40 10 7 40 17 14 40 12 28

[0129] In some embodiments, the present disclosure provides a high dose regimen, wherein the first dose is about 15 to 40 µg, the second dose is about 30 to 80 µg, and the maintenance dose is about 140 to 200 µg. Examples of these low dose regimens can be found below in Table B.Atty. Docket No.0282-0001PR1 Table B First dose Second dose Maintenance dose First phase Second phase Third phase 7 15- 1-2 6-8 30- 1-2 6-8 140- 10-14 6-8 140- 15- 12- 140 10- 26-30 40 80 200 200 25 16 - 14 200 8 15- 1 6-8 30- 1 6-8 140- 10-14 6-8 140- 15- 12- 140 10- 26-30 40 80 200 200 25 16 - 14 200 9 15- 1 7 30- 1 7 140- 10-14 7 140- 15- 14 140 10- 28 40 80 200 200 25 - 14 200 10 20 1-2 6-8 40 1-2 6-8 160 10-14 6-8 160 15- 12- 160 10- 26-30 25 16 14 11 20 1 6-8 40 1 6-8 160 10-14 6-8 160 15- 12- 160 >10 26-30 25 16 12 20 1 NA 40 1 NA 160 10 7 160 17 14 160 12 28

[0130] In some embodiments, the methods described herein are a first line treatment. The term “first line treatment” refers to the initial, or first treatment recommended for a disease or illness, e.g., cutaneous melanoma. In some embodiments, this can also be referred to as primary treatment, initial treatment, or induction therapy. In some embodiments, the first line treatment can include a combination treatment, e.g., treatment with another method, e.g., another active agent and / or surgery.

[0131] In some embodiments, the methods described herein are a second line treatment. In some embodiments, the methods described herein are third line, fourth line, fifth line, sixth line, etc. treatment. Second line or further lines of therapy (third line, fourth line, seventh line, etc.) may be used for a few different reasons, e.g., the first-line treatment doesn't work, the first-line treatment worked but has since stopped working, the first-line treatment has side effects that are not tolerated, and / or new treatment becomes available that appears to be more effective than the present treatment

[0132] In some embodiments, the methods described herein are a first line mono-therapy treatment, i.e., the only active agent used for the treatment of the cutaneous melanoma is theAtty. Docket No.0282-0001PR1 heterodimeric TCR-anti-CD3 antibody fusion molecule. In some embodiments, no corticosteroids are administered before administration of the heterodimeric TCR-anti-CD3 antibody fusion molecule. In some embodiments, no corticosteroids are administered 3 months, 2 months, 1 month, 3 weeks, 2 weeks or 1 week before administration of the heterodimeric TCR-anti-CD3 antibody fusion molecule. In some embodiments, no corticosteroids are administered 3 weeks before administration of the heterodimeric TCR- anti-CD3 antibody fusion molecule.

[0133] In some embodiments, the methods described herein are a monotherapy, e.g., a first line treatment monotherapy wherein no other anti-cancer drugs are co-administered during the dosing regimen. In some embodiments, the methods described herein are a monotherapy, e.g., a first line treatment monotherapy, wherein no other anti-cancer drugs are co- administered 3 months, 2 months, 1 month, 3 weeks, 2 weeks or 1 week before administration of the heterodimeric TCR-anti-CD3 antibody fusion molecule. In some embodiments, the methods described herein are a monotherapy, e.g., a first line treatment monotherapy, wherein no other anti-cancer drugs are co-administered 3 months, 2 months, 1 month, 3 weeks, 2 weeks or 1 week after completion of the dosage regimen.

[0134] In some embodiments, the methods described herein are not a monotherapy, i.e., the method comprises administering the heterodimeric TCR-anti-CD3 antibody fusion molecule and one or more additional anti-cancer therapeutic agents. In some embodiments, the dosing regimen further comprises administering a second therapeutic agent, a third therapeutic agent, a fourth therapeutic agent, or greater than four therapeutic agents. In some embodiments, the dosing regimen further comprises administering a second therapeutic agent. In some embodiments, the second therapeutic agent, a third therapeutic agent, a fourth therapeutic agent, or greater than four therapeutic agents can be an anti-cancer therapeutic agent. In some embodiments, the second therapeutic agent is a chemotherapeutic agent. In some embodiments, the second therapeutic is a biologic agent, e.g., a PD-1 inhibitor, a T-cell inactivator, a BRAF inhibitor, or a MEK inhibitor. In some embodiments, the second therapeutic agent comprises a checkpoint inhibitor. In some embodiments, the checkpoint inhibitor comprises of: Aldesleukin, Atezolizumab (TECENTRIQ®), Avelumab, Bevacizumab (Avastin®) Binimetinib, Braftovi (Encorafenib), Cobimetinib Fumarate, Cotellic (Cobimetinib Fumarate), Dabrafenib Mesylate, Dacarbazine, Durvalumab. Encorafenib, Galunisertib, Gemcitabine, IL-2 (Aldesleukin), Interferon, Imlygic (Talimogene Laherparepvec), Intron A (Recombinant Interferon Alfa-2b), Ipilimumab, Keytruda (Pembrolizumab), Kimmtrak (Tebentafusp-tebn), Mekinist (Trametinib Dimethyl Sulfoxide),Atty. Docket No.0282-0001PR1 Mektovi (Binimetinib), Merestinib, Nivolumab, Nivolumab and Relatlimab-rmbw, Opdivo (Nivolumab), Opdualag (Nivolumab and Relatlimab-rmbw), Pembrolizumab, Proleukin (Aldesleukin), Recombinant Interferon Alfa-2b, Tafinlar (Dabrafenib Mesylate), Talimogene Laherparepvec, Tebentafusp-tebn, Trametinib Dimethyl Sulfoxide, Tremelimumab, Vemurafenib, Yervoy (Ipilimumab), Zelboraf (Vemurafenib) or a combination thereof. In some embodiments, the checkpoint inhibitor comprises Nivolumab, Relatimab, or a combination thereof. In some embodiments, the checkpoint inhibitor comprises Nivolumab. Any of the listed checkpoint inhibitors can be administered as outlined in their Prescribing Information. For example, the Prescribing Information for Nivolumab (OPDIVO®, Bristol Myers Squibb), or the Prescribing Information for the combination of Nivolumab and Relatimab (OPDUALAG®, Bristol Myers Squibb) can be used to administer those therapeutic agents.

[0135] In some embodiments, the second (or third or fourth, etc.) therapeutic agent is co- administered with the heterodimeric TCR-anti-CD3 antibody fusion molecule, i.e., is administered on the same days as the heterodimeric TCR-anti-CD3 antibody fusion molecule. In some embodiments, a second therapeutic agent is administered according to a dosing amount and regimen known for that specific therapeutic agent. In some embodiments, the second therapeutic agent is administered once a week to once a month. In some embodiments, the second therapeutic agent is administered once a week. In some embodiments, the second therapeutic agent can be administered at a reduced frequency or reduced dosage amounts when co-administered with the heterodimeric TCR-anti-CD3 antibody fusion molecule.

[0136] In some embodiments, the second therapeutic agent is administered for the duration of the dosing regimen. In some embodiments, the second therapeutic agent is administered for a period of time known for that specific therapeutic agent. In some embodiments, the second therapeutic agent is administered for at least 10 weeks.. In some embodiments, the second therapeutic agent is 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 therapeutic agent is administered for at least 40 weeks. In some embodiments, the second therapeutic agent is administered for a shorter duration when co-administered with the heterodimeric TCR-anti-CD3 antibody fusion molecule relative to the second therapeutic being administered as a monotherapy.Atty. Docket No.0282-0001PR1

[0137] In a specific embodiment to treatment with heterodimeric TCR-anti-CD3 antibody fusion molecule and a second therapeutic agent, the disclosure provides a method of treating a cutaneous melanoma in a subject comprising administering to the subject (A) a composition comprising a heterodimeric TCR-anti-CD3 antibody fusion molecule in a dosing regimen, wherein the heterodimeric TCR-anti-CD3 antibody fusion molecule comprises (a) an alpha chain amino acid sequence that has at least 90%, at least 95% or 100% identity to the amino acid sequence of SEQ ID NO: 14, and (b) a beta chain amino acid sequence that has at least 90%, at least 95% or 100% identity to the amino acid sequence of SEQ ID NO: 16, wherein the alpha chain comprises complementarity-determining regions (CDRs) comprising alpha chain CDRs 1, 2 and 3 having the amino acid sequences of SEQ ID NOs: 3, 4 and 5 respectively and the beta chain comprises beta chain CDRs 1, 2 and 3 having the amino acid sequences of SEQ ID NOs: 9, 10 and 11 respectively, wherein the dosing regimen comprises administering: (i) a first dose comprising 2 to 30 ug of the TCR-anti-CD3 antibody fusion molecule, (ii) a second dose comprising 5 to 60 µg of the TCR-anti-CD3 antibody fusion molecule, and (iii) a maintenance dose comprising 15 to 200 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 once every 6 to 8 days, the second phase is administered once every 12 to 16 days, and the third phase is administered once every 26 to 30 days; and wherein the second dose comprises a larger amount of the TCR-anti-CD3 antibody fusion molecule than the first dose, and the maintenance dose comprises a larger amount of the TCR-anti-CD3 antibody fusion molecule than the second dose; and (B) a second therapeutic agent, wherein the second therapeutic agent is administered once every week to once every month; wherein the method is a first line treatment.

[0138] In some embodiments, the second therapeutic agent comprises a checkpoint inhibitor. In some embodiments, the checkpoint inhibitor comprises Nivolumab, Relatimab, or a combination thereof. In some embodiments, the checkpoint inhibitor comprises Nivolumab.

[0139] Administration of the heterodimeric TCR-anti-CD3 antibody fusion molecule can be by any conventional means known for administering anti-cancer biologics, e.g., intravenousAtty. Docket No.0282-0001PR1 (e.g., intravenous cannula (IVC) or central venous access device (CVAD)), intramuscular, subcutaneous, etc. In some embodiments, the heterodimeric TCR-anti-CD3 antibody fusion molecule can be administered by intravenous infusion.

[0140] In specific embodiments, the disclosure provides a method of treating a cutaneous melanoma in a subject comprising administering to the subject a composition comprising a heterodimeric TCR-anti-CD3 antibody fusion molecule in a dosing regimen, wherein the heterodimeric TCR-anti-CD3 antibody fusion molecule comprises (a) an alpha chain amino acid sequence that has at least 90%, at least 95% or 100% identity to the amino acid sequence of SEQ ID NO: 14, and (b) a beta chain amino acid sequence that has at least 90%, at least 95% or 100% identity to the amino acid sequence of SEQ ID NO: 16, wherein the alpha chain comprises complementarity-determining regions (CDRs) comprising alpha chain CDRs 1, 2 and 3 having the amino acid sequences of SEQ ID NOs: 3, 4 and 5 respectively and the beta chain comprises beta chain CDRs 1, 2 and 3 having the amino acid sequences of SEQ ID NOs: 9, 10 and 11 respectively, wherein the dosing regimen comprises: (i) administering one first dose comprising 3 µg of the TCR-anti-CD3 antibody fusion molecule, (ii) administering one second dose comprising 10 µg of the TCR-anti-CD3 antibody fusion molecule one week after the first dose, and (iii) administering a maintenance dose comprising 40 µg of the TCR-anti-CD3 antibody fusion molecule one week after the second dose, and every week during the first phase, wherein the first phase is about 9 to 10 weeks, (iv) administering the maintenance dose every 2 weeks during the second phase, wherein the second phase is about 38 weeks, and (v) administering the maintenance dose every 4 weeks during the third phase, wherein the third phase is about 48 weeks, wherein the method is a first line treatment.

[0141] In another specific embodiments, the disclosure provides a method of treating a cutaneous melanoma in a subject comprising administering to the subject a composition comprising a heterodimeric TCR-anti-CD3 antibody fusion molecule in a dosing regimen, wherein the heterodimeric TCR-anti-CD3 antibody fusion molecule comprises (a) an alpha chain amino acid sequence that has at least 90%, at least 95% or 100% identity to the amino acid sequence of SEQ ID NO: 14, and (b) a beta chain amino acid sequence that has atAtty. Docket No.0282-0001PR1 least 90%, at least 95% or 100% identity to the amino acid sequence of SEQ ID NO: 16, wherein the alpha chain comprises complementarity-determining regions (CDRs) comprising alpha chain CDRs 1, 2 and 3 having the amino acid sequences of SEQ ID NOs: 3, 4 and 5 respectively and the beta chain comprises beta chain CDRs 1, 2 and 3 having the amino acid sequences of SEQ ID NOs: 9, 10 and 11 respectively, wherein the dosing regimen comprises: (i) administering one first dose comprising 20 µg of the TCR-anti-CD3 antibody fusion molecule, (ii) administering one second dose comprising 40 µg of the TCR-anti-CD3 antibody fusion molecule one week after the first dose, and (iii) administering a maintenance dose comprising 160 µg of the TCR-anti-CD3 antibody fusion molecule one week after the second dose, and then every week during the first phase, wherein the first phase is about 9 to 10 weeks, (iv) administering the maintenance dose every 2 weeks during the second phase, wherein the second phase is about 38 weeks, and (v) administering the maintenance dose every 4 weeks during the third phase, wherein the third phase is about 48 weeks, wherein the method is a first line treatment.

[0142] In some embodiments, the methods provided herein can be directed to a PRAME- positive cancer, e.g., cancers including, but not limited to, 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, the disclosure provides a method of treating a PRAME- positive cancer in a subject comprising administering to the subject a composition comprising a heterodimeric TCR-anti-CD3 antibody fusion molecule in a dosing regimen, wherein the heterodimeric TCR-anti-CD3 antibody fusion molecule comprises (a) an alpha chain amino acid sequence that has at least 90%, at least 95% or 100% identity to the amino acid sequence of SEQ ID NO: 14, and (b) a beta chain amino acid sequence that has at least 90%, at least 95% or 100% identity to the amino acid sequence of SEQ ID NO: 16, wherein the alpha chain comprises complementarity-determining regions (CDRs) comprising alpha chain CDRs 1, 2 and 3 having the amino acid sequences of SEQ ID NOs: 3, 4 and 5 respectively and the beta chain comprises beta chain CDRs 1, 2 and 3 having the amino acid sequences of SEQ ID NOs: 9, 10 and 11 respectively,Atty. Docket No.0282-0001PR1 wherein the dosing regimen comprises administering: (i) a first dose comprising 2 to 30 ug of the TCR-anti-CD3 antibody fusion molecule, (ii) a second dose comprising 5 to 60 µg of the TCR-anti-CD3 antibody fusion molecule, and (iii) a maintenance dose comprising 15 to 200 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 once every 6 to 8 days, the second phase is administered once every 12 to 16 days, and the third phase is administered once every 26 to 30 days; and wherein the second dose comprises a larger amount of the heterodimeric TCR-anti-CD3 antibody fusion molecule than the first dose, and the maintenance dose comprises a larger amount of the heterodimeric TCR-anti-CD3 antibody fusion molecule than the second dose, wherein the method is a first line treatment. In some embodiments, the PRAME-positive cancer is an unresectable Stage III melanoma. Composition / kit

[0143] For administration to subjects, the heterodimeric TCR-anti-CD3 antibody fusion molecule of the disclosure, can be provided as part of a composition together with one or more pharmaceutically acceptable carriers or excipients. This pharmaceutical composition can be in any suitable form, (e.g., depending upon the desired method of administering it to a subject). It can be provided in unit dosage form and will generally be provided in a sealed container and can be provided as part of a kit. Such a kit would normally (although not necessarily) include instructions for use, and specifically can include detailed instructions related to the dosage amounts and the timing of administration. The kit can include a plurality of containers.

[0144] In some embodiments, the kit can comprise more than one container, wherein each container comprises a specific amount of lyophilized heterodimeric TCR-anti-CD3 antibody fusion molecule. The lyophilized heterodimeric TCR-anti-CD3 antibody fusion molecule in one container can be reconstituted in a liquid vehicle, e.g., a buffered saline solution, prior to administration to the subject, while the remaining containers remain lyophilized. In some embodiments, the kit can comprise more than one container, wherein each container comprises a specific amount of heterodimeric TCR-anti-CD3 antibody fusion molecule, wherein the heterodimeric TCR-anti-CD3 antibody fusion molecule is solubilized in a liquid vehicle, e.g., a buffer system and / or one or more excipients.Atty. Docket No.0282-0001PR1

[0145] Thus, for example, the kit can comprise (i) one to five first containers, each first container comprising a first dose comprising 2 to 4 µg of a heterodimeric TCR-anti-CD3 antibody fusion molecule, (ii) one to five second containers, each second container comprising a second dose comprising 5 to 14 µg of the heterodimeric TCR-anti-CD3 antibody fusion molecule, and (iii) three or more maintenance containers, each maintenance container comprising a maintenance dose comprising 15 to 55 µg of the heterodimeric TCR- anti-CD3 antibody fusion molecule, and (iv) instructions for administering the heterodimeric TCR-anti-CD3 antibody fusion molecule in a dosing regimen. In some embodiments, the instructions specify a dosing regimen comprising (a) administering the first dose weekly, (b) administering the second dose weekly, (c) administering the maintenance dose weekly during the first phase, (d) administering the maintenance dose every 2 weeks during the second phase, and (e) administering the maintenance dose every 4 weeks during the third phase.

[0146] In some embodiments, the disclosure provides a kit comprising: (i) one first container comprising a first dose comprising 2 to 4 µg of a heterodimeric TCR-anti-CD3 antibody fusion molecule, (ii) one second container comprising a second dose comprising 5 to 14 µg of the heterodimeric TCR-anti-CD3 antibody fusion molecule, (iii) one or more maintenance containers, each maintenance container comprising a maintenance dose comprising 15 to 55 µg of the heterodimeric TCR-anti-CD3 antibody fusion molecule, and (iv) instructions for administering the heterodimeric TCR-anti-CD3 antibody fusion molecule in a dosing regimen, wherein the dosing regimen is specified as (a) administering the first dose on week 1, (b) administering the second dose on week 2, (c) administering a maintenance dose weekly during the first phase for 10 weeks, (d) administering the maintenance dose every 2 weeks during the second phase for 38 weeks, and (e) administering the maintenance dose every 4 weeks during the third phase for 48 weeks.

[0147] In some embodiments, the disclosure provides a kit comprising: (i) one first container comprising a first dose comprising 20 µg of a heterodimeric TCR-anti-CD3 antibody fusion molecule, (ii) one second container comprising a second dose comprising 40 µg of the heterodimeric TCR-anti-CD3 antibody fusion molecule,Atty. Docket No.0282-0001PR1 (iii) one or more maintenance containers, each maintenance container comprising a maintenance dose comprising 160 µg of the heterodimeric TCR-anti-CD3 antibody fusion molecule, and (iv) instructions for administering the heterodimeric TCR-anti-CD3 antibody fusion molecule in a dosing regimen, wherein the dosing regimen is specified as (a) administering the first dose on week 1, (b) administering the second dose on week 2, (c) administering a maintenance dose weekly during the first phase for 10 weeks, (d) administering the maintenance dose every 2 weeks during the second phase for 38 weeks, and (e) administering the maintenance dose every 4 weeks during the third phase for 48 weeks.

[0148] In some embodiments, the heterodimeric TCR-anti-CD3 antibody fusion molecule in any of the kits described herein comprise (a) an alpha chain amino acid sequence that has at least 90%, at least 95% or 100% identity to the amino acid sequence of SEQ ID NO: 14, and (b) a beta chain amino acid sequence that has at least 90%, at least 95% or 100% identity to the amino acid sequence of SEQ ID NO: 16, wherein the alpha chain comprises complementarity-determining regions (CDRs) comprising alpha chain CDRs 1, 2 and 3 having the amino acid sequences of SEQ ID NOs: 3, 4 and 5 respectively and the beta chain comprises beta chain CDRs 1, 2 and 3 having the amino acid sequences of SEQ ID NOs: 9, 10 and 11 respectively.

[0149] In some embodiments, the disclosure provides a kit for the ascending dose regimen, the kit comprising: (i) one first container comprising a first dose comprising 3 µg of a heterodimeric TCR-anti-CD3 antibody fusion molecule, (ii) one second container comprising a second dose comprising 10µg of the heterodimeric TCR-anti-CD3 antibody fusion molecule, (iii) three or more maintenance containers, each maintenance container comprising a maintenance dose comprising 4 µg of the heterodimeric TCR-anti-CD3 antibody fusion molecule.

[0150] In some embodiments, the disclosure provides a kit for the ascending dose regimen, the kit comprising: (i) one first container comprising a first dose comprising 20 µg of a heterodimeric TCR-anti-CD3 antibody fusion molecule,Atty. Docket No.0282-0001PR1 (ii) one second container comprising a second dose comprising 40 µg of the heterodimeric TCR-anti-CD3 antibody fusion molecule, (iii) three or more maintenance containers, each maintenance container comprising a maintenance dose comprising 160 µg of the heterodimeric TCR-anti-CD3 antibody fusion molecule.

[0151] In some embodiments, the disclosure provides a kit for the maintenance dose regimen, the kit comprising: three or more maintenance containers, each maintenance container comprising a maintenance dose comprising 40 µg of the heterodimeric TCR-anti-CD3 antibody fusion molecule. In some embodiments, the kit for the maintenance dose regimen comprises enough maintenance containers for one month,2 months, three months, 4 months, 5 months, or 6 months.

[0152] In some embodiments, the disclosure provides a kit for the maintenance dose regimen, the kit comprising: three or more maintenance containers, each maintenance container comprising a maintenance dose comprising 160 µg of the heterodimeric TCR-anti-CD3 antibody fusion molecule.

[0153] In some embodiments, the kit for the maintenance dose regimen comprises enough maintenance containers for one month,2 months, three months, 4 months, 5 months, or 6 months.

[0154] The disclosure provides that the kit can comprise 1 or 2 first containers, 1 or 2 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,

[0155] In some embodiments, the first dose, the second dose and / or the maintenance dose further comprise(s) a pharmaceutically acceptable excipient to form a first pharmaceutical formulation, second pharmaceutical formulation, and / or maintenance pharmaceutical formulation, respectively. In some embodiments, the pH of the first, second and / or maintenance pharmaceutical formulation is pH adjusted. In some embodiments, the first, second and / or maintenance pharmaceutical formulation has a pH of about 6.5 to about 7.5.

[0156] In some embodiments, the kit comprises only one anti-cancer therapeutic agent. In some embodiments, the kit comprises the heterodimeric TCR-anti-CD3 antibody fusionAtty. Docket No.0282-0001PR1 molecule and further comprises a second therapeutic agent. In some embodiments, the second therapeutic agent in the kit comprises a checkpoint inhibitor. In some embodiments, the checkpoint inhibitor in the kit comprises Nivolumab, Relatimab, or a combination thereof. In some embodiments, the checkpoint inhibitor in the kit comprises Nivolumab.

[0157] The kits described herein contain one or more containers, e.g., a first container comprising a first dose, a second container comprising a second dose, and / or a maintenance container comprising 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 container holds or contains a composition that comprises the heterodimeric TCR- anti-CD3 antibody fusion molecule the disclosure and can have a sterile access port (for example, the container can be an intravenous solution bag or a vial having a stopper pierceable by a hypodermic injection needle). The instructions, e.g., label or package insert ,indicates that the heterodimeric TCR-anti-CD3 antibody fusion molecule is used for treating a subject eligible for treatment, e.g., one having or predisposed to developing a disease described herein, with specific guidance regarding dosing amounts and intervals of the composition and any other medicament being provided. In some embodiments, the kit can further comprise an additional container comprising a pharmaceutically acceptable diluent buffer, such as bacteriostatic water for injection (BWFI), phosphate-buffered saline, Ringer's solution, and / or dextrose solution. The kit can further include other materials desirable from a commercial and user standpoint, including other buffers, diluents, filters, needles, and syringes.

[0158] Various modifications of the disclosure in addition to 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 the disclosure are as for each of the other aspects mutatis mutandis. The documents referred to herein are incorporated by reference to the fullest extent permitted by law. EXAMPLE Example 1. IMCF106C-301 Clinical Trial Design

[0159] A randomized, controlled study is conducted to evaluate the efficacy of IMC-F106C + nivolumab compared with nivolumab regimens in HLA-A*02:01-positive adult participants with previously untreated advanced (unresectable or metastatic) cutaneous melanoma. Participants are randomized 1:1:1 to (i) IMC-F106C (40 µg maintenance dose) + nivolumabAtty. Docket No.0282-0001PR1 (Arm A), (ii) IMC-F106C (160 µg maintenance dose) + nivolumab (Arm B), or (iii) a nivolumab regimen (Arm C) (FIG.1).

[0160] The control Arm C will include: • Nivolumab + relatlimab, an emerging standard of care (SoC) in many countries, independent of programmed death-ligand 1 (PD-L1) status. • Nivolumab monotherapy, which is a global SoC, in all other countries.

[0161] The use of either nivolumab monotherapy or nivolumab + relatlimab in Arm C is country-specific. Investigators cannot select the control arm. The target randomization to nivolumab + relatlimab is approximately 20% of the control arm.

[0162] After the first participants are dosed (approximately 30 per arm), an independent committee will compare safety and limited efficacy data from Arms A and B and select the IMC-F106C dose with which to continue randomization for the primary efficacy analysis.

[0163] Randomization will be stratified by 1) American Joint Committee on Cancer (AJCC; 8th Edition) M stage with lactate dehydrogenase (LDH; M0 or M1 with normal LDH levels vs M1 with elevated LDH levels); 2) prior anti-PD-1 / PD-L1 (PD-[L]1) adjuvant therapy (yes vs no); and 3) BRAF V600 mutation status (positive vs negative). i. Dose Amounts.

[0164] The optimal regimen for IMC-F106C is an intra-subject dose escalation that induces rapid tachyphylaxis in cytokine release syndrome (CRS), requires minimal corticosteroids, and is well tolerated at the target dose and clinically active. IMC-F106C was previously studied over a thousand-fold dose range (0.2 to 320 µg). See, e.g., WO 2023 / 099622, incorporated by reference herein.

[0165] The threshold target dose for clinical activity is 20 µg. At this dose and through 320 µg, durable partial responses (PRs) are observed across several solid tumors including cutaneous melanoma (CM), uveal melanoma (UM), and ovarian carcinoma (FIG.2).

[0166] Repeat dosing up to 320 µg is tolerable. While 320 µg is tolerable, 160 µg is chosen as the highest dose for further testing because 1) this is the lowest dose where exposure was detected throughout the dosing period, and 2) compared to the next higher dose of 320 µg, 160 µg has a lower incidence of cytokine release syndrome (CRS) and requires less dexamethasone premedication (4 mg) than 320 µg (8 mg).

[0167] The impact of corticosteroid premedication on the efficacy of T-cell engagers (TCEs) is currently not completely established. IMC-F106C regimens with corticosteroidAtty. Docket No.0282-0001PR1 premedication are clinically active. Other TCEs targeting B-cell maturation antigen (BCMA) or CD20 use even higher premedication doses of dexamethasone (e.g., 16-20 mg) and still have demonstrated clinical activity (Tecvayli 2022, Epkinly 2023, Columvi 2023). However, immunotherapy regimens of the captioned disclosure strive to minimize premedication with corticosteroids. The 20 and 40 µg IMC-F106C cohorts do not include any premedication prior to the initial dose and also do not mandate corticosteroid premedication at subsequent doses.

[0168] Because of this range of well tolerated and clinically active doses, this study explores two separateIMC-F106C regimens that span a 4-fold dose range and differentially require corticosteroid premedication: 1.40 µg regimen (Arm A ): 3 µg (Week 1), 10 µg (Week 2), and 40 µg (Week 3+); no premedication is indicated for initial infusion and is not mandated at subsequent infusions. 2.160 µg regimen (Arm B): 20 µg (Week 1), 40 µg (Week 2), and 160 µg (Week 3+); corticosteroid premedication is mandated prior to the initial 3 infusions. ii. Dose Frequency.

[0169] IMC-F106C (both Arm A and Arm B) is administered weekly (QW) for the first 12 weeks, then every 2 weeks (Q2W) from week 13 to Week 51 (Month 12), and then every 4 weeks (Q4W) from week 52 to Week 101 (Month 24). Nivolumab is administered every 4 weeks (Q4W) until Week 101 (Month 24). Both therapies are administered until disease progression, unacceptable toxicity, withdrawal of consent, or for two years, whichever occurs first.

[0170] As a monotherapy, IMC-F106C can be administered QW. However, when combined with an active backbone therapy, in some embodiments IMC-F106C is dosed less frequently. For this clinical study, IMC-F106C has the same initial 3 intra-subject escalation doses as for monotherapy and is administered QW until Week 12, Q2W until Week 51, and then Q4W until Week 101 (end of Year 2) (Table 1).

[0171] First 12 weeks: QW dosing of IMC-F106C through the first tumor assessment ensures the highest IMC-F106C exposure when tumor progression on nivolumab is most likely to occur and when the greatest tumor decrease is observed for IMC-F106C. In this cutaneous melanoma population, the steepest decline in the Kaplan-Meier estimate of progression-free survival (PFS) for nivolumab is the initial 12 weeks, when approximatelyAtty. Docket No.0282-0001PR1 40% of all subjects’ progress (~60% of all progression events; Wolchok et al., N Engl J Med. 377(14): 1345-56, 2017).

[0172] Weeks 13-51: Q2W dosing of IMC-F106C from week 13 to Week 51 covers the period where progression on nivolumab dramatically slows down; approximately 15% of all subjects’ progress over these 9 months (Wolchok et al., N Engl J Med.377(14): 1345-56, 2017). Therefore, the Q2W regimen results in IMCF106C exposure for up to half of the dosing period and adds only 1 additional clinic visit per month beyond the Q4W nivolumab regimen.

[0173] Weeks 53-101: Q4W dosing of IMC-F106C coincides with the remaining 1 year of Q4W dosing of nivolumab for participant convenience. This is the period where progression on nivolumab plateaus, and thus, more frequent IMC-F106C dosing is not required (Wolchok et al., N Engl J Med.377(14): 1345-56, 2017). Table 1. Intervention IMC-F106C IMC-F106C Nivolumab Nivolumab + Label 40 µg Regimen 160 µg Regimen Relatlimab Biologic (sTCR: scFv Biologic (sTCR: Biologic Biologics Type fusion scFv fusion (monoclonal (monoclonal protein) protein) antibody) antibodies) Dose Concentrate for Concentrate for Concentrate for Concentrate for Formulation solution for infusion solution for solution for solution for infusion infusion infusion 12 mg / mL Unit Dose nivolumab and 4 Strength(s) 0.2 mg / mL 0.2 mg / mL 10 mg / mL mg / mL relatlimab (total dose, 16 mg / mL) Dosage 3 µg on Week 1, 20 µg on Week 1, 480 mg Q4W Nivolumab Regimen 10 µg on Week 2, 40 µg on Week 2, until Week 101 480 mg and 40 µg on Week 3, and 160 µg on Week 3, relatlimab QW doses of 40 µg and QW doses of 160 mg (total dose until 160 µg until 640 mg) Q4W until Week 13 and then Week 13 and then Week 101 Q2W until Week 53 Q2W until Week and then Q4W until 53 and then Q4W Week 101 until Week 101 Route of IV infusion IV infusion IV infusion IV infusion Administration Use Experimental Experimental Combination Comparator Partner or Comparator iii. PopulationAtty. Docket No.0282-0001PR1

[0174] Participants are eligible to be included in the study only if all the following criteria apply: 1. Participants must be ≥ 18 years of age at the time of signing the ICF. 2. Participants must be HLA-A*02:01-positive (testing by central laboratory). 3. Participants must have histologically confirmed Stage IV or unresectable Stage III cutaneous melanoma, per the AJCC 8th edition staging criteria (Gershenwald et al., CA Cancer J Clin.67(6):472-92, 2017; Keung et al., Expert Rev Anticancer Ther.18(8): 775-84, 2018). 4. An archived (within 6 months of randomization) or fresh tumor tissue sample from an unresectable or metastatic lesion must be confirmed as adequate. 5. Participants must have measurable disease per Response Evaluation Criteria in Solid Tumors (RECIST) 1.1 as assessed locally. Lesions situated in a previously irradiated area are considered measurable if progression has been shown in such lesions. 6. Participant must have BRAF V600 mutation status determined per local institutional standards or if not available locally then centrally prior to randomization. 8. Participant must have a life expectancy of at least three months. iv. Clinical Analysis

[0175] At each disease assessment, a computed tomography (CT) scan or magnetic resonance imaging (MRI) with contrast enhancement is used to assess all known and suspected sites of disease, including the neck, chest, abdomen, and / or pelvis, as appropriate based on extent of disease.

[0176] A brain MRI with contrast (or a CT scan with contrast if MRI contrast is not tolerated) is required at Screening and is required at subsequent assessments if disease was detected at Screening or as clinically indicated. All tumor measurements are taken and recorded in metric notation (mm) using a ruler or calipers. Imaging-based evaluation is preferred to evaluation by clinical examination when both methods have been used to assess the antitumor effect of a treatment. For optimal evaluation of participants, the same methods of assessment and technique are used to assess each reported lesion at baseline and during follow-up. Contrast-enhanced CT of the chest, abdomen, and pelvis are performed using a 5 mm slice thickness.

[0177] Additional scans of any nature are performed if clinically indicated.Atty. Docket No.0282-0001PR1

[0178] Tumor response will be determined according to RECIST 1.1. Investigator assessment of efficacy response data are collected and used for treatment decision making during the study. Participants who continue to receive treatment beyond progressive disease (PD) per RECIST 1.1 are to continue efficacy assessments until treatment is discontinued.

[0179] Cytology and histology: Cytology and histology are used to differentiate between PR and CR in rare cases (i.e., after treatment to differentiate between residual benign lesions and residual malignant lesions in tumor types such as germ cell tumors).

[0180] Clinical examination: Clinical lesions are considered measurable when they are superficial (i.e., skin nodules and palpable lymph nodes) and ≥ 10 mm in diameter. For the case of skin lesions being followed as target lesions, documentation by color photography, including a ruler to estimate the size of the lesion, is required. Target lesions response criteria are summarized in Table 2. Table 2 Response Criteria Evaluation of Target Lesions Complete Response Disappearance of all non-nodal target lesions. In addition, any pathological lymph nodes assigned as target lesions must have a reduction in short axis to <10 mm Partial Response At least a 30% decrease in the sum of diameter of all target lesions, taking as reference the baseline sum of diameters Progressive Disease At least a 20% increase in the sum of diameter of all measured target lesions, taking as reference the smallest sum of diameter of all target lesions recorded at or after baseline. In addition to the relative increase of 20%, the sum must also demonstrate an absolute increase of at least 5 mm Stable Disease Neither sufficient shrinkage to qualify for partial response or complete response nor an increase in lesions that would qualify for progressive disease

[0181] Clinical disease progression is acceptable and, wherever possible, is confirmed by imaging studies. The appearance of a new lesion is always associated with PD and has to be recorded as a new lesion. If a new lesion is equivocal, for example, because of its small size, continued therapy and follow-up evaluation will clarify if it represents a truly new disease. If repeat scans confirm there is definitely a new lesion, then progression should be declared using the date of the first observation of the lesion.

[0182] If new disease is observed in a region that was not scanned at baseline or where the particular baseline scan is not available for some reason, then this should be considered PD.Atty. Docket No.0282-0001PR1

[0183] A lymph node is considered as a “new lesion” and is therefore indicative of PD if the short axis increases in size to ≥ 10 mm for the first time in the study plus a 5 mm absolute increase.

[0184] Serum samples will be collected for measurement of concentrations of IMC-F106C and analyzed using a validated pharmacokinetic (PK) assay. The PK analysis set will be used in all PK data analysis and PK summary statistics. The noncompartmental PK parameters that will be assessed are presented in Table 3. Table 3. Parameter Definition CmaxMaximum (peak) observed drµg concentration (mass × volume-1) TmaxTime to reach maximum (peak) drµg concentration (time) ClastLast measurable time point of the concentration-time curve CavgAverage concentration t1 / 2Elimination half-life

[0185] Pharmacodynamics are evaluated using blood-derived samples for all participants. Whole blood samples are collected for measurement of messenger RNA (mRNA) levels of, for example, tumor markers, immune markers, and evaluation of the TCR repertoire.

[0186] Whole blood samples are collected and processed to isolate peripheral blood mononuclear cells (PBMCs), which are used for evaluation of the TCR repertoire.

[0187] Blood samples are collected for measurement of ctDNA and other plasma biomarkers. Reduction in ctDNA in peripheral blood is an emerging pharmacodynamic marker of tumor burden reduction in numerous solid tumors.

[0188] Tumor biopsy from unresectable or metastatic lesion (fresh or archival biopsy taken within 6 months of randomization) are evaluated for PD-L1, CD3, CD163, and PRAME expression to evaluate the association with efficacy including progression-free survival (PFS), objective response rate (ORR), and overall survival (OS).

[0189] The primary endpoint is PFS, defined as the time from randomization to the date of first documented PD (per RECIST 1.1 per Blinded Independent Central Review, BICR) or death due to any cause, whichever occurs first, regardless of whether the participant withdraws from randomized therapy.

[0190] The primary estimand of PFS is based on full analysis set (FAS), analyzed according to the randomized treatment. The set of intercurrent events consists of starting subsequentAtty. Docket No.0282-0001PR1 anticancer therapy, which are accounted for by censoring at the last evaluable tumor assessment on or prior to the date of starting subsequent therapy for the primary estimand. The point estimate of the treatment effect (hazard ratio, HR), as well as the associated 2-sided 95% CI, are estimated using a Cox-proportional-hazards model stratified by the stratification factors using the Efron approach for handling ties.

[0191] One of the key secondary endpoints is OS, which is defined as time between the date of randomization and the date of death from any cause.

[0192] OS is analyzed based on the FAS according to the randomized treatment. The point estimate of the treatment effect (HR), as well as the associated 2-sided 95% CI, is estimated using a Cox-proportional-hazards model stratified by stratification factors using the Ephron approach for handling ties. The null hypothesis of no difference in OS is tested using a 2- sided log-rank test stratified by the stratification factors.

[0193] One of the key secondary endpoints is the ORR, which is defined as the proportion of participants with a best overall response (BOR; per BICR) of complete response (CR) or PR, based on RECIST 1.1.

[0194] ORR id analyzed based on the FAS according to the randomized treatment. Estimatesof ORR, along with its exact 2-sided 95% CI by Clopper and Person id presented by treatment group. The point estimate of the treatment effect (odds ratios), as well as the associated 2-sided95% CIs and p-value, are generated using a 2-sided Cochran-Mantel- Haenszel (CMH) test stratified by the stratification factors. The null hypothesis of no difference in ORR is judged based on the p-value.

[0195] The objectives and endpoints of this study are presented in Table 4. Table 4 Type Objectives Endpoints Primary EfficacyF106TCo+conmivpoaluremPaFbStofonrivIMoluCm- ab RECPISFTS1a.s1assessed by the BICR according toregimens SecondaryEfficacyforTo compare overall survival OSIMC-F106C + nivolumab to nivolumab regimensrate fToor IcMomCp-Fa1re06oCve+ranllivreoslupmonasbeRECOISRTR1.a1s assessed by BICR according toto nivolumab regimens

Claims

Atty. Docket No.0282-0001PR1 CLAIMS What is claimed is:

1. A method of treating a cutaneous melanoma in a subject comprising administering to the subject a composition comprising a heterodimeric TCR-anti-CD3 antibody fusion molecule in a dosing regimen, wherein the heterodimeric TCR-anti-CD3 antibody fusion molecule comprises (a) an alpha chain amino acid sequence that has at least 90%, at least 95% or 100% identity to the amino acid sequence of SEQ ID NO: 14, and (b) a beta chain amino acid sequence that has at least 90%, at least 95% or 100% identity to the amino acid sequence of SEQ ID NO: 16, wherein the alpha chain comprises complementarity-determining regions (CDRs) comprising alpha chain CDRs 1, 2 and 3 having the amino acid sequences of SEQ ID NOs: 3, 4 and 5 respectively and the beta chain comprises beta chain CDRs 1, 2 and 3 having the amino acid sequences of SEQ ID NOs: 9, 10 and 11 respectively, wherein the dosing regimen comprises administering: (i) a first dose comprising 2 to 30 ug of the TCR-anti-CD3 antibody fusion molecule, (ii) a second dose comprising 5 to 60 µg of the TCR-anti-CD3 antibody fusion molecule, and (iii) a maintenance dose comprising 15 to 200 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 once every 6 to 8 days, the second phase is administered once every 12 to 16 days, and the third phase is administered once every 26 to 30 days; and wherein the second dose comprises a larger amount of the TCR-anti-CD3 antibody fusion molecule than the first dose, and the maintenance dose comprises a larger amount of the TCR-anti-CD3 antibody fusion molecule than the second dose, wherein the method is a first line treatment.

2. The method of claim 1, wherein the first dose comprises 2 to 4 ug of the TCR-anti- CD3 antibody fusion molecule, the second dose comprises 5 to 14 µg of the TCR-anti-CD3 antibody fusion molecule, and the maintenance dose comprises 15 to 55 µg of the TCR-anti- CD3 antibody fusion molecule.

3. The method of claim 2, wherein the first dose comprises 3 ug of the TCR-anti-CD3 antibody fusion molecule, the second dose comprises 10 µg of the TCR-anti-CD3 antibodyAtty. Docket No.0282-0001PR1 fusion molecule, and the maintenance dose comprises 40 µg of the TCR-anti-CD3 antibody fusion molecule.

4. The method of claim 1, wherein the first dose comprises 15 to 40 µg of the TCR-anti- CD3 antibody fusion molecule, the second dose comprises 30 to 80 µg of the TCR-anti-CD3 antibody fusion molecule, and the maintenance dose comprises 140 to 200 µg of the TCR- anti-CD3 antibody fusion molecule.

5. The method of claim 4, wherein the first dose comprises 20 µg of the TCR-anti-CD3 antibody fusion molecule, the second dose comprises 40 µg of the TCR-anti-CD3 antibody fusion molecule, and the maintenance dose comprises 160 µg of the TCR-anti-CD3 antibody fusion molecule.

6. The method of any one of claims 1 to 5, wherein the alpha chain amino acid sequence has 100% identity to the amino acid sequence of SEQ ID NO:

14.

7. The method of any one of claims 1 to 6, wherein the beta chain amino acid sequence has 100% identity to the amino acid sequence of SEQ ID NO:

16.

8. The method of any one of claims 1 to7, wherein the first dose is administered once every 7 days.

9. The method of any one of claims 1 to 8, wherein the first dose is administered for 1 week to about 3 weeks.

10. The method of claim 9, wherein the first dose is administered for 1 week.

11. The method of any one of claims 1 to 10 wherein the second dose is administered 6 to 8 days following the first dose.

12. The method of any one of claims 1 to 11, wherein the second dose is administered once every 7 days.

13. The method of any one of claims 1 to 12, wherein the second dose is administered for 1 week to about 3 weeks.

14. The method of claim 13, wherein the second dose is administered for 1 week.

15. The method of any one of claims 1 to 14, wherein the maintenance dose in the first phase is administered 6 to 8 days following the second dose.Atty. Docket No.0282-0001PR1 16. The method of any one of claims 1 to 15, wherein the maintenance dose in the first phase is administered once in a week.

17. The method of any one of claims 1 to 16, wherein the first phase is about 2 weeks to about 101 weeks.

18. The method of claim 17, wherein the first phase is about 8 weeks to about 14 weeks.

19. The method of claim 18, wherein the first phase is 10 weeks.

20. The method of any one of claims 1 to 19, wherein the maintenance dose in the second phase is administered once every 2 weeks.

21. The method of any one of claims 1 to 20, wherein the second phase is about 30 weeks to 50 weeks.

22. The method of claim 21, wherein the second phase is 38 weeks.

23. The method of any one of claims 1 to 22, wherein the maintenance dose in the third phase is administered every 4 weeks.

24. The method of any one of claims 1 to 23, wherein the third phase is about 40 weeks to about 60 weeks.

25. The method of claim 24, wherein the third phase is about 48 weeks.

26. The method of any one of claims 1 to 25, wherein the first phase is 10 weeks, the second phase is 38 weeks, and the third phase is 48 weeks.

27. The method of any one of claims 1 to 26, 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.

28. The method of any one of claims 1 to 27, wherein the dosing regimen is about 90 weeks to about 110 weeks.

29. The method of any one of claims 1 to 28, wherein no corticosteroids are administered within three weeks prior to the dosing regimen and throughout the dosing regimen.

30. The method of any one of claims 1 to 29, wherein no other anti-cancer drugs are co- administered during the dosing regimen.Atty. Docket No.0282-0001PR1 31. The method of any one of claims 1 to 30, wherein the subject is HLA-A*02 positive.

32. The method of any one of claims 1 to 31, wherein the subject is HLA-A*02:01 positive.

33. The method of any one of claims 1 to 32, wherein the dosing regimen further comprises administering a second therapeutic agent.

34. The method of claim 33, wherein the second therapeutic agent comprises a checkpoint inhibitor.

35. The method of claim 34, wherein the checkpoint inhibitor comprises of: Aldesleukin, Atezolizumab (TECENTRIQ®), Avelumab, Bevacizumab (Avastin®) Binimetinib, Braftovi (Encorafenib), Cobimetinib Fumarate, Cotellic (Cobimetinib Fumarate), Dabrafenib Mesylate, Dacarbazine, Durvalumab. Encorafenib, Galunisertib, Gemcitabine, IL-2 (Aldesleukin), Interferon, Imlygic (Talimogene Laherparepvec), Intron A (Recombinant Interferon Alfa-2b), Ipilimumab, Keytruda (Pembrolizumab), Kimmtrak (Tebentafusp-tebn), Mekinist (Trametinib Dimethyl Sulfoxide), Mektovi (Binimetinib), Merestinib, Nivolumab, Nivolumab and Relatlimab-rmbw, Opdivo (Nivolumab), Opdualag (Nivolumab and Relatlimab-rmbw), Pembrolizumab, Proleukin (Aldesleukin), Recombinant Interferon Alfa- 2b, Tafinlar (Dabrafenib Mesylate), Talimogene Laherparepvec, Tebentafusp-tebn, Trametinib Dimethyl Sulfoxide, Tremelimumab, Vemurafenib, Yervoy (Ipilimumab), Zelboraf (Vemurafenib) or a combination thereof.

36. The method of claim 34, wherein the checkpoint inhibitor comprises Nivolumab, Relatimab, or a combination thereof.

37. The method of claim 34, wherein the checkpoint inhibitor comprises Nivolumab.

38. The method of any one of claims 33 to 37, wherein the second therapeutic agent is administered once a week to once a month.

39. The method of claim 38, wherein the second therapeutic agent is administered once a week.

40. The method of any one of claims 33 to 39, wherein the second therapeutic agent is administered for at least 10 weeks.Atty. Docket No.0282-0001PR1 41. The method of claim 40, wherein the second therapeutic agent is administered for at least 40 weeks.

42. The method of any one of claims 34 to 41, wherein the second therapeutic agent is administered for the duration of the dosing regimen.

43. The method of any one of claims 1 to 42, wherein the cutaneous melanoma is a Stage IV melanoma.

44. The method of claim 43, wherein Stage IV melanoma is histologically confirmed.

45. The method of any one of claims 1 to 44, wherein the cutaneous melanoma is an unresectable Stage III melanoma .

46. The method of any one of claims 1 to 45, wherein the subject has BRAF V600 mutation.

47. The method of any one of claims 1 to 46, wherein the subject has been diagnosed with a life expectancy greater than 3 months.

48. The method of any one of claims 1 to 47, wherein the composition is administered by intravenous (IV) infusion.

49. A method of treating a cutaneous melanoma in a subject comprising administering to the subject a composition comprising a heterodimeric TCR-anti-CD3 antibody fusion molecule in a dosing regimen, wherein the heterodimeric TCR-anti-CD3 antibody fusion molecule comprises (a) an alpha chain amino acid sequence that has at least 90%, at least 95% or 100% identity to the amino acid sequence of SEQ ID NO: 14, and (b) a beta chain amino acid sequence that has at least 90%, at least 95% or 100% identity to the amino acid sequence of SEQ ID NO: 16, wherein the alpha chain comprises complementarity-determining regions (CDRs) comprising alpha chain CDRs 1, 2 and 3 having the amino acid sequences of SEQ ID NOs: 3, 4 and 5 respectively and the beta chain comprises beta chain CDRs 1, 2 and 3 having the amino acid sequences of SEQ ID NOs: 9, 10 and 11 respectively, wherein the dosing regimen comprises: (i) administering one first dose comprising 3 µg of the TCR-anti-CD3 antibody fusion molecule,Atty. Docket No.0282-0001PR1 (ii) administering one second dose comprising 10 µg of the TCR-anti-CD3 antibody fusion molecule one week after the first dose, and (iii) administering a maintenance dose comprising 40 µg of the TCR-anti-CD3 antibody fusion molecule one week after the second dose, and every week during the first phase, wherein the first phase is about 9 to 10 weeks, (iv) administering the maintenance dose every 2 weeks during the second phase, wherein the second phase is about 38 weeks, and (v) administering the maintenance dose every 4 weeks during the third phase, wherein the third phase is about 48 weeks, wherein the method is a first line treatment.

50. A method of treating a cutaneous melanoma in a subject comprising administering to the subject a composition comprising a heterodimeric TCR-anti-CD3 antibody fusion molecule in a dosing regimen, wherein the heterodimeric TCR-anti-CD3 antibody fusion molecule comprises (a) an alpha chain amino acid sequence that has at least 90%, at least 95% or 100% identity to the amino acid sequence of SEQ ID NO: 14, and (b) a beta chain amino acid sequence that has at least 90%, at least 95% or 100% identity to the amino acid sequence of SEQ ID NO: 16, wherein the alpha chain comprises complementarity-determining regions (CDRs) comprising alpha chain CDRs 1, 2 and 3 having the amino acid sequences of SEQ ID NOs: 3, 4 and 5 respectively and the beta chain comprises beta chain CDRs 1, 2 and 3 having the amino acid sequences of SEQ ID NOs: 9, 10 and 11 respectively, wherein the dosing regimen comprises: (i) administering one first dose comprising 20 µg of the TCR-anti-CD3 antibody fusion molecule, (ii) administering one second dose comprising 40 µg of the TCR-anti-CD3 antibody fusion molecule one week after the first dose, and (iii) administering a maintenance dose comprising 160 µg of the TCR-anti-CD3 antibody fusion molecule one week after the second dose, and then every week during the first phase, wherein the first phase is about 9 to 10 weeks, (iv) administering the maintenance dose every 2 weeks during the second phase, wherein the second phase is about 38 weeks, and (v) administering the maintenance dose every 4 weeks during the third phase, wherein the third phase is about 48 weeks, wherein the method is a first line treatment.Atty. Docket No.0282-0001PR1 51. A kit comprising: (i) one first container comprising a first dose comprising 2 to 4 µg of a heterodimeric TCR-anti-CD3 antibody fusion molecule, (ii) one second container comprising a second dose comprising 5 to 14 µg of the heterodimeric TCR-anti-CD3 antibody fusion molecule, (iii) one or more maintenance containers, each maintenance container comprising a maintenance dose comprising 15 to 55 µg of the heterodimeric TCR-anti-CD3 antibody fusion molecule, and (iv) instructions for administering the heterodimeric TCR-anti-CD3 antibody fusion molecule in a dosing regimen, wherein the dosing regimen is specified as (a) administering the first dose on week 1, (b) administering the second dose on week 2, (c) administering a maintenance dose weekly during the first phase for 10 weeks, (d) administering the maintenance dose every 2 weeks during the second phase for 38 weeks, and (e) administering the maintenance dose every 4 weeks during the third phase for 48 weeks.

52. A kit comprising: (i) one first container comprising a first dose comprising 20 µg of a heterodimeric TCR-anti-CD3 antibody fusion molecule, (ii) one second container comprising a second dose comprising 40 µg of the heterodimeric TCR-anti-CD3 antibody fusion molecule, (iii) one or more maintenance containers, each maintenance container comprising a maintenance dose comprising 160 µg of the heterodimeric TCR-anti-CD3 antibody fusion molecule, and (iv) instructions for administering the heterodimeric TCR-anti-CD3 antibody fusion molecule in a dosing regimen, wherein the dosing regimen is specified as (a) administering the first dose on week 1, (b) administering the second dose on week 2, (c) administering a maintenance dose weekly during the first phase for 10 weeks, (d) administering the maintenance dose every 2 weeks during the second phase for 38 weeks, and (e) administering the maintenance dose every 4 weeks during the third phase for 48 weeks.

53. The kit according to claim 51 or 52, wherein the heterodimeric TCR-anti-CD3 antibody fusion molecule comprises (a) an alpha chain amino acid sequence that has at least 90%, at least 95% or 100% identity to the amino acid sequence of SEQ ID NO: 14, and (b) a beta chain amino acid sequence that has at least 90%, at least 95% or 100% identity to theAtty. Docket No.0282-0001PR1 amino acid sequence of SEQ ID NO: 16, wherein the alpha chain comprises complementarity-determining regions (CDRs) comprising alpha chain CDRs 1, 2 and 3 having the amino acid sequences of SEQ ID NOs: 3, 4 and 5 respectively and the beta chain comprises beta chain CDRs 1, 2 and 3 having the amino acid sequences of SEQ ID NOs: 9, 10 and 11 respectively.

54. The kit according to claim 51 or 52, wherein the kit comprises 2 to 10 maintenance containers.

55. The kit according to claim 51 or 52, wherein the first dose, the second dose and / or the maintenance dose further comprise(s) a pharmaceutically acceptable excipient to form a first pharmaceutical formulation, second pharmaceutical formulation, and / or maintenance pharmaceutical formulation, respectively.

56. The kit according to claim 55 wherein the first, second and / or maintenance pharmaceutical formulation has a pH of about 6.5 to about 7.

5.

57. The kit according to claim 51 or 52, wherein the kit further comprises a second therapeutic agent.

58. The kit according to claim 57, wherein the second therapeutic agent comprises a checkpoint inhibitor.

59. The kit according to claim 58, wherein the checkpoint inhibitor comprises of: Aldesleukin, Atezolizumab (TECENTRIQ®), Avelumab, Bevacizumab (Avastin®) Binimetinib, Braftovi (Encorafenib), Cobimetinib Fumarate, Cotellic (Cobimetinib Fumarate), Dabrafenib Mesylate, Dacarbazine, Durvalumab. Encorafenib, Galunisertib, Gemcitabine, IL-2 (Aldesleukin), Interferon, Imlygic (Talimogene Laherparepvec), Intron A (Recombinant Interferon Alfa-2b), Ipilimumab, Keytruda (Pembrolizumab), Kimmtrak (Tebentafusp-tebn), Mekinist (Trametinib Dimethyl Sulfoxide), Mektovi (Binimetinib), Merestinib, Nivolumab, Nivolumab and Relatlimab-rmbw, Opdivo (Nivolumab), Opdualag (Nivolumab and Relatlimab-rmbw), Pembrolizumab, Proleukin (Aldesleukin), Recombinant Interferon Alfa-2b, Tafinlar (Dabrafenib Mesylate), Talimogene Laherparepvec, Tebentafusp-tebn, Trametinib Dimethyl Sulfoxide, Tremelimumab, Vemurafenib, Yervoy (Ipilimumab), Zelboraf (Vemurafenib) or a combination thereof.

60. The kit according to claim 58, wherein the checkpoint inhibitor comprises Nivolumab, Relatimab, or a combination thereof.Atty. Docket No.0282-0001PR1 61. The kit according to claim 58, wherein the checkpoint inhibitor comprises Nivolumab.

62. A method of treating a PRAME-positive cancer in a subject comprising administering to the subject a composition comprising a heterodimeric TCR-anti-CD3 antibody fusion molecule in a dosing regimen, wherein the heterodimeric TCR-anti-CD3 antibody fusion molecule comprises (a) an alpha chain amino acid sequence that has at least 90%, at least 95% or 100% identity to the amino acid sequence of SEQ ID NO: 14, and (b) a beta chain amino acid sequence that has at least 90%, at least 95% or 100% identity to the amino acid sequence of SEQ ID NO: 16, wherein the alpha chain comprises complementarity-determining regions (CDRs) comprising alpha chain CDRs 1, 2 and 3 having the amino acid sequences of SEQ ID NOs: 3, 4 and 5 respectively and the beta chain comprises beta chain CDRs 1, 2 and 3 having the amino acid sequences of SEQ ID NOs: 9, 10 and 11 respectively, wherein the dosing regimen comprises administering: (i) a first dose comprising 2 to 30 ug of the TCR-anti-CD3 antibody fusion molecule, (ii) a second dose comprising 5 to 60 µg of the TCR-anti-CD3 antibody fusion molecule, and (iii) a maintenance dose comprising 15 to 200 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 once every 6 to 8 days, the second phase is administered once every 12 to 16 days, and the third phase is administered once every 26 to 30 days; and wherein the second dose comprises a larger amount of the TCR-anti-CD3 antibody fusion molecule than the first dose, and the maintenance dose comprises a larger amount of the TCR-anti-CD3 antibody fusion molecule than the second dose, wherein the method is a first line treatment.

63. The method of claim 62, wherein the PRAME-positive cancer is an unresectable Stage III melanoma.

64. A method of treating a cutaneous melanoma in a subject comprising administering to the subject (A) a composition comprising a heterodimeric TCR-anti-CD3 antibody fusion molecule in a dosing regimen,Atty. Docket No.0282-0001PR1 wherein the heterodimeric TCR-anti-CD3 antibody fusion molecule comprises (a) an alpha chain amino acid sequence that has at least 90%, at least 95% or 100% identity to the amino acid sequence of SEQ ID NO: 14, and (b) a beta chain amino acid sequence that has at least 90%, at least 95% or 100% identity to the amino acid sequence of SEQ ID NO: 16, wherein the alpha chain comprises complementarity-determining regions (CDRs) comprising alpha chain CDRs 1, 2 and 3 having the amino acid sequences of SEQ ID NOs: 3, 4 and 5 respectively and the beta chain comprises beta chain CDRs 1, 2 and 3 having the amino acid sequences of SEQ ID NOs: 9, 10 and 11 respectively, wherein the dosing regimen comprises administering: (i) a first dose comprising 2 to 30 ug of the TCR-anti-CD3 antibody fusion molecule, (ii) a second dose comprising 5 to 60 µg of the TCR-anti-CD3 antibody fusion molecule, and (iii) a maintenance dose comprising 15 to 200 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 once every 6 to 8 days, the second phase is administered once every 12 to 16 days, and the third phase is administered once every 26 to 30 days; and wherein the second dose comprises a larger amount of the TCR-anti-CD3 antibody fusion molecule than the first dose, and the maintenance dose comprises a larger amount of the TCR-anti-CD3 antibody fusion molecule than the second dose; and (A) a second therapeutic agent, wherein the second therapeutic agent is administered once every week to once every month; wherein the method is a first line treatment.

65. The method of claim 64, wherein the second therapeutic agent comprises a checkpoint inhibitor.

66. The method of claim 65, wherein the checkpoint inhibitor comprises of: Aldesleukin, Atezolizumab (TECENTRIQ®), Avelumab, Bevacizumab (Avastin®) Binimetinib, Braftovi (Encorafenib), Cobimetinib Fumarate, Cotellic (Cobimetinib Fumarate), Dabrafenib Mesylate, Dacarbazine, Durvalumab. Encorafenib, Galunisertib, Gemcitabine, IL-2 (Aldesleukin), Interferon, Imlygic (Talimogene Laherparepvec), Intron A (Recombinant Interferon Alfa-2b), Ipilimumab, Keytruda (Pembrolizumab), Kimmtrak (Tebentafusp-tebn), Mekinist (Trametinib Dimethyl Sulfoxide), Mektovi (Binimetinib), Merestinib, Nivolumab, Nivolumab and Relatlimab-rmbw, Opdivo (Nivolumab), Opdualag (Nivolumab and Relatlimab-rmbw), Pembrolizumab, Proleukin (Aldesleukin), Recombinant Interferon Alfa-Atty. Docket No.0282-0001PR1 2b, Tafinlar (Dabrafenib Mesylate), Talimogene Laherparepvec, Tebentafusp-tebn, Trametinib Dimethyl Sulfoxide, Tremelimumab, Vemurafenib, Yervoy (Ipilimumab), Zelboraf (Vemurafenib) or a combination thereof.

67. The method of claim 66, wherein the checkpoint inhibitor comprises Nivolumab, Relatimab, or a combination thereof.

68. The method of claim 66, wherein the checkpoint inhibitor comprises Nivolumab.

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