LAG-3 antagonists for the treatment of melanoma

Abstract

To provide an improved method for treating unresectable or metastatic melanoma.SOLUTION: The present invention provides methods of treating unresectable or metastatic melanoma in a human patient with a lymphocyte activation gene-3 (LAG-3) antagonist. In one aspect, the methods include a combination of a LAG-3 antagonist and a cytotoxic T lymphocyte antigen-4 (CTLA-4) inhibitor. In one aspect, the method comprises one or more additional therapeutic agents and / or anti-cancer therapies.SELECTED DRAWING: None

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This PCT application claims the benefit of priority to U.S. Provisional Patent Application No. 62 / 932,916, filed November 8, 2019, the entire contents of which are incorporated herein by reference.

[0002] FIELD OF THE INVENTION The invention described herein relates to methods of treating unresectable or metastatic melanoma in human patients comprising lymphocyte activation gene 3 (LAG-3) antagonists. [Background technology]

[0003] Background of the Invention Human cancers harbor numerous genetic mutations and epigenetic changes, generating neoantigens that can be recognized by the immune system (Sjoblom et al. (2006) Science 314:268-74). The adaptive immune system, comprised of T and B lymphocytes, has broad capabilities and excellent specificity in responding to diverse tumor antigens, potentially providing powerful anticancer effects. Furthermore, the immune system exhibits considerable plasticity and possesses memory components. By effectively utilizing these characteristics of the adaptive immune system, immunotherapy could become unique among all cancer therapies.

[0004] Lymphocyte activation gene-3 (LAG-3; CD223) is a ubiquitous marker of activated CD4 + and CD8 +LAG-3 is a type I transmembrane protein expressed on the cell surface of T cells, as well as a subset of NK cells and dendritic cells (Triebel F, et al., J. Exp. Med. 1990; 171:1393-1405; Workman CJ, et al., J. Immunol. 2009; 182(4):1885-91). LAG-3 is closely related to CD4, a coreceptor for T helper cell activation. Both molecules have four extracellular Ig-like domains and bind to major histocompatibility complex (MHC) class II. In contrast to CD4, LAG-3 is expressed only on the cell surface of activated T cells, and its cleavage from the cell surface terminates LAG-3 signaling. LAG-3 is also found as a soluble protein, the function of which is unknown.

[0005] Cytotoxic T-lymphocyte antigen-4 (CTLA-4) is an immunoinhibitory receptor belonging to the CD28 family. CTLA-4 is expressed exclusively on T cells in vivo and binds to two ligands, CD80 and CD86 (also known as B7-1 and B7-2, respectively).

[0006] It is an object of the present invention to provide improved methods for treating unresectable or metastatic melanoma. Summary of the Invention

[0007] Summary of the Invention The present invention relates to a method of treating unresectable or metastatic melanoma in a human patient, comprising administering to the patient (a) a lymphocyte-activation gene-3 (LAG-3) antagonist, and (b) a cytotoxic T-lymphocyte antigen 4 (CTLA-4) inhibitor, wherein the patient has a susceptibility mutation to targeted inhibitor therapy.

[0008] The present invention relates to a method of treating unresectable or metastatic melanoma in a human patient, comprising administering to the patient (a) a LAG-3 antagonist, and (b) a CTLA-4 inhibitor, wherein the patient has previously received a PD-1 pathway inhibitor as treatment for the melanoma.

[0009] In one aspect, the method is a first line therapy.

[0010] In some aspects, the method is a second line therapy.

[0011] In some aspects, the method is a third line therapy.

[0012] In some aspects, the patient is progressing on prior therapy.

[0013] In some aspects, the patient has not received prior systemic therapy for cancer, the patient has not received prior systemic therapy for melanoma, or the patient has not received prior systemic therapy for unresectable or metastatic melanoma.

[0014] In some aspects, the patient is naive to a previous immuno-oncology therapy, the patient is naive to a previous immuno-oncology therapy for the melanoma, or the melanoma is naive to a previous immuno-oncology therapy.

[0015] In one aspect, the patient has histologically confirmed unresectable Stage III or Stage IV melanoma.

[0016] In some aspects, the patient has an Eastern Cooperative Oncology Group (ECOG) performance status of 0 or 1.

[0017] In some aspects, the patient has a type B aggressive fibrosarcoma proto-oncogene (BRAF), mitogen-activated extracellular signal-regulated kinase kinase (MEK), neuroblastoma RAS viral oncogene homolog (NRAS), and / or proto-oncogene c-KIT (KIT) mutation that is susceptible to targeted inhibitor therapy. In some aspects, the patient has a BRAF mutation that is susceptible to targeted inhibitor therapy.

[0018] In one aspect, one or more immune cells in tumor tissue from a patient express LAG-3. In one aspect, at least about 1%, at least about 3%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or about 100% of the immune cells express LAG-3. In one aspect, the immune cells are tumor-infiltrating lymphocytes. In one aspect, the tumor-infiltrating lymphocytes are CD8 + It is a cell.

[0019] In certain aspects, one or more tumor cells in a tumor tissue from a patient express PD-L1. In certain aspects, at least about 1%, at least about 3%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or about 100% of the tumor cells express PD-L1. In certain aspects, at least about 1% of the tumor cells express PD-L1.

[0020] In one aspect, the LAG-3 antagonist is an anti-LAG-3 antibody.

[0021] In one aspect, the anti-LAG-3 antibody is a full-length antibody.

[0022] In some aspects, the anti-LAG-3 antibody is a monoclonal antibody, a human antibody, a humanized antibody, a chimeric antibody, or a multispecific antibody. In some aspects, the multispecific antibody is a dual affinity retargeting antibody (DART), a DVD-Ig, or a bispecific antibody.

[0023] In one aspect, the anti-LAG-3 antibody is a F(ab')2 fragment, a Fab' fragment, a Fab fragment, an Fv fragment, an scFv fragment, a dsFv fragment, a dAb fragment, or a single-chain binding polypeptide peptide.

[0024] In one aspect, the anti-LAG-3 antibody is BMS-986016 (leratolimab), IMP731 (H5L7BW), MK-4280 (28G-10), REGN3767 (fianlimab), GSK2831781, humanized BAP050, IMP-701 (LAG-5250, yeramilimab), aLAG3(0414), aLAG3(0416), Sym022, TSR-033, TSR-075, XmAb22841, MGD013, BI754111, FS118, P 13B02-30, AVA-017, 25F7, AGEN1746, or an antigen-binding portion thereof.

[0025] In one aspect, the anti-LAG-3 antibody comprises the CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO:3, and the CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO:5.

[0026] In one aspect, the anti-LAG-3 antibody comprises: (a) a heavy chain variable region, CDR1, comprising the sequence set forth in SEQ ID NO:7; (b) a heavy chain variable region, CDR2, comprising the sequence set forth in SEQ ID NO:8; (c) a heavy chain variable region, CDR3, comprising the sequence set forth in SEQ ID NO:9; (d) a light chain variable region, CDR1, comprising the sequence set forth in SEQ ID NO:10; (e) a light chain variable region, CDR2, comprising the sequence set forth in SEQ ID NO:11; and (f) a light chain variable region CDR3 comprising the sequence set forth in SEQ ID NO: 12.

[0027] In one aspect, the anti-LAG-3 antibody comprises a heavy chain variable region and a light chain variable region comprising the sequences set forth in SEQ ID NOs: 3 and 5, respectively.

[0028] In one aspect, the anti-LAG-3 antibody comprises a heavy chain and a light chain comprising the sequences set forth in SEQ ID NOs: 1 and 2, respectively.

[0029] In one aspect, the anti-LAG-3 antibody comprises a heavy chain and a light chain comprising the sequences set forth in SEQ ID NOs: 30 and 2, respectively.

[0030] In one aspect, the LAG-3 antagonist is a soluble LAG-3 polypeptide. In one aspect, the soluble LAG-3 polypeptide is a fusion polypeptide. In one aspect, the soluble LAG-3 polypeptide comprises a ligand-binding fragment of the LAG-3 extracellular domain. In one aspect, the ligand-binding fragment of the LAG-3 extracellular domain comprises an amino acid sequence having at least about 90%, at least about 95%, at least about 98%, at least about 99%, or about 100% sequence identity to SEQ ID NO: 41. In one aspect, the soluble LAG-3 polypeptide further comprises a half-life extending moiety. In one aspect, the half-life extending moiety comprises an immunoglobulin constant region or portion thereof, an immunoglobulin-binding polypeptide, immunoglobulin G (IgG), albumin-binding polypeptide (ABP), a PAS-modified moiety, a HES-modified moiety, XTEN, a PEGylated moiety, an Fc region, or any combination thereof. In one aspect, the soluble LAG-3 polypeptide is IMP321 (eftilagimod alfa).

[0031] In one aspect, the CTLA-4 inhibitor is an anti-CTLA-4 antibody.

[0032] In one aspect, the anti-CTLA-4 antibody is a full-length antibody.

[0033] In some aspects, the anti-CTLA-4 antibody is a monoclonal antibody, a human antibody, a humanized antibody, a chimeric antibody, or a multispecific antibody. In some aspects, the multispecific antibody is a DART, a DVD-Ig, or a bispecific antibody.

[0034] In one aspect, the anti-CTLA-4 antibody is a F(ab')2 fragment, a Fab' fragment, a Fab fragment, an Fv fragment, an scFv fragment, a dsFv fragment, a dAb fragment, or a single-chain binding polypeptide peptide.

[0035] In one aspect, the anti-CTLA-4 antibody is ipilimumab, tremelimumab, MK-1308, AGEN-1884, or an antigen-binding portion thereof.

[0036] In one aspect, the anti-CTLA-4 antibody comprises the CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO:34, and the CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO:32.

[0037] In one aspect, the anti-CTLA-4 antibody comprises: (a) a heavy chain variable region, CDR1, comprising the sequence set forth in SEQ ID NO: 35; (b) a heavy chain variable region, CDR2, comprising the sequence set forth in SEQ ID NO: 36; (c) a heavy chain variable region, CDR3, comprising the sequence set forth in SEQ ID NO: 37; (d) a light chain variable region, CDR1, comprising the sequence set forth in SEQ ID NO: 38; (e) a light chain variable region, CDR2, comprising the sequence set forth in SEQ ID NO: 39; and (f) a light chain variable region, CDR3, comprising the sequence set forth in SEQ ID NO: 40. Includes:

[0038] In one aspect, the anti-CTLA-4 antibody comprises a heavy chain variable region and a light chain variable region comprising the sequences set forth in SEQ ID NOs: 34 and 32, respectively.

[0039] In one aspect, the anti-CTLA-4 antibody comprises the heavy and light chains of ipilimumab.

[0040] In one aspect, the LAG-3 antagonist and the CTLA-4 inhibitor are formulated for intravenous administration.

[0041] In one aspect, the LAG-3 antagonist and the CTLA-4 inhibitor are formulated separately.

[0042] In one aspect, the LAG-3 antagonist and the CTLA-4 inhibitor are formulated as a single agent.

[0043] In one aspect, the LAG-3 antagonist is administered prior to the CTLA-4 inhibitor.

[0044] In one aspect, the CTLA-4 inhibitor is administered prior to the LAG-3 antagonist.

[0045] In one aspect, the LAG-3 antagonist and the CTLA-4 inhibitor are administered simultaneously.

[0046] In one aspect, the LAG-3 antagonist and / or CTLA-4 inhibitor is administered in a fixed dose.

[0047] In one aspect, the LAG-3 antagonist and / or CTLA-4 inhibitor is administered in an amount of at least about 0.25 mg to about 2000 mg, about 0.25 mg to about 1600 mg, about 0.25 mg to about 1200 mg, about 0.25 mg to about 800 mg, about 0.25 mg to about 400 mg, about 0.25 mg to about 100 mg, about 0.25 mg to about 50 mg, about 0.25 mg to about 40 mg, about 0.25 mg to about 30 mg, about 0.25 mg to about 20 mg, about 20 mg to about 2000 mg, about 20 mg to about 1600 mg, about 20 mg to about 1200 mg, about 20 mg to about 800 mg, about 20 mg to about 1000 mg, about 0.25 mg to about 500 mg, about 0.25 mg to about 40 mg, about 0.25 mg to about 30 mg, about 0.25 mg to about 20 mg, about 20 mg to about 2000 mg, about 20 mg to about 1600 mg, about 20 mg to about 1200 mg, about 20 mg to about 800 mg, about 20 mg to about 1000 mg, about 0.25 ... It is administered at a dosage of about 400 mg, about 20 mg to about 100 mg, about 100 mg to about 2000 mg, about 100 mg to about 1800 mg, about 100 mg to about 1600 mg, about 100 mg to about 1400 mg, about 100 mg to about 1200 mg, about 100 mg to about 1000 mg, about 100 mg to about 800 mg, about 100 mg to about 600 mg, about 100 mg to about 400 mg, 400 mg to about 2000 mg, about 400 mg to about 1800 mg, about 400 mg to about 1600 mg, about 400 mg to about 1400 mg, about 400 mg to about 1200 mg, or about 400 mg to about 1000 mg.

[0048] In one aspect, the LAG-3 antagonist and / or CTLA-4 inhibitor is administered in an amount of about 0.25 mg, about 0.5 mg, about 0.75 mg, about 1 mg, about 1.25 mg, about 1.5 mg, about 1.75 mg, about 2 mg, 2.25 mg, about 2.5 mg, about 2.75 mg, about 3 mg, about 3.25 mg, about 3.5 mg, about 3.75 mg, about 4 mg, about 4.2 5mg, about 4.5mg, about 4.75mg, about 5mg, about 5.25mg, about 5.5mg, about 5.75mg, about 6mg, about 6.25mg, about 6.5mg, about 6.75mg, about 7 mg, about 7.25 mg, about 7.5 mg, about 7.75 mg, about 8 mg, about 8.25 mg, about 8.5 mg, about 8.75 mg, about 9 mg, about 9.25 mg, about 9.5 mg, about 9.75mg, about 10mg, about 20mg, about 30mg, about 40mg, about 50mg, about 60mg, about 70mg, about 80mg, about 90mg, about 100mg, about 110mg, about 120mg, about 130mg, about 140mg, about 150mg, about 160mg, about 170mg, about 18 0mg, about 190mg, about 200mg, about 210mg, about 220mg, about 230mg, about 240mg, about 250mg, about 260mg, about 270mg, about 280mg, about 290mg, about 300mg, about 310mg, about 320mg, about 330mg, about 340mg, about 35 0mg, approx. 360mg, approx. 370mg, approx. 380mg, approx. 390mg, approx. 400mg, approx. 410mg, approx. 420mg, approx. 430mg, approx. 440mg, approx. 450mg, approx. 20mg, about 530mg, about 540mg, about 550mg, about 560mg, about 570mg, about 580mg, about 590mg, about 600mg, about 610mg, about 620mg, about 630mg, about 640mg, about 650mg, about 660mg, about 670mg, about 680mg, about 6 90mg, about 700mg, about 710mg, about 720mg, about 730mg, about 740mg, about 750mg, about 760mg, about 770mg, about 780mg, about 790mg, about 800mg, about 810mg, about 820mg, about 830mg, about 840mg, about 850mg, about 860mg, about 870mg, about 880mg, about 890mg, about 900mg, about 910mg, about 920mg, about 930mg, about 940mg, about 950mg, about 960mg, about 970mg, about 980mg, about 990mg, about 1000mg, about 1040mg, about 1080m g, about 1100 mg, about 1140 mg, about 1180 mg, about 1200 mg, about 1240 mg, about 1280 mg, about 1300 mg, about 1340 mg, about 1380 mg, about 1400 mg, about 1440 mg, about 1480 mg, about 1500 mg, about 1540 mg, about 1580 mg, about 1600 mg, about 1640 mg, about 1680 mg, about 1700 mg, about 1740 mg, about 1780 mg, about 1800 mg, about 1840 mg, about 1880 mg, about 1900 mg, about 1940 mg, about 1980 mg, or about 2000 mg.

[0049] In one aspect, the LAG-3 antagonist and / or CTLA-4 inhibitor is administered in a weight-dependent dose.

[0050] In some aspects, the LAG-3 antagonist and / or CTLA-4 inhibitor is administered at a dose of from about 0.003 mg / kg to about 25 mg / kg, from about 0.003 mg / kg to about 20 mg / kg, from about 0.003 mg / kg to about 15 mg / kg, from about 0.003 mg / kg to about 10 mg / kg, from about 0.003 mg / kg to about 5 mg / kg, from about 0.003 mg / kg to about 1 mg / kg, from about 0.003 mg / kg to about 0.9 mg / kg, from about 0.003 mg / kg to about 1 mg / kg, ...1 mg / kg, from about 0.003 mg / kg to about 1 mg / kg, from about 0.003 mg / kg to about 1 mg / kg, from about 0.003 mg / kg to about 1 mg / kg, from about 0.003 mg / kg to about 1 mg / kg, from about 0.003 mg / kg to about 1 mg / kg, from about 0.003 mg / kg to about 1 mg / kg, from about 0.003 mg / kg to about 1 mg / kg, from about 0.003 mg / kg to about 1 mg / kg .003mg / kg~about 0.8mg / kg, about 0.003mg / kg~about 0.7mg / kg, about 0.003mg / kg~about 0.6mg / kg, about 0.003mg / kg~about 0.5mg / kg, about 0.003m g / kg~about 0.4mg / kg, about 0.003mg / kg~about 0.3mg / kg, about 0.003mg / kg~about 0.2mg / kg, about 0.003mg / kg~about 0.1mg / kg, about 0.1mg / kg~about 2 5mg / kg, approximately 0.1mg / kg to approximately 20mg / kg, approximately 0.1mg / kg to approximately 15mg / kg, approximately 0.1mg / kg to approximately 10mg / kg, approximately 0.1mg / kg to approximately 5mg / kg, approximately 0.1mg / kg to approximately 1mg / kg, about 1mg / kg to about 25mg / kg, about 1mg / kg to about 20mg / kg, about 1mg / kg to about 15mg / kg, about 1mg / kg to about 10mg / kg, about 1mg / kg to about 5mg / kg, about 5 The drug is administered at a dose of about 10 mg / kg to about 25 mg / kg, about 5 mg / kg to about 20 mg / kg, about 5 mg / kg to about 15 mg / kg, about 5 mg / kg to about 10 mg / kg, about 10 mg / kg to about 25 mg / kg, about 10 mg / kg to about 20 mg / kg, about 10 mg / kg to about 15 mg / kg, about 15 mg / kg to about 25 mg / kg, about 15 mg / kg to about 20 mg / kg, or about 20 mg / kg to about 25 mg / kg.

[0051] In one aspect, the LAG-3 antagonist and / or CTLA-4 inhibitor is administered at a dose of about 0.003 mg / kg, about 0.004 mg / kg, about 0.005 mg / kg, about 0.006 mg / kg, about 0.007 mg / kg, about 0.008 mg / kg, about 0.009 mg / kg, about 0.01 mg / kg, about 0.02 mg / kg, about 0.03 mg / kg, about 0.04 mg / kg, about 0.05 mg / kg, about 0.06 mg / kg, about 0.07 mg / kg, about 0.08 mg / kg, about 0.09 mg / kg, about 0.1 mg / kg, about 0.2 mg / kg, about 0.3 mg / kg, about 0.4 mg / kg, about 0.5 mg / kg, about 0.6 mg / kg, about 0.7 mg / kg, about 0. The compound is administered at a dose of about 8 mg / kg, about 0.9 mg / kg, about 1.0 mg / kg, about 2.0 mg / kg, about 3.0 mg / kg, about 4.0 mg / kg, about 5.0 mg / kg, about 6.0 mg / kg, about 7.0 mg / kg, about 8.0 mg / kg, about 9.0 mg / kg, about 10.0 mg / kg, about 11.0 mg / kg, about 12.0 mg / kg, about 13.0 mg / kg, about 14.0 mg / kg, about 15.0 mg / kg, about 16.0 mg / kg, about 17.0 mg / kg, about 18.0 mg / kg, about 19.0 mg / kg, about 20.0 mg / kg, about 21.0 mg / kg, about 22.0 mg / kg, about 23.0 mg / kg, about 24.0 mg / kg, or about 25.0 mg / kg.

[0052] In some aspects, the LAG-3 antagonist and / or CTLA-4 inhibitor is administered about once every week, about once every 2 weeks, about once every 3 weeks, about once every 4 weeks, about once every 5 weeks, about once every 6 weeks, about once every 7 weeks, about once every 8 weeks, about once every 9 weeks, about once every 10 weeks, about once every 11 weeks, or about once every 12 weeks.

[0053] In some aspects, the method further comprises administering to the patient an additional therapeutic agent. In some aspects, the additional therapeutic agent comprises an anti-cancer agent. In some aspects, the anti-cancer agent comprises a tyrosine kinase inhibitor, an anti-angiogenic agent, a checkpoint inhibitor, a checkpoint stimulator, a chemotherapeutic agent, an immunotherapeutic agent, a platinum agent, an alkylating agent, a taxane, a nucleoside analog, an antimetabolite, a topisomerase inhibitor, an anthracycline, a vinca alkaloid, or any combination thereof.

[0054] In some aspects, the tyrosine kinase inhibitor comprises dabrafenib, vemurafenib, encorafenib, trametinib, cobimetinib, binimetinib, or any combination thereof.

[0055] In some aspects, anti-angiogenic agents include inhibitors of vascular endothelial growth factor (VEGF), VEGF receptor (VEGFR), platelet-derived growth factor (PDGF), PDGF receptor (PDGFR), angiopoietin (Ang), tyrosine kinase with Ig-like and EGF-like domains (Tie) receptor, hepatocyte growth factor (HGF), tyrosine-protein kinase Met (c-MET), C-type lectin family 14 member A (CLEC14A), multimerin 2 (MMRN2), shock protein 70-1A (HSP70-1A), epidermal growth factor (EGF), EGF receptor (EGFR), or combinations thereof.

[0056] In some aspects, anti-angiogenic agents include bevacizumab, ramucirumab, aflibercept, tanibirumab, olaratumab, nesbacumab, AMG780, MEDI3617, vanucizumab, rilotumab, ficlatuzumab, TAK-701, onartuzumab, emibetuzumab, or any combination thereof.

[0057] In some aspects, the checkpoint inhibitor is a programmed cell death-1 (PD-1) pathway inhibitor, a T-cell immunoglobulin and ITIM domain (TIGIT) inhibitor, a T-cell immunoglobulin and mucin-domain containing-3 (TIM-3) inhibitor, a TIM-1 inhibitor, a TIM-4 inhibitor, a B7-H3 inhibitor, a B7-H4 inhibitor, a B- and T-cell lymphocyte reducing agent (BTLA) inhibitor, a V-domain Ig inhibitor of T-cell activation inhibitor (VISTA), an indoleamine 2,3-dioxygenase (IDO) inhibitors, nicotinamide adenine dinucleotide phosphooxidase isoform 2 (NOX2) inhibitors, killer cell immunoglobulin-like receptor (KIR) inhibitors, adenosine A2a receptor (A2aR) inhibitors, transforming growth factor beta (TGF-β) inhibitors, phosphoinositide 3-kinase (PI3K) inhibitors, CD47 inhibitors, CD48 inhibitors, CD73 inhibitors, CD113 inhibitors, sialic acid-binding immunoglobulin-like lectin 7 (SIGLEC-7) inhibitors, SIGL EC-9 inhibitors, SIGLEC-15 inhibitors, glucocorticoid-induced TNFR-related protein (GITR) inhibitors, galectin-1 inhibitors, galectin-9 inhibitors, carcinoembryonic antigen-related cell adhesion molecule-1 (CEACAM-1) inhibitors, G protein-coupled receptor 56 (GPR56) inhibitors, glycoprotein A repeat dominant (GARP) inhibitors, 2B4 inhibitors, programmed cell death-1 homolog (PD1H) inhibitors, leukocyte-associated immunoglobulin-like receptor 1 (LAIR1) inhibitors, or any combination thereof.

[0058] In one aspect, the checkpoint inhibitor comprises a PD-1 pathway inhibitor.

[0059] In one aspect, the PD-1 pathway inhibitor is an anti-PD-1 antibody and / or an anti-PD-L1 antibody.

[0060] In one aspect, the PD-1 pathway inhibitor is an anti-PD-1 antibody.

[0061] In one aspect, the anti-PD-1 antibody is a full-length antibody.

[0062] In some aspects, the anti-PD-1 antibody is a monoclonal antibody, a human antibody, a humanized antibody, a chimeric antibody, or a multispecific antibody. In some aspects, the multispecific antibody is a DART, a DVD-Ig, or a bispecific antibody.

[0063] In one aspect, the anti-PD-1 antibody is a F(ab')2 fragment, a Fab' fragment, a Fab fragment, an Fv fragment, an scFv fragment, a dsFv fragment, a dAb fragment, or a single-chain binding polypeptide peptide.

[0064] In one aspect, the anti-PD-1 antibody is nivolumab, pembrolizumab, PDR001 (spartalizumab), MEDI-0680, TSR-042, cemiplimab, JS001, PF-06801591, BGB-A317, BI 754091, INCSHR1210, GLS-010, AM-001, STI-1110, AGEN2034, MGA012, BCD-100, IBI308, SSI-361, or an antigen-binding portion thereof.

[0065] In one aspect, the PD-1 pathway inhibitor is a soluble PD-L2 polypeptide. In one aspect, the soluble PD-L2 polypeptide is a fusion polypeptide. In one aspect, the soluble PD-L2 polypeptide comprises a ligand-binding fragment of the PD-L2 extracellular domain. In one aspect, the soluble PD-L2 polypeptide further comprises a half-life extending moiety. In one aspect, the half-life extending moiety comprises an immunoglobulin constant region or portion thereof, an immunoglobulin-binding polypeptide, immunoglobulin G (IgG), an albumin-binding polypeptide (ABP), a PAS site, a HES site, XTEN, a PEGylation site, an Fc region, or any combination thereof. In one aspect, the soluble PD-L2 polypeptide is AMP-224.

[0066] In one aspect, the PD-1 pathway inhibitor is an anti-PD-L1 antibody.

[0067] In one aspect, the anti-PD-L1 antibody is a full-length antibody.

[0068] In some aspects, the anti-PD-L1 antibody is a monoclonal antibody, a human antibody, a humanized antibody, a chimeric antibody, or a multispecific antibody. In some aspects, the multispecific antibody is a DART, DVD-Ig, or a bispecific antibody.

[0069] In one aspect, the anti-PD-L1 antibody is a F(ab')2 fragment, a Fab' fragment, a Fab fragment, an Fv fragment, an scFv fragment, a dsFv fragment, a dAb fragment, or a single-chain binding polypeptide peptide.

[0070] In one aspect, the anti-PD-L1 antibody is BMS-936559, atezolizumab, durvalumab, avelumab, STI-1014, CX-072, KN035, LY3300054, BGB-A333, ICO 36, CK-301, or an antigen-binding portion thereof.

[0071] In one aspect, the PD-1 pathway inhibitor is BMS-986189.

[0072] The present invention relates to a method of treating unresectable or metastatic melanoma in a human patient, comprising administering to the patient (a) an approximately 360 mg dose of an anti-LAG-3 antibody, and (b) an approximately 3 mg / kg dose of an anti-CTLA-4 antibody.

[0073] The present invention relates to a method of treating unresectable or metastatic melanoma in a human patient, comprising administering to the patient (a) an approximately 720 mg dose of an anti-LAG-3 antibody, and (b) an approximately 3 mg / kg dose of an anti-CTLA-4 antibody.

[0074] The present invention relates to a method of treating unresectable or metastatic melanoma in a human patient, comprising administering to the patient (a) an approximately 1080 mg dose of an anti-LAG-3 antibody, and (b) an approximately 3 mg / kg dose of an anti-CTLA-4 antibody.

[0075] The present invention relates to a method of treating unresectable or metastatic melanoma in a human patient, comprising administering to the patient (a) an approximately 1200 mg dose of an anti-LAG-3 antibody, and (b) an approximately 3 mg / kg dose of an anti-CTLA-4 antibody.

[0076] The present invention relates to a method of treating unresectable or metastatic melanoma in a human patient, comprising administering to the patient (a) an approximately 360 mg dose of an anti-LAG-3 antibody, and (b) an approximately 3 mg / kg dose of an anti-CTLA-4 antibody, wherein the patient has previously received a PD-1 pathway inhibitor as a treatment for the melanoma.

[0077] The present invention relates to a method of treating unresectable or metastatic melanoma in a human patient, comprising administering to the patient (a) an approximately 720 mg dose of an anti-LAG-3 antibody, and (b) an approximately 3 mg / kg dose of an anti-CTLA-4 antibody, wherein the patient has previously received a PD-1 pathway inhibitor as a treatment for the melanoma.

[0078] The present invention relates to a method of treating unresectable or metastatic melanoma in a human patient, comprising administering to the patient (a) an approximately 1080 mg dose of an anti-LAG-3 antibody, and (b) an approximately 3 mg / kg dose of an anti-CTLA-4 antibody, wherein the patient has previously received a PD-1 pathway inhibitor as a treatment for the melanoma.

[0079] The present invention relates to a method of treating unresectable or metastatic melanoma in a human patient, comprising administering to the patient (a) an approximately 1200 mg dose of an anti-LAG-3 antibody, and (b) an approximately 3 mg / kg dose of an anti-CTLA-4 antibody, wherein the patient has previously received a PD-1 pathway inhibitor as a treatment for the melanoma.

[0080] In one aspect, the anti-LAG-3 antibody and the anti-CTLA-4 antibody are administered about once every week, about once every 2 weeks, about once every 3 weeks, about once every 4 weeks, about once every 5 weeks, about once every 6 weeks, about once every 7 weeks, about once every 8 weeks, about once every 9 weeks, about once every 10 weeks, about once every 11 weeks, or about once every 12 weeks.

[0081] In one aspect, the patient has histologically confirmed unresectable Stage III or Stage IV melanoma.

[0082] In one aspect, the patient has an Eastern Cooperative Oncology Group (ECOG) performance status of 0 or 1.

[0083] In one aspect, one or more immune cells in tumor tissue from a patient express LAG-3. In one aspect, at least about 1%, at least about 3%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or about 100% of the immune cells express LAG-3. In one aspect, at least about 1% of the immune cells express LAG-3. In one aspect, the immune cells are tumor-infiltrating lymphocytes. In one aspect, the tumor-infiltrating lymphocytes are CD8 + In one aspect, greater than about 1% of the patient's tumor-infiltrating lymphocyte cells express LAG-3.

[0084] In some aspects, one or more tumor cells in a tumor tissue from a patient express PD-L1. In some aspects, at least about 1%, at least about 3%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or about 100% of the tumor cells express PD-L1. In some aspects, at least about 1% of the tumor cells express PD-L1. In some aspects, more than about 1% of the patient's tumor cells express PD-L1.

[0085] In one aspect, the patient's tumor cells comprise a BRAF V600 mutation.

[0086] In one aspect, the anti-LAG-3 antibody and / or the anti-CTLA-4 antibody is a full-length antibody.

[0087] In certain aspects, the anti-LAG-3 antibody and / or anti-CTLA-4 antibody is a monoclonal antibody, a human antibody, a humanized antibody, a chimeric antibody, or a multispecific antibody. In certain aspects, the multispecific antibody is a dual affinity retargeting antibody (DART), a DVD-Ig, or a bispecific antibody.

[0088] In one aspect, the anti-LAG-3 antibody and / or anti-CTLA-4 antibody is a F(ab')2 fragment, a Fab' fragment, a Fab fragment, an Fv fragment, an scFv fragment, a dsFv fragment, a dAb fragment, or a single-chain binding polypeptide peptide.

[0089] In one aspect, the anti-LAG-3 antibody is BMS-986016 (leratolimab), IMP731 (H5L7BW), MK4280 (28G-10), REGN3767 (fianlimab), GSK2831781, humanized BAP050, IMP-701 (LAG-525, yeramilimab), aLAG3(0414), aLAG3(0416), Sym022, TSR-033, TSR-075, XmAb22841, MGD013, BI754111, FS118, P 13B02-30, AVA-017, 25F7, AGEN1746, or an antigen-binding portion thereof.

[0090] In one aspect, the anti-LAG-3 antibody comprises the CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO:3, and the CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO:5.

[0091] In one aspect, the anti-LAG-3 antibody comprises: (a) a heavy chain variable region, CDR1, comprising the sequence set forth in SEQ ID NO:7; (b) a heavy chain variable region, CDR2, comprising the sequence set forth in SEQ ID NO:8; (c) a heavy chain variable region, CDR3, comprising the sequence set forth in SEQ ID NO:9; (d) a light chain variable region, CDR1, comprising the sequence set forth in SEQ ID NO:10; (e) a light chain variable region, CDR2, comprising the sequence set forth in SEQ ID NO:11; and (f) a light chain variable region, CDR3, comprising the sequence set forth in SEQ ID NO:12.

[0092] In one aspect, the anti-LAG-3 antibody comprises a heavy chain variable region and a light chain variable region comprising the sequences set forth in SEQ ID NO:3 and SEQ ID NO:5, respectively.

[0093] In one aspect, the anti-LAG-3 antibody comprises a heavy chain and a light chain comprising the sequences set forth in SEQ ID NO:1 and SEQ ID NO:2, respectively.

[0094] In one aspect, the anti-LAG-3 antibody comprises a heavy chain and a light chain comprising the sequences set forth in SEQ ID NO:30 and SEQ ID NO:2, respectively.

[0095] In one aspect, the anti-CTLA-4 antibody is ipilimumab, tremelimumab, MK-1308, AGEN-1884, or an antigen-binding portion thereof.

[0096] In one aspect, the anti-CTLA-4 antibody comprises the CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO:34, and the CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO:32.

[0097] In one aspect, the anti-CTLA-4 antibody comprises: (a) a heavy chain variable region, CDR1, comprising the sequence set forth in SEQ ID NO: 35; (b) a heavy chain variable region, CDR2, comprising the sequence set forth in SEQ ID NO: 36; (c) a heavy chain variable region, CDR3, comprising the sequence set forth in SEQ ID NO: 37; (d) a light chain variable region, CDR1, comprising the sequence set forth in SEQ ID NO: 38; (e) a light chain variable region, CDR2, comprising the sequence set forth in SEQ ID NO: 39; and (f) a light chain variable region, CDR3, comprising the sequence set forth in SEQ ID NO: 40.

[0098] In one aspect, the anti-CTLA-4 antibody comprises a heavy chain variable region and a light chain variable region comprising the sequences set forth in SEQ ID NOs: 34 and 32, respectively.

[0099] In one aspect, the anti-CTLA-4 antibody comprises the heavy and light chains of ipilimumab.

[0100] In some aspects, the method further comprises administering a PD-1 pathway inhibitor. In some aspects, the PD-1 pathway inhibitor is an anti-PD-1 antibody and / or an anti-PD-L1 antibody. In some aspects, the anti-PD-1 antibody is nivolumab, pembrolizumab, PDR001 (spartalizumab), MEDI-0680, TSR-042, REGN2810 (cemiplimab), JS001, PF-06801591, BGB-A317, BI 754091, INCSHR1210, GLS-010, AM-001, STI-1110, AGEN2034, MGA012, BCD-100, IBI308, SSI-361, or an antigen-binding portion thereof.

[0101] In one aspect, the anti-LAG-3 antibody and the anti-CTLA-4 antibody are formulated for intravenous administration.

[0102] In one aspect, the anti-LAG-3 antibody and the anti-CTLA-4 antibody are formulated separately.

[0103] In one aspect, the anti-LAG-3 antibody and the anti-CTLA-4 antibody are formulated as a single agent.

[0104] In one aspect, the anti-LAG-3 antibody and the anti-CTLA-4 antibody are co-administered.

[0105] In one aspect, the anti-LAG-3 antibody and the anti-CTLA-4 antibody are administered separately.

[0106] In one aspect, the anti-LAG-3 antibody is administered simultaneously with the anti-CTLA-4 antibody.

[0107] In one aspect, the anti-LAG-3 antibody is administered prior to administration of the anti-CTLA-4 antibody.

[0108] In one aspect, the anti-LAG-3 antibody is administered after administration of the anti-CTLA-4 antibody.

[0109] The present invention relates to a method of treating unresectable or metastatic melanoma in a human patient, comprising administering to the patient (a) an anti-LAG-3 antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 3, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO: 5, and (b) an anti-CTLA-4 antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 34, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO: 32, wherein the patient has previously received a PD-1 pathway inhibitor for treatment of the melanoma.

[0110] The present invention relates to a method for treating unresectable or metastatic melanoma in a human patient, comprising administering to the patient (a) an approximately 360 mg dose of an anti-LAG-3 antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 3, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO: 5, and (b) an approximately 3 mg / kg dose of an anti-CTLA-4 antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 34, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO: 32.

[0111] The present invention relates to a method for treating unresectable or metastatic melanoma in a human patient, comprising administering to the patient (a) an approximately 720 mg dose of an anti-LAG-3 antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 3, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO: 5, and (b) an approximately 3 mg / kg dose of an anti-CTLA-4 antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 34, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO: 32.

[0112] The present invention relates to a method for treating unresectable or metastatic melanoma in a human patient, comprising administering to the patient (a) an approximately 1080 mg dose of an anti-LAG-3 antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 3, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO: 5, and (b) an approximately 3 mg / kg dose of an anti-CTLA-4 antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 34, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO: 32.

[0113] The present invention relates to a method for treating unresectable or metastatic melanoma in a human patient, comprising administering to the patient (a) an approximately 1200 mg dose of an anti-LAG-3 antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 3, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO: 5, and (b) an approximately 3 mg / kg dose of an anti-CTLA-4 antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 34, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO: 32.

[0114] The present invention relates to a method of treating unresectable or metastatic melanoma in a human patient, comprising administering to the patient (a) an approximately 360 mg dose of an anti-LAG-3 antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 3, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO: 5, and (b) an approximately 3 mg / kg dose of an anti-CTLA-4 antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 34, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO: 32, wherein the patient has previously received a PD-1 pathway inhibitor for treatment of the melanoma.

[0115] The present invention relates to a method of treating unresectable or metastatic melanoma in a human patient, comprising administering to the patient (a) an approximately 720 mg dose of an anti-LAG-3 antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 3, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO: 5, and (b) an approximately 3 mg / kg dose of an anti-CTLA-4 antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 34, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO: 32, wherein the patient has previously received a PD-1 pathway inhibitor as treatment for the melanoma.

[0116] The present invention relates to a method of treating unresectable or metastatic melanoma in a human patient, comprising administering to the patient (a) an approximately 1080 mg dose of an anti-LAG-3 antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 3, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO: 5, and (b) an approximately 3 mg / kg dose of an anti-CTLA-4 antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 34, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO: 32, wherein the patient has previously received a PD-1 pathway inhibitor for treatment of the melanoma.

[0117] The present invention relates to a method of treating unresectable or metastatic melanoma in a human patient, comprising administering to the patient (a) an approximately 1200 mg dose of an anti-LAG-3 antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 3, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO: 5, and (b) an approximately 3 mg / kg dose of an anti-CTLA-4 antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 34, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO: 32, wherein the patient has previously received a PD-1 pathway inhibitor for treatment of the melanoma.

[0118] In one aspect, the anti-LAG-3 antibody and the anti-CTLA-4 antibody are administered about once every week, about once every 2 weeks, about once every 3 weeks, about once every 4 weeks, about once every 5 weeks, about once every 6 weeks, about once every 7 weeks, about once every 8 weeks, about once every 9 weeks, about once every 10 weeks, about once every 11 weeks, or about once every 12 weeks.

[0119] In one aspect, the anti-LAG-3 antibody and the anti-CTLA-4 antibody are administered about once every three weeks.

[0120] In one aspect, the patient has histologically confirmed unresectable Stage III or Stage IV melanoma.

[0121] In one aspect, the patient has an Eastern Cooperative Oncology Group (ECOG) performance status of 0 or 1.

[0122] In one aspect, one or more immune cells in tumor tissue from a patient express LAG-3. In one aspect, at least about 1%, at least about 3%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or about 100% of the immune cells express LAG-3. In one aspect, at least about 1% of the immune cells express LAG-3. In one aspect, the immune cells are tumor-infiltrating lymphocytes. In one aspect, the tumor-infiltrating lymphocytes are CD8 + In one aspect, greater than about 1% of the patient's tumor-infiltrating lymphocyte cells express LAG-3.

[0123] In some aspects, one or more tumor cells in a tumor tissue from a patient express PD-L1. In some aspects, at least about 1%, at least about 3%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or about 100% of the tumor cells express PD-L1. In some aspects, at least about 1% of the tumor cells express PD-L1. In some aspects, more than 1% of the patient's tumor cells express PD-L1.

[0124] In one aspect, the patient's tumor cells comprise a BRAF V600 mutation.

[0125] The present invention provides a method for treating unresectable or metastatic melanoma in a human patient, comprising: (a) an about 360 mg, about 720 mg, about 1080 mg, or about 1200 mg dose of an anti-LAG-3 antibody comprising: (i) a heavy chain variable region CDR1 comprising the sequence set forth in SEQ ID NO:7; (ii) a heavy chain variable region CDR2 comprising the sequence set forth in SEQ ID NO:8; (iii) a heavy chain variable region CDR3 comprising the sequence set forth in SEQ ID NO:9; (iv) a light chain variable region CDR1 comprising the sequence set forth in SEQ ID NO:10; (v) a light chain variable region CDR2 comprising the sequence set forth in SEQ ID NO:11; and (vi) a light chain variable region CDR3 comprising the sequence set forth in SEQ ID NO:12; and (b) an about 3 mg / kg dose of: (i) a heavy chain variable region CDR1 comprising the sequence set forth in SEQ ID NO:35; (ii) a heavy chain variable region CDR2 comprising the sequence set forth in SEQ ID NO:36; (iii) a heavy chain variable region CDR3 comprising the sequence set forth in SEQ ID NO:37; (iv) a light chain variable region CDR1 comprising the sequence set forth in SEQ ID NO:38; and (vi) a light chain variable region CDR3 comprising the sequence set forth in SEQ ID NO:40.

[0126] The present invention provides a method for treating unresectable or metastatic melanoma in a human patient, comprising: (a) an about 360 mg, about 720 mg, about 1080 mg, or about 1200 mg dose of an anti-LAG-3 antibody comprising: (i) a heavy chain variable region CDR1 comprising the sequence set forth in SEQ ID NO:7; (ii) a heavy chain variable region CDR2 comprising the sequence set forth in SEQ ID NO:8; (iii) a heavy chain variable region CDR3 comprising the sequence set forth in SEQ ID NO:9; (iv) a light chain variable region CDR1 comprising the sequence set forth in SEQ ID NO:10; (v) a light chain variable region CDR2 comprising the sequence set forth in SEQ ID NO:11; and (vi) a light chain variable region CDR3 comprising the sequence set forth in SEQ ID NO:12; and (b) an about 3 mg / kg dose of: (i) a heavy chain variable region CDR1 comprising the sequence set forth in SEQ ID NO:35; (ii) a heavy chain variable region CDR2 comprising the sequence set forth in SEQ ID NO:36; (iii) a heavy chain variable region CDR3 comprising the sequence set forth in SEQ ID NO:37; (iv) a light chain variable region CDR1 comprising the sequence set forth in SEQ ID NO:38; and (vi) a light chain variable region CDR3 comprising the sequence set forth in SEQ ID NO: 40; wherein the patient has previously received a PD-1 pathway inhibitor as a treatment for melanoma.

[0127] The present invention relates to a method for treating unresectable or metastatic melanoma in a human patient, comprising administering to the patient an effective amount of each of: (a) about a 360 mg, about a 720 mg, about a 1080 mg, or about a 1200 mg dose of an anti-LAG-3 antibody comprising a heavy chain variable region and a light chain variable region comprising the sequences set forth in SEQ ID NOs: 3 and 5, respectively; and (b) about a 3 mg / kg dose of an anti-CTLA-4 antibody comprising a heavy chain variable region and a light chain variable region comprising the sequences set forth in SEQ ID NOs: 34 and 32, respectively.

[0128] The present invention relates to a method of treating unresectable or metastatic melanoma in a human patient, comprising administering to the patient (a) about a 360 mg, about a 720 mg, about a 1080 mg, or about a 1200 mg dose of an anti-LAG-3 antibody comprising a heavy chain variable region and a light chain variable region comprising the sequences set forth in SEQ ID NOs: 3 and 5, respectively, and (b) about a 3 mg / kg dose of an anti-CTLA-4 antibody comprising a heavy chain variable region and a light chain variable region comprising the sequences set forth in SEQ ID NOs: 34 and 32, respectively, wherein the patient has previously received a PD-1 pathway inhibitor as treatment for the melanoma.

[0129] In one aspect, the anti-LAG-3 antibody and the anti-CTLA-4 antibody are administered about once every week, about once every 2 weeks, about once every 3 weeks, about once every 4 weeks, about once every 5 weeks, about once every 6 weeks, about once every 7 weeks, about once every 8 weeks, about once every 9 weeks, about once every 10 weeks, about once every 11 weeks, or about once every 12 weeks.

[0130] In one aspect, the anti-LAG-3 antibody and the anti-CTLA-4 antibody are administered about once every three weeks.

[0131] In one aspect, the anti-LAG-3 antibody is leratolimab.

[0132] In one aspect, the anti-CTLA-4 antibody is ipilimumab.

[0133] The present invention relates to a method of treating unresectable or metastatic melanoma in a human patient, comprising administering to the patient an anti-LAG-3 antibody comprising: (a) a heavy chain variable region CDR1 comprising the sequence set forth in SEQ ID NO:7; (b) a heavy chain variable region CDR2 comprising the sequence set forth in SEQ ID NO:8; (c) a heavy chain variable region CDR3 comprising the sequence set forth in SEQ ID NO:9; (d) a light chain variable region CDR1 comprising the sequence set forth in SEQ ID NO:10; (e) a light chain variable region CDR2 comprising the sequence set forth in SEQ ID NO:11; and (f) a light chain variable region CDR3 comprising the sequence set forth in SEQ ID NO:12, wherein the patient has previously received a PD-1 pathway inhibitor as a treatment for the melanoma; and wherein at least one dose of the anti-LAG-3 antibody is administered at a dose of about 360 mg, about 720 mg, about 1080 mg, or about 1200 mg.

[0134] In some aspects, the patient is further administered chemotherapy.

[0135] In one aspect, the patient's tumor cells express fibrinogen-like protein 1 (FGL1).

[0136] In one aspect, the presence of a BRAF V600E mutation in a tumor specimen is confirmed prior to initiation of treatment. In one aspect, the presence of a BRAF V600E mutation is confirmed using the cobas® 4800 BRAF V600 Mutation Test.

[0137] In one aspect, the patient is not further administered a PD-1 pathway inhibitor. DETAILED DESCRIPTION OF THE INVENTION

[0138] Detailed Description of the Invention The present invention provides methods of treating unresectable or metastatic melanoma in a human patient, comprising administering to the patient an LAG-3 antagonist (e.g., an anti-LAG-3 antibody). In some aspects, the patient has a mutation that sensitizes to targeted inhibitor therapy (e.g., a type B aggressive fibrosarcoma proto-oncogene (BRAF) mutation). In some aspects, the method is a first-line therapy, a second-line therapy, or a third-line therapy. In some aspects, the patient has previously been administered a programmed cell death-1 (PD-1) pathway inhibitor (e.g., an anti-PD-1 antibody) as a treatment for the melanoma. In some aspects, the patient has histologically confirmed unresectable Stage III or Stage IV melanoma. The present invention also relates to methods of treating unresectable or metastatic melanoma in a human patient, comprising a combination of an LAG-3 antagonist (e.g., an anti-LAG-3 antibody) and a CTLA-4 inhibitor (e.g., an anti-CTLA-4 antibody). In certain aspects, the method further comprises administering one or more additional therapeutic agents (e.g., a PD-1 pathway inhibitor such as an anti-PD-1 antibody) and / or anti-cancer therapy (e.g., chemotherapy) in combination with the LAG-3 antagonist, or in combination with the LAG-3 antagonist and the CTLA-4 inhibitor. In certain aspects, the method further comprises determining LAG-3 and / or PD-L1 expression in tumor tissue from the patient and / or administering to the patient a LAG-3 antagonist or a combination of a LAG-3 antagonist and a CTLA-4 inhibitor based on LAG-3 and / or PD-L1 expression in the patient's tumor tissue.

[0139] I. Terminology In order that the present description may be more readily understood, certain terms are first defined. As used herein, unless expressly defined differently herein, each of the following terms may have the meaning set forth below. Additional definitions are set forth throughout the specification.

[0140] It is noted that the term "a" or "an" entity means one or more of that entity. For example, "a nucleotide sequence" is understood to refer to one or more nucleotide sequences. Thus, the terms "a" (or "an"), "one or more," and "at least one" can be used interchangeably herein.

[0141] As used herein, the term "and / or" should be construed as meaning that each of the two specified features or components is specifically disclosed, regardless of the presence or absence of the other. Thus, as used herein, the term "and / or" in phrases such as "A and / or B" is intended to include "A and B," "A or B," "A" alone, and "B" alone. Similarly, the term "and / or" in phrases such as "A, B, and / or C" is intended to encompass each of the following embodiments: A, B, and C; A, B, or C; A or C; A or B; B, or C; A and C; A and B; B, and C; A alone; B alone; and C alone.

[0142] Where an aspect or aspects are described herein with the term "comprising," it is understood that other similar aspects described with the terms "consisting of" and / or "consisting essentially of" are also provided.

[0143] The terms "about" or "essentially comprising" refer to a value or composition that is within an acceptable error range of a particular value or composition as determined by one of ordinary skill in the art, which may depend in part on how the value or composition is measured or determined, i.e., the limitations of the measurement system. For example, "about" or "essentially comprising" can mean within one or more standard deviations per practice in the art. Alternatively, "about" or "essentially comprising" can mean a range of up to 10% or 20% (i.e., ±10% or ±20%). For example, about 3 mg can include any number between 2.7 mg and 3.3 mg (for 10%) or 2.4 mg and 3.6 mg (for 20%). Furthermore, particularly with respect to biological systems or processes, these terms can mean up to an order of magnitude higher or up to five times the value. When a particular value or composition is provided in the specification and claims, unless otherwise specified, the meaning of "about" or "essentially comprising" should be assumed to be within an acceptable error range of that particular value or composition.

[0144] As described herein, any concentration range, percentage range, ratio range, or integer range should be understood to include any integer value within the stated range, and fractions thereof, where appropriate (e.g., tenths and hundredths of integers, etc.), unless otherwise specified.

[0145] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this disclosure pertains.For example, the Concise Dictionary of Biomedicine and Molecular Biology, Juo, Pei-Show, 2nd ed., 2002, CRC Press; The Dictionary of Cell and Molecular Biology, 5th ed., 2013, Academic Press; and the Oxford Dictionary of Biochemistry and Molecular Biology, 2006, Oxford University Press provide those skilled in the art with a general dictionary of many terms used in this disclosure.

[0146] Units, prefixes, and symbols are expressed in the form accepted by the International System of Units (SI) as their base units. Numeric ranges include the numbers that define the range.

[0147] The headings provided herein are not intended to be limiting of the various aspects described herein, but rather may be incorporated by reference to the entire specification, and accordingly, the terms defined immediately below are more fully defined by reference to the specification as a whole.

[0148] "Antagonist" includes, but is not limited to, any molecule that can block, reduce, or otherwise limit the interaction or activity of a target molecule (e.g., LAG-3). In some aspects, the antagonist is an antibody. In other aspects, the antagonist comprises a small molecule. The terms "inhibitor" and "antagonist" are used interchangeably herein.

[0149] "Antibody" (Ab) includes, but is not limited to, a glycoprotein immunoglobulin that specifically binds to an antigen and comprises at least two heavy (H) chains and two light (L) chains inter-connected by disulfide bonds. Each H chain contains a heavy chain variable region (herein referred to as VH ) and a heavy chain constant region (abbreviated as C H The heavy chain constant region contains C H1 , C H2 and C H3 Each light chain comprises three constant regions: a light chain variable region (herein referred to as V L ) and a light chain constant region (abbreviated as C L The light chain constant region contains one constant domain, C L Includes V H and V L The regions can be further subdivided into regions of hypervariability, called complementarity-determining regions (CDRs), interspersed with more conserved regions, called framework regions (FRs). H and V L contains three CDRs and four FRs, arranged from the amino terminus to the carboxy terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable regions of the heavy and light chains contain binding domains that interact with antigens. The constant region of the antibody may mediate the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (C1q) of the classical complement system. The heavy chain may or may not have a C-terminal lysine. Unless otherwise specified herein, amino acids in the variable regions are numbered using the Kabat numbering system, and amino acids in the constant regions are numbered using the EU system.

[0150] Immunoglobulins can be derived from any of the commonly known isotypes, including, but not limited to, IgA, secretory IgA, IgG, and IgM. IgG subclasses are also well known to those skilled in the art and include, but are not limited to, human IgG1, IgG2, IgG3, and IgG4. "Isotype" refers to the antibody class or subclass (e.g., IgM or IgG1) encoded by heavy chain constant region genes. The term "antibody" includes, by way of example, both natural and non-natural antibodies; monoclonal and polyclonal antibodies; chimeric and humanized antibodies; human or non-human antibodies; fully synthetic antibodies; single-chain antibodies; monospecific antibodies; bispecific antibodies; and multispecific antibodies. Non-human antibodies can be humanized by recombinant methods to reduce their immunogenicity in humans. Unless expressly stated otherwise, and unless the context dictates otherwise, the term "antibody" also includes antigen-binding fragments or portions of any of the above immunoglobulins, including monovalent and divalent fragments or portions, that retain the ability to specifically bind to the antigen to which the whole immunoglobulin binds. Examples of "antigen-binding portions" or "antigen-binding fragments" include: (1) V L , V H , L C and C H1 (2) a Fab fragment (a fragment obtained by papain cleavage) or similar monovalent fragment, consisting of two Fab fragments linked by a disulfide bridge in the hinge region; (3) a Fd fragment, consisting of a VH domain and a CH1 domain; (4) a single-arm V L and V H(5) single-domain antibody (dAb) fragments consisting of a VH domain (Ward et al., (1989) Nature 341:544-46); (6) bi-single-domain antibodies (dual-affinity re-targeting antibodies (DART)) consisting of two VH domains linked by a hinge; or (7) dual-variable domain immunoglobulins. Furthermore, the two domains of the Fv fragment, VH, L and V H Although the V and V are encoded by separate genes, they can be joined by a synthetic linker using recombinant methods, L Area and V H The domains can be produced as single protein chains that pair to form monovalent molecules (known as single-chain Fvs (scFvs); see, e.g., Bird et al. (1988) Science 242:423-426; and Huston et al. (1988) Proc. Natl. Acad. Sci. USA 85:5879-5883).

[0151] By "isolated antibody" is meant an antibody that is substantially free of other antibodies having different antigenic specificities (e.g., an isolated antibody that specifically binds to LAG-3 is substantially free of antibodies that do not specifically bind to LAG-3). However, an isolated antibody that specifically binds to LAG-3 may have cross-reactivity to other antigens, such as LAG-3 molecules from different species. Furthermore, an isolated antibody may be substantially free of other cellular material and / or chemicals.

[0152] The term "monoclonal antibody" ("mAb") refers to antibody molecules of single molecular composition, i.e., a non-naturally occurring preparation of antibody molecules which are essentially identical in primary sequence and which display a single binding specificity and affinity for a particular epitope. A mAb is an example of an isolated antibody. MAbs may be produced by hybridoma, recombinant, transgenic, or other techniques known to those skilled in the art.

[0153] A "human" antibody (HuMAb) refers to an antibody having variable regions in which both the framework and CDR regions are derived from human germline immunoglobulin sequences. Furthermore, if the antibody contains a constant region, the constant region also is derived from human germline immunoglobulin sequences. The human antibodies of the invention may include amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutation in vivo). However, as used herein, the term "human antibody" is not intended to include antibodies in which CDR sequences derived from the germline of other mammalian species, such as a mouse, have been grafted onto human framework sequences. The terms "human" antibody and "fully human" antibody are used interchangeably.

[0154] "Humanized antibody" refers to an antibody in which some, most, or all of the amino acids outside the CDR domains of a non-human antibody have been substituted with corresponding amino acids from a human immunoglobulin. In one aspect of a humanized form of an antibody, some, most, or all of the amino acids outside the CDR domains have been substituted with amino acids from a human immunoglobulin, while some, most, or all of the amino acids within one or more CDR regions remain unchanged. Minor additions, deletions, insertions, substitutions, or modifications of amino acids are permissible as long as they do not impair the ability of the antibody to bind to a specific antigen. A "humanized" antibody retains the same antigen specificity as the original antibody.

[0155] By "chimeric antibody" is meant an antibody in which the variable region is derived from one species and the constant region is derived from another species, such as an antibody in which the variable region is derived from a mouse antibody and the constant region is derived from a human antibody.

[0156] An "anti-antigen" antibody refers to an antibody that specifically binds to an antigen. For example, an anti-LAG-3 antibody specifically binds to LAG-3.

[0157] "LAG-3" refers to lymphocyte activation gene-3. The term "LAG-3" includes variants, isoforms, homologs, orthologs, and paralogs. For example, an antibody specific for human LAG-3 protein may, in certain cases, cross-react with LAG-3 protein from species other than human. In other aspects, an antibody specific for human LAG-3 protein is completely specific for human LAG-3 protein and does not exhibit species or other types of cross-reactivity, or may cross-react with LAG-3 from certain other species, but not with all other species (e.g., cross-reacts with monkey LAG-3 but not with mouse LAG-3). The term "human LAG-3" refers to the human sequence LAG-3, e.g., the complete amino acid sequence of human LAG-3 in GenBank Accession No. NP_002277. The term "mouse LAG-3" refers to the mouse sequence LAG-3, e.g., the complete amino acid sequence of mouse LAG-3 in GenBank Accession No. NP_032505. LAG-3 is also known in the art, e.g., as CD223. A human LAG-3 sequence may differ from human LAG-3 in GenBank Accession No. NP_002277, e.g., by having conserved mutations or mutations in non-conserved regions, and the LAG-3 has substantially the same biological function as human LAG-3 in GenBank Accession No. NP_002277. For example, the biological function of human LAG-3 is to have an epitope within the extracellular domain of LAG-3 that is specifically bound by an antibody of the invention or to bind to an MHC class II molecule.

[0158] A particular human LAG-3 sequence will generally be at least about 90% identical in amino acid sequence to the human LAG-3 of GenBank Accession No. NP_002277 and will contain amino acid residues that identify the amino acid sequence as human when compared to LAG-3 amino acid sequences of other species (e.g., mouse). In some cases, the human LAG-3 may be at least about 95%, or even at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical in amino acid sequence to the LAG-3 of GenBank Accession No. NP_002277. In certain aspects, the human LAG-3 sequence will not exhibit more than 10 amino acid sequence differences from the LAG-3 sequence of GenBank Accession No. NP_002277. In certain aspects, the human LAG-3 exhibits no more than 5 amino acid differences, or no more than 4 amino acid differences, no more than 3 amino acid differences, no more than 2 amino acid differences, or no more than 1 amino acid difference from the LAG-3 sequence of GenBank Accession No. NP_002277.

[0159] "Programmed Death-1 (PD-1)" refers to an immune inhibitory receptor belonging to the CD28 family. PD-1 is expressed in vivo primarily on preactivated T cells and binds to two ligands, PD-L1 and PD-L2. As used herein, the term "PD-1" includes human PD-1 (hPD-1), variants, isoforms, and species homologs of hPD-1, as well as analogs that share at least one epitope with hPD-1. The complete hPD-1 sequence can be found in GenBank Accession No. U64863. "PD-1" and "PD-1 receptor" are used interchangeably herein.

[0160] "Cytotoxic T-lymphocyte antigen-4 (CTLA-4)" refers to an immune inhibitory receptor belonging to the CD28 family. CTLA-4 is expressed exclusively on T cells in vivo and binds to two ligands, CD80 and CD86 (also known as B7-1 and B7-2, respectively). As used herein, the term "CTLA-4" includes human CTLA-4 (hCTLA-4), its variants, isoforms, and species homologs, as well as analogs that share at least one epitope with hCTLA-4. The complete hCTLA-4 sequence can be found in GenBank Accession No. AAB59385.

[0161] "Programmed death ligand-1 (PD-L1)" is one of two cell surface glycoprotein ligands for PD-1 (the other is PD-L2) that downregulates T cell activation and cytokine secretion upon binding to PD-1. As used herein, the term "PD-L1" includes human PD-L1 (hPD-L1), variants, isoforms, and species homologs of hPD-L1, and analogs that share at least one epitope with hPD-L1. The complete hPD-L1 sequence can be found in GenBank Accession No. Q9NZQ7.

[0162] As used herein, the term "programmed death-ligand-2 (PD-L2)" includes human PD-L2 (hPD-L2), variants, isoforms, and species homologs of hPD-L2, and analogs that share at least one epitope with hPD-L2. The complete hPD-L2 sequence can be found in GenBank Accession No. Q9BQ51.

[0163] As used herein, a "patient" includes a patient suffering from unresectable or metastatic melanoma. The terms "subject" and "patient" are used interchangeably.

[0164] "Administering" refers to physically introducing a therapeutic agent (e.g., a composition or formulation comprising a therapeutic agent) into a subject using any of a variety of methods and delivery systems known to those of skill in the art. Examples of routes of administration include intravenous, intramuscular, subcutaneous, intraperitoneal, spinal, or other parenteral routes of administration, for example, by injection or infusion. As used herein, the term "parenteral administration" refers to methods of administration other than enteral and topical administration, usually by injection, and includes, but is not limited to, intravenous, intramuscular, intraarterial, intrathecal, intrathoracic, intralesional, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, intrathecal, intraspinal, epidural, and intrasternal injection and infusion, and in vivo electroporation. In some aspects, the Therapeutic Agent is administered by a non-parenteral route, which in some aspects is orally administered. Other non-parenteral routes include topical, epithelial, or mucosal routes of administration, such as intranasal, intravaginal, intrarectal, sublingual, or topical administration. Administration can also be, for example, single, multiple, and / or one or more times over an extended period of time.

[0165] "Treatment" or "treatment" of a subject means any therapeutic intervention or method performed on a subject, or administration of an active ingredient to a subject, for the purpose of reversing, alleviating, ameliorating, inhibiting, or delaying the progression, onset, severity, or recurrence of symptoms, complications, or conditions, or biochemical signs associated with a disease. Response Evaluation Criteria in Solid Tumors (RECIST) are a measure of treatment effectiveness and established criteria that define when a tumor responds, stabilizes, or progresses during treatment. RECIST 1.1 is the current guideline for solid tumor measurement and definition to objectively assess changes in tumor size for use in clinical trials of adult and pediatric cancers.

[0166] As used herein, "effective treatment" refers to treatment that produces a beneficial effect, e.g., an improvement in at least one symptom of a disease or disorder. A beneficial effect can take the form of an improvement relative to baseline, i.e., an improvement relative to a measurement or observation made before initiating treatment according to a method. A beneficial effect can also take the form of halting, slowing, delaying, or stabilizing the adverse progression of markers of a solid tumor. Effective treatment can refer to the alleviation of at least one symptom of a solid tumor. Such effective treatment can, for example, relieve a patient's pain, reduce the size and / or number of lesions, reduce or prevent tumor metastasis, and / or slow tumor growth.

[0167] The term "effective amount" refers to an amount of an agent that provides a desired biological, therapeutic, and / or prophylactic result. That result can be a reduction, amelioration, alleviation, regression, delay, and / or remission of one or more signs, symptoms, or causes of a disease, or any other desired alteration of a biological system. With respect to solid tumors, an effective amount includes an amount sufficient to shrink a tumor and / or reduce the rate of tumor growth (e.g., inhibit tumor growth), or prevent or delay other undesirable cell proliferation. In one aspect, an effective amount is an amount sufficient to prevent or delay tumor development. An effective amount can be administered in one or more administrations. An effective amount of a drug or composition is an amount that can (i) reduce the number of cancer cells; (ii) reduce tumor size; (iii) inhibit, delay, to some extent, slow, or stop cancer cell invasion of peripheral organs; (iv) inhibit, i.e., to some extent, slow or stop tumor metastasis; (v) inhibit tumor growth; (vi) prevent or delay tumor onset and / or recurrence; and / or (vii) alleviate to some extent one or more symptoms associated with cancer. In one example, an "effective amount" is an amount of anti-LAG-3 antibody, an amount of anti-LAG-3 antibody alone and an amount of an additional therapeutic agent (e.g., anti-CTLA-4), or a combination thereof, that has been clinically proven to affect a significant reduction in cancer or delay in the progression of cancer, such as solid tumors.

[0168] As used herein, the terms "fixed dose," "constant dose," and "constant fixed dose" are used interchangeably and refer to a dose administered to a patient regardless of the patient's weight or body surface area (BSA). Thus, a fixed dose or constant dose is provided as an absolute amount of drug (e.g., μg or mg amount) rather than as a mg / kg dose.

[0169] The use of the term "fixed dose combination," with respect to compositions of the invention, means that two or more different inhibitors described herein (e.g., an anti-LAG-3 antibody and an anti-CTLA-4 antibody) in a single composition are present in the composition in a specific (fixed) ratio to each other. In certain aspects, the fixed dose is based on the weight (e.g., mg) of the inhibitors. In certain aspects, the fixed dose is based on the concentration (e.g., mg / ml) of the inhibitors. In certain aspects, the ratio is at least about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, about 1:30, about 1:40, about 1:50, about 1:60, about 1:70, about 1:80, about 1:90, about 1:100, about 1:120, about 1:140, about 1: 1:160, about 1:180, about 1:200, about 200:1, about 180:1, about 160:1, about 140:1, about 120:1, about 100:1, about 90:1, about 80:1, about 70:1, about 60:1, about 50:1, about 40:1, about 30:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1 or about 2:1.

[0170] As used herein, the term "weight-based dose" means that the dose administered to a patient is calculated based on the patient's weight.

[0171] As used herein, "dosing interval" refers to the time lapse between administration of multiple doses of a formulation described herein to a subject. Thus, a dosing interval can be expressed as a time range.

[0172] As used herein, the term "dosing frequency" refers to the frequency with which multiple doses of the formulations described herein are administered over a given period of time. Dosing frequency can be expressed as the number of doses per given period of time, such as once per week or once every two weeks.

[0173] As used herein, the terms "about once per week," "about once per week," "about once every two weeks," or other similar administration interval terms refer to approximate times, and "about once per week" or "about once per week" can include every 7 days ± 2 days, i.e., every 5 to 9 days. Thus, an administration frequency of "once per week" can be every 5 days, every 6 days, every 7 days, every 8 days, or every 9 days. "About once per 3 weeks" can include once every 21 days ± 3 days, i.e., once every 25 to 31 days. Similar approximations apply, for example, to about once per 2 weeks, about once per 4 weeks, about once per 5 weeks, about once per 6 weeks, about once per 7 weeks, about once per 8 weeks, about once per 9 weeks, about once per 10 weeks, about once per 11 weeks, and about once per 12 weeks. In one aspect, a dosing interval of about once every 6 weeks or about once every 12 weeks means that the first dose can be administered on any day in week 1, followed by any day in week 6 or week 12, respectively. In other aspects, a dosing interval of about once every 6 weeks or about once every 12 weeks means that the first dose is administered on a particular day (e.g., a Monday) in week 1, followed by the next dose on the same day (e.g., a Monday) in week 6 or week 12, respectively.

[0174] As used herein, the term "tumor" refers to any mass of tissue resulting from excessive cell growth or proliferation, either benign (non-cancerous) or malignant (cancerous), including pre-cancerous lesions.

[0175] As used herein, the term "biological sample" refers to a biological material isolated from a subject. Biological samples can include any biological material, for example, by sequencing nucleic acids in a tumor (or circulating tumor cells) and identifying genomic alterations in the sequenced nucleic acids. The biological sample can be any suitable biological tissue or body fluid, such as, for example, tumor tissue, blood, plasma, and serum. The biological sample can be a test tissue sample (e.g., a tissue sample containing tumor cells and tumor-infiltrating inflammatory cells). In one aspect, the sample is a tumor tissue biopsy, such as formalin-fixed, paraffin-embedded (FFPE) tumor tissue or fresh-frozen tumor tissue. In another aspect, the biological sample is a liquid biopsy containing one or more of blood, serum, plasma, circulating tumor cells, exoRNA, ctDNA, and cfDNA, in some embodiments.

[0176] As an example, an "anti-cancer agent" promotes cancer regression in a subject. In a preferred aspect, a therapeutically effective amount of the agent promotes cancer regression to the point of eliminating the cancer. "Promoting cancer regression" means that administration of an effective amount of an anti-cancer agent, alone or in combination with another agent, results in a reduction in tumor growth or size, tumor necrosis, a decrease in the severity of at least one disease symptom, an increase in the frequency and duration of disease-free periods, or prevention of functional impairment or disability due to disease affliction. Additionally, the terms "effective" and "efficacy" with respect to treatment include both pharmacological effectiveness and physiological safety. Pharmacological effectiveness refers to the ability of a drug to promote cancer regression in a patient. Physiological safety refers to the level of toxicity or other adverse physiological effects (side effects) at the cellular, organ, and / or organismal level resulting from the administration of a drug.

[0177] As an example of tumor treatment, a therapeutically effective amount of an anti-cancer agent can inhibit cell proliferation or tumor growth by at least about 20%, at least about 40%, at least about 60%, or at least about 80% compared to untreated subjects. In other aspects of the invention, tumor regression can be observed, which lasts for at least about 20 days, more preferably at least about 40 days, or at least about 60 days. Apart from these ultimate measures of therapeutic efficacy, evaluation of immunotherapeutic agents must also consider immune-related response patterns.

[0178] As used herein, "immuno-oncology" therapy or "IO" or "IO" therapy refers to therapy that involves harnessing the immune response to target and treat tumors in a subject. Thus, as used herein, IO therapy is a type of anti-cancer therapy. In some aspects, IO therapy involves administering to the subject an antibody. In some aspects, IO therapy involves administering to the subject immune cells, e.g., T cells, e.g., modified T cells, e.g., T cells modified to express a chimeric antigen receptor or a specific T cell receptor. In some aspects, IO therapy involves administering to the subject a therapeutic vaccine. In some aspects, IO therapy involves administering to the subject a cytokine or chemokine. In some aspects, IO therapy involves administering to the subject an interleukin. In some aspects, IO therapy involves administering to the subject an interferon. In some aspects, IO therapy involves administering to the subject a colony-stimulating factor.

[0179] An "immune response" is one mediated by the action of one or more cells of the immune system (e.g., T lymphocytes, B lymphocytes, natural killer (NK) cells, macrophages, eosinophils, mast cells, dendritic cells, or neutrophils) and soluble macromolecules (including antibodies, cytokines, and complement) produced by any of these cells or by the liver, resulting in the selective targeting, binding, damaging, destroying, and / or eliminating from the vertebrate body, invading pathogens, pathogen-infected cells or tissues, cancerous or other abnormal cells, or, in the case of autoimmune or pathological inflammation, normal human cells or tissues.

[0180] The terms "LAG-3 positive" or "LAG-3 expression positive" with respect to LAG-3 expression refer to immune cells (e.g., CD8 + This refers to tumor tissue (e.g., a test tissue sample) that is scored as expressing LAG-3 based on the proportion (i.e., percentage) of tumor-infiltrating lymphocytes (e.g., T cells) in the tumor tissue (e.g., tumor-infiltrating lymphocytes, e.g., T cells) (e.g., 1% or more expression).

[0181] "LAG-3 negative" or "LAG-3 expression negative" means tumor tissue (e.g., a test tissue sample) that is not scored as expressing LAG-3 (e.g., LAG-3 expression less than 1%).

[0182] The terms "PD-L1 positive" or "positive PD-L1 expression," with respect to cell surface PD-L1 expression, refer to tumor tissue (e.g., a test tissue sample) that is scored as expressing PD-L1 based on the proportion (i.e., percentage) of tumor cells that express PD-L1 (e.g., 1% or greater expression).

[0183] The terms "PD-L1 negative" or "PD-L1 expression negative" refer to tumor tissue (e.g., a test tissue sample) that is not scored as expressing PD-L1 (e.g., less than 1% expression).

[0184] As used herein, "Eastern Cooperative Oncology Group (ECOG) performance status (PS)" is a numbering scale used to define the patient population studied in a trial so that it is uniformly reproducible among the physicians enrolling the patients.

[0185] Various aspects of the invention are described in further detail in the following subsections.

[0186] II. The Methods of the Invention The present invention provides methods of treating unresectable or metastatic melanoma in a human patient, comprising administering to the patient a LAG-3 antagonist (e.g., an anti-LAG-3 antibody), alone or in combination with a CTLA-4 inhibitor (e.g., an anti-CTLA-4 antibody). In some aspects, the method further comprises administering one or more additional therapeutic agents (e.g., a PD-1 inhibitor, such as an anti-PD-1 antibody) and / or therapies (e.g., chemotherapy). .

[0187] In one aspect, the method comprises administering a LAG-3 antagonist and a CTLA-4 inhibitor, wherein the patient has a susceptibility mutation to targeted inhibitor therapy (e.g., a BRAF mutation).

[0188] In one aspect, the method comprises administering a LAG-3 antagonist and a CTLA-4 inhibitor, wherein the patient has previously received a PD-1 pathway inhibitor for the treatment of melanoma.

[0189] In some aspects, this method is a first-line (1L) therapy.

[0190] In some aspects, this method is a second line (2L) therapy.

[0191] In some aspects, this method is a third line (3L) therapy.

[0192] In some aspects, the patient has progressed on prior therapy (e.g., standard of care therapy). Standard of care therapy for different types of cancer is well known to those skilled in the art. For example, the National Comprehensive Cancer Network (NCCN), a coalition of 21 major cancer centers in the United States, publishes the NCCN Clinical Practice Guidelines in Oncology (NCCN GUIDELINES®), which provide detailed, up-to-date information on standard therapies for many types of cancer. See https: / / www.nccn.org / professionals / physician_gls / default.aspx, last accessed November 4, 2020.

[0193] In some aspects, the patient has not received prior systemic therapy for cancer, the patient has not received prior systemic therapy for melanoma, or the patient has not received prior systemic therapy for unresectable or metastatic melanoma.

[0194] In some aspects, the patient is naive to prior immuno-oncology (IO) therapy. In some aspects, the patient has not received IO therapy, has received IO therapy for a cancer other than melanoma, or has received prior IO therapy for melanoma but not for the current melanoma. In some aspects, the patient is naive to prior IO therapy, the patient is naive to prior IO therapy for melanoma, or the melanoma is naive to prior IO therapy. In some aspects, the prior IO therapy is an antibody. In some aspects, the antibody binds to a checkpoint inhibitor. In some aspects, the prior IO therapy is an anti-PD-1 antibody.

[0195] In some aspects, the methods described herein prolong progression-free survival (PFS), objective response rate (ORR), overall survival (OS), or any combination thereof, as compared to standard of care and / or prior therapies described herein.

[0196] In certain aspects, the methods of the present invention extend progression-free survival by more than about 12 months, about 13 months, about 14 months, about 15 months, about 16 months, about 17 months, about 18 months, about 2 years, about 3 years, about 4 years, about 5 years, about 6 years, about 7 years, about 8 years, about 9 years, or about 10 years relative to standard of care therapy.

[0197] In certain aspects, the methods of the invention reduce tumor size, inhibit tumor growth, eliminate tumor from the patient, prevent recurrence of melanoma, cause remission of melanoma, provide a complete or partial response, or any combination thereof.

[0198] In some aspects, the methods of the invention reduce tumor size by at least about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, or about 100% compared to the tumor size before administration.

[0199] In one aspect, the methods of the invention comprise administering a LAG-3 antagonist to a patient based on the stage and / or performance status of the patient's cancer, which may be indicated by any one or more systems known in the art.

[0200] In some aspects, melanoma is staged according to a tumor / node / metastasis (TNM) staging system, such as the American Joint Committee on Cancer (AJCC) classification.

[0201] In one aspect, the patient has Stage I melanoma, also known as in situ melanoma. In Stage I, the cancer is limited to the epidermis. There has been no metastasis to nearby lymph nodes or distant parts of the body. In one aspect, the patient has histologically confirmed unresectable Stage I melanoma.

[0202] In one aspect, the patient has stage II melanoma. In stage II, the tumor is at least 1 mm thick and may be thicker than 4 mm. It may or may not be ulcerated. The cancer has not spread to nearby lymph nodes or distant parts of the body. In one aspect, the patient has histologically confirmed unresectable stage II melanoma.

[0203] In one aspect, the patient has stage III melanoma. In one aspect, the patient has histologically confirmed unresectable stage III melanoma. Stage III is divided into stage IIIA, stage IIIB, stage IIIC, and stage IIID.

[0204] In Stage IIIA, the tumor is 2 mm or less thick and may or may not be ulcerated. The cancer has spread to one to three nearby lymph nodes, but they are very small and visible under a microscope. It has not spread to distant parts of the body. In some cases, a patient has histologically confirmed unresectable Stage IIIA melanoma.

[0205] In Stage IIIB, (1) there is no sign of a primary tumor, and (a) cancer has spread to only one nearby lymph node, or (b) cancer has spread to very small areas of nearby skin (satellite tumors) or to the cutaneous lymphatic vessels around the tumor (without reaching nearby lymph nodes), or (2) the tumor is 4 mm or less thick and may or may not be ulcerated, and (a) cancer has spread to only one nearby lymph node, (b) cancer has spread to very small areas of nearby skin (satellite tumors) or to the cutaneous lymphatic vessels around the tumor (without reaching nearby lymph nodes), or (c) cancer has spread to two to three nearby lymph nodes. In Stage IIIB, cancer has not spread to distant parts of the body. In one aspect, a patient has histologically confirmed unresectable Stage IIIB melanoma.

[0206] In Stage IIIC, (1) there is no sign of the primary tumor and (a) cancer has spread to two or more nearby lymph nodes, at least one of which can be seen or felt, (b) cancer has spread to a very small area of ​​nearby skin (satellite tumor) or to the cutaneous lymphatic channels around the tumor and has reached nearby lymph nodes, or (c) cancer has spread to nearby lymph nodes as a mass; (2) the tumor is 4 mm thick or less and may or may not be ulcerated, and (a) cancer has spread to a very small area of ​​nearby skin (satellite tumor) or to the cutaneous lymphatic channels around the tumor and has reached nearby lymph nodes, (b) cancer has spread to four or more nearby lymph nodes or or (3) the tumor is 2 to 4 mm thick and ulcerated, or thicker than 4 mm but not ulcerated, and (a) the cancer has spread to one or more nearby lymph nodes, and / or (b) the cancer has spread to very small areas of nearby skin (satellite tumors) or to the cutaneous lymphatic ducts around the tumor, or (4) the tumor is more than 4 mm thick and ulcerated, and (a) the cancer has spread to 1 to 3 nearby lymph nodes, but these are not tightly packed, or (b) the cancer has spread to very small areas of nearby skin (satellite tumors) or to the cutaneous lymphatic ducts around the tumor, which may or may not have reached one nearby lymph node. In Stage IIIC, the cancer has not spread to distant parts of the body. In one aspect, a patient has histologically confirmed unresectable Stage IIIC melanoma.

[0207] In Stage IIID, the tumor is thicker than 4 mm and ulcerated, and (a) cancer has spread to four or more nearby lymph nodes, (b) cancer has spread to nearby lymph nodes as a mass, or (c) cancer has spread to very small areas of nearby skin (satellite tumors) or to the lymphatic vessels in the skin around the tumor and cancer has spread to at least two nearby lymph nodes or to lymph nodes as a mass. In Stage IIID, cancer has not spread to distant parts of the body.

[0208] In Stage IV, the tumor may be of any thickness, may or may not be ulcerated, and may or may not have spread to nearby lymph nodes. In Stage IV, the cancer has spread to distant lymph nodes or to organs such as the lungs, liver, or brain. In one aspect, the patient has histologically confirmed Stage IV melanoma.

[0209] In one aspect, the patient has histologically confirmed unresectable Stage III or Stage IV melanoma.

[0210] In some respects, performance status is indicated by the Eastern Cooperative Oncology Group Performance Status (ECOG PS), which utilizes standardized criteria for determining how the disease affects a patient's ability to perform daily activities. Example ECOG PS definitions include: "0" for a patient who is fully active and able to perform all pre-disease performance without limitation; "1" for a patient who is limited in physically strenuous activity but is ambulatory and able to perform light or sedentary tasks; "2" for a patient who is ambulatory and able to perform all self-care for more than 50% of the waking hours but is unable to perform any work activities; "3" for a patient who is only able to perform limited self-care and is bed- or chair-bound for more than 50% of the waking hours; and "4" for a patient who is completely disabled, unable to perform any self-care, and completely bed- or chair-bound.

[0211] In some aspects, the patient has an ECOG PS of 0, 1, 2, 3, or 4. In some aspects, the patient has an ECOG PS of 3 or less. In some aspects, the patient has an ECOG PS of 2 or less. In some aspects, the patient has an ECOG PS of 1 or greater. In some aspects, the patient has an ECOG PS of 0 or 1.

[0212] In one aspect, the patient has type B aggressive fibrosarcoma proto-oncogene (BRAF, e.g., a BRAF V600 mutation such as BRAF V600E or BRAF V600K), mitogen-activated extracellular signal-regulated kinase kinase (MEK), neuroblastoma RAS viral oncogene homolog (NRAS), and / or proto-oncogene c-KIT (KIT) mutations that are susceptible to targeted inhibitor therapy.

[0213] In some aspects, the patient has a BRAF mutation. In some aspects, the BRAF mutation is a BRAF V600 mutation. In some aspects, the BRAF mutation is a BRAF V600E mutation. In some aspects, the BRAF mutation is a BRAF V600K mutation.

[0214] In one aspect, the patient does not have a BRAF, MEK, NRAS and / or KIT mutation that is susceptible to targeted inhibitor therapy.

[0215] In some aspects, the targeted inhibitor therapy comprises a BRAF and / or MEK tyrosine kinase inhibitor, hi some aspects, the targeted inhibitor therapy comprises dabrafenib, vemurafenib, encorafenib, trametinib, cobimetinib, and / or binimetinib.

[0216] In one aspect, the invention includes a method of selecting unresectable or metastatic melanoma in a human patient for immunotherapy, comprising determining the level of LAG-3 and / or PD-L1 expression in a tumor sample.

[0217] In one aspect, the invention includes a method of treating unresectable or metastatic melanoma in a human patient, comprising: (a) determining the level of LAG-3 expression, the level of PD-L1 expression, and / or the levels of LAG-3 and PD-L1 expression in a tumor sample; and (b) administering to the patient a therapeutically effective amount of a LAG-3 antagonist, alone or in combination with a therapeutically effective amount of a CTLA-4 inhibitor. In some aspects, the method further comprises administering one or more additional therapeutic agents and / or therapies (e.g., chemotherapy).

[0218] In certain aspects, one or more immune cells in the tumor tissue from the patient express LAG-3 (i.e., the tumor tissue from the patient is LAG-3 positive), and / or one or more tumor cells in the tumor tissue from the patient express PD-L1 (i.e., the tumor tissue from the patient is PD-L1 positive). In certain aspects, one or more immune cells in the tumor tissue from the patient express LAG-3. In certain aspects, at least about 1%, at least about 2%, at least about 3%, at least about 4%, at least about 5%, at least about 7%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or about 100% of the immune cells express LAG-3. In one aspect, at least about 1% of immune cells express LAG-3. In one aspect, more than about 1% of immune cells express LAG-3. In one aspect, the immune cells are tumor-infiltrating lymphocytes. In one aspect, the tumor-infiltrating lymphocytes are CD8 +In one aspect, one or more tumor cells in a tumor tissue from a patient express PD-L1. In one aspect, at least about 1%, at least about 2%, at least about 3%, at least about 4%, at least about 5%, at least about 7%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or about 100% of the tumor cells express PD-L1. In one aspect, at least about 1% of the tumor cells express PD-L1. In one aspect, more than about 1% of the tumor cells express PD-L1. In one aspect, any of the values ​​for "at least about X%" is "≧X%".

[0219] In one aspect, one or more immune cells in the tumor tissue from the patient do not express LAG-3 (i.e., the tumor tissue from the patient is LAG-3 negative). In one aspect, the tumor tissue is LAG-3 negative when less than about 1% of the immune cells express LAG-3.

[0220] In one aspect, one or more tumor cells in the tumor tissue from the patient do not express PD-L1 (i.e., the tumor tissue from the patient is PD-L1 negative). In one aspect, tumor tissue is PD-L1 negative when less than about 1% of the tumor cells express PD-L1.

[0221] In some aspects, the methods of the invention comprise identifying a patient as having immune cells (e.g., tumor-infiltrating lymphocytes) and / or tumor cells that express or contain particular markers. For example, in some embodiments, the melanoma is LAG-3 positive. In some embodiments, the melanoma is PD-L1 positive. In some embodiments, the melanoma is LAG-3 positive and PD-L1 positive. In some embodiments, the melanoma contains a BRAF V600 mutation. In some embodiments, the melanoma is LAG-3 positive and expresses a BRAF V600 mutation. In some embodiments, the melanoma is LAG-3 positive and contains tumor cells that express wild-type BRAF. In some embodiments, the melanoma is LAG-3 positive, PD-L1 positive, and contains wild-type BRAF. In some embodiments, the melanoma is LAG-3 positive, PD-L1 positive, and contains a BRAF V600 mutation. In some embodiments, the melanoma is PD-L1 positive and contains wild-type BRAF. In some embodiments, the melanoma is PD-L1 positive and contains a BRAF V600 mutation.

[0222] The present invention may also include methods of preventing relapse and / or inducing remission in a patient, comprising administering to the patient an immunotherapy described herein.

[0223] In other aspects, each patient in the method experiences (i) an increase in progression-free survival of 12 months or more, (ii) a reduction in tumor size of at least about 10%, about 20%, about 30%, about 40%, or about 50% or more compared to the tumor size prior to administration, or (iii) both.

[0224] The methods of the present invention can treat unresectable or metastatic melanoma, reduce tumor size, inhibit tumor growth, eliminate tumor from the patient, prevent tumor recurrence, induce remission in the patient, or any combination thereof, as a result of administration of the immunotherapies described herein. In certain aspects, administration of the immunotherapies described herein induces a complete response. In other aspects, administration of the immunotherapies described herein induces a partial response.

[0225] In one aspect, LAG-3 and / or PD-L1 expression is determined by receiving the results of an assay that can determine LAG-3 expression and / or PD-L1 expression.

[0226] Methods for determining PD-L1 expression in tumor samples, methods for identifying patients as having PD-L1-positive malignancies, and methods for determining PD-L1 expression in malignancies are described in PCT / US2016 / 029878, the disclosure of which is incorporated herein by reference.

[0227] In one aspect, a test tissue sample is obtained from a patient to assess LAG-3 and / or PD-L1 expression and / or whether the cancer contains a BRAF V600 mutation. In one aspect, the test tissue sample includes any clinically relevant tissue sample, such as, but not limited to, a tumor biopsy, a core biopsy tissue sample, an incisional biopsy, an excision biopsy, a surgical specimen, a fine needle aspirate, or a bodily fluid sample, such as blood, plasma, serum, lymph, ascites, cyst fluid, urine, etc. In one aspect, the test tissue sample is from a primary tumor. In one aspect, the test tissue sample is from a metastasis. In one aspect, test tissue samples are obtained from a patient at multiple time points, e.g., before, during, and / or after treatment. In one aspect, the test tissue samples are obtained from different locations on the patient, e.g., a sample from the primary tumor and a sample from a distant metastasis.

[0228] In certain aspects, the test tissue sample is a paraffin-embedded, fixed tissue sample. In certain embodiments, the test tissue sample is a formalin-fixed, paraffin-embedded (FFPE) tissue sample. In certain aspects, the test tissue sample is a fresh tissue (e.g., tumor) sample. In certain aspects, the test tissue sample is a frozen tissue sample. In certain aspects, the test tissue sample is a fresh frozen (FF) tissue (e.g., tumor) sample. In certain aspects, the test tissue sample is cells isolated from a bodily fluid. In certain aspects, the test tissue sample comprises circulating tumor cells (CTCs). In certain aspects, the test tissue sample comprises tumor-infiltrating lymphocytes (TILs). In certain aspects, the test tissue sample comprises tumor cells and tumor-infiltrating lymphocytes (TILs). In certain aspects, the test tissue sample comprises circulating lymphocytes. In certain aspects, the test tissue sample is an archival tissue sample. In certain aspects, the test tissue sample is an archival tissue sample with a known history of diagnosis, treatment, and / or outcome. In some aspects, the sample is a tissue mass. In some aspects, the biopsy tissue sample is a collection of dispersed cells. In some aspects, the sample size ranges from about 1 cell to about 1 x 10 6 In some aspects, the sample size ranges from about 1 cell to about 1 x 10 cells. 5 In some aspects, the sample size is from about 1 cell to about 10,000 cells. In some aspects, the sample size is from about 1 cell to about 1,000 cells. In some aspects, the sample size is from about 1 cell to about 100 cells. In some aspects, the sample size is from about 1 cell to about 10 cells. In some aspects, the sample size is a single cell. In some aspects, assessment of LAG-3, PD-L1 and / or BRAF V600 status is based on circulating tumor DNA.

[0229] In another aspect, assessing LAG-3, PD-L1 and / or BRAF V600 status can be accomplished without obtaining a test tissue sample. In one aspect, selecting a suitable patient includes (i) optionally providing a test tissue sample obtained from a patient with cancer of a tissue, wherein the test tissue sample includes tumor cells and / or tumor-infiltrating inflammatory cells; and (ii) assessing the proportion of cells in the test tissue sample that express LAG-3, PD-L1 and / or BRAF V600 based on assessing that the proportion of cells in the test tissue sample that express LAG-3, PD-L1 and / or BRAF V600 is greater than a predetermined threshold level.

[0230] However, it should be understood that in methods involving measuring LAG-3, PD-L1 and / or BRAF V600 status in a test tissue sample, the step involving providing a test tissue sample obtained from the patient is an optional step. That is, in certain aspects, the method includes this step, while in other embodiments, this step is not included in the method. It should also be understood that in certain aspects, the "measuring" or "assessing" step to identify or determine the number or percentage of cells in the test tissue sample that express LAG-3 and / or PD-L1 is performed by a transformation method that assays for LAG-3 and / or PD-L1, e.g., by performing a reverse transcriptase polymerase chain reaction (RT-PCR) assay or an IHC assay. In certain other aspects, no transformation step is included, and LAG-3 and / or PD-L1 expression is assessed, e.g., by reviewing test result reports from a laboratory. In certain aspects, LAG-3 and / or PD-L1 expression is assessed by reviewing immunohistochemistry assay results from a laboratory. In certain aspects, the steps of the method up to and including assessing LAG-3 and / or PD-L1 expression provide intermediate results that can be provided to a physician or healthcare provider for use in selecting suitable candidates for the methods of the invention. In certain aspects, the steps of providing the intermediate results are performed by a physician or other person working under the guidance of a physician. In other aspects, these steps are performed by an independent laboratory or an independent person, such as a laboratory technician. In one aspect, the presence of a BRAF V600 mutation is determined using a parallel approach of LAG-3 and / or PD-L1.

[0231] In certain aspects of the methods of the invention, LAG-3 expression and PD-L1 expression, and / or the proportion of cells containing a BRAF V600 mutation, are assessed by performing an assay to detect the presence of LAG-3, PD-L1 and / or BRAF RNA. In further aspects, the presence of LAG-3, PD-L1 and / or BRAF RNA is detected by RT-PCR, in situ hybridization, or RNase protection. In some aspects, the presence of LAG-3, PD-L1 and / or BRAF RNA is detected by an RT-PCR-based assay. In some aspects, scoring the RT-PCR-based assay comprises assessing the level of LAG-3, PD-L1 and / or BRAF RNA expression in the test tissue sample relative to a predetermined level. In some aspects, the expression of one or more of LAG-3, PD-L1, and BRAF V600 is assessed using gene expression profiling.

[0232] In other aspects, LAG-3 expression and / or PD-L1 expression, and / or the proportion of cells containing a BRAF V600 mutation is assessed by performing an assay to detect the presence of LAG-3 and PD-L1 and / or the inclusion of a BRAF V600 mutant polypeptide. In further aspects, the presence of LAG-3, PD-L1 and / or BRAF V600 polypeptide is determined by IHC, enzyme-linked immunosorbent assay (ELISA), in vivo imaging, or flow cytometry. In some aspects, LAG-3 and / or PD-L1 expression and / or BRAF V600 status is assayed by IHC. In all other aspects of these methods, cell surface expression of LAG-3 and / or PD-L1 and / or the presence of a BRAF V600 mutation is assayed using, for example, IHC or in vivo imaging.

[0233] In other aspects, the percentage of cells in the test tissue sample that express LAG-3 and / or PD-L1 and / or contain a BRAF V600 mutation is assessed by flow cytometry. In one aspect, the test tissue sample assayed by flow cytometry contains tumor-infiltrating immune cells. In one aspect, the flow cytometry is a multiplex assay. In one aspect, flow cytometry scoring comprises detecting expression of markers including LAG-3, CD4, CD8, FOXP3, and any combination thereof. In one aspect, flow cytometry scoring comprises assessing the percentage of T cells in the test tissue sample that express LAG-3 and / or PD-L1 and / or contain a BRAF V600 mutation. In one aspect, flow cytometry scoring comprises assessing the percentage of CD8 T cells in the test tissue sample that express LAG-3 and / or PD-L1 and / or contain a BRAF V600 mutation. + In one aspect, flow cytometry scoring includes assessing the proportion of CD4 T cells in the test tissue sample that express LAG-3 and / or PD-L1, and / or contain a BRAF V600 mutation. + In one aspect, flow cytometry scoring includes assessing the proportion of T cells in the test tissue sample that express LAG-3 and / or PD-L1, and / or contain a BRAF V600 mutation, FOXP3 + This includes assessing the proportion of T cells.

[0234] In certain aspects of any of the present invention, the proportion of cells in a test tissue sample that express LAG-3, express PD-L1, and / or contain a BRAF V600 mutation is assessed by performing an assay to detect the presence of LAG-3, PD-L1, and / or BRAF V600 mutant polypeptides. In some aspects, the presence of LAG-3, PD-L1, and / or BRAF V600 polypeptides is detected by immunohistochemistry. In some aspects, the test tissue sample is a tumor biopsy. In some aspects, the test tissue sample is a formalin-fixed, paraffin-embedded (FFPE) sample.

[0235] In some aspects, the immunohistochemistry assay is a monoplex assay. In some aspects, the immunohistochemistry assay is a multiplex assay. In some aspects, the multiplex immunohistochemistry assay can detect the presence of CD4, CD8, FOXP3, or any combination thereof.

[0236] In one aspect, the immunohistochemistry assay comprises contacting the tumor sample with 17B4 mouse anti-human LAG-3 IgG1 monoclonal antibody. In one aspect, the immunohistochemistry assay comprises contacting the tumor sample with an anti-LAG-3 antibody comprising heavy and light chain variable regions comprising the sequences set forth in SEQ ID NOs: 3 and 5, respectively. In one aspect, the immunohistochemistry assay comprises contacting the tumor sample with SP346 rabbit anti-human LAG-3 IgG monoclonal antibody. In one aspect, the immunohistochemistry assay comprises contacting the tumor sample with 11E3 (Novusbio), 874501 (Novusbio), or EPR4392(2) (Abcam) anti-human LAG-3 monoclonal antibody. In one aspect, the immunohistochemistry assay comprises contacting the tumor sample with reagents in a Dako PD-L1 IHC 28-8 kit to assay PD-L1 expression.

[0237] In one aspect, the immunohistochemistry assay is scored at low magnification. In one aspect, low magnification is about 20x. In one aspect, the immunohistochemistry assay is scored at high magnification. In one aspect, high magnification is about 40x.

[0238] In some aspects, the immunohistochemistry assay is scored by image analysis software. In some aspects, the immunohistochemistry assay is scored by a pathologist's visual immunoscore. In some aspects, the immunohistochemistry assay is scored manually.

[0239] In one aspect, scoring the immunohistochemistry assay in the test tissue sample comprises assessing the percentage of cells that express LAG-3 and / or PD-L1 and / or contain a BRAF V600 mutation. In one aspect, scoring the immunohistochemistry assay comprises assessing the percentage of immune cells in the test tissue sample that express LAG-3 and / or PD-L1 and / or contain a BRAF V600 mutation. In one aspect, scoring the immunohistochemistry assay comprises assessing the percentage of T cells in the test tissue sample that express LAG-3 and / or PD-L1 and / or contain a BRAF V600 mutation. In one aspect, scoring the immunohistochemistry assay comprises assessing the percentage of CD8 T cells in the test tissue sample that express LAG-3. + In one aspect, the scoring of the immunohistochemistry assay involves assessing the proportion of CD4 T cells in the test tissue sample that express LAG-3. + In one aspect, the scoring of the immunohistochemistry assay involves assessing the proportion of T cells in the live test tissue sample that express LAG-3. + This includes assessing the proportion of T cells.

[0240] In one aspect, the immunohistochemistry assay is a multiplex assay that further comprises detecting expression of MHC class II by tumor cells. In one aspect, scoring the immunohistochemistry assay comprises assessing the proportion of cells in the test tissue sample that express MHC class II. In one aspect, scoring the immunohistochemistry assay comprises assessing the proportion of non-immune cells in the test tissue sample that express MHC class II.

[0241] In certain aspects, the expression of fibrinogen-like protein 1 (FGL1) by tumor cells is measured.

[0242] Imaging technologies provide important tools in cancer research and treatment. Recent developments in molecular imaging systems, including positron emission tomography (PET), single-photon emission computed tomography (SPECT), fluorescence reflectance imaging (FRI), fluorescence-mediated tomography (FMT), bioluminescence imaging (BLI), laser scanning confocal microscopy (LSCM), and multiphoton microscopy (MPM), may foresee further applications of these technologies in cancer research. Some of these molecular imaging systems allow clinicians not only to see where tumors are located in the body, but also to visualize the expression and activity of specific molecules, cells, and biological processes that affect tumor behavior and / or responsiveness to therapeutic agents (Condeelis and Weissleder, Cold Spring Harb. Perspect. Biol. 2(12):a003848 (2010)). Combining the sensitivity and resolution of PET with antibody specificity, immunoPET imaging allows for the monitoring and assaying of antigen expression, particularly in tissue samples (McCabe and Wu, Cancer Biother. Radiopharm. 25(3):253-61 (2010); Olafsen et al., Protein Eng. Des. Sel. 23(4):243-9 (2010)). In certain aspects of the methods of the invention, LAG-3, PD-L1 and / or BRAF V600 expression is assayed by immunoPET imaging. In certain aspects of the methods of the invention, the proportion of cells in a test tissue sample that express LAG-3 is assessed by performing an assay to determine the presence of LAG-3, PD-L1 and / or BRAF V600 polypeptides on the surface of cells in the test tissue sample. In certain aspects, the test tissue sample is an FFPE tissue sample. In other aspects, the presence of LAG-3, PD-L1 and / or BRAF V600 polypeptides is determined by an IHC assay. In further aspects, the IHC assay is performed using automated methods.

[0243] In some aspects, the presence of a BRAF V600E mutation in a tumor specimen is confirmed prior to initiation of treatment. In particular aspects, the BRAF V600E mutation is confirmed by an FDA-approved test. In particular aspects, the test to confirm the presence of a BRAF V600E mutation is the cobas® 4800 BRAF V600 Mutation test.

[0244] II.A. Assaying LAG-3 and / or PD-L1 Expression and / or Presence of BRAF V600 Mutations by Automated IHC In one aspect of the methods of the invention, automated IHC methods are used to assay for the expression of LAG-3 and / or PD-L1 and / or the presence of BRAF V600 mutations in FFPE tissue specimens. The invention provides methods for detecting the presence of human LAG-3, PD-L1 and / or BRAF V600 in a test tissue sample, or quantifying the level of human LAG-3, PD-L1 and / or BRAF V600 antigen or the proportion of cells expressing the antigen in the sample, the methods comprising contacting a test sample and a negative control sample with a monoclonal antibody (mAb) that specifically binds to human LAG-3, PD-L1 and / or BRAF V600 under conditions that allow the formation of a complex between the antibody, or a portion thereof, and human LAG-3, PD-L1 and / or BRAF V600. In certain aspects, the test and control tissue samples are FFPE samples. The formation of complexes is then detected, with a difference in complex formation between the test sample and the negative control sample indicating the presence of human LAG-3, PD-L1 and / or BRAF V600 antigen in the sample. Various methods are used to quantitate LAG-3, PD-L1 and / or BRAF V600.

[0245] In certain aspects, the automated IHC method includes the steps of (a) deparaffinizing and rehydrating tissue sections mounted in an automated stainer; (b) retrieving antigens in the automated stainer; (c) loading reagents into the automated stainer; and (d) running the automated stainer and neutralizing endogenous peroxidase in the tissue specimen; blocking nonspecific protein binding sites on the slide; incubating the slide with a primary Ab; incubating with a post-primary blocking agent; incubating with a post-primary antibody detection agent, such as another antibody that may or may not be conjugated to a detection enzyme; incubating with a polymeric enzyme detection reagent; adding a chromogenic substrate to develop color; and counterstaining with hematoxylin. In some aspects, retrieving antigens includes using any heat-based antigen retrieval device.

[0246] In one aspect, to assess the presence of LAG-3, PD-L1, and / or BRAF V600 in tumor tissue samples, a pathologist examines the number of LAG-3+ tumor-infiltrating lymphocytes, PD-L1+ tumor cells, and / or BRAF V600+ tumor cells in each field under a microscope, mentally estimates the percentage of cells that are positive, and then averages the estimates for each field to obtain a final percentage value. Different staining intensities are defined as 0 / negative, 1+ / weak, 2+ / intermediate, and 3+ / strong. Generally, percentage values ​​are first assigned to the 0 and 3+ buckets, and then the intermediate 1+ and 2+ intensities are considered. For highly heterogeneous tissues, the sample is divided into zones, each zone is scored separately, and then combined into a single set of percentage values. The percentages of negative and positive cells for different staining intensities are determined from each region, and the median value is assigned to each zone. Final percentage values ​​are assigned to the tissue for each staining intensity category: negative, 1+, 2+, and 3+. The sum of all staining intensities should equal 100%.

[0247] In some aspects, staining is also assessed in tumor-infiltrating inflammatory cells, such as macrophages and lymphocytes. Macrophages and lymphocytes are assessed for LAG-3, PD-L1, and / or BRAF V600 staining, with each cell category scored as positive or negative for all samples. Staining is also characterized according to the designation of immune cells inside or outside the tumor. "Inside" refers to immune cells present on the border of the tumor area without being physically intercalated within the tumor tissue and / or between tumor cells. "Outside" refers to the absence of physical association with the tumor, with immune cells found in the periphery or adjacent tissues associated with connective tissue.

[0248] In certain aspects of these scoring methods, samples are scored by two or more pathologists working independently, and the scores are then combined. In certain other aspects, the identification of positive and negative cells is scored using appropriate software.

[0249] The histology score (H-score) is used as a more quantitative measure of the IHC data. The histology score is calculated as follows: Histology score = [(%tumor x1 (low intensity)) + (%tumor x2 (intermediate intensity)) + (%tumor x3 (high intensity)].

[0250] To determine the tissue score, the pathologist estimates the percentage of cells staining in each intensity category within the sample. Because expression of many biomarkers is heterogeneous, the tissue score is a more accurate representation of overall expression. The final tissue score ranges from 0 (minimum score, no expression) to 300 (maximum score, maximum expression and inclusive).

[0251] II.B.LAG-3 Antagonists LAG-3 antagonists for use in the methods of the present invention include, but are not limited to, LAG-3 binding agents and soluble LAG-3 polypeptides. LAG-3 binding agents include antibodies that specifically bind to LAG-3 (i.e., "anti-LAG-3 antibodies"). As used herein, the term "LAG-3 antagonist" is used interchangeably with the term "LAG-3 inhibitor."

[0252] In one aspect, the LAG-3 antagonist is an anti-LAG-3 antibody.

[0253] Antibodies that bind to LAG-3 are described, for example, in International Publication No. WO / 2015 / 042246 and U.S. Patent Publication Nos. 2014 / 0093511 and 2011 / 0150892, each of which is incorporated herein by reference in its entirety.

[0254] An exemplary LAG-3 antibody useful in the invention is 25F7 (described in U.S. Publication No. 2011 / 0150892). A further exemplary LAG-3 antibody useful in the invention is BMS-986016 (relatolimab). In certain aspects, anti-LAG-3 antibodies useful in the invention cross-compete with 25F7 or BMS-986016. In certain aspects, anti-LAG-3 antibodies useful in the invention bind to the same epitope as 25F7 or BMS-986016. In certain aspects, anti-LAG-3 antibodies comprise the six CDRs of 25F7 or BMS-986016.

[0255] Other art-recognized anti-LAG-3 antibodies that can be used in the methods of the present invention include IMP731 (H5L7BW) described in US2011 / 007023, MK-4280 (28G-10) described in WO2016028672 and U.S. Application Publication No. 2020 / 0055938, Burova E, et al., J. Immunother, Cancer (2016); 4(Supp. 1):P195 and REGN3767 (fianlimab), described in U.S. Pat. No. 10,358,495; humanized BAP050, described in WO2017 / 019894; GSK2831781; IMP-701 (LAG525; yeramilimab), described in U.S. Pat. No. 10,711,060 and U.S. Application Publication No. 2020 / 0172617; aLAG3(0414); aLAG3(0416); Sym022; TSR-033; TSR-075; XmAb22841; MGD013; BI754111; FS118; P13B02-30; AVA-017; and AGEN1746. These and other anti-LAG-3 antibodies useful in the present invention are described in, e.g., US 10,188,730, WO2016 / 028672, WO2017 / 106129, WO2017 / 062888, WO2009 / 044273, WO2018 / 069500, WO2016 / 126858, WO201 4 / 179664, WO2016 / 200782, WO2015 / 200119, WO2017 / 019846, WO2017 / 198741, W O2017 / 220555, WO2017 / 220569, WO2018 / 071500, WO2017 / 015560, WO2017 / 0254 98, WO2017 / 087589, WO2017 / 087901, WO2018 / 083087, WO2017 / 149143, WO2017 / 219995, US2017 / 0260271, WO2017 / 086367, WO2017 / 086419, WO2018 / 034227, W WO2018 / 185046, WO2018 / 185043, WO2018 / 217940, WO19 / 011306, WO2018 / 208868, WO2014 / 140180, WO2018 / 201096, WO2018 / 204374 and WO2019 / 018730.The entire contents of each of these documents are incorporated herein by reference.

[0256] Anti-LAG-3 antibodies that can be used in the methods of the invention also include isolated antibodies that specifically bind to human LAG-3 and cross-compete with any of the anti-LAG-3 antibodies described herein, e.g., leratolimab, for binding to human LAG-3. In one aspect, the anti-LAG-3 antibody binds to the same epitope as any of the anti-LAG-3 antibodies described herein, e.g., leratolimab.

[0257] In one aspect, an antibody that cross-competes with any of the anti-LAG-3 antibodies described herein, e.g., leratolimab, for binding to human LAG-3 or binds to the same epitope as any of the anti-LAG-3 antibodies described herein, e.g., leratolimab, is a monoclonal antibody. For administration to a human subject, these cross-competing antibodies can be chimeric, engineered, humanized, or human. Such chimeric, engineered, humanized, or human monoclonal antibodies can be prepared and isolated by methods well known in the art.

[0258] The ability of antibodies to cross-compete for binding to an antigen indicates that these antibodies bind to the same epitope region of the antigen and sterically hinder the binding of other cross-competing antibodies to a specific epitope region. Because of their binding to the same epitope region, these cross-competing antibodies are expected to have significantly similar functional properties to a control antibody, such as leratolimab. Cross-competing antibodies can be easily identified based on their ability to cross-compete in standard binding assays, such as Biacore analysis, ELISA assays, or flow cytometry (see, for example, WO2013 / 173223).

[0259] Anti-LAG-3 antibodies that can be used in the methods of the invention also include antigen-binding portions of any of the above full-length antibodies. It has been shown that the antigen-binding function of an antibody can be performed by fragments of a full-length antibody.

[0260] In one aspect, the anti-LAG-3 antibody is a full-length antibody.

[0261] In some aspects, the anti-LAG-3 antibody is a monoclonal antibody, a human antibody, a humanized antibody, a chimeric antibody, or a multispecific antibody. In some aspects, the multispecific antibody is a dual affinity retargeting antibody (DART), a DVD-Ig, or a bispecific antibody.

[0262] In one aspect, the anti-LAG-3 antibody is a F(ab')2 fragment, a Fab' fragment, a Fab fragment, an Fv fragment, an scFv fragment, a dsFv fragment, a dAb fragment, or a single-chain binding polypeptide peptide.

[0263] In one aspect, the anti-LAG-3 antibody is BMS-986016 (leratolimab), IMP731 (H5L7BW), MK4280 (28G-10), REGN3767 (fianlimab), GSK2831781, humanized BAP050, IMP-701 (LAG525, yeramilimab), aLAG3(0414), aLAG3(0416), Sym022, TSR-033, TSR-075, XmAb22841, MGD013, BI754111, FS118, P13B02-30, AVA-017, 25F7, AGEN1746, or an antigen-binding portion thereof.

[0264] In one aspect, the anti-LAG-3 antibody is leratolimab.

[0265] In one aspect, the anti-LAG-3 antibody comprises the heavy and light chain CDRs, heavy and light chain variable regions, or heavy and light chain sequences of an anti-LAG-3 antibody described herein, such as the sequences provided in the documents described herein, or known in the art.

[0266] In certain aspects, the methods described herein include anti-LAG-3 antibodies having at least about 90% sequence identity to an anti-LAG-3 antibody described herein or known in the art (e.g., having at least about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identity to the sequence of an anti-LAG-3 antibody, such as the heavy chain variable region and / or light chain variable region of the anti-LAG-3 antibody, or the heavy chain and / or light chain of the anti-LAG-3 antibody).

[0267] In one aspect, the anti-LAG-3 antibody comprises the CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO:3, and the CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO:5.

[0268] In one aspect, the anti-LAG-3 antibody comprises: (a) a heavy chain variable region, CDR1, comprising the sequence set forth in SEQ ID NO:7; (b) a heavy chain variable region, CDR2, comprising the sequence set forth in SEQ ID NO:8; (c) a heavy chain variable region, CDR3, comprising the sequence set forth in SEQ ID NO:9; (d) a light chain variable region, CDR1, comprising the sequence set forth in SEQ ID NO:10; (e) a light chain variable region, CDR2, comprising the sequence set forth in SEQ ID NO:11; and (f) a light chain variable region, CDR3, comprising the sequence set forth in SEQ ID NO:12.

[0269] In one aspect, the anti-LAG-3 antibody comprises a heavy chain variable region and a light chain variable region comprising the sequences set forth in SEQ ID NOs: 3 and 5, respectively.

[0270] In one aspect, the anti-LAG-3 antibody comprises a heavy chain and a light chain comprising the sequences set forth in SEQ ID NOs: 1 and 2, respectively.

[0271] In one aspect, the anti-LAG-3 antibody comprises a heavy chain and a light chain comprising the sequences set forth in SEQ ID NOs: 30 and 2, respectively.

[0272] In one aspect, the anti-LAG-3 antibody is REGN3767 (fianlimab). In one aspect, fianlimab is administered intravenously at a dose of about 1 mg / kg, about 3 mg / kg, about 10 mg / kg, or about 20 mg / kg about once every three weeks.

[0273] In one aspect, the anti-LAG-3 antibody is LAG525 (Ielamirimab). In one aspect, Ielamirimab is administered intravenously at a dose of about 300 mg, about 400 mg, about 500 mg, about 600 mg, about 700 mg, about 800 mg, about 900 mg, about 1000 mg, about 1100 mg, about 1200 mg, or about 1300 mg about once every 2, 3, or 4 weeks.

[0274] In one aspect, the anti-LAG-3 antibody is MK4280. In one aspect, MK4280 is administered intravenously at a dose of about 7 mg, 21 mg, 70 mg, 210 mg, or 700 mg about once every three weeks.

[0275] In one aspect, the LAG-3 antagonist is a soluble LAG-3 polypeptide. In one aspect, the soluble LAG-3 polypeptide is a fusion polypeptide, e.g., a fusion protein comprising the extracellular portion of LAG-3. In one aspect, the soluble LAG-3 polypeptide is a LAG-3-Fc fusion polypeptide capable of binding to MHC class II. In one aspect, the soluble LAG-3 polypeptide comprises a ligand-binding fragment of the LAG-3 extracellular domain. In one aspect, the ligand-binding fragment of the LAG-3 extracellular domain comprises an amino acid sequence having at least about 90%, at least about 95%, at least about 98%, at least about 99%, or about 100% sequence identity to SEQ ID NO:41. In one aspect, the soluble LAG-3 polypeptide further comprises a half-life extending moiety. In one aspect, the half-life extending moiety comprises an immunoglobulin constant region or portion thereof, an immunoglobulin-binding polypeptide, immunoglobulin G (IgG), an albumin-binding polypeptide (ABP), a PAS-modified moiety, a HES-modified moiety, XTEN, a PEGylated moiety, an Fc region, or any combination thereof. In one aspect, the soluble LAG-3 polypeptide is IMP321 (eftilagimod alfa). See, e.g., Brignone C, et al., J. Immunol. (2007); 179:4202-4211 and WO2009 / 044273.

[0276] In one aspect, an anti-LAG-3 antibody is used to determine LAG-3 expression. In one aspect, an anti-LAG-3 antibody is selected for its ability to bind to LAG-3 in formalin-fixed, paraffin-embedded (FFPE) tissue biopsies. In one aspect, an anti-LAG-3 antibody can bind to LAG-3 in frozen tissue. In one aspect, an anti-LAG-3 antibody can distinguish between membrane-bound, cytoplasmic, and / or soluble forms of LAG-3.

[0277] In one aspect, an anti-LAG-3 antibody useful for assaying, detecting, and / or quantitating LAG-3 expression in accordance with the methods described herein is the 17B4 mouse IgG1 anti-human LAG-3 monoclonal antibody. See, e.g., Matsuzaki, J et al., PNAS (2010);107:7875.

[0278] In one aspect, the LAG-3 antagonist is formulated for intravenous administration.

[0279] In one aspect, the LAG-3 antagonist is administered in a fixed dose.

[0280] In some aspects, the LAG-3 antagonist is administered in an amount of at least about 0.25 mg to about 2000 mg, about 0.25 mg to about 1600 mg, about 0.25 mg to about 1200 mg, about 0.25 mg to about 800 mg, about 0.25 mg to about 400 mg, about 0.25 mg to about 100 mg, about 0.25 mg to about 50 mg, about 0.25 mg to about 40 mg, about 0.25 mg to about 30 mg, about 0.25 mg to about 20 mg, about 20 mg to about 2000 mg, about 20 mg to about 1600 mg, about 20 mg to about 1200 mg, about 20 mg to about 800 mg, about 20 mg to about 400 mg, about 20 mg to about 2 The compound is administered in a dose of 0 mg to about 100 mg, about 100 mg to about 2000 mg, about 100 mg to about 1800 mg, about 100 mg to about 1600 mg, about 100 mg to about 1400 mg, about 100 mg to about 1200 mg, about 100 mg to about 1000 mg, about 100 mg to about 800 mg, about 100 mg to about 600 mg, about 100 mg to about 400 mg, about 400 mg to about 2000 mg, about 400 mg to about 1800 mg, about 400 mg to about 1600 mg, about 400 mg to about 1400 mg, about 400 mg to about 1200 mg, or about 400 mg to about 1000 mg.

[0281] In one aspect, the LAG-3 antagonist is administered in an amount of about 0.25 mg, about 0.5 mg, about 0.75 mg, about 1 mg, about 1.25 mg, about 1.5 mg, about 1.75 mg, about 2 mg, about 2.25 mg, about 2.5 mg, about 2.75 mg, about 3 mg, about 3.25 mg, about 3.5 mg, about 3.75 mg, about 4 mg, about 4.25 mg, about 4.5 mg, about 4.75 mg, about 5 mg, about 5.25 mg, about 5.5 mg, about 5.75 mg, about 6 mg, about 6.25 mg, about 6.5 mg, about 6.75 mg, about 7 mg, about 7.25 mg, about 7.5 mg, about 7.75 mg, about 8 mg, about 8.25 mg, about 8.5 mg, about 8.75 mg, about 9 mg, about 9.25 mg, about 9.5 mg, or about 9.75mg, about 10mg, about 20mg, about 30mg, about 40mg, about 50mg, about 60mg, about 70mg, about 80mg, about 90mg, about 100mg, about 110mg, about 120mg, about 130mg, about 140mg, about 150mg, about 160mg, about 170mg, about 18 0mg, about 190mg, about 200mg, about 210mg, about 220mg, about 230mg, about 240mg, about 250mg, about 260mg, about 270mg, about 280mg, about 290mg, about 300mg, about 310mg, about 320mg, about 330mg, about 340mg, about 35 0mg, approx. 360mg, approx. 370mg, approx. 380mg, approx. 390mg, approx. 400mg, approx. 410mg, approx. 420mg, approx. 430mg, approx. 440mg, approx. 450mg, approx. 20mg, about 530mg, about 540mg, about 550mg, about 560mg, about 570mg, about 580mg, about 590mg, about 600mg, about 610mg, about 620mg, about 630mg, about 640mg, about 650mg, about 660mg, about 670mg, about 680mg, about 6 90mg, about 700mg, about 710mg, about 720mg, about 730mg, about 740mg, about 750mg, about 760mg, about 770mg, about 780mg, about 790mg, about 800mg, about 810mg, about 820mg, about 830mg, about 840mg, about 850mg, about 860mg, about 870mg, about 880mg, about 890mg, about 900mg, about 910mg, about 920mg, about 930mg, about 940mg, about 950mg, about 960mg, about 970mg, about 980mg, about 990mg, about 1000mg, about 1040mg, about 1080m g, about 1100 mg, about 1140 mg, about 1180 mg, about 1200 mg, about 1240 mg, about 1280 mg, about 1300 mg, about 1340 mg, about 1380 mg, about 1400 mg, about 1440 mg, about 1480 mg, about 1500 mg, about 1540 mg, about 1580 mg, about 1600 mg, about 1640 mg, about 1680 mg, about 1700 mg, about 1740 mg, about 1780 mg, about 1800 mg, about 1840 mg, about 1880 mg, about 1900 mg, about 1940 mg, about 1980 mg, or about 2000 mg.

[0282] In some aspects, the LAG-3 antagonist is administered at a dose of about 360 mg, about 720 mg, about 1080 mg, or about 1200 mg.

[0283] In one aspect, the LAG-3 antagonist is administered in a weight-based dose.

[0284] In some aspects, the LAG-3 antagonist is administered at a dose of from about 0.003 mg / kg to about 25 mg / kg, from about 0.003 mg / kg to about 20 mg / kg, from about 0.003 mg / kg to about 15 mg / kg, from about 0.003 mg / kg to about 10 mg / kg, from about 0.003 mg / kg to about 5 mg / kg, from about 0.003 mg / kg to about 1 mg / kg, from about 0.003 mg / kg to about 0.9 mg / kg, from about 0.003 mg / kg to about 0 .8mg / kg, about 0.003mg / kg to about 0.7mg / kg, about 0.003mg / kg to about 0.6mg / kg, about 0.003mg / kg to about 0.5mg / kg, about 0.003mg / kg to about 0.4m g / kg, about 0.003mg / kg to about 0.3mg / kg, about 0.003mg / kg to about 0.2mg / kg, about 0.003mg / kg to about 0.1mg / kg, about 0.1mg / kg to about 25mg / kg, about 0.1mg / kg to about 20mg / kg, about 0.1mg / kg to about 15mg / kg, about 0.1mg / kg to about 10mg / kg, about 0.1mg / kg to about 5mg / kg, about 0.1mg / kg to about 1mg / kg , about 1 mg / kg to about 25 mg / kg, about 1 mg / kg to about 20 mg / kg, about 1 mg / kg to about 15 mg / kg, about 1 mg / kg to about 10 mg / kg, about 1 mg / kg to about 5 mg / kg, about 5 mg / k The dose is administered at a range of about 10 mg / kg to about 25 mg / kg, about 5 mg / kg to about 20 mg / kg, about 5 mg / kg to about 15 mg / kg, about 5 mg / kg to about 10 mg / kg, about 10 mg / kg to about 25 mg / kg, about 10 mg / kg to about 20 mg / kg, about 10 mg / kg to about 15 mg / kg, about 15 mg / kg to about 25 mg / kg, about 15 mg / kg to about 20 mg / kg, or about 20 mg / kg to about 25 mg / kg.

[0285] In one aspect, the LAG-3 antagonist is administered at a concentration of about 0.003 mg / kg, about 0.004 mg / kg, about 0.005 mg / kg, about 0.006 mg / kg, about 0.007 mg / kg, about 0.008 mg / kg, about 0.009 mg / kg, about 0.01 mg / kg, about 0.02 mg / kg, about 0.03 mg / kg, about 0.04 mg / kg, about 0.05 mg / kg, about 0.06 mg / kg, about 0.07 mg / kg, about 0.08 mg / kg, about 0.09 mg / kg, about 0.1 mg / kg, about 0.2 mg / kg, about 0.3 mg / kg, about 0.4 mg / kg, about 0.5 mg / kg, about 0.6 mg / kg, about 0.7 mg / kg, about 0.8 mg / kg, about The compound is administered at a dose of 0.9 mg / kg, about 1.0 mg / kg, about 2.0 mg / kg, about 3.0 mg / kg, about 4.0 mg / kg, about 5.0 mg / kg, about 6.0 mg / kg, about 7.0 mg / kg, about 8.0 mg / kg, about 9.0 mg / kg, about 10.0 mg / kg, about 11.0 mg / kg, about 12.0 mg / kg, about 13.0 mg / kg, about 14.0 mg / kg, about 15.0 mg / kg, about 16.0 mg / kg, about 17.0 mg / kg, about 18.0 mg / kg, about 19.0 mg / kg, about 20.0 mg / kg, about 21.0 mg / kg, about 22.0 mg / kg, about 23.0 mg / kg, about 24.0 mg / kg, or about 25.0 mg / kg.

[0286] In some aspects, doses are administered about once every week, about once every 2 weeks, about once every 3 weeks, about once every 4 weeks, about once every 5 weeks, about once every 6 weeks, about once every 7 weeks, about once every 8 weeks, about once every 9 weeks, about once every 10 weeks, about once every 11 weeks, or about once every 12 weeks.

[0287] In some aspects, the LAG-3 antagonists described herein are administered as monotherapy, i.e., the LAG-3 antagonist is not administered in combination with another therapeutic agent.

[0288] In certain aspects, a LAG-3 antagonist or a combination of a LAG-3 antagonist and a CTLA-4 inhibitor described herein is administered with one or more additional therapeutic and / or anti-cancer therapies.

[0289] II.C. CTLA-4 Inhibitors CTLA-4 inhibitors that can be used in the methods described herein are known in the art. In certain aspects, CTLA-4 inhibitors for use in the methods described herein include, but are not limited to, CTLA-4 binding agents. In certain aspects, the CTLA-4 binding agent binds to human CTLA-4 and disrupts the interaction between CTLA-4 and the human B7 receptor. Because the interaction between CTLA-4 and B7 transmits a signal that leads to the inactivation of T cells that bear the CTLA-4 receptor, disruption of the interaction effectively induces, enhances, or prolongs the activation of such T cells, thereby inducing, enhancing, or prolonging an immune response.

[0290] In one aspect, the CTLA-4 inhibitor is an anti-CTLA-4 antibody.

[0291] A human monoclonal antibody that specifically binds to CTLA-4 with high affinity is described in U.S. Patent No. 6,984,720. Other anti-CTLA-4 monoclonal antibodies are described in, for example, U.S. Patent Nos. 5,977,318, 6,051,227, 6,682,736 and 7,034,121 and International Publication Nos. WO2012 / 122444, WO2007 / 113648, WO2016 / 196237 and WO2000 / 037504 (each of which is incorporated herein by reference in its entirety). The anti-CTLA-4 human monoclonal antibody described in U.S. Patent No. 6,984,720 has been demonstrated to exhibit one or more of the following characteristics: (a) a specific binding affinity of at least about 10, as measured by Biacore analysis; 7 M -1 or about 10 9 M -1 or about 10 10 M -1 From 10 11 M -1 or higher equilibrium binding constant (K a (b) specifically binds human CTLA-4 with a binding affinity of at least about 10 3 m -1s -1 , about 10 4 m -1 s -1 or about 10 5 m -1 s -1 The dynamic binding constant (k a );(c) at least about 10 3 m -1 s -1 , about 10 4 m -1 s -1 or about 10 5 m -1 s -1 The dynamic dissociation constant (k d and (d) inhibit the binding of CTLA-4 to B7-1 (CD80) and B7-2 (CD86). Anti-CTLA-4 antibodies useful in the present invention include monoclonal antibodies that specifically bind to human CTLA-4 and exhibit at least one, at least two, or at least three of the above characteristics.

[0292] Anti-CTLA-4 antibodies that may be used in the methods described herein include ipilimumab (YERVOY®, also known as MDX-010, 10D1; see U.S. Patent No. 6,984,720), MK-1308 (Merck, also known as quavonlimab), AGEN-1884 (Agenus Inc., also known as zalifrelimab; see WO 2016 / 196237), tremelimumab (AstraZeneca; also known as ticilimumab, CP-675,206; see WO 2000 / 037504 and Ribas, Update Cancer Ther. 2(3): 133-39 (2007)), REGN4659 (Regeneron Pharmaceuticals, Inc.; see U.S. Application Publication No. 2019 / 0048096), and CTLA-4 probody and BMS-986249 (BMS, an anti-CTLA-4 antibody with a cleaved peptide mask; see WO18 / 085555).

[0293] In one aspect, the anti-CTLA-4 antibody specifically binds to human CTLA-4 and cross-competes for binding to human CTLA-4 with any of the anti-CTLA-4 antibodies described herein, e.g., ipilimumab and / or tremelimumab. In one aspect, the anti-CTLA-4 antibody binds to the same epitope as any of the anti-CTLA-4 antibodies described herein, e.g., ipilimumab and / or tremelimumab.

[0294] In one aspect, antibodies that cross-compete with, or bind to the same epitope as, any of the anti-CTLA-4 antibodies described herein, e.g., ipilimumab and / or tremelimumab, for binding to human CTLA-4 are monoclonal antibodies. For administration to human subjects, these cross-competing antibodies are chimeric, modified, or humanized or human antibodies.

[0295] Anti-CTLA-4 antibodies that can be used in the methods described herein also include antigen-binding portions of any of the above full-length antibodies.

[0296] In some aspects, the anti-CTLA-4 antibody is a full-length antibody. In some aspects, the anti-CTLA-4 antibody is a monoclonal antibody, a human antibody, a humanized antibody, a chimeric antibody, or a multispecific antibody. In some aspects, the multispecific antibody is a DART, a DVD-Ig, or a bispecific antibody.

[0297] In one aspect, the anti-CTLA-4 antibody is a F(ab')2 fragment, a Fab' fragment, a Fab fragment, an Fv fragment, an scFv fragment, a dsFv fragment, a dAb fragment, or a single-chain binding polypeptide peptide.

[0298] In one aspect, the anti-CTLA-4 antibody is ipilimumab, tremelimumab, MK1308, AGEN1884, REGN4659, or an antigen-binding portion thereof.

[0299] In one aspect, the CTLA-4 antibody is tremelimumab (also known as CP-675,206). Tremelimumab is a human IgG2 monoclonal anti-CTLA-4 antibody. Tremelimumab is described in WO / 2012 / 122444, U.S. Application Publication No. 2012 / 263677, or WO2007 / 113648 A2.

[0300] In one aspect, the anti-CTLA-4 antibody is ipilimumab. Ipilimumab is a fully human, IgG1 monoclonal antibody that blocks the binding of CTLA-4 to its B7 ligand, thereby stimulating T cell activation.

[0301] In one aspect, the anti-CTLA-4 antibody comprises the heavy and light chain CDRs, heavy and light chain variable regions, or heavy and light chain sequences of the anti-CTLA-4 antibodies described herein, such as the sequences provided in the documents described herein, or known in the art.

[0302] In certain aspects, the methods described herein include anti-CTLA-4 antibodies having at least about 90% sequence identity to an anti-CTLA-4 antibody described herein or known in the art (e.g., having at least about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identity to the sequence of an anti-CTLA-4 antibody, such as the heavy chain variable region and / or light chain variable region of the anti-CTLA-4 antibody, or the heavy chain and / or light chain of the anti-CTLA-4 antibody).

[0303] In one aspect, the anti-CTLA-4 antibody comprises the complementarity-determining regions (CDRs) of ipilimumab, identified as 10D1 in U.S. Patent Nos. 6,984,720 and 7,605,238, each of which is incorporated herein by reference in its entirety. Ipilimumab (formerly known as IvtDX-010 and BMS-734016) is sold as YERVOY® and is approved for the treatment of metastatic melanoma and is undergoing clinical trials in other cancers. See Hoos et al. (2010) Semin. Oncol. 37:533; Hodi et al. (2010) New Engl J. Med. 363:711; Pardoll (2012) Nat. Immunol. 13(12):1129.

[0304] In one aspect, the anti-CTLA-4 antibody comprises the heavy and light chain CDRs of ipilimumab, the heavy chain and light chain variable region of ipilimumab, or the heavy chain and light chain of ipilimumab.

[0305] In one aspect, the anti-CTLA-4 antibody comprises a heavy chain variable region CDR1, CDR2, and CDR3 domains having the sequence set forth in SEQ ID NO:34, and a light chain variable region CDR1, CDR2, and CDR3 domains having the sequence set forth in SEQ ID NO:32.

[0306] In one aspect, the anti-CTLA-4 antibody comprises: (a) a heavy chain variable region, CDR1, comprising the sequence set forth in SEQ ID NO: 35; (b) a heavy chain variable region, CDR2, comprising the sequence set forth in SEQ ID NO: 36; (c) a heavy chain variable region, CDR3, comprising the sequence set forth in SEQ ID NO: 37; (d) a light chain variable region, CDR1, comprising the sequence set forth in SEQ ID NO: 38; (e) a light chain variable region, CDR2, comprising the sequence set forth in SEQ ID NO: 39; and (f) a light chain variable region, CDR3, comprising the sequence set forth in SEQ ID NO: 40.

[0307] In one aspect, the anti-CTLA-4 antibody comprises a heavy chain variable region and a light chain variable region comprising the sequences set forth in SEQ ID NO:34 and / or SEQ ID NO:32, respectively.

[0308] In one aspect, the anti-CTLA-4 antibody comprises a heavy chain variable region and a light chain variable region comprising the sequences set forth in SEQ ID NO:34 and SEQ ID NO:32, respectively.

[0309] In one aspect, the anti-CTLA-4 antibody comprises the heavy and light chains of ipilimumab.

[0310] In one aspect, the CTLA-4 inhibitor (e.g., an anti-CTLA-4 antibody) is formulated for intravenous administration.

[0311] In one aspect, the CTLA-4 inhibitor is administered in a fixed dose.

[0312] In some aspects, the CTLA-4 inhibitor is administered in an amount of at least about 0.25 mg to about 2000 mg, about 0.25 mg to about 1600 mg, about 0.25 mg to about 1200 mg, about 0.25 mg to about 800 mg, about 0.25 mg to about 400 mg, about 0.25 mg to about 100 mg, about 0.25 mg to about 50 mg, about 0.25 mg to about 40 mg, about 0.25 mg to about 30 mg, about 0.25 mg to about 20 mg, about 20 mg to about 2000 mg, about 20 mg to about 1600 mg, about 20 mg to about 1200 mg, about 20 mg to about 800 mg, about 20 mg to about 400 mg, about 20 mg to about 5 ... mg to about 100 mg, about 100 mg to about 2000 mg, about 100 mg to about 1800 mg, about 100 mg to about 1600 mg, about 100 mg to about 1400 mg, about 100 mg to about 1200 mg, about 100 mg to about 1000 mg, about 100 mg to about 800 mg, about 100 mg to about 600 mg, about 100 mg to about 400 mg, about 400 mg to about 2000 mg, about 400 mg to about 1800 mg, about 400 mg to about 1600 mg, about 400 mg to about 1400 mg, about 400 mg to about 1200 mg, or about 400 mg to about 1000 mg.

[0313] In one aspect, the CTLA-4 inhibitor is about 0.25 mg, about 0.5 mg, about 0.75 mg, about 1 mg, about 1.25 mg, about 1.5 mg, about 1.75 mg, about 2 mg, about 2.25 mg, about 2.5 mg, about 2.75 mg, about 3 mg, about 3.25 mg, about 3.5 mg, about 3.75 mg, about 4 mg, about 4.25 mg, about 4.5 mg, about 4.75 mg, about 5 mg, about 5.25 mg, about 5.5 mg, about 5.75 mg, about 6 mg, about 6.25 mg, about 6.5 mg, about 6.75 mg, about 7 mg, about 7.25 mg, about 7.5 mg, about 7.75 mg, about 8 mg, about 8.25 mg, about 8.5 mg, about 8.75 mg, about 9 mg, about 9.25 mg, about 9.5 mg, about 9.75mg, about 10mg, about 20mg, about 30mg, about 40mg, about 50mg, about 60mg, about 70mg, about 80mg, about 90mg, about 100mg, about 110mg, about 120mg, about 130mg, about 140mg, about 150mg, about 160mg, about 170mg, about 18 0mg, about 190mg, about 200mg, about 210mg, about 220mg, about 230mg, about 240mg, about 250mg, about 260mg, about 270mg, about 280mg, about 290mg, about 300mg, about 310mg, about 320mg, about 330mg, about 340mg, about 35 0mg, approx. 360mg, approx. 370mg, approx. 380mg, approx. 390mg, approx. 400mg, approx. 410mg, approx. 420mg, approx. 430mg, approx. 440mg, approx. 450mg, approx. 20mg, about 530mg, about 540mg, about 550mg, about 560mg, about 570mg, about 580mg, about 590mg, about 600mg, about 610mg, about 620mg, about 630mg, about 640mg, about 650mg, about 660mg, about 670mg, about 680mg, about 6 90mg, about 700mg, about 710mg, about 720mg, about 730mg, about 740mg, about 750mg, about 760mg, about 770mg, about 780mg, about 790mg, about 800mg, about 810mg, about 820mg, about 830mg, about 840mg, about 850mg, about 860mg, about 870mg, about 880mg, about 890mg, about 900mg, about 910mg, about 920mg, about 930mg, about 940mg, about 950mg, about 960mg, about 970mg, about 980mg, about 990mg, about 1000mg, about 1040mg, about 1080m g, about 1100 mg, about 1140 mg, about 1180 mg, about 1200 mg, about 1240 mg, about 1280 mg, about 1300 mg, about 1340 mg, about 1380 mg, about 1400 mg, about 1440 mg, about 1480 mg, about 1500 mg, about 1540 mg, about 1580 mg, about 1600 mg, about 1640 mg, about 1680 mg, about 1700 mg, about 1740 mg, about 1780 mg, about 1800 mg, about 1840 mg, about 1880 mg, about 1900 mg, about 1940 mg, about 1980 mg, or about 2000 mg.

[0314] In one aspect, the CTLA-4 inhibitor is administered in a weight-based dose.

[0315] In some aspects, the CTLA-4 inhibitor is administered in an amount of from about 0.003 mg / kg to about 25 mg / kg, from about 0.003 mg / kg to about 20 mg / kg, from about 0.003 mg / kg to about 15 mg / kg, from about 0.003 mg / kg to about 10 mg / kg, from about 0.003 mg / kg to about 5 mg / kg, from about 0.003 mg / kg to about 1 mg / kg, from about 0.003 mg / kg to about 0.9 mg / kg, from about 0.003 mg / kg to about 0.8 mg / kg. mg / kg, about 0.003mg / kg to about 0.7mg / kg, about 0.003mg / kg to about 0.6mg / kg, about 0.003mg / kg to about 0.5mg / kg, about 0.003mg / kg to about 0.4mg / kg, about 0.003mg / kg to about 0.3mg / kg, about 0.003mg / kg to about 0.2mg / kg, about 0.003mg / kg to about 0.1mg / kg, about 0.1mg / kg to about 25mg / kg, about 0 .1mg / kg~about 20mg / kg, about 0.1mg / kg~about 15mg / kg, about 0.1mg / kg~about 10mg / kg, about 0.1mg / kg~about 5mg / kg, about 0.1mg / kg~about 1mg / kg , about 1 mg / kg to about 25 mg / kg, about 1 mg / kg to about 20 mg / kg, about 1 mg / kg to about 15 mg / kg, about 1 mg / kg to about 10 mg / kg, about 1 mg / kg to about 5 mg / kg, about 5 mg / k The dose is administered at a range of about 10 mg / kg to about 25 mg / kg, about 5 mg / kg to about 20 mg / kg, about 5 mg / kg to about 15 mg / kg, about 5 mg / kg to about 10 mg / kg, about 10 mg / kg to about 25 mg / kg, about 10 mg / kg to about 20 mg / kg, about 10 mg / kg to about 15 mg / kg, about 15 mg / kg to about 25 mg / kg, about 15 mg / kg to about 20 mg / kg, or about 20 mg / kg to about 25 mg / kg.

[0316] In one aspect, the CTLA-4 inhibitor is administered at a concentration of about 0.003 mg / kg, about 0.004 mg / kg, about 0.005 mg / kg, about 0.006 mg / kg, about 0.007 mg / kg, about 0.008 mg / kg, about 0.009 mg / kg, about 0.01 mg / kg, about 0.02 mg / kg, about 0.03 mg / kg, about 0.04 mg / kg, about 0.05 mg / kg, about 0.06 mg / kg, about 0.07 mg / kg, about 0.08 mg / kg, about 0.09 mg / kg, about 0.1 mg / kg, about 0.2 mg / kg, about 0.3 mg / kg, about 0.4 mg / kg, about 0.5 mg / kg, about 0.6 mg / kg, about 0.7 mg / kg, about 0.8 mg / kg, or about 0. 9 mg / kg, about 1.0 mg / kg, about 2.0 mg / kg, about 3.0 mg / kg, about 4.0 mg / kg, about 5.0 mg / kg, about 6.0 mg / kg, about 7.0 mg / kg, about 8.0 mg / kg, about 9.0 mg / kg, about 10.0 mg / kg, about 11.0 mg / kg, about 12.0 mg / kg, about 13.0 mg / kg, about 14.0 mg / kg, about 15.0 mg / kg, about 16.0 mg / kg, about 17.0 mg / kg, about 18.0 mg / kg, about 19.0 mg / kg, about 20.0 mg / kg, about 21.0 mg / kg, about 22.0 mg / kg, about 23.0 mg / kg, about 24.0 mg / kg, or about 25.0 mg / kg.

[0317] In some aspects, the dose is administered about once every week, about once every 2 weeks, about once every 3 weeks, about once every 4 weeks, about once every 5 weeks, about once every 6 weeks, about once every 7 weeks, about once every 8 weeks, about once every 9 weeks, about once every 10 weeks, about once every 11 weeks, or about once every 12 weeks.

[0318] In one aspect, the CTLA-4 inhibitor is ipilimumab and is administered at a dose of about 3 mg / kg about once every 3 weeks, at a dose of about 10 mg / kg about once every 3 weeks, or at a dose of about 10 mg / kg about once every 12 weeks. In one aspect, ipilimumab is administered in 4 doses.

[0319] In one aspect, the LAG-3 antagonist and the CTLA-4 inhibitor are formulated separately.

[0320] In one aspect, the LAG-3 antagonist and the CTLA-4 inhibitor are formulated as a single agent.

[0321] In one aspect, the LAG-3 antagonist is administered before the CTLA-4 inhibitor.

[0322] In one aspect, the CTLA-4 inhibitor is administered before the LAG-3 antagonist.

[0323] In one aspect, the LAG-3 antagonist and the CTLA-4 inhibitor are administered simultaneously.

[0324] II.D. Anti-cancer Therapies and Therapeutic Agents In certain aspects, a LAG-3 antagonist described herein, or a combination of a LAG-3 antagonist and a CTLA-4 inhibitor described herein, is administered with one or more additional anti-cancer therapies and / or therapeutic agents.

[0325] The additional therapeutic agent and / or anti-cancer therapy may include any known therapeutic agent or anti-cancer therapy, including the standard of care in the art for the treatment of patients with melanoma.

[0326] In some aspects, the additional anti-cancer therapy comprises surgery, radiation therapy, chemotherapy, immunotherapy, or any combination thereof. In some aspects, the additional anti-cancer therapy comprises chemotherapy comprising any chemotherapeutic agent described herein. In some aspects, the chemotherapy comprises platinum-doublet chemotherapy.

[0327] In some aspects, the additional therapeutic agent comprises an anti-cancer agent, hi some aspects, the anti-cancer agent comprises a tyrosine kinase inhibitor, an anti-angiogenic agent, a checkpoint inhibitor, a checkpoint stimulator, a chemotherapeutic agent, an immunotherapeutic agent, a platinum agent, an alkylating agent, a taxane, a nucleoside analog, an antimetabolite, a topisomerase inhibitor, an anthracycline, a vinca alkaloid, or any combination thereof.

[0328] In some aspects, the tyrosine kinase inhibitor (TKI) is sorafenib (e.g., sorafenib tosylate, also known as NEXAVAR®), lenvatinib (e.g., lenvatinib mesylate, also known as LENVIMA®), regorafenib (e.g., STIVARGA®), cabozantinib (e.g., cabozantinib S-malate, also known as CABOMETYX®), sunitinib (e.g., sunitinib malate, also known as SUTENT®), Brivanib, linifanib, pemigatinib (also known as PEMAZYRE™), everolimus (also known as AFINITOR® or ZORTRESS®), gefitinib (IRESSA®, a small molecule TKI for EGFR), imatinib (e.g., imatinib mesylate), lapatinib (e.g., lapatinib ditosylate, also known as TYKERB®), nilotinib (nilotinib hydrochloride, also known as TASIGNA®), pazopanib (pazopanib hydrochloride), salt, also known as VOTRIENT®), temsirolimus (also known as TORISEL®), erlotinib (e.g., erlotinib hydrochloride, also known as Tarceva®, a small molecule TKI for EGFR), afatinib (Gilotrif®, a small molecule TKI for EGFR), dacomitinib (VIZIMPRO®, a small molecule TKI for EGFR), osimeritinib (TAGRISSO®, a small molecule TKI for EGFR), alectinib (ALECENSA®, a small molecule TKI for ALK), small molecule TKI), ceritinib (ZYKADIA®, a small molecule TKI for ALK and ROS-1), brigatinib (ALUNBRIG®, a small molecule TKI for ALK), crizotinib (XALKORI®, a small molecule TKI for ALK and ROS-1), loratinib (LORBRENA®, a small molecule TKI for ALK and ROS-1), entrectinib (ROZLYTREK®, a small molecule TKI for ROS-1 and NTRK), larotrectinib (ROZLYTREK®, a small molecule TKI for NTRK),Dabrafenib (TAFINLAR®, a BRAF-targeting small molecule TKI), vemurafenib (ZELBORAF®, a BRAF-targeting small molecule TKI), encorafenib (BRAFTOVI®, a BRAF-targeting small molecule TKI), trametinib (MEKINIST®, a MEK-targeting small molecule TKI), cobimetinib (COTELLIC®, a MEK-targeting small molecule TKI), binimetinib (MEKTOVI®, a MEK-targeting small molecule TKI), or any combination thereof.

[0329] In some aspects, the TKI is an inhibitor of BRAF. In some aspects, the TKI is dabrafenib, vemurafenib, and / or encorafenib.

[0330] In some aspects, the TKI is an inhibitor of MEK. In some aspects, the TKI is trametinib, cobimetinib, and / or binimetinib.

[0331] In some aspects, the patient has a BRAF and / or MEK mutation, and the methods described herein further comprise administering a TKI that is an inhibitor of BRAF and / or a TKI that is an inhibitor of MEK as the targeted inhibitor therapy.

[0332] In some aspects, anti-angiogenic agents include inhibitors of vascular endothelial growth factor (VEGF), VEGF receptor (VEGFR), platelet-derived growth factor (PDGF), PDGF receptor (PDGFR), angiopoietin (Ang), tyrosine kinase with Ig-like and EGF-like domains (Tie) receptor, hepatocyte growth factor (HGF), tyrosine-protein kinase Met (c-MET), C-type lectin family 14 member A (CLEC14A), multimerin 2 (MMRN2), shock protein 70-1A (HSP70-1A), epidermal growth factor (EGF), EGFR, or any combination thereof. In some aspects, the anti-angiogenic agent comprises bevacizumab (also known as AVASTIN®), ramucirumab (also known as CYRAMZA®), aflibercept (also known as EYLEA® or ZALTRAP®), tanibirumab, olaratumab (also known as LARTRUVO™), nesbacumab, AMG780, MEDI3617, vanucizumab, rilotumab, ficlatuzumab, TAK-701, onartuzumab, emibetuzumab, or any combination thereof. In some aspects, the anti-angiogenic agent is bevacizumab.

[0333] In one aspect, checkpoint stimulators comprise an agonist of B7-1, B7-2, CD28, 4-1BB (CD137), 4-1BBL, GITR, inducible T cell costimulator (ICOS), ICOS-L, OX40, OX40L, CD70, CD27, CD40, death receptor 3 (DR3), CD28H, or any combination thereof.

[0334] In some aspects, the chemotherapeutic agent comprises an alkylating agent, antimetabolite, antitumor antibiotic, antimitotic, hormone or hormone-modulating agent, protein tyrosine kinase inhibitor, epidermal growth factor inhibitor, proteasome inhibitor, other neoplastic agent, or any combination thereof.

[0335] In some aspects, the immunotherapeutic agent comprises an antibody that specifically binds to EGFR (e.g., cetuximab (ERBITUX®)), ALK, ROS-1, NTRK, BRAF, ICOS, CD137 (4-1BB), CD134 (OX40), NKG2A, CD27, CD96, GITR, herpes virus entry mediator (HVEM), PD-1, PD-L1, CTLA-4, BTLA, TIM-3, A2aR, killer cell lectin-like receptor G1 (KLRG-1), natural killer cell receptor 2B4 (CD244), CD160, TIGIT, VISTA, KIR, TGFβ, IL-10, IL-8, B7-H4, Fas ligand, CSF1R, CXCR4, mesothelin, CEACAM-1, CD52, HER2, MICA, MICB, or any combination thereof.

[0336] In some aspects, the platinum agent comprises cisplatin, carboplatin, oxaliplatin, satraplatin, picoplatin, nedaplatin, triplatin (e.g., triplatin tetraacetate), lipoplatin, phenanthriplatin, or any combination thereof.

[0337] In some aspects, alkylating agents comprise altretamine, bendamustine, busulfan, carboplatin, carmustine, chlorambucil, cisplatin, cyclophosphamide, dacarbazine, ifosfamide, lomustine, mechlorethamine, melphalan, oxaliplatin, procarbazine, streptozocin, temozolomide, thiotepa, or any combination thereof.

[0338] In some aspects, the taxane comprises paclitaxel, albumin-bound paclitaxel (i.e., nab-paclitaxel), docetaxel, cabazitaxel, or any combination thereof.

[0339] In some aspects, the nucleoside analog comprises cytarabine, gemcitabine, lamivudine, entecavir, telbivudine, or any combination thereof.

[0340] In some aspects, antimetabolites comprise capecitabine, cladribine, clofarabine, cytarabine, floxuridine, fludarabine, fluorouracil, mercaptopurine, methotrexate, pemetrexed, pentostatin, pralatrexate, thioguanine, or any combination thereof.

[0341] In some embodiments, the topoisomerase inhibitor comprises etoposide, mitoxantrone, doxorubicin, irinotecan, topotecan, camptothecin, or any combination thereof.

[0342] In some aspects, the anthracycline comprises doxorubicin, daunorubicin, epirubicin, idarubicin, or any combination thereof.

[0343] In some aspects, the vinca alkaloids include vinblastine, vincristine, vinorelbine, vindesine, vincaminol, vineridine, vinburnine, or any combination thereof.

[0344] II.D.1. Checkpoint Inhibitors In some aspects, the anti-cancer agent administered as an additional therapeutic agent in the methods described herein is a checkpoint inhibitor.

[0345] In some aspects, the checkpoint inhibitor is a programmed cell death-1 (PD-1) pathway inhibitor, a cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) inhibitor, a T-cell immunoglobulin and ITIM domain (TIGIT) inhibitor, a T-cell immunoglobulin and mucin domain-containing 3 (TIM-3) inhibitor, a TIM-1 inhibitor, a TIM-4 inhibitor, a B7-H3 inhibitor, a B7-H4 inhibitor, a B-cell and T-cell lymphodepleting agent (BTLA) inhibitor, a V-domain Ig inhibitor of T-cell activation inhibitor (VISTA), an indoleamine 2,3-dioxygenase (IDO) inhibitors, nicotinamide adenine dinucleotide phosphooxidase isoform 2 (NOX2) inhibitors, killer cell immunoglobulin-like receptor (KIR) inhibitors, adenosine A2a receptor (A2aR) inhibitors, transforming growth factor beta (TGF-β) inhibitors, phosphoinositide 3-kinase (PI3K) inhibitors, CD47 inhibitors, CD48 inhibitors, CD73 inhibitors, CD113 inhibitors, sialic acid-binding immunoglobulin-like lectin 7 (SIGLEC-7) inhibitors, SIGL EC-9 inhibitors, SIGLEC-15 inhibitors, glucocorticoid-induced TNFR-related protein (GITR) inhibitors, galectin-1 inhibitors, galectin-9 inhibitors, carcinoembryonic antigen-related cell adhesion molecule-1 (CEACAM-1) inhibitors, G protein-coupled receptor 56 (GPR56) inhibitors, glycoprotein A repeat dominant (GARP) inhibitors, 2B4 inhibitors, programmed cell death-1 homolog (PD1H) inhibitors, leukocyte-associated immunoglobulin-like receptor 1 (LAIR1) inhibitors, or any combination thereof.

[0346] In one aspect, the checkpoint inhibitor is formulated for intravenous administration.

[0347] In some aspects, the checkpoint inhibitor is formulated separately from the LAG-3 antagonist and / or the CTLA-4 inhibitor. In some aspects, when the checkpoint inhibitor comprises two or more checkpoint inhibitors, each checkpoint inhibitor is formulated separately.

[0348] In one aspect, the checkpoint inhibitor is administered prior to the LAG-3 antagonist and / or the CTLA-4 inhibitor.

[0349] In one aspect, the LAG-3 antagonist and / or CTLA-4 inhibitor is administered prior to the checkpoint inhibitor.

[0350] In some aspects, the checkpoint inhibitor is formulated as a co-formulation with the LAG-3 antagonist and / or the CTLA-4 inhibitor. In some aspects, when the checkpoint inhibitor comprises two or more checkpoint inhibitors, the two or more checkpoint inhibitors are formulated as a co-formulation.

[0351] In one aspect, the checkpoint inhibitor is administered simultaneously with the LAG-3 antagonist and / or the CTLA-4 inhibitor.

[0352] In some aspects, the checkpoint inhibitor is administered in a fixed dose.

[0353] In some aspects, the checkpoint inhibitor is administered in an amount of at least about 0.25 mg to about 2000 mg, about 0.25 mg to about 1600 mg, about 0.25 mg to about 1200 mg, about 0.25 mg to about 800 mg, about 0.25 mg to about 400 mg, about 0.25 mg to about 100 mg, about 0.25 mg to about 50 mg, about 0.25 mg to about 40 mg, about 0.25 mg to about 30 mg, about 0.25 mg to about 20 mg, about 20 mg to about 2000 mg, about 20 mg to about 1600 mg, about 20 mg to about 1200 mg, about 20 mg to about 800 mg, about 20 mg to about 400 mg, about 20 mg to about 2 The compound is administered in a dose of 0 mg to about 100 mg, about 100 mg to about 2000 mg, about 100 mg to about 1800 mg, about 100 mg to about 1600 mg, about 100 mg to about 1400 mg, about 100 mg to about 1200 mg, about 100 mg to about 1000 mg, about 100 mg to about 800 mg, about 100 mg to about 600 mg, about 100 mg to about 400 mg, about 400 mg to about 2000 mg, about 400 mg to about 1800 mg, about 400 mg to about 1600 mg, about 400 mg to about 1400 mg, about 400 mg to about 1200 mg, or about 400 mg to about 1000 mg.

[0354] In some aspects, the checkpoint inhibitor is about 0.25 mg, about 0.5 mg, about 0.75 mg, about 1 mg, about 1.25 mg, about 1.5 mg, about 1.75 mg, about 2 mg, about 2.25 mg, about 2.5 mg, about 2.75 mg, about 3 mg, about 3.25 mg, about 3.5 mg, about 3.75 mg, about 4 mg, about 4.25 mg, about 4.5 mg, about 4.75 mg, about 5 mg, about 5.25 mg, about 5.5 mg, about 5.75 mg, about 6 mg, about 6.25 mg, about 6.5 mg, about 6.75 mg, about 7 mg, about 7.25 mg, about 7.5 mg, about 7.75 mg, about 8 mg, about 8.25 mg, about 8.5 mg, about 8.75 mg, about 9 mg, about 9.25 mg, about 9.5 mg, or about 9.75mg, about 10mg, about 20mg, about 30mg, about 40mg, about 50mg, about 60mg, about 70mg, about 80mg, about 90mg, about 100mg, about 110mg, about 120mg, about 130mg, about 140mg, about 150mg, about 160mg, about 170mg, about 18 0mg, about 190mg, about 200mg, about 210mg, about 220mg, about 230mg, about 240mg, about 250mg, about 260mg, about 270mg, about 280mg, about 290mg, about 300mg, about 310mg, about 320mg, about 330mg, about 340mg, about 35 0mg, approx. 360mg, approx. 370mg, approx. 380mg, approx. 390mg, approx. 400mg, approx. 410mg, approx. 420mg, approx. 430mg, approx. 440mg, approx. 450mg, approx. 20mg, about 530mg, about 540mg, about 550mg, about 560mg, about 570mg, about 580mg, about 590mg, about 600mg, about 610mg, about 620mg, about 630mg, about 640mg, about 650mg, about 660mg, about 670mg, about 680mg, about 6 90mg, about 700mg, about 710mg, about 720mg, about 730mg, about 740mg, about 750mg, about 760mg, about 770mg, about 780mg, about 790mg, about 800mg, about 810mg, about 820mg, about 830mg, about 840mg, about 850mg, about 860mg, about 870mg, about 880mg, about 890mg, about 900mg, about 910mg, about 920mg, about 930mg, about 940mg, about 950mg, about 960mg, about 970mg, about 980mg, about 990mg, about 1000mg, about 1040mg, about 1080m g, about 1100 mg, about 1140 mg, about 1180 mg, about 1200 mg, about 1240 mg, about 1280 mg, about 1300 mg, about 1340 mg, about 1380 mg, about 1400 mg, about 1440 mg, about 1480 mg, about 1500 mg, about 1540 mg, about 1580 mg, about 1600 mg, about 1640 mg, about 1680 mg, about 1700 mg, about 1740 mg, about 1780 mg, about 1800 mg, about 1840 mg, about 1880 mg, about 1900 mg, about 1940 mg, about 1980 mg, or about 2000 mg.

[0355] In some aspects, the checkpoint inhibitor is administered in a weight-based dose.

[0356] In some aspects, the checkpoint inhibitor is administered in an amount of from about 0.003 mg / kg to about 25 mg / kg, from about 0.003 mg / kg to about 20 mg / kg, from about 0.003 mg / kg to about 15 mg / kg, from about 0.003 mg / kg to about 10 mg / kg, from about 0.003 mg / kg to about 5 mg / kg, from about 0.003 mg / kg to about 1 mg / kg, from about 0.003 mg / kg to about 0.9 mg / kg, from about 0.003 mg / kg to about 0 .8mg / kg, about 0.003mg / kg to about 0.7mg / kg, about 0.003mg / kg to about 0.6mg / kg, about 0.003mg / kg to about 0.5mg / kg, about 0.003mg / kg to about 0.4m g / kg, about 0.003mg / kg to about 0.3mg / kg, about 0.003mg / kg to about 0.2mg / kg, about 0.003mg / kg to about 0.1mg / kg, about 0.1mg / kg to about 25mg / kg, about 0.1mg / kg to about 20mg / kg, about 0.1mg / kg to about 15mg / kg, about 0.1mg / kg to about 10mg / kg, about 0.1mg / kg to about 5mg / kg, about 0.1mg / kg to about 1mg / kg , about 1 mg / kg to about 25 mg / kg, about 1 mg / kg to about 20 mg / kg, about 1 mg / kg to about 15 mg / kg, about 1 mg / kg to about 10 mg / kg, about 1 mg / kg to about 5 mg / kg, about 5 mg / k The dose is administered at a range of about 10 mg / kg to about 25 mg / kg, about 5 mg / kg to about 20 mg / kg, about 5 mg / kg to about 15 mg / kg, about 5 mg / kg to about 10 mg / kg, about 10 mg / kg to about 25 mg / kg, about 10 mg / kg to about 20 mg / kg, about 10 mg / kg to about 15 mg / kg, about 15 mg / kg to about 25 mg / kg, about 15 mg / kg to about 20 mg / kg, or about 20 mg / kg to about 25 mg / kg.

[0357] In some aspects, the checkpoint inhibitor is administered at a concentration of about 0.003 mg / kg, about 0.004 mg / kg, about 0.005 mg / kg, about 0.006 mg / kg, about 0.007 mg / kg, about 0.008 mg / kg, about 0.009 mg / kg, about 0.01 mg / kg, about 0.02 mg / kg, about 0.03 mg / kg, about 0.04 mg / kg, about 0.05 mg / kg, about 0.06 mg / kg, about 0.07 mg / kg, about 0.08 mg / kg, about 0.09 mg / kg, about 0.1 mg / kg, about 0.2 mg / kg, about 0.3 mg / kg, about 0.4 mg / kg, about 0.5 mg / kg, about 0.6 mg / kg, about 0.7 mg / kg, about 0.8 mg / kg, about 0. ...9 mg 0.9 mg / kg, about 1.0 mg / kg, about 2.0 mg / kg, about 3.0 mg / kg, about 4.0 mg / kg, about 5.0 mg / kg, about 6.0 mg / kg, about 7.0 mg / kg, about 8.0 mg / kg, about 9.0 mg / kg, about 10.0 mg / kg, about 11.0 mg / kg, about 12.0 mg / kg, about 13.0 mg / kg, about 14.0 mg / kg, about 15.0 mg / kg, about 16.0 mg / kg, about 17.0 mg / kg, about 18.0 mg / kg, about 19.0 mg / kg, about 20.0 mg / kg, about 21.0 mg / kg, about 22.0 mg / kg, about 23.0 mg / kg, about 24.0 mg / kg, or about 25.0 mg / kg.

[0358] In one aspect, a dose of checkpoint inhibitor is administered once every week, once every 2 weeks, once every 3 weeks, once every 4 weeks, once every 5 weeks, once every 6 weeks, once every 7 weeks, once every 8 weeks, once every 9 weeks, once every 10 weeks, once every 11 weeks, or once every 12 weeks.

[0359] In some aspects, each of the LAG-3 antagonists, CTLA-4 inhibitors and / or checkpoint inhibitors described herein is administered in a fixed dose.

[0360] In certain aspects, each of the LAG-3 antagonists, CTLA-4 inhibitors, and / or checkpoint inhibitors described herein is administered in varying amounts. For example, in certain aspects, the maintenance (or continuing) dose of the LAG-3 antagonist, CTLA-4 inhibitor, and / or checkpoint inhibitor may be higher than or the same as the loading dose initially administered to the patient. In certain aspects, the maintenance dose of the LAG-3 antagonist, CTLA-4 inhibitor, and / or checkpoint inhibitor may be lower than or the same as the loading dose.

[0361] II.D.1.a.PD-1 Pathway Inhibitors In one aspect, checkpoint inhibitors for use in the methods of the invention comprise PD-1 pathway inhibitors.

[0362] In one aspect, the PD-1 pathway inhibitor is a PD-1 inhibitor and / or a PD-L1 inhibitor.

[0363] In one aspect, the PD-1 inhibitor and / or the PD-L1 inhibitor is a small molecule.

[0364] In one aspect, the PD-1 inhibitor and / or PD-L1 inhibitor is a millamolecule.

[0365] In one aspect, the PD-1 inhibitor and / or the PD-L1 inhibitor is a macrocyclic peptide.

[0366] In a particular aspect, the PD-1 inhibitor and / or PD-L1 inhibitor is BMS-986189.

[0367] In one aspect, the PD-1 inhibitor is an inhibitor described in International Publication WO2014 / 151634, which is incorporated herein by reference in its entirety.

[0368] In one aspect, the PD-1 inhibitor is INCMGA00012 Insight Pharmaceuticals.

[0369] In one aspect, the PD-1 inhibitor comprises a combination of an anti-PD-1 antibody and a PD-1 small molecule inhibitor described herein.

[0370] In one aspect, the PD-L1 inhibitor is a small molecule having the formula shown in Formula (I) (including miramolecules): [ka] In the formula, R 1 -R 13 is the amino acid side chain, and R a -R nは、 hydrogen, methyl, or forms a ring with adjacent R groups, and R 14 -C(O)NHR 15 where R 15 is a glycine residue that may be substituted with hydrogen or additional glycine residues and / or tails that may improve pharmacokinetic properties. In one aspect, PD-L1 inhibitors include compounds described in International Publication No. WO2014 / 151634, the entire contents of which are incorporated herein by reference. In some aspects, PD-L1 inhibitors include compounds described in International Publication Nos. WO2016 / 039749, WO2016 / 149351, WO2016 / 077518, WO2016 / 100285, WO2016 / 100608, WO2016 / 126646, WO2016 / 057624, WO2017 / 151830, WO2017 / 176608, WO2018 / 085750, WO2018 / 237153, or WO2019 / 070643, the contents of each of which are incorporated herein by reference in their entirety.

[0371] In some aspects, the PD-L1 inhibitor includes a small molecule PD-L1 inhibitor described in International Publication Nos. WO2015 / 034820, WO2015 / 160641, WO2018 / 044963, WO2017 / 066227, WO2018 / 009505, WO2018 / 183171, WO2018 / 11848, WO2019 / 147662, or WO2019 / 169123, the contents of each of which are incorporated herein by reference in their entirety.

[0372] In one aspect, the PD-1 pathway inhibitor is a soluble PD-L2 polypeptide. In one aspect, the soluble PD-L2 polypeptide is a fusion polypeptide. In one aspect, the soluble PD-L2 polypeptide comprises a ligand-binding fragment of the PD-L2 extracellular domain. In one aspect, the soluble PD-L2 polypeptide further comprises a half-life extending moiety. In one aspect, the half-life extending moiety comprises an immunoglobulin constant region or portion thereof, an immunoglobulin-binding polypeptide, immunoglobulin G (IgG), an albumin-binding polypeptide (ABP), a PAS-modified moiety, a HES-modified moiety, XTEN, a PEGylated moiety, an Fc region, or any combination thereof. In one aspect, the soluble PD-L2 polypeptide is AMP-224 (see, e.g., US 2013 / 0017199).

[0373] In one aspect, the PD-1 pathway inhibitor is an anti-PD-1 antibody and / or an anti-PD-L1 antibody.

[0374] II.D.1.ai anti-PD-1 antibody Anti-PD-1 antibodies known in the art can be used in the methods described herein. Various human monoclonal antibodies that specifically bind to PD-1 with high affinity are disclosed in U.S. Patent No. 8,008,449. The anti-PD-1 human antibodies disclosed in U.S. Patent No. 8,008,449 have been demonstrated to exhibit one or more of the following properties: (a) a 1×10 -antibody activity determined by surface plasmon resonance using a Biacore biosensor system; -7 M or lower K D(b) binds to human PD-1 in a specific manner; (b) does not substantially bind to human CD28, CTLA-4, or ICOS; (c) increases T-cell proliferation in a mixed lymphocyte reaction (MLR) assay; (d) increases interferon-γ production in an MLR assay; (e) increases IL-2 secretion in an MLR assay; (f) binds to human PD-1 and cynomolgus PD-1; (g) inhibits the binding of PD-L1 and / or PD-L2 to PD-1; (h) stimulates an antigen-specific memory response; (i) stimulates an antibody response; and (j) inhibits tumor cell proliferation in vivo. Anti-PD-1 antibodies useful in the present invention include monoclonal antibodies that specifically bind to human PD-1 and exhibit at least one, and in some aspects at least five, of the foregoing characteristics.

[0375] Other anti-PD-1 monoclonal antibodies that may be used in the methods described herein are described in, e.g., U.S. Patent Nos. 6,808,710, 7,488,802, 8,168,757, and 8,354,509; U.S. Application Publication No. 2016 / 0272708; and PCT Publication No. WO2012 / 145493. WO2008 / 156712, WO2015 / 112900, WO2012 / 145493, WO2015 / 112800, WO2014 / 206107 , WO2015 / 35606, WO2015 / 085847, WO2014 / 179664, WO2017 / 020291, WO2017 / 020858 , WO2016 / 197367, WO2017 / 024515, WO2017 / 025051, WO2017 / 123557, WO2016 / 10615 9, WO2014 / 194302, WO2017 / 040790, WO2017 / 133540, WO2017 / 132827, WO2017 / 0244 65, WO2017 / 025016, WO2017 / 106061, WO2017 / 19846, WO2017 / 024465, WO2017 / 025016, WO2017 / 132825, and WO2017 / 133540 (each of which is incorporated herein by reference in its entirety).

[0376] Anti-PD-1 antibodies that may be used in the methods described herein include nivolumab (also known as OPDIVO®, 5C4, BMS-936558, MDX-1106, and ONO-4538), pembrolizumab (Merck; also known as KEYTRUDA®, lambrolizumab, and MK-3475; see WO2008 / 156712), PDR001 (Novartis; also known as spartalizumab; see WO2015 / 112900), MEDI-0680 (AstraZeneca; also known as AMP-514; see WO2012 / 145493), TSR-042 (Tesaro Biopharmaceutical; also known as ANB011 or dostallimab; see WO2014 / 179664), cemiplimab (Regeneron; also known as LIBTAYO® or REGN-2810; see WO2015 / 112800 and U.S. Patent No. 9,987,500), JS001 (TAIZHOU JUNSHI PHARMA; also known as toripalimab; see Si-Yang Liu et al., J. Hematol. Oncol. 10:136 (2017)), PF-06801591 (Pfizer; also known as southernlimab; see US2016 / 0159905), BGB-A317 (Beigene; also known as tislelizumab; see WO2015 / 35606 and US2015 / 0079109), BI 754091 (Boehringer Ingelheim; see Zettl M et al., Cancer. Res. (2018);78(13 Suppl):Abstract 4558), INCSHR1210 (Jiangsu Hengrui Medicine; also known as SHR-1210 or camrelizumab; see WO2015 / 085847; see Si-Yang Liu et al., J. Hematol. Oncol. 10:136 (2017)), GLS-010 (Wuxi / Harbin Gloria Pharmaceuticals; also known as WBP3055; Si-Yang Liu et al., J. Hematol. Oncol. 10:136 (2017)), AM-0001 (Armo), STI-1110 (Sorrento Therapeutics; WO 2014 / 194302), AGEN2034 (Agenus; see WO2017 / 040790), MGA012 (Macrogenics, see WO2017 / 19846), BCD-100 (Biocad; Kaplon et al., mAbs 10(2):183-203 (2018)), IBI308 (Innovent; also known as sintilimab; see WO2017 / 024465, WO2017 / 025016, WO2017 / 132825, and WO2017 / 133540), and SSI-361 (Lyvgen Biopharma Holdings Limited, US2018 / 0346569).

[0377] In one aspect, the anti-PD-1 antibody comprises the heavy and light chain CDRs, heavy and light chain variable regions, or heavy and light chain sequences of an anti-PD-1 antibody described herein, such as the sequences provided in the documents described herein, or known in the art.

[0378] In certain aspects, the methods described herein include anti-PD-1 antibodies that have at least about 90% sequence identity to an anti-PD-1 antibody described herein or known in the art (e.g., at least about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identity to the sequence of an anti-PD-1 antibody, such as the heavy chain variable region and / or light chain variable region of the anti-PD-1 antibody, or the heavy chain and / or light chain of the anti-PD-1 antibody).

[0379] Anti-PD-1 antibodies that can be used in the methods described herein include isolated antibodies that specifically bind to human PD-1 and cross-compete with any of the anti-PD-1 antibodies described herein, e.g., nivolumab, for binding to human PD-1 (e.g., U.S. Patent Nos. 8,008,449 and 8,779,105; WO 2013 / 173223). In one aspect, the anti-PD-1 antibody binds to the same epitope as any of the anti-PD-1 antibodies described herein, e.g., nivolumab.

[0380] In one aspect, an antibody that cross-competes with any of the anti-PD-1 antibodies described herein, e.g., nivolumab, for binding to human PD-1 or that binds to the same epitope as any of the anti-PD-1 antibodies described herein, e.g., nivolumab, is a monoclonal antibody. For administration to a human subject, these cross-competing antibodies can be chimeric, engineered, humanized, or human. Such chimeric, engineered, humanized, or human monoclonal antibodies can be prepared and isolated by methods well known in the art.

[0381] Anti-PD-1 antibodies that can be used in the methods described herein also include antigen-binding portions of any of the full-length antibodies described above.

[0382] Anti-PD-1 antibodies suitable for use in the methods of the present invention are antibodies that bind to PD-1 with high specificity and affinity, inhibit binding of PD-L1 and / or PD-L2, and inhibit the immunosuppressive effects of the PD-1 signaling pathway. In any of the compositions or methods described herein, an anti-PD-1 "antibody" includes an antigen-binding portion or fragment that exhibits functional properties similar to those of a full antibody in binding to the PD-1 receptor, inhibiting ligand binding, and upregulating the immune system. In a particular aspect, the anti-PD-1 antibody, or antigen-binding portion thereof, cross-competes with nivolumab for binding to human PD-1.

[0383] In some aspects, the anti-PD-1 antibody is a full-length antibody. In some aspects, the anti-PD-1 antibody is a monoclonal antibody, a human antibody, a humanized antibody, a chimeric antibody, or a multispecific antibody. In some aspects, the multispecific antibody is a DART, DVD-Ig, or a bispecific antibody.

[0384] In one aspect, the anti-PD-1 antibody is a F(ab')2 fragment, a Fab' fragment, a Fab fragment, an Fv fragment, an scFv fragment, a dsFv fragment, a dAb fragment, or a single-chain binding polypeptide peptide.

[0385] In one aspect, the anti-PD-1 antibody is nivolumab, pembrolizumab, PDR001 (spartalizumab), MEDI-0680, TSR-042, cemiplimab, JS001, PF-06801591, BGB-A317, BI 754091, INCSHR1210, GLS-010, AM-001, STI-1110, AGEN2034, MGA012, BCD-100, IBI308, SSI-361, or an antigen-binding portion thereof.

[0386] In one aspect, the anti-PD-1 antibody is formulated for intravenous administration.

[0387] In one aspect, the anti-PD-1 antibody is administered intravenously over about 30 minutes.

[0388] In one aspect, the anti-PD-1 antibody is nivolumab, a fully human IgG4(S228P) PD-1 immune checkpoint inhibitor antibody that selectively blocks interaction with PD-1 ligands (PD-L1 and PD-L2), thereby preventing downregulation of anti-tumor T-cell function (U.S. Patent No. 8,008,449; Wang et al., 2014 Cancer Immunol Res. 2(9):846-56).

[0389] In one aspect, nivolumab is administered at a dose of about 240 mg once every two weeks.

[0390] In one aspect, nivolumab is administered at a dose of about 480 mg once every four weeks.

[0391] In one aspect, nivolumab is administered at a dose of about 1 mg / kg, followed on the same day by ipilimumab administered about once every three weeks for about four doses, followed by nivolumab at about 240 mg once every two weeks or about 480 mg once every four weeks.

[0392] In one aspect, the anti-PD-1 antibody is pembrolizumab. Pembrolizumab is a humanized monoclonal IgG4 (S228P) antibody directed against the human cell surface receptor PD-1. Pembrolizumab is described, for example, in U.S. Patent Nos. 8,354,509 and 8,900,587.

[0393] In one aspect, pembrolizumab is administered at a dose of about 200 mg once about every two weeks. In one aspect, pembrolizumab is administered at a dose of about 200 mg once about every three weeks. In one aspect, pembrolizumab is administered at a dose of about 400 mg once about every six weeks. In one aspect, pembrolizumab is administered at a dose of about 300 mg once about every four to five weeks.

[0394] In one aspect, the anti-PD-1 antibody is cemiplimab (REGN2810). Cemiplimab is described, e.g., in WO2015 / 112800 and U.S. Patent No. 9,987,500.

[0395] In one aspect, cemiplimab is administered intravenously at a dose of about 3 mg / kg or about 350 mg about once every three weeks.

[0396] In one aspect, the anti-PD-1 antibody is spartalizumab (PDR001). Spartalizumab is described, e.g., in WO2015 / 112900 and U.S. Patent No. 9,683,048.

[0397] In one aspect, spartalizumab is administered intravenously at a dose of about 300 mg about once every three weeks or at a dose of about 400 mg about once every four weeks.

[0398] II.D.1.a.ii. Anti-PD-L1 antibody Anti-PD-L1 antibodies known in the art can be used in the methods of the invention. Examples of anti-PD-L1 antibodies useful in the compositions and methods of the invention include the antibodies described in U.S. Patent No. 9,580,507. The anti-PD-L1 human monoclonal antibody described in U.S. Patent No. 9,580,507 has been shown to exhibit one or more of the following characteristics: (a) a PD-L1 activity of 1 x 10 as determined by surface plasmon resonance using a Biacore biosensor system; -7 M or lower K D (b) binds to human PD-L1 in a mixed lymphocyte reaction (MLR) assay; (c) increases interferon-γ production in an MLR assay; (d) increases IL-2 secretion in an MLR assay; (e) stimulates antibody responses; and (f) reverses the effects of T regulatory cells on T cell effector cells and / or dendritic cells. Anti-PD-L1 antibodies that may be used in the present invention include monoclonal antibodies that specifically bind to human PD-L1 and exhibit at least one, and in certain aspects at least five, of the above characteristics.

[0399] Anti-PD-L1 antibodies that may be used in the methods of the invention include BMS-936559 (12A4, also known as MDX-1105; see, e.g., U.S. Pat. No. 7,943,743 and WO 2013 / 173223), atezolizumab (Roche; TECENTRIQ®; MPDL3280A, also known as RG7446; see U.S. Pat. No. 8,217,149; see also Herbst et al. (2013) J Clin Oncol 31(suppl):3000), durvalumab (AstraZeneca; IMFINZ). (商標), also known as MEDI-4736; see WO2011 / 066389), avelumab (Pfizer; also known as BAVENCIO®, MSB-0010718C; see WO2013 / 079174), STI-1014 (Sorrento; see WO2013 / 181634), CX-072 (Cytomx; see WO2016 / 149201), KN035 (3D Med / Alphamab; see Zhang et al., Cell Discov. 7:3 (March 2017)), LY3300054 (Eli Lilly Co.; see, e.g., WO2017 / 034916), BGB-A333 (BeiGene; Desai et al., JCO 36 (15suppl):TPS3113 (2018)), ICO 36, and CK-301 (Checkpoint Therapeutics; see Gorelik et al., AACR:Abstract 4606 (Apr 2016)).

[0400] In one aspect, the anti-PD-L1 antibody comprises the heavy and light chain CDRs, heavy chain variable regions, or heavy and light chain sequences of an anti-PD-L1 antibody described herein, such as the sequences provided in the documents referenced herein, or known in the art.

[0401] In some aspects, the methods described herein include anti-PD-L1 antibodies that have at least about 90% sequence identity to an anti-PD-L1 antibody described herein or known in the art (e.g., at least about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identity to the sequence of an anti-PD-L1 antibody, such as a heavy chain variable region and / or a light chain variable region of an anti-PD-L1 antibody, or a heavy chain and / or a light chain of an anti-PD-L1 antibody).

[0402] Anti-PD-L1 antibodies that may be used in the methods of the present invention also include isolated antibodies that specifically bind to human PD-L1 and cross-compete with any of the anti-PD-L1 antibodies described herein, e.g., atezolizumab, durvalumab, and / or avelumab, for binding to human PD-L1. In one aspect, the anti-PD-L1 antibody binds to the same epitope as any of the anti-PD-L1 antibodies described herein, e.g., atezolizumab, durvalumab, and / or avelumab. In a particular aspect, an antibody that cross-competes with any of the anti-PD-L1 antibodies described herein, e.g., atezolizumab, durvalumab, and / or avelumab, for binding to human PD-L1, or binds to the same epitope region as any of the anti-PD-L1 antibodies described herein, e.g., atezolizumab, durvalumab, and / or avelumab, is a monoclonal antibody. For administration to human subjects, these cross-competing antibodies are chimeric, modified, or humanized or human antibodies. Such chimeric, modified, humanized, or human monoclonal antibodies can be prepared and isolated by methods well known in the art.

[0403] Anti-PD-L1 antibodies that may be used in the methods of the invention also include antigen-binding portions of any of the full-length antibodies described above.

[0404] Anti-PD-L1 antibodies that may be used in the methods of the present invention are antibodies that bind to PD-L1 with high specificity and affinity, inhibit PD-1 binding, and inhibit the immunosuppressive effects of the PD-1 signaling pathway. In any of the compositions or methods described herein, an anti-PD-L1 "antibody" includes an antigen-binding portion or fragment that binds to PD-L1 and exhibits functional properties similar to those of the whole antibody in inhibiting receptor binding and upregulating the immune system. In a particular aspect, the anti-PD-L1 antibody, or antigen-binding portion thereof, cross-competes with atezolizumab, durvalumab, and / or avelumab for binding to human PD-L1.

[0405] In one aspect, an anti-PD-L1 antibody is substituted for the anti-PD-1 antibody in any of the methods described herein.

[0406] In one aspect, the anti-PD-L1 antibody is a full-length antibody.

[0407] In some aspects, the anti-PD-L1 antibody is a monoclonal antibody, a human antibody, a humanized antibody, a chimeric antibody, or a multispecific antibody. In some aspects, the multispecific antibody is a DART, DVD-Ig, or a bispecific antibody.

[0408] In one aspect, the anti-PD-L1 antibody is a F(ab')2 fragment, a Fab' fragment, a Fab fragment, an Fv fragment, an scFv fragment, a dsFv fragment, a dAb fragment, or a single-chain binding polypeptide peptide.

[0409] In one aspect, the anti-PD-L1 antibody is BMS-936559, atezolizumab, durvalumab, Aveluma, STI-1014, CX-072, KN035, LY3300054, BGB-A333, ICO 36, CK-301, or an antigen-binding portion thereof.

[0410] In one aspect, the PD-L1 antibody is atezolizumab. Atezolizumab is a fully humanized IgG1 monoclonal anti-PD-L1 antibody. In one aspect, atezolizumab is administered at a fixed dose of about 800 mg about once every two weeks. In one aspect, atezolizumab is administered at a fixed dose of about 840 mg about once every two weeks.

[0411] In one aspect, the PD-L1 antibody is durvalumab. Durvalumab is a human IgG1 kappa monoclonal anti-PD-L1 antibody. In one aspect, durvalumab is administered at a dose of about 10 mg / kg about once every two weeks. In one aspect, durvalumab is administered at a dose of about 10 mg / kg about once every two weeks for up to 12 months. In one aspect, durvalumab is administered at a flat dose of about 800 mg / kg about once every two weeks. In one aspect, durvalumab is administered at a flat dose of about 1200 mg / kg about once every three weeks.

[0412] In one aspect, the PD-L1 antibody is avelumab. Avelumab is a human IgG1 lambda monoclonal anti-PD-L1 antibody. In one aspect, avelumab is administered at a fixed dose of about 800 mg approximately once every two weeks.

[0413] III. Pharmaceutical Compositions The LAG-3 antagonists (e.g., anti-LAG-3 antibodies), CTLA-4 inhibitors (e.g., anti-CTLA-4 antibodies), PD-1 pathway inhibitors (e.g., anti-PD-1 antibodies), and / or other therapeutic agents described herein can be comprised in compositions, e.g., one or more pharmaceutical compositions comprising the LAG-3 antagonists, CTLA-4 inhibitors, PD-1 pathway inhibitors, and / or other therapeutic agents described herein, and a pharmaceutically acceptable carrier. The LAG-3 antagonists, CTLA-4 inhibitors, PD-1 pathway inhibitors, and / or other therapeutic agents can be formulated together in any combination or separately. As used herein, "pharmaceutically acceptable carrier" includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like that are physiologically compatible.

[0414] In certain aspects, carriers for compositions comprising a LAG-3 antagonist, CTLA-4 inhibitor, PD-1 pathway inhibitor, and / or other therapeutic agent described herein are suitable for intravenous, intramuscular, subcutaneous, parenteral, spinal, or epidermal administration (e.g., by injection or infusion). In certain aspects, the carrier is suitable for parenteral administration, e.g., oral administration. In certain aspects, subcutaneous injection is based on Halozyme Therapeutics' ENHANZE® drug delivery technology (see U.S. Patent No. 7,767,429, the entire contents of which are incorporated herein by reference). ENHANZE® uses a co-formulation of an antibody with a recombinant human hyaluronidase enzyme (rHuPH20), which removes previous limitations on the amount of biologics and drugs that can be delivered subcutaneously via an extracellular matrix (see U.S. Patent No. 7,767,429). Pharmaceutical compositions of the invention may include one or more pharmaceutically acceptable salts, antioxidants, aqueous and non-aqueous carriers, and / or adjuvants, such as preservatives, wetting agents, emulsifying agents, and dispersing agents. In some aspects, pharmaceutical compositions of the invention may further include a recombinant human hyaluronidase enzyme, such as rHuPH20.

[0415] Treatment continues as long as clinical benefit is observed or until unacceptable toxicity or disease progression occurs. Dosage and frequency vary depending on the half-life of the LAG-3 antagonist, CTLA-4 inhibitor, PD-1 pathway inhibitor, and / or other therapeutic agent in the patient. Generally, for the antibodies described herein, human antibodies exhibit the longest half-life, followed by humanized antibodies, chimeric antibodies, and non-human antibodies. Dosage and frequency vary depending on whether the treatment is preventive or therapeutic. In preventive applications, relatively low doses are generally administered at relatively infrequent intervals over a long period of time. Some patients continue to receive treatment for the rest of their lives. In therapeutic applications, relatively high doses are often required at relatively short intervals until disease progression is suppressed or stopped, preferably until the patient shows partial or complete improvement in disease symptoms. The patient can then receive a preventive regimen.

[0416] The actual dosage level of the active ingredient (e.g., LAG-3 antagonist, CTLA-4 inhibitor, PD-1 pathway inhibitor, and / or other therapeutic agent) in the pharmaceutical compositions of the present invention can be varied to provide an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition, and administration method without being excessively toxic to the patient. The selected dosage level can vary depending on various pharmacokinetic factors, including the activity of the particular composition of the present invention used, the route of administration, the time of administration, the excretion rate of the particular compound used, the duration of treatment, other drugs, compounds, and / or substances used in combination with the particular composition used, the age, sex, weight, condition, general health, and previous medical history of the patient being treated, and similar factors well known in the medical field. The compositions of the present invention can be administered by one or more routes of administration using one or more of a variety of methods well known in the art. As will be understood by those skilled in the art, the route and / or method of administration can vary depending on the desired results.

[0417] IV. Patient Population The present invention provides clinical methods for treating unresectable or metastatic melanoma in human patients comprising an immunotherapeutic agent described herein, e.g., a LAG-3 antagonist (e.g., an anti-LAG-3 antibody), or a combination of a LAG-3 antagonist and a CTLA-4 inhibitor (e.g., an anti-CTLA-4 antibody). In certain aspects, the patient receiving the immunotherapy described herein has a sensitizing mutation to targeted inhibitor therapy. In certain aspects, the patient receiving the immunotherapy described herein has previously received a PD-1 pathway inhibitor for the treatment of melanoma. In one aspect, the prior PD-1 pathway inhibitor is an anti-PD-1 antibody. In one aspect, the patient receives the prior PD-1 pathway inhibitor if the period from the last dose to the date of recurrence is 6 months or more. In another aspect, the patient has unresectable or metastatic melanoma that is refractory to chemotherapy treatment.

[0418] In one aspect, the patient has unresectable or metastatic melanoma that is refractory to treatment with an immune checkpoint inhibitor. In another aspect, the patient has unresectable or metastatic melanoma that is refractory to treatment with a PD-1 inhibitor. In another aspect, the patient has metastatic or unresectable melanoma that is refractory to treatment with an anti-PD-1 antibody. In one aspect, the patient has unresectable or metastatic melanoma that is predicted to be refractory to treatment with an anti-PD-1 antibody. In one aspect, the unresectable or metastatic melanoma is deemed refractory to anti-PD-1 therapy based on biomarker analysis. In a particular aspect, the unresectable or metastatic melanoma is refractory to anti-PD-1 antibody monotherapy. In another aspect, the patient has unresectable or metastatic melanoma that is refractory to treatment with an anti-PD-L1 antibody.

[0419] Patients may be tested or selected for one or more of the above clinical characteristics before, during, or after treatment.

[0420] V. Immunotherapy In one aspect, the immunotherapy provided herein comprises administering a LAG-3 antagonist (e.g., an anti-LAG-3 antibody) or a combination of a LAG-3 antagonist and a CTLA-4 inhibitor (e.g., an anti-CTLA-4 antibody) to treat unresectable or metastatic melanoma.

[0421] In one aspect, the present invention provides an anti-LAG-3 antibody and an anti-CTLA-4 antibody according to a defined clinical dosing regimen for treating patients with metastatic or unresectable melanoma. In a particular aspect, the anti-LAG-3 antibody is BMS-986016 (lilatolimab). In another aspect, the anti-CTLA-4 antibody is ipilimumab. In some aspects, the patient is further administered an anti-PD-1 antibody. In a further aspect, the patient is additionally administered chemotherapy.

[0422] As used herein, adjunctive or combined administration (coadministration) includes simultaneous administration of the compounds in the same or different dosage forms, or separate administration of the compounds (e.g., sequential administration). Thus, for example, an anti-LAG-3 antibody and an anti-CTLA-4 antibody can be administered simultaneously in a single formulation. Alternatively, an anti-LAG-3 antibody and an anti-CTLA-4 antibody can be formulated for separate administration and administered simultaneously or sequentially (e.g., a first antibody is administered within about 30 minutes before the administration of a second antibody).

[0423] For example, the anti-CTLA-4 antibody can be administered first, followed (e.g., immediately after) by the anti-LAG-3 antibody, or vice versa. In one aspect, the anti-CTLA-4 antibody is administered before the administration of the anti-LAG-3 antibody. In another aspect, the anti-CTLA-4 antibody is administered after the administration of the anti-LAG-3 antibody. In another aspect, the anti-LAG-3 antibody and the anti-CTLA-4 antibody are administered simultaneously. Such simultaneous or sequential administration preferably results in both antibodies being present in the treated patient at the same time.

[0424] VI. Treatment Protocol Suitable treatment protocols for the methods described herein include administering to the patient an effective amount of a LAG-3 antagonist (e.g., an anti-LAG-3 antibody) or an effective amount of a LAG-3 antagonist and an effective amount of a CTLA-4 inhibitor (e.g., an anti-CTLA-4 antibody).

[0425] In one aspect, a suitable protocol involves, for example, administering to a patient an effective amount of any anti-LAG-3 antibody of the invention in combination with an effective amount of any anti-CTLA-4 antibody of the invention.

[0426] In one aspect, a suitable treatment protocol includes, for example, administering to a patient an effective amount of each of the following: (a) an anti-LAG-3 antibody, e.g., an antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO:3, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO:5, and (b) an anti-CTLA-4 antibody, e.g., an antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO:34, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO:32.

[0427] In one aspect, a suitable treatment protocol comprises any of the doses described herein for the LAG-3 antagonist and / or CTLA-4 inhibitor.

[0428] In one aspect, a suitable treatment protocol comprises any dosing period (e.g., any dosing cycle) described herein.

[0429] In one aspect, a suitable treatment protocol comprises an anti-LAG-3 antibody at a dose of about 360 mg and an anti-CTLA-4 antibody at a dose of about 3 mg / kg.

[0430] In one aspect, a suitable treatment protocol comprises an anti-LAG-3 antibody at a dose of about 720 mg and an anti-CTLA-4 antibody at a dose of about 3 mg / kg.

[0431] In one aspect, a suitable treatment protocol comprises an anti-LAG-3 antibody at a dose of about 1080 mg and an anti-CTLA-4 antibody at a dose of about 3 mg / kg.

[0432] In one aspect, a suitable treatment protocol comprises an about 1200 mg dose of an anti-LAG-3 antibody and an about 3 mg / kg dose of an anti-CTLA-4 antibody.

[0433] In one aspect, a preferred treatment protocol involves patients who have previously received a PD-1 pathway inhibitor as a treatment for melanoma.

[0434] The present invention provides a method of inhibiting the growth of unresectable or metastatic melanoma in a human patient, the method comprising administering to the patient: (a) an anti-LAG-3 antibody at a dose of about 360 mg, about 720 mg, about 1080 mg, or about 1200 mg; and (b) an anti-CTLA-4 antibody at a dose of about 3 mg / kg. In one aspect, the patient has previously received a PD-1 pathway inhibitor as a treatment for the melanoma. In one aspect, the anti-LAG-3 antibody comprises CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO:3, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO:5, and the anti-CTLA-4 antibody comprises CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO:34, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO:32. In one aspect, the anti-LAG-3 antibody is liratolimab and the anti-CTLA-4 antibody is ipilimumab. In one aspect, the administration cycle is every three weeks (Q3W) and can be repeated as needed.

[0435] The present invention provides a method of treating unresectable or metastatic melanoma in a human patient, the method comprising administering to the patient: (a) an anti-LAG-3 antibody at a dose of about 360 mg, about 720 mg, about 1080 mg, or about 1200 mg; and (b) an anti-CTLA-4 antibody at a dose of about 3 mg / kg. In one aspect, the patient has previously received a PD-1 pathway inhibitor as treatment for the melanoma. In one aspect, the anti-LAG-3 antibody comprises CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO:3, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO:5, and the anti-CTLA-4 antibody comprises CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO:34, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO:32. In one aspect, the anti-LAG-3 antibody is liratolimab and the anti-CTLA-4 antibody is ipilimumab. In one aspect, the administration cycle is every three weeks (Q3W) and can be repeated as needed.

[0436] The present invention provides a method for treating unresectable or metastatic melanoma in a human patient, the method comprising administering to the patient (a) an approximately 360 mg dose of an anti-LAG-3 antibody, and (b) an approximately 3 mg / kg dose of an anti-CTLA-4 antibody.

[0437] The present invention provides a method for treating unresectable or metastatic melanoma in a human patient, the method comprising administering to the patient (a) an approximately 720 mg dose of an anti-LAG-3 antibody, and (b) an approximately 3 mg / kg dose of an anti-CTLA-4 antibody.

[0438] The present invention provides a method for treating unresectable or metastatic melanoma in a human patient, the method comprising administering to the patient (a) an approximately 1080 mg dose of an anti-LAG-3 antibody, and (b) an approximately 3 mg / kg dose of an anti-CTLA-4 antibody.

[0439] The present invention provides a method for treating unresectable or metastatic melanoma in a human patient, the method comprising administering to the patient (a) an approximately 1200 mg dose of an anti-LAG-3 antibody, and (b) an approximately 3 mg / kg dose of an anti-CTLA-4 antibody.

[0440] The present invention provides a method of treating unresectable or metastatic melanoma in a human patient, the method comprising administering to the patient (a) about a 360 mg dose of an anti-LAG-3 antibody, and (b) about a 3 mg / kg dose of an anti-CTLA-4 antibody, wherein the patient has previously received a PD-1 pathway inhibitor as treatment for the melanoma.

[0441] The present invention provides a method of treating unresectable or metastatic melanoma in a human patient, the method comprising administering to the patient (a) about a 720 mg dose of an anti-LAG-3 antibody, and (b) about a 3 mg / kg dose of an anti-CTLA-4 antibody, wherein the patient has previously received a PD-1 pathway inhibitor as a treatment for the melanoma.

[0442] The present invention provides a method of treating unresectable or metastatic melanoma in a human patient, the method comprising administering to the patient (a) about a 1080 mg dose of an anti-LAG-3 antibody, and (b) about a 3 mg / kg dose of an anti-CTLA-4 antibody, wherein the patient has previously received a PD-1 pathway inhibitor as treatment for the melanoma.

[0443] The present invention provides a method of treating unresectable or metastatic melanoma in a human patient, the method comprising administering to the patient (a) about a 1200 mg dose of an anti-LAG-3 antibody, and (b) about a 3 mg / kg dose of an anti-CTLA-4 antibody, wherein the patient has previously received a PD-1 pathway inhibitor as a treatment for the melanoma.

[0444] The present invention provides a method for treating unresectable or metastatic melanoma in a human patient, the method comprising administering to the patient: (a) an anti-LAG-3 antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 3, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO: 5; and (b) an anti-CTLA-4 antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 34, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO: 32, wherein the patient has previously received a PD-1 pathway inhibitor for the treatment of the melanoma.

[0445] The present invention provides a method for treating unresectable or metastatic melanoma in a human patient, the method comprising administering to the patient: (a) an approximately 360 mg dose of an anti-LAG-3 antibody comprising CDR1, CDR2 and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 3, and CDR1, CDR2 and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO: 5; and (b) an approximately 3 mg / kg dose of an anti-CTLA-4 antibody comprising CDR1, CDR2 and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 34, and CDR1, CDR2 and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO: 32.

[0446] The present invention provides a method for treating unresectable or metastatic melanoma in a human patient, the method comprising administering to the patient: (a) an approximately 720 mg dose of an anti-LAG-3 antibody comprising CDR1, CDR2 and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 3, and CDR1, CDR2 and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO: 5; and (b) an approximately 3 mg / kg dose of an anti-CTLA-4 antibody comprising CDR1, CDR2 and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 34, and CDR1, CDR2 and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO: 32.

[0447] The present invention provides a method for treating unresectable or metastatic melanoma in a human patient, the method comprising administering to the patient: (a) an approximately 1080 mg dose of an anti-LAG-3 antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 3, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO: 5; and (b) an approximately 3 mg / kg dose of an anti-CTLA-4 antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 34, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO: 32.

[0448] The present invention provides a method for treating unresectable or metastatic melanoma in a human patient, the method comprising administering to the patient: (a) an approximately 1200 mg dose of an anti-LAG-3 antibody comprising CDR1, CDR2 and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 3, and CDR1, CDR2 and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO: 5; and (b) an approximately 3 mg / kg dose of an anti-CTLA-4 antibody comprising CDR1, CDR2 and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 34, and CDR1, CDR2 and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO: 32.

[0449] The present invention provides a method for treating unresectable or metastatic melanoma in a human patient, the method comprising administering to the patient: (a) an approximately 360 mg dose of an anti-LAG-3 antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 3, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO: 5; and (b) an approximately 3 mg / kg dose of an anti-CTLA-4 antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 34, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO: 32, wherein the patient has previously received a PD-1 pathway inhibitor for treatment of the melanoma.

[0450] The present invention provides a method for treating unresectable or metastatic melanoma in a human patient, the method comprising administering to the patient: (a) an approximately 720 mg dose of an anti-LAG-3 antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 3, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO: 5; and (b) an approximately 3 mg / kg dose of an anti-CTLA-4 antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 34, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO: 32, wherein the patient has previously received a PD-1 pathway inhibitor for the treatment of the melanoma.

[0451] The present invention provides a method for treating unresectable or metastatic melanoma in a human patient, the method comprising administering to the patient: (a) an approximately 1080 mg dose of an anti-LAG-3 antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 3, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO: 5; and (b) an approximately 3 mg / kg dose of an anti-CTLA-4 antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 34, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO: 32, wherein the patient has previously received a PD-1 pathway inhibitor for the treatment of melanoma.

[0452] The present invention provides a method for treating unresectable or metastatic melanoma in a human patient, the method comprising administering to the patient: (a) an approximately 1200 mg dose of an anti-LAG-3 antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 3, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO: 5; and (b) an approximately 3 mg / kg dose of an anti-CTLA-4 antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 34, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO: 32, wherein the patient has previously received a PD-1 pathway inhibitor for treatment of the melanoma.

[0453] The present invention provides a method of treating unresectable or metastatic melanoma in a human patient, the method comprising: (a) administering to a human patient an anti-LAG-3 antibody comprising: (i) a heavy chain variable region CDR1 comprising the sequence set forth in SEQ ID NO:7; (ii) a heavy chain variable region CDR2 comprising the sequence set forth in SEQ ID NO:8; (iii) a heavy chain variable region CDR3 comprising the sequence set forth in SEQ ID NO:9; (iv) a light chain variable region CDR1 comprising the sequence set forth in SEQ ID NO:10; (v) a light chain variable region CDR2 comprising the sequence set forth in SEQ ID NO:11; and (vi) a light chain variable region CDR3 comprising the sequence set forth in SEQ ID NO:12, at a dose of about 360 mg, about 720 mg, about 1080 mg, or about 1200 mg. The method includes administering to the patient an anti-CTLA-4 antibody comprising: (v) a light chain variable region CDR1 comprising the sequence set forth in SEQ ID NO: 38; (v) a light chain variable region CDR2 comprising the sequence set forth in SEQ ID NO: 39; and (vi) a light chain variable region CDR3 comprising the sequence set forth in SEQ ID NO: 40.

[0454] The present invention provides a method of treating unresectable or metastatic melanoma in a human patient, the method comprising: (a) administering to a human patient an anti-LAG-3 antibody comprising: (i) a heavy chain variable region CDR1 comprising the sequence set forth in SEQ ID NO:7; (ii) a heavy chain variable region CDR2 comprising the sequence set forth in SEQ ID NO:8; (iii) a heavy chain variable region CDR3 comprising the sequence set forth in SEQ ID NO:9; (iv) a light chain variable region CDR1 comprising the sequence set forth in SEQ ID NO:10; (v) a light chain variable region CDR2 comprising the sequence set forth in SEQ ID NO:11; and (vi) a light chain variable region CDR3 comprising the sequence set forth in SEQ ID NO:12, at a dose of about 360 mg, about 720 mg, about 1080 mg, or about 1200 mg. and (vi) a light chain variable region CDR3 comprising the sequence set forth in SEQ ID NO: 40, wherein the patient has previously received a PD-1 pathway inhibitor as a treatment for melanoma.

[0455] The present invention provides a method for treating unresectable or metastatic melanoma in a human patient, the method comprising administering to the patient an effective amount of each of: (a) an about 360 mg, about 720 mg, about 1080 mg, or about 1200 mg dose of an anti-LAG-3 antibody comprising a heavy chain variable region and a light chain variable region comprising the sequences set forth in SEQ ID NOs: 3 and 5, respectively; and (b) an about 3 mg / kg dose of an anti-CTLA-4 antibody comprising a heavy chain variable region and a light chain variable region comprising the sequences set forth in SEQ ID NOs: 34 and 32, respectively.

[0456] The present invention provides a method of treating unresectable or metastatic melanoma in a human patient, the method comprising administering to the patient (a) about a 360 mg, about a 720 mg, about a 1080 mg, or about a 1200 mg dose of an anti-LAG-3 antibody comprising a heavy chain variable region and a light chain variable region comprising the sequences set forth in SEQ ID NOs: 3 and 5, respectively, and (b) about a 3 mg / kg dose of an anti-CTLA-4 antibody comprising a heavy chain variable region and a light chain variable region comprising the sequences set forth in SEQ ID NOs: 34 and 32, respectively, wherein the patient has previously received a PD-1 pathway inhibitor for treatment of the melanoma.

[0457] In one aspect, the anti-LAG-3 antibody and the anti-CTLA-4 antibody are administered about once every week, about once every 2 weeks, about once every 3 weeks, about once every 4 weeks, about once every 5 weeks, about once every 6 weeks, about once every 7 weeks, about once every 8 weeks, about once every 9 weeks, about once every 10 weeks, about once every 11 weeks, or about once every 12 weeks.

[0458] In one aspect, the anti-LAG-3 antibody and the anti-CTLA-4 antibody are administered about once every three weeks.

[0459] The present invention provides a method for treating unresectable or metastatic melanoma in a human patient, the method comprising administering to the patient an anti-LAG-3 antibody comprising: (a) a heavy chain variable region CDR1 comprising the sequence set forth in SEQ ID NO:7; (b) a heavy chain variable region CDR2 comprising the sequence set forth in SEQ ID NO:8; (c) a heavy chain variable region CDR3 comprising the sequence set forth in SEQ ID NO:9; (d) a light chain variable region CDR1 comprising the sequence set forth in SEQ ID NO:10; (e) a light chain variable region CDR2 comprising the sequence set forth in SEQ ID NO:11; and (f) a light chain variable region CDR3 comprising the sequence set forth in SEQ ID NO:12, wherein the patient has previously received a PD-1 pathway inhibitor as a treatment for the melanoma; and at least one dose of the anti-LAG-3 antibody is administered at a dose of about 360 mg, about 720 mg, about 1080 mg, or about 1200 mg.

[0460] In one aspect, the anti-LAG-3 antibody is administered about once every week, about once every 2 weeks, about once every 3 weeks, about once every 4 weeks, about once every 5 weeks, about once every 6 weeks, about once every 7 weeks, about once every 8 weeks, about once every 9 weeks, about once every 10 weeks, about once every 11 weeks, or about once every 12 weeks.

[0461] In one aspect, the anti-LAG-3 antibody is administered about once every three weeks.

[0462] The present invention provides a method for treating unresectable or metastatic melanoma in a human patient, the method comprising administering to the patient (a) one dose of an anti-LAG-3 antibody, wherein the anti-LAG-3 antibody is AGEN1746, and (b) one dose of an anti-CTLA-4 antibody, wherein the anti-CTLA-4 antibody is AGEN1884, wherein the patient has previously received a PD-1 pathway inhibitor as treatment for the melanoma.

[0463] The present invention provides a method of treating unresectable or metastatic melanoma in a human patient, the method comprising administering to the patient (a) one dose of an anti-LAG-3 antibody, wherein the anti-LAG-3 antibody is MK4280, and (b) one dose of an anti-CTLA-4 antibody, wherein the anti-CTLA-4 antibody is MK1308, wherein the patient has previously received a PD-1 pathway inhibitor as treatment for the melanoma.

[0464] The present invention provides a method of treating unresectable or metastatic melanoma in a human patient, the method comprising administering to the patient (a) one dose of an anti-LAG-3 antibody, where the anti-LAG-3 antibody is REGN3767, and (b) one dose of an anti-CTLA-4 antibody, where the anti-CTLA-4 antibody is REGN4659, wherein the patient has previously received a PD-1 pathway inhibitor as treatment for the melanoma.

[0465] In one aspect, the anti-LAG-3 antibody and the anti-CTLA-4 antibody are formulated for intravenous administration.

[0466] In one aspect, the administration cycle is about once every three weeks and can be repeated as needed.

[0467] In one aspect, the anti-LAG-3 antibody is BMS-986016 and the anti-CTLA-4 antibody is ipilimumab.

[0468] In one aspect, the anti-LAG-3 antibody is MK4280.

[0469] In one aspect, the anti-LAG-3 antibody is REGN3767.

[0470] In one aspect, the anti-LAG-3 antibody is LAG525.

[0471] In one aspect, the anti-CTLA-4 antibody is a bispecific antibody.

[0472] In one aspect, the anti-LAG-3 antibody is a bispecific antibody.

[0473] In one aspect, the bispecific antibody binds to both CTLA-4 and LAG-3. In one aspect, the CTLA-4 / LAG-3 bispecific antibody is XmAb22841.

[0474] In one aspect, the bispecific antibody binds to both PD-1 and LAG-3. In one aspect, the PD-1 / LAG-3 bispecific antibody is TSR-075. In one aspect, the PD-1 / LAG-3 bispecific antibody is MGD013.

[0475] In one aspect, the antibody is a PD-L1 / LAG-3 bispecific antibody. In one aspect, the PD-L1 / LAG-3 bispecific antibody is FS-118.

[0476] In some aspects, in any of the methods described herein, the patient is not further administered an anti-PD-1 antibody.

[0477] VII. Results Patients treated according to the methods described herein preferably experience an improvement in at least one symptom of melanoma. In one aspect, the improvement is measured by a decrease in the amount and / or size of measurable tumor lesions. In another aspect, the lesions can be measured by chest X-ray or CT or MRI film. In another aspect, cytology or histology can be used to evaluate response to treatment.

[0478] In one aspect, treated patients exhibit a complete response (CR), partial response (PR), stable disease (SD), immune-related complete response (irCR), immune-related partial response (irPR), or immune-related stable disease (irSD). In another aspect, treated patients experience tumor shrinkage and / or a reduced growth rate, i.e., suppression of tumor growth. In another aspect, unwanted cell proliferation is reduced or inhibited. In yet another aspect, one or more of the following may occur: the number of cancer cells may be reduced; tumor size may be reduced; cancer cell invasion to peripheral organs may be inhibited, delayed, slowed, or halted; tumor metastasis may be delayed or inhibited; tumor growth may be suppressed; tumor recurrence may be prevented or delayed; or one or more symptoms associated with cancer may be alleviated to some extent.

[0479] In another aspect, administration of an effective amount of an anti-LAG-3 antibody or an anti-LAG-3 antibody and an anti-CTLA-4 antibody according to any of the methods of the invention results in at least one therapeutic effect selected from the group consisting of a reduction in tumor size, a reduction in the number of metastatic lesions appearing over time, a complete remission, a partial remission, or stable disease.

[0480] In yet another aspect, the treatment method results in a clinical benefit rate (CBR=CR+PR+SD≧6 months) that is superior to that achieved by a treatment method that does not include (i) determining LAG-3, PD-L1 and / or BRAF V600 expression levels in a tumor sample prior to treatment, and (ii) treating the tumor. In other aspects, the improvement in clinical benefit rate is about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80% or more compared to a treatment method that does not include (i) determining LAG-3, PD-L1 and / or BRAF V600 expression levels in a tumor sample prior to treatment, and (ii) treating the tumor. In yet other aspects, the treatment method results in an objective response rate (ORR=CR+PR) of at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or about 100%.

[0481] VIII. Kits and Unit Dosage Forms Also within the scope of the present invention are diagnostic kits comprising an anti-LAG-3 antibody for assaying LAG-3 expression as a biomarker for screening patients for immunotherapy or predicting the efficacy of immunotherapy. The kit typically includes a label and instructions indicating the intended use of the contents of the kit. The term "label" includes any written or recorded material that accompanies or is included with the kit, or that is otherwise contained in the kit. In certain aspects of the diagnostic kit, a first anti-LAG-3 antibody for assaying, detecting, and / or quantifying LAG-3 expression and at least one therapeutic antibody for the treatment of unresectable or metastatic melanoma (e.g., a second anti-LAG-3 antibody, or a second anti-LAG-3 antibody and an anti-CTLA-4 antibody) are packaged together in a kit. In one aspect, the kit further includes an anti-PD-L1 antibody for assaying, detecting, and / or quantifying PD-L1 expression as a biomarker for predicting the efficacy of immunotherapy. In one aspect, the immunotherapy comprises administering to the patient therapeutically effective amounts of a LAG-3 antagonist (e.g., an anti-LAG-3 antibody and, e.g., an anti-PD1 antibody) and a CTLA-4 inhibitor (e.g., an anti-CTLA-4 antibody).

[0482] In certain aspects, the diagnostic kit comprises an anti-human LAG-3 monoclonal antibody for assaying, detecting, and / or quantifying LAG-3 expression. See, e.g., J. Matsuzaki, et al.; PNAS 107, 7875 (2010).

[0483] The present invention also provides a therapeutic kit comprising an anti-LAG-3 antibody, e.g., BMS-986016, or a pharmaceutical composition comprising an anti-LAG-3 antibody, e.g., BMS-986016, and an anti-CTLA-4 antibody, e.g., ipilimumab, in a therapeutically effective amount suitable for use in any of the methods of the present invention. In certain aspects of the therapeutic kit, the anti-LAG-3 antibody is packaged together with the anti-CTLA-4 antibody in a unit dosage form. The kit may also optionally include instructions, including, for example, an administration schedule, to enable a practitioner (e.g., a doctor, a nurse, or a patient) to administer the composition contained therein to a patient with cancer (e.g., a solid tumor). The kit may also include a syringe.

[0484] Optionally, the diagnostic and / or therapeutic kit includes multiple packages of single-dose pharmaceutical compositions each containing an effective amount of an anti-LAG-3 antibody or anti-CTLA-4 antibody for a single administration according to any of the methods of the present invention. Devices or instruments necessary for administering the pharmaceutical composition(s) may also be included in the kit. For example, the kit may provide one or more pre-filled syringes containing a quantity of an anti-LAG-3 antibody or anti-CTLA-4 antibody.

[0485] In one aspect, the present invention provides a kit for treating a patient with unresectable or metastatic melanoma, comprising: (a) a dose of an anti-LAG-3 antibody, e.g., an antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 3, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO: 5; (b) a dose of an anti-CTLA-4 antibody, e.g., an antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 34, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO: 32; and (c) instructions for using the anti-LAG-3 antibody and the anti-CTLA-4 antibody in any of the methods described herein.

[0486] This invention is further illustrated by the following examples, which should not be construed as further limiting. The contents of all references cited herein are expressly incorporated by reference. [Example]

[0487] Example Example 1 Efficacy of anti-lymphocyte-activation gene-3 antibody (anti-LAG-3; BMS-986016) in combination with ipilimumab in patients with metastatic or unresectable melanoma The purpose of this trial is to evaluate the combination of BMS-986016 (leratolimab) and ipilimumab in the treatment of metastatic or unresectable melanoma.

[0488] Patients will be selected based on the following eligibility criteria: (1) documented progression on prior anti-programmed cell death protein 1 (PD-1)-containing regimens limited to nivolumab or pembrolizumab; (2) women of childbearing potential (WOCBP) must have a negative serum or urine pregnancy test; (3) patients must have histologically confirmed advanced unresectable (stage III) or metastatic (stage IV) melanoma according to the AJCC staging system; (4) must provide tumor tissue from unresectable or metastatic disease sites for biomarker analysis; (5) must provide tumor tissue from unresectable or metastatic disease sites for biomarker analysis; (6) have an Eastern Cooperative Oncology Group (ECOG) performance status of 0-1; and (7) be able to comply with treatment, patient-reported outcomes (PROs), PK, pharmacodynamic sample collection, and required study follow-up. Patients will be selected based on the following exclusion criteria: (1) a history of uveal melanoma; (2) a positive human immunodeficiency virus (HIV) test or a history of acquired immunodeficiency syndrome; (3) a history of treatment with ipilimumab, latolimab, or other CTLA-4 or LAG-3 targeted agents; and (4) a positive blood screen for hepatitis C antibodies, hepatitis B surface antigen, or antibodies to HIV-1 and HIV-2.

[0489] During treatment, patients will receive BMS-986016 (lilatolimab) at doses of 360 mg, 720 mg, 1080 mg, or 1200 mg and ipilimumab at a dose of 3 mg / kg every 3 weeks in each treatment cycle.

[0490] Sequence Listing SEQ ID NO: 1: Heavy chain amino acid sequence; anti-LAG-3 mAb (BMS-986016) QVQLQQWGAGLLKPSETLSLTCAVYGGSFSDYYWNWIRQPPGKGLEWIGEINHRGSTNSNPSLKSRVTLSLDTSKNQFSLKLRSVTAADTAVYYCAFGYSDYEYNWFDPWG QGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYG PPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKT ISKAKGQPREPQVYTLPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK SEQ ID NO: 2 Light chain amino acid sequence; anti-LAG-3 mAb (BMS-986016) EIVLTQSPATLSLSPGERATLSCRASQSISSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPLTFGQGTNLEIK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 3: Heavy chain variable region (VH) amino acid sequence; anti-LAG-3 mAb (BMS-986016) QVQLQQWGAGLLKPSETLSLTCAVYGGSFSDYYWNWIRQPPGKGLEWIGEINHRGSTNSNPSLKSRVTLSLDTSKNQFSLKLRSVTAADTAVYYCAFGYSDYEYNWFDPWGQGTLVTVSS SEQ ID NO: 4: Heavy chain variable region (VH) nucleotide sequence; anti-LAG-3 mAb (BMS-986016) caggtgcagctacagcagtggggcgcaggactgttgaagccttcggagaccctgtccctcacctgcgctgtctatggtgggtccttcagtgattactactggaactggatccgccagcccccagggaaggggctggagtggattggggaaatcaatcatcgtggaagcaccaactccaac ccgtccctcaagagtcgagtcaccctatcactagacacgtccaagaaccagttctccctgaagctgaggtctgtgaccgccgcggacacggctgtgtattactgtgcgtttggatatagtgactacgagtacaactggttcgacccctggggccagggaaccctggtcaccgtctcctca SEQ ID NO: 5: Light chain variable region (VL) amino acid sequence; anti-LAG-3 mAb (BMS-986016) EIVLTQSPATLSLSPGERATLSCRASQSISSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPLTFGQGTNLEIK SEQ ID NO: 6: Light chain variable region (VL) nucleotide sequence; anti-LAG-3 mAb (BMS-986016) gaaattgtgttgacacagtctccagccaccctgtctttgtctccaggggaaagagccaccctctcctgcagggccagtcagagtattagcagctacttagcctggtaccaacagaaacctggccaggctcccaggctcctcatctatgatgcatccaaca gggccactggcatcccagccaggttcagtggcagtgggtctgggacagacttcactctcaccatcagcagcctagagcctgaagattttgcagtttattactgtcagcagcgtagcaactggcctctcacttttggccaggggaccaacctggagatcaaa SEQ ID NO: 7: Heavy chain CDR1 amino acid sequence; anti-LAG-3 mAb (BMS-986016) DYYWN SEQ ID NO: 8: Heavy chain CDR2 amino acid sequence; anti-LAG-3 mAb (BMS-986016) EINHRGSTNSNPSLKS SEQ ID NO: 9 Heavy chain CDR3 amino acid sequence; anti-LAG-3 mAb (BMS-986016) GYSDYEYNWFDP SEQ ID NO: 10 Light chain CDR1 amino acid sequence; anti-LAG-3 mAb (BMS-986016) RASQSISSYLA SEQ ID NO: 11 Light chain CDR2 amino acid sequence; anti-LAG-3 mAb (BMS-986016) DASNRAT SEQ ID NO: 12 Light chain CDR3 amino acid sequence; anti-LAG-3 mAb (BMS-986016) QQRSNWPLT SEQ ID NO: 13: Heavy chain amino acid sequence; anti-PD-1 mAb (BMS936558) QVQLVESGGGVVQPGRSLRLDCKASGITFSNSGMHWVRQAPGKGLEWVAVIWYDGSKRYYADSVKGRFTISRDNSKNTLFLQMNSLRAEDTAVYYCATNDDYWGQGTLVT VSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCP PCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTIS KAKGQPREPQVYTLPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK SEQ ID NO: 14 Light chain amino acid sequence; anti-PD-1 mAb (BMS936558) EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQSSNWPRTFGQGTKVEI KRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 15: Heavy chain variable region (VH) amino acid sequence; anti-PD-1 mAb (BMS936558) QVQLVESGGGVVQPGRSLRLDCKASGITFSNSGMHWVRQAPGKGLEWVAVIWYDGSKRYYADSVKGRFTISRDNSKNTLFLQMNSLRAEDTAVYYCATNDDYWGQGTLVTVSS SEQ ID NO: 16: Heavy chain variable region (VH) nucleotide sequence; anti-PD-1 mAb (BMS936558) caggtgcagctggtggagtctgggggaggcgtggtccagcctgggaggtccctgagactcgactgtaaagcgtctggaatcaccttcagtaactctggcatgcactgggtccgccaggctccaggcaaggggctggagtgggtggcagttatttggtatgatggaagta aaagatactatgcagactccgtgaagggccgattcaccatctccagagacaattccaagaacacgctgtttctgcaaatgaacagcctgagagccgaggacacggctgtgtattactgtgcgacaaacgacgactactggggccagggaaccctggtcaccgtctcctca SEQ ID NO: 17: Light chain variable region (VL) amino acid sequence; anti-PD-1 mAb (BMS936558) EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQSSNWPRTFGQGTKVEIK SEQ ID NO: 18: Light chain variable region (VL) nucleotide sequence; anti-PD-1 mAb (BMS936558) gaaattgtgttgacacagtctccagccaccctgtctttgtctccaggggaaagagccaccctctcctgcagggccagtcagagtgttagtagttacttagcctggtaccaacagaaacctggccaggctcccaggctcctcatctatgatgcatccaaca gggccactggcatcccagccaggttcagtggcagtgggtctgggacagacttcactctcaccatcagcagcctagagcctgaagattttgcagtttattactgtcagcagagtagcaactggcctcggacgttcggccaagggaccaaggtggaaatcaaa SEQ ID NO: 19: Heavy chain CDR1 amino acid sequence; anti-PD-1 mAb (BMS936558) NSGMH SEQ ID NO: 20: Heavy chain CDR2 amino acid sequence; anti-PD-1 mAb (BMS936558) VIWYDGSKRYYADSVKG SEQ ID NO: 21: Heavy chain CDR3 amino acid sequence; anti-PD-1 mAb (BMS936558) NDDY SEQ ID NO: 22 Light chain CDR1 amino acid sequence; anti-PD-1 mAb (BMS936558) RASQSVSSYLA SEQ ID NO: 23 Light chain CDR2 amino acid sequence; anti-PD-1 mAb (BMS936558) DASNRAT SEQ ID NO: 24 Light chain CDR3 amino acid sequence; anti-PD-1 mAb (BMS936558) QQSSNWPRT SEQ ID NO: 25 Heavy chain nucleotide sequence; anti-LAG-3 mAb (BMS-986016) SEQ ID NO: 26 Light chain nucleotide sequence; anti-LAG-3 mAb (BMS-986016) gaaattgtgttgacacagtctccagccaccctgtctttgtctccaggggaaagagccaccctctcctgcagggccagtcagagtattagcagctacttagcctggtaccaacagaaacctggccaggctcccaggctcctcatctatgatgcatccaacag ggccactggcatcccagccaggttcagtggcagtgggtctgggacagacttcactctcaccatcagcagcctagagcctgaagattttgcagtttattactgtcagcagcgtagcaactggcctctcacttttggccaggggaccaacctggagatcaaac gtacggtggctgcaccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggag agtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgttag SEQ ID NO: 27 LAG-3 epitope PGHPLAPG SEQ ID NO: 28 LAG-3 epitope HPAAPSSW SEQ ID NO: 29 LAG-3 epitope PAAPSSWG SEQ ID NO: 30 Heavy chain amino acid sequence; anti-LAG-3 mAb lacking terminal lysine (BMS-986016) QVQLQQWGAGLLKPSETLSLTCAVYGGSFSDYYWNWIRQPPGKGLEWIGEINHRGSTNSNPSLKSRVTLSLDTSKNQFSLKLRSVTAADTAVYYCAFGYSDYEYNWFDPWG QGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYG PPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEK TISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 31 Light chain variable region (Vk), 10D1 from Vk A-27 gaaattgtgttgacgcagtctccaggcaccctgtctttgtctccaggggaaagagccaccctctcctgcagggccagtcagagtgttggcagcagctacttagcctggtaccagcagaaacctggccaggctcccaggctcctcatctatggtgcattcagc agggccactggcatcccagacaggttcagtggcagtgggtctgggacagacttcactctcaccatcagcagactggagcctgaagattttgcagtgtattactgtcagcagtatggtagctcaccgtggacgttcggccaagggaccaaggtggaaatcaaac SEQ ID NO: 32 Predicted sequence of the light chain variable region of 10D1 EIVLTQSPGTLSLSPGERATLSCRASQSVGSSYLAWYQQKPGQAPRLLIYGAFSRATGIP DRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSSPWTFGQGTKVEIK SEQ ID NO: 33 Heavy chain variable region 10D1 caggtgcagctggtggagtctgggggaggcgtggtccagcctgggaggtccctgagactctcctgtgcagcctctggattcaccttcagtagctatactatgcactgggtccgccaggctccaggcaaggggctggagtgggtgacatttatatcatatgatggaaacaataaatac tacgcagactccgtgaagggccgattcaccatctccagagacaattccaagaacacgctgtatctgcaaatgaacagcctgagagctgaggacacggctatatattactgtgcgaggaccggctggctggggccctttgactactggggccagggaaccctggtcaccgtctcctcag SEQ ID NO: 34 Predicted sequence of the heavy chain variable region of 10D1 from VH 3-30.3 QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYTMHWVRQAPGKGLEWVTFISYDGNNKYY ADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAIYYCARTGWLGPFDYWGQGTLVTVSS SEQ ID NO: 35 10D1 HC CDR1 SYTMH SEQ ID NO: 36 10D1 HC CDR2 FISYDGNNKYYADSVKG SEQ ID NO: 37 10D1 HC CDR3 TGWLGPFDY SEQ ID NO: 38 10D1 LC CDR1 RASQSVGSSYLA SEQ ID NO: 39 10D1 LC CDR2 GAFSRAT SEQ ID NO: 40 10D1 LC CDR3 QQYGSSPWT SEQ ID NO: 41 Lymphocyte activation gene 3 protein amino acid sequence (human, NP_002277) MWEAQFLGLLFLQPLWVAPVKPLQPGAEVPVVWAQEGAPAQLPCSPTIPLQDLSLLRRAGVTWQHQPDSGPPAAAPGHPLAPGPHPAAPSSWGPRPRRYTVLSVGPGGLRSGRLPLQPRVQLDERGRQRGDFSLWLRPARRADAGEYRAAVHLRDRALSCRLRLRLGQASMTASPPGSLRASDWVILNCSFSRPDRPASVHWFRNRGQGRVPVRESPHHHLAESFLFLPQVSPMDSGPWGCILTYRDGFNVSIMYNLTVLGLEPPTPLTVYAGAGSRVGLPCRLPAGVGTRSFLTAKWTPPGGGPDLLVTGDNGDFTLRLEDVSQAQAGTYTCHIHLQEQQLNATVTLAIITVTPKSFGSPGSLGKLLCEVTPVSGQERFVWSSLDTPSQRSFSGPWLEAQEAQLLSQPWQCQLYQGERLLGAAVYFTELSSPGAQRSGRAPGALPAGHLLLFLILGVLSLLLLVTGAFGFHLWRRQWRPRRFSALEQGIHPPQAQSKIEELEQEPEPEPEPEPEPEPEPEPEQL

Claims

1. 1. A method of treating unresectable or metastatic melanoma in a human patient, comprising: (a) lymphocyte activation gene-3 (LAG-3) antagonists, and (b) Cytotoxic T lymphocyte antigen-4 (CTLA-4) inhibitor to said patient, wherein the patient has a mutation that sensitizes to targeted inhibitor therapy.

2. 1. A method for treating unresectable or metastatic melanoma in a human patient, comprising: (a) a LAG-3 antagonist, and (b) CTLA-4 inhibitor to said patient, wherein the patient has previously been administered a PD-1 pathway inhibitor as a treatment for melanoma.

3. The method of claim 1, which is a first-line therapy.

4. 3. The method of claim 1 or 2, which is a second-line therapy.

5. 3. The method of claim 1 or 2, which is a third line therapy.

6. 6. The method of claim 2, 4 or 5, wherein the patient has had progression on prior therapy.

7. 7. The method of any one of claims 1-6, wherein the patient has not received prior systemic therapy for cancer, the patient has not received prior systemic therapy for melanoma, or the patient has not received prior systemic therapy for unresectable or metastatic melanoma.

8. 8. The method of any one of claims 1 to 7, wherein the patient is naive to a previous immuno-oncology therapy, the patient is naive to a previous immuno-oncology therapy for the melanoma, or the melanoma is naive to a previous immuno-oncology therapy.

9. 9. The method of any one of claims 1 to 8, wherein the patient has histologically confirmed unresectable stage III or stage IV melanoma.

10. 10. The method of any one of claims 1 to 9, wherein the patient has an Eastern Cooperative Oncology Group (ECOG) performance status of 0 or 1.

11. 11. The method of any one of claims 1 to 10, wherein the patient has a B-type aggressive fibrosarcoma proto-oncogene (BRAF), mitogen-activated extracellular signal-regulated kinase kinase (MEK), neuroblastoma RAS viral oncogene homolog (NRAS), and / or proto-oncogene c-KIT (KIT) mutation that is susceptible to targeted inhibitor therapy.

12. 12. The method of any one of claims 1 to 11, wherein the patient has a BRAF mutation that is susceptible to targeted inhibitor therapy.

13. The method of any one of claims 1 to 12, wherein one or more immune cells in the tumor tissue from the patient express LAG-3.

14. 14. The method of claim 13, wherein at least about 1%, at least about 3%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or about 100% of the immune cells express LAG-3.

15. The method of claim 13 or 14, wherein at least about 1% of immune cells express LAG-3.

16. 16. The method of any one of claims 13 to 15, wherein the immune cells are tumor-infiltrating lymphocytes.

17. Tumor-infiltrating lymphocytes are CD8 ＋ The method of claim 16, wherein the cell is a cell.

18. The method of any one of claims 1 to 17, wherein one or more tumor cells in the tumor tissue from the patient express PD-L1.

19. 19. The method of claim 18, wherein at least about 1%, at least about 3%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or about 100% of the tumor cells express PD-L1.

20. 20. The method of claim 18 or 19, wherein at least about 1% of the tumor cells express PD-L1.

21. The method of any one of claims 1 to 20, wherein the LAG-3 antagonist is an anti-LAG-3 antibody.

22. The method of claim 21, wherein the anti-LAG-3 antibody is a full-length antibody.

23. The method of claim 21 or 22, wherein the anti-LAG-3 antibody is a monoclonal antibody, a human antibody, a humanized antibody, a chimeric antibody, or a multispecific antibody.

24. 24. The method of claim 23, wherein the multispecific antibody is a dual affinity retargeting antibody (DART), a DVD-binding protein (DVD-Ig), or a bispecific antibody.

25. The anti-LAG-3 antibody is F(ab') ２ 22. The method of claim 21, wherein the antibody is a fragment, a Fab' fragment, a Fab fragment, an Fv fragment, a scFv fragment, a dsFv fragment, a dAb fragment or a single chain binding polypeptide peptide.

26. 26. The method of any one of claims 21 to 25, wherein the anti-LAG-3 antibody is BMS-986016 (leratolimab), IMP731 (H5L7BW), MK-4280 (28G-10), REGN3767 (fianlimab), GSK2831781, humanized BAP050, IMP-701 (LAG-525, yelamirimab), aLAG3(0414), aLAG3(0416), Sym022, TSR-033, TSR-075, XmAb22841, MGD013, BI754111, FS118, P 13B02-30, AVA-017, 25F7, AGEN1746, or an antigen-binding portion thereof.

27. 27. The method of any one of claims 21 to 26, wherein the anti-LAG-3 antibody comprises the CDR1 domain, CDR2 domain, and CDR3 domain of a heavy chain variable region having the sequence set forth in SEQ ID NO:3, and the CDR1 domain, CDR2 domain, and CDR3 domain of a light chain variable region having the sequence set forth in SEQ ID NO:

5.

28. The anti-LAG-3 antibody (a) a heavy chain variable region CDR1 comprising the sequence set forth in SEQ ID NO:7; (b) a heavy chain variable region CDR2 comprising the sequence set forth in SEQ ID NO:8; (c) a heavy chain variable region CDR3 comprising the sequence set forth in SEQ ID NO:9; (d) a light chain variable region CDR1 comprising the sequence set forth in SEQ ID NO: 10; (e) a light chain variable region CDR2 comprising the sequence set forth in SEQ ID NO: 11; and (f) a light chain variable region CDR3 comprising the sequence set forth in SEQ ID NO: 12 28. The method of any one of claims 21 to 27, comprising:

29. 29. The method of any one of claims 21 to 28, wherein the anti-LAG-3 antibody comprises a heavy chain variable region and a light chain variable region comprising the sequences set forth in SEQ ID NOs: 3 and 5, respectively.

30. 30. The method of any one of claims 21 to 24 and 26 to 29, wherein the anti-LAG-3 antibody comprises a heavy chain and a light chain comprising the sequences set forth in SEQ ID NOs: 1 and 2, respectively.

31. 30. The method of any one of claims 21 to 24 and 26 to 29, wherein the anti-LAG-3 antibody comprises a heavy chain and a light chain comprising the sequences set forth in SEQ ID NOs: 30 and 2, respectively.

32. 21. The method of any one of claims 1 to 20, wherein the LAG-3 antagonist is a soluble LAG-3 polypeptide.

33. 33. The method of claim 32, wherein the soluble LAG-3 polypeptide is a fusion polypeptide.

34. The method of claim 32 or 33, wherein the soluble LAG-3 polypeptide comprises a ligand-binding fragment of the LAG-3 extracellular domain.

35. 35. The method of claim 34, wherein the ligand-binding fragment of the LAG-3 extracellular domain comprises an amino acid sequence having at least about 90%, at least about 95%, at least about 98%, at least about 99%, or about 100% sequence identity to SEQ ID NO:

41.

36. 36. The method of any one of claims 32 to 35, wherein the soluble LAG-3 polypeptide further comprises a half-life extending moiety.

37. 37. The method of claim 36, wherein the half-life extending moiety comprises an immunoglobulin constant region or a portion thereof, an immunoglobulin-binding polypeptide, immunoglobulin G (IgG), an albumin-binding polypeptide (ABP), a PAS-modified moiety, a HES-modified moiety, XTEN, a PEGylated moiety, an Fc region, or any combination thereof.

38. 38. The method of any one of claims 32 to 37, wherein the soluble LAG-3 polypeptide is IMP321 (eftilagimod alfa).

39. 39. The method of any one of claims 1 to 38, wherein the CTLA-4 inhibitor is an anti-CTLA-4 antibody.

40. 40. The method of claim 39, wherein the anti-CTLA-4 antibody is a full-length antibody.

41. The method of claim 39 or 40, wherein the anti-CTLA-4 antibody is a monoclonal antibody, a human antibody, a humanized antibody, a chimeric antibody, or a multispecific antibody.

42. 42. The method of claim 41, wherein the multispecific antibody is a DART, a DVD-Ig, or a bispecific antibody.

43. The anti-CTLA-4 antibody is an F(ab') ２ 40. The method of claim 39, wherein the antibody is a fragment, a Fab' fragment, a Fab fragment, an Fv fragment, a scFv fragment, a dsFv fragment, a dAb fragment or a single chain binding polypeptide peptide.

44. 44. The method of any one of claims 39 to 43, wherein the anti-CTLA-4 antibody is ipilimumab, tremelimumab, MK-1308, AGEN-1884, or an antigen-binding portion thereof.

45. 45. The method of any one of claims 39 to 44, wherein the anti-CTLA-4 antibody comprises the CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 34, and the CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO:

32.

46. The anti-CTLA-4 antibody (a) a heavy chain variable region CDR1 comprising the sequence set forth in SEQ ID NO: 35; (b) a heavy chain variable region CDR2 comprising the sequence set forth in SEQ ID NO: 36; (c) a heavy chain variable region CDR3 comprising the sequence set forth in SEQ ID NO: 37; (d) a light chain variable region CDR1 comprising the sequence set forth in SEQ ID NO: 38; (e) a light chain variable region CDR2 comprising the sequence set forth in SEQ ID NO: 39; and (f) a light chain variable region CDR3 comprising the sequence set forth in SEQ ID NO: 40 46. ​​The method of any one of claims 39 to 45, comprising:

47. 47. The method of any one of claims 39 to 46, wherein the anti-CTLA-4 antibody comprises a heavy chain variable region and a light chain variable region comprising the sequences set forth in SEQ ID NOs: 34 and 32, respectively.

48. 48. The method of any one of claims 39-42 and 44-47, wherein the anti-CTLA-4 antibody comprises the heavy and light chains of ipilimumab.

49. 49. The method of any one of claims 1 to 48, wherein the LAG-3 antagonist and the CTLA-4 inhibitor are formulated for intravenous administration.

50. 50. The method of any one of claims 1 to 49, wherein the LAG-3 antagonist and the CTLA-4 inhibitor are formulated separately.

51. 50. The method of any one of claims 1 to 49, wherein the LAG-3 antagonist and the CTLA-4 inhibitor are formulated as a single agent.

52. 51. The method of any one of claims 1 to 50, wherein the LAG-3 antagonist is administered prior to the CTLA-4 inhibitor.

53. 51. The method of any one of claims 1 to 50, wherein the CTLA-4 inhibitor is administered prior to the LAG-3 antagonist.

54. 51. The method of any one of claims 1 to 50, wherein the LAG-3 antagonist and the CTLA-4 inhibitor are administered simultaneously.

55. 55. The method of any one of claims 1 to 54, wherein the LAG-3 antagonist and / or CTLA-4 inhibitor is administered in a fixed dose.

56. and / or the LAG-3 antagonist and / or CTLA-4 inhibitor is at least about 0.25 mg to about 2000 mg, about 0.25 mg to about 1600 mg, about 0.25 mg to about 1200 mg, about 0.25 mg to about 800 mg, about 0.25 mg to about 400 mg, about 0.25 mg to about 100 mg, about 0.25 mg to about 50 mg, about 0.25 mg to about 40 mg, about 0.25 mg to about 30 mg, about 0.25 mg to about 20 mg, about 20 mg to about 2000 mg, about 20 mg to about 1600 mg, about 20 mg to about 1200 mg, about 20 mg to about 800 mg, about 20 mg to about 400 mg, about 20 mg to about 1 56. The method of any one of claims 1 to 55, wherein the compound is administered at a dose of about 100 mg, about 100 mg to about 2000 mg, about 100 mg to about 1800 mg, about 100 mg to about 1600 mg, about 100 mg to about 1400 mg, about 100 mg to about 1200 mg, about 100 mg to about 1000 mg, about 100 mg to about 800 mg, about 100 mg to about 600 mg, about 100 mg to about 400 mg, 400 mg to about 2000 mg, about 400 mg to about 1800 mg, about 400 mg to about 1600 mg, about 400 mg to about 1400 mg, about 400 mg to about 1200 mg, or about 400 mg to about 1000 mg.

57. A LAG-3 antagonist and / or a CTLA-4 inhibitor is about 0.25 mg, about 0.5 mg, about 0.75 mg, about 1 mg, about 1.25 mg, about 1.5 mg, about 1.75 mg, about 2 mg, 2.25 mg, about 2.5 mg, about 2.75 mg, about 3 mg, about 3.25 mg, about 3.5 mg, about 3.75 mg, about 4 mg, about 4.25 mg, about 4.5 mg, about 4.75 mg, about 5 mg, about 5.25 mg, about 5.5 mg, about 5.75 mg, about 6 mg, about 6.25 mg, about 6.5 mg, about 6.75 mg, about 7 mg, about 7.25 mg, about 7.5 mg, about 7.75 mg, about 8 mg, about 8.25 mg, about 8.5 mg, about 8.75 mg, about 9 mg, about 9.25 mg, about 9.5 mg, about 9.75 mg, about 10 mg, about 20 mg, about 30 mg, about 40 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, about 100 mg, about 110 mg, about 120 mg, about 130 mg, about 140 mg, about 150 mg, about 160 mg, about 170 mg, about 180 mg, about 190 mg, about 200 mg, about 210 mg, about 220 mg, about 230 mg, about 240 mg, about 250 mg, about 260 mg, about 270 mg, about 280 mg, about 290 mg, about 300 mg, about 310 mg, about 320 mg, about 330 mg, about 340 mg, about 350 mg, about 360 mg, about 370 mg, about 380 mg, about 390 mg, about 400 mg, about 410 mg, about 420 mg, about 430 mg, about 440 mg, about 450 mg, about 460 mg, about 470 mg, about 480 mg, about 490 mg, about 500 mg, about 510 mg, about 520 mg, about 530 mg, about 540 mg, about 550 mg, about 560 mg, about 570 mg, about 580 mg, about 590 mg, about 600 mg, about 610 mg, about 620 mg, about 630 mg, about 640 mg, about 650 mg, about 660 mg, about 670 mg, about 680 mg, about 690 mg, about 700 mg, about 710 mg, about 720 mg, about 730 mg, about 740 mg, about 750 mg, about 760 mg, about 770 mg, about 780 mg, about 790 mg, about 800 mg, about 810 mg, about 820 mg, about 830 mg, about 840 mg, about 850 mg, about 860 mg, about 870 mg, about 880 mg, about 890 mg, about 900 mg, about 910 mg, about 920 mg, about 930 mg, about 940 mg, about 950 mg, about 960 mg, about 970 mg, about 980 mg, about 990 mg, about 1000 mg,57. The method of any one of claims 1 to 56, wherein the compound is administered at a dose of about 1040 mg, about 1080 mg, about 1100 mg, about 1140 mg, about 1180 mg, about 1200 mg, about 1240 mg, about 1280 mg, about 1300 mg, about 1340 mg, about 1380 mg, about 1400 mg, about 1440 mg, about 1480 mg, about 1500 mg, about 1540 mg, about 1580 mg, about 1600 mg, about 1640 mg, about 1680 mg, about 1700 mg, about 1740 mg, about 1780 mg, about 1800 mg, about 1840 mg, about 1880 mg, about 1900 mg, about 1940 mg, about 1980 mg, or about 2000 mg.

58. 55. The method of any one of claims 1 to 54, wherein the LAG-3 antagonist and / or CTLA-4 inhibitor is administered in a weight-dependent dose.

59. and the LAG-3 antagonist and / or CTLA-4 inhibitor is administered at a dose of about 0.003 mg / kg to about 25 mg / kg, about 0.003 mg / kg to about 20 mg / kg, about 0.003 mg / kg to about 15 mg / kg, about 0.003 mg / kg to about 10 mg / kg, about 0.003 mg / kg to about 5 mg / kg, about 0.003 mg / kg to about 1 mg / kg, about 0.003 mg / kg to about 0.9 mg / kg, about 0.003 mg / kg to about 0 .. 8 mg / kg, about 0.003 mg / kg to about 0.7 mg / kg, about 0.003 mg / kg to about 0.6 mg / kg, about 0.003 mg / kg to about 0.5 mg / kg, about 0.003 mg / kg to about 0.4 mg / kg, About 0.003 mg / kg to about 0.3 mg / kg, about 0.003 mg / kg to about 0.2 mg / kg, about 0.003 mg / kg to about 0.1 mg / kg, about 0.1 mg / kg to about 25 mg / kg, about 0.1 mg / kg to About 20 mg / kg, about 0.1 mg / kg to about 15 mg / kg, about 0.1 mg / kg to about 10 mg / kg, about 0.1 mg / kg to about 5 mg / kg, about 0.1 mg / kg to about 1 mg / kg, about 1 mg / kg to about 25 m g / kg, about 1 mg / kg to about 20 mg / kg, about 1 mg / kg to about 15 mg / kg, about 1 mg / kg to about 10 mg / kg, about 1 mg / kg to about 5 mg / kg, about 5 mg / kg to about 25 mg / kg, about 5 mg / kg 59. The method of any one of claims 1 to 54 and 58, wherein the patient is administered at a dose of about 20 mg / kg, about 5 mg / kg to about 15 mg / kg, about 5 mg / kg to about 10 mg / kg, about 10 mg / kg to about 25 mg / kg, about 10 mg / kg to about 20 mg / kg, about 10 mg / kg to about 15 mg / kg, about 15 mg / kg to about 25 mg / kg, about 15 mg / kg to about 20 mg / kg, or about 20 mg / kg to about 25 mg / kg.

60. The LAG-3 antagonist and / or CTLA-4 inhibitor is administered at a dose of about 0.003 mg / kg, about 0.004 mg / kg, about 0.005 mg / kg, about 0.006 mg / kg, about 0.007 mg / kg, about 0.008 mg / kg, about 0.009 mg / kg, about 0.01 mg / kg, about 0.02 mg / kg, about 0.03 mg / kg, about 0.04 mg / kg, about 0.05 mg / kg, about 0.06 mg / kg, about 0.07 mg / kg, about 0.08 mg / kg, about 0.09 mg / kg, about 0.1 mg / kg, about 0.2 mg / kg, about 0.3 mg / kg, about 0.4 mg / kg, about 0.5 mg / kg, about 0.6 mg / kg, about 0.7 mg / kg, about 0.8 mg / kg, about 0.9 mg / kg, about 1 60. The method of any one of claims 1 to 54, 58 and 59, wherein the agonist is administered at a dose of about 0 mg / kg, about 2.0 mg / kg, about 3.0 mg / kg, about 4.0 mg / kg, about 5.0 mg / kg, about 6.0 mg / kg, about 7.0 mg / kg, about 8.0 mg / kg, about 9.0 mg / kg, about 10.0 mg / kg, about 11.0 mg / kg, about 12.0 mg / kg, about 13.0 mg / kg, about 14.0 mg / kg, about 15.0 mg / kg, about 16.0 mg / kg, about 17.0 mg / kg, about 18.0 mg / kg, about 19.0 mg / kg, about 20.0 mg / kg, about 21.0 mg / kg, about 22.0 mg / kg, about 23.0 mg / kg, about 24.0 mg / kg, or about 25.0 mg / kg.

61. 61. The method of any one of claims 1 to 60, wherein the LAG-3 antagonist and / or CTLA-4 inhibitor is administered about once every week, about once every two weeks, about once every three weeks, about once every four weeks, about once every five weeks, about once every six weeks, about once every seven weeks, about once every eight weeks, about once every nine weeks, about once every ten weeks, about once every eleven weeks, or about once every twelve weeks.

62. 62. The method of any one of claims 1 to 61, further comprising administering to the patient an additional therapeutic agent.

63. 63. The method of claim 62, wherein the additional therapeutic agent comprises an anti-cancer agent.

64. 64. The method of claim 63, wherein the anti-cancer agent comprises a tyrosine kinase inhibitor, an anti-angiogenic agent, a checkpoint inhibitor, a checkpoint stimulator, a chemotherapeutic agent, an immunotherapeutic agent, a platinum agent, an alkylating agent, a taxane, a nucleoside analog, an antimetabolite, a topisomerase inhibitor, an anthracycline, a vinca alkaloid, or any combination thereof.

65. 65. The method of claim 64, wherein the tyrosine kinase inhibitor comprises dabrafenib, vemurafenib, encorafenib, trametinib, cobimetinib, binimetinib, or any combination thereof.

66. 65. The method of claim 64, wherein the anti-angiogenic agent comprises an inhibitor of vascular endothelial growth factor (VEGF), VEGF receptor (VEGFR), platelet-derived growth factor (PDGF), PDGF receptor (PDGFR), angiopoietin (Ang), tyrosine kinase with Ig-like and EGF-like domains (Tie) receptor, hepatocyte growth factor (HGF), tyrosine-protein kinase Met (c-MET), C-type lectin family 14 member A (CLEC14A), multimerin 2 (MMRN2), shock protein 70-1A (HSP70-1A), epidermal growth factor (EGF), EGF receptor (EGFR), or a combination thereof.

67. 67. The method of claim 64 or 66, wherein the anti-angiogenic agent comprises bevacizumab, ramucirumab, aflibercept, tanibirumab, olaratumab, nesbacumab, AMG780, MEDI3617, vanucizumab, rilotumumab, ficlatuzumab, TAK-701, onartuzumab, emibetuzumab, or any combination thereof.

68. Checkpoint inhibitors include programmed cell death-1 (PD-1) pathway inhibitors, T-cell immunoglobulin and ITIM domain (TIGIT) inhibitors, T-cell immunoglobulin and mucin-domain containing-3 (TIM-3) inhibitors, TIM-1 inhibitors, TIM-4 inhibitors, B7-H3 inhibitors, B7-H4 inhibitors, B and T cell lymphocyte reduction (BTLA) inhibitors, V domain Ig inhibitors of T cell activation (VISTA) inhibitors, and indoleamines. 2,3-dioxygenase (IDO) inhibitors, nicotinamide adenine dinucleotide phosphate oxidase isoform 2 (NOX2) inhibitors, killer cell immunoglobulin-like receptor (KIR) inhibitors, adenosine A2a receptor (A2aR) inhibitors, transforming growth factor beta (TGF-β) inhibitors, phosphoinositide 3-kinase (PI3K) inhibitors, CD47 inhibitors, CD48 inhibitors, CD73 inhibitors, CD113 inhibitors, sialic acid-binding immunoglobulin-like lectin 7 (SIGLEC-7) inhibitors, SIGLEC-9 inhibitors 65. The method of claim 64, comprising an anti-cancer agent, a SIGLEC-15 inhibitor, a glucocorticoid-induced TNFR-related protein (GITR) inhibitor, a galectin-1 inhibitor, a galectin-9 inhibitor, a carcinoembryonic antigen-related cell adhesion molecule-1 (CEACAM-1) inhibitor, a G protein-coupled receptor 56 (GPR56) inhibitor, a glycoprotein A repeat dominant (GARP) inhibitor, a 2B4 inhibitor, a programmed cell death-1 homolog (PD1H) inhibitor, a leukocyte-associated immunoglobulin-like receptor 1 (LAIR1) inhibitor, or any combination thereof.

69. 69. The method of any one of claims 64 and 68, wherein the checkpoint inhibitor comprises a PD-1 pathway inhibitor.

70. The method of claim 68 or 69, wherein the PD-1 pathway inhibitor is an anti-PD-1 antibody and / or an anti-PD-L1 antibody.

71. 71. The method of any one of claims 68 to 70, wherein the PD-1 pathway inhibitor is an anti-PD-1 antibody.

72. The method of claim 70 or 71, wherein the anti-PD-1 antibody is a full-length antibody.

73. The method of any one of claims 70 to 72, wherein the anti-PD-1 antibody is a monoclonal antibody, a human antibody, a humanized antibody, a chimeric antibody, or a multispecific antibody.

74. 74. The method of claim 73, wherein the multispecific antibody is a DART, a DVD-Ig, or a bispecific antibody.

75. The anti-PD-1 antibody is F(ab') ２ 72. The method of claim 70 or 71, wherein the antibody is a fragment, a Fab' fragment, a Fab fragment, an Fv fragment, a scFv fragment, a dsFv fragment, a dAb fragment or a single chain binding polypeptide peptide.

76. 76. The method of any one of claims 70 to 75, wherein the anti-PD-1 antibody is nivolumab, pembrolizumab, PDR001 (spartalizumab), MEDI-0680, TSR-042, cemiplimab, JS001, PF-06801591, BGB-A317, BI 754091, INCSHR1210, GLS-010, AM-001, STI-1110, AGEN2034, MGA012, BCD-100, IBI308, SSI-361, or an antigen-binding portion thereof.

77. 70. The method of claim 68 or 69, wherein the PD-1 pathway inhibitor is a soluble PD-L2 polypeptide.

78. 78. The method of claim 77, wherein the soluble PD-L2 polypeptide is a fusion polypeptide.

79. 79. The method of claim 77 or 78, wherein the soluble PD-L2 polypeptide comprises a ligand-binding fragment of the PD-L2 extracellular domain.

80. 80. The method of any one of claims 77-79, wherein the soluble PD-L2 polypeptide further comprises a half-life extending moiety.

81. 81. The method of claim 80, wherein the half-life extending moiety comprises an immunoglobulin constant region or a portion thereof, an immunoglobulin-binding polypeptide, immunoglobulin G (IgG), an albumin-binding polypeptide (ABP), a PASylation site, a HESylation site, XTEN, a PEGylation site, an Fc region, or any combination thereof.

82. 82. The method of any one of claims 77 to 81, wherein the soluble PD-L2 polypeptide is AMP-224.

83. 71. The method of any one of claims 68 to 70, wherein the PD-1 pathway inhibitor is an anti-PD-L1 antibody.

84. The method of claim 70 or 83, wherein the anti-PD-L1 antibody is a full-length antibody.

85. The method of any one of claims 70 and 83 to 84, wherein the anti-PD-L1 antibody is a monoclonal antibody, a human antibody, a humanized antibody, a chimeric antibody, or a multispecific antibody.

86. 86. The method of claim 85, wherein the multispecific antibody is a DART, a DVD-Ig, or a bispecific antibody.

87. The anti-PD-L1 antibody is F(ab') ２ 84. The method of claim 70 or 83, wherein the antibody is a fragment, a Fab' fragment, a Fab fragment, an Fv fragment, a scFv fragment, a dsFv fragment, a dAb fragment or a single chain binding polypeptide peptide.

88. 88. The method of any one of claims 70 and 83-87, wherein the anti-PD-L1 antibody is BMS-936559, atezolizumab, durvalumab, avelumab, STI-1014, CX-072, KN035, LY3300054, BGB-A333, ICO 36, CK-301, or an antigen-binding portion thereof.

89. 70. The method of claim 68 or 69, wherein the PD-1 pathway inhibitor is BMS-986189.

90. 1. A method of treating unresectable or metastatic melanoma in a human patient, comprising: (a) an anti-LAG-3 antibody in an amount of about 360 mg; and (b) an anti-CTLA-4 antibody at a dose of about 3 mg / kg; to said patient.

91. 1. A method of treating unresectable or metastatic melanoma in a human patient, comprising: (a) an anti-LAG-3 antibody at a dose of about 720 mg; and (b) an anti-CTLA-4 antibody at a dose of about 3 mg / kg; to said patient.

92. 1. A method of treating unresectable or metastatic melanoma in a human patient, comprising: (a) an anti-LAG-3 antibody at a dose of about 1080 mg; and (b) an anti-CTLA-4 antibody at a dose of about 3 mg / kg; to said patient.

93. 1. A method of treating unresectable or metastatic melanoma in a human patient, comprising: (a) an anti-LAG-3 antibody at a dose of about 1200 mg; and (b) an anti-CTLA-4 antibody at a dose of about 3 mg / kg; to said patient.

94. 1. A method of treating unresectable or metastatic melanoma in a human patient, comprising: (a) an anti-LAG-3 antibody at a dose of about 360 mg; and (b) an anti-CTLA-4 antibody at a dose of about 3 mg / kg; to said patient, wherein the patient has previously received a PD-1 pathway inhibitor as a treatment for melanoma.

95. 1. A method of treating unresectable or metastatic melanoma in a human patient, comprising: (a) an anti-LAG-3 antibody at a dose of about 720 mg; and (b) an anti-CTLA-4 antibody at a dose of about 3 mg / kg; to said patient, wherein the patient has previously received a PD-1 pathway inhibitor as a treatment for melanoma.

96. 1. A method of treating unresectable or metastatic melanoma in a human patient, comprising: (a) an anti-LAG-3 antibody in an amount of about 1080 mg; and (b) an anti-CTLA-4 antibody at a dose of about 3 mg / kg; to said patient, wherein the patient has previously received a PD-1 pathway inhibitor as a treatment for melanoma.

97. 1. A method of treating unresectable or metastatic melanoma in a human patient, comprising: (a) an anti-LAG-3 antibody at a dose of about 1200 mg; and (b) an anti-CTLA-4 antibody at a dose of about 3 mg / kg; to said patient, wherein the patient has previously received a PD-1 pathway inhibitor as a treatment for melanoma.

98. 98. The method of any one of claims 90 to 97, wherein the anti-LAG-3 antibody and the anti-CTLA-4 antibody are administered about once every week, about once every two weeks, about once every three weeks, about once every four weeks, about once every five weeks, about once every six weeks, about once every seven weeks, about once every eight weeks, about once every nine weeks, about once every ten weeks, about once every eleven weeks, or about once every twelve weeks.

99. 99. The method of any one of claims 90 to 98, wherein the patient has histologically confirmed unresectable stage III or stage IV melanoma.

100. 100. The method of any one of claims 90-99, wherein the patient has an Eastern Cooperative Oncology Group (ECOG) performance status of 0 or 1.

101. 101. The method of any one of claims 90 to 100, wherein one or more immune cells in the tumor tissue from the patient express LAG-3.

102. 102. The method of claim 101, wherein at least about 1%, at least about 3%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or about 100% of the immune cells express LAG-3.

103. 103. The method of claim 101 or 102, wherein at least about 1% of immune cells express LAG-3.

104. 104. The method of any one of claims 101 to 103, wherein the immune cells are tumor-infiltrating lymphocytes.

105. Tumor-infiltrating lymphocytes are CD8 ＋ The method of claim 104, wherein the cell is a cell.

106. 106. The method of claim 104 or 105, wherein more than about 1% of the patient's tumor-infiltrating lymphocyte cells express LAG-3.

107. The method of any one of claims 90 to 106, wherein one or more tumor cells in the tumor tissue from the patient express PD-L1.

108. 108. The method of claim 107, wherein at least about 1%, at least about 3%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or about 100% of the tumor cells express PD-L1.

109. 109. The method of claim 107 or 108, wherein at least about 1% of the tumor cells express PD-L1.

110. 110. The method of any one of claims 107 to 109, wherein more than about 1% of the patient's tumor cells express PD-L1.

111. 111. The method of any one of claims 90 to 110, wherein the patient's tumor cells contain a BRAF V600 mutation.

112. The method of any one of claims 90 to 111, wherein the anti-LAG-3 antibody and / or the anti-CTLA-4 antibody is a full-length antibody.

113. The method of any one of claims 90 to 112, wherein the anti-LAG-3 antibody and / or anti-CTLA-4 antibody is a monoclonal antibody, a human antibody, a humanized antibody, a chimeric antibody, or a multispecific antibody.

114. 114. The method of claim 113, wherein the multispecific antibody is a dual affinity retargeting antibody (DART), a DVD-binding protein, or a bispecific antibody.

115. The anti-LAG-3 antibody and / or the anti-CTLA-4 antibody may be an F(ab') ２ 112. The method of any one of claims 90 to 111, wherein the antibody is a fragment, a Fab' fragment, a Fab fragment, an Fv fragment, a scFv fragment, a dsFv fragment, a dAb fragment or a single chain binding polypeptide peptide.

116. 116. The method of any one of claims 90 to 115, wherein the anti-LAG-3 antibody is BMS-986016 (leratolimab), IMP731 (H5L7BW), MK4280 (28G-10), REGN3767 (fianlimab), GSK2831781, humanized BAP050, IMP-701 (LAG-525, yelamirimab), aLAG3(0414), aLAG3(0416), Sym022, TSR-033, TSR-075, XmAb22841, MGD013, BI754111, FS118, P 13B02-30, AVA-017, 25F7, AGEN1746, or an antigen-binding portion thereof.

117. 117. The method of any one of claims 90 to 116, wherein the anti-LAG-3 antibody comprises CDR1, CDR2 and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO:3, and CDR1, CDR2 and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO:

5.

118. Anti-LAG-3 antibody (a) a heavy chain variable region CDR1 comprising the sequence set forth in SEQ ID NO:7; (b) a heavy chain variable region CDR2 comprising the sequence set forth in SEQ ID NO:8; (c) a heavy chain variable region CDR3 comprising the sequence set forth in SEQ ID NO:9; (d) a light chain variable region CDR1 comprising the sequence set forth in SEQ ID NO: 10; (e) a light chain variable region CDR2 comprising the sequence set forth in SEQ ID NO: 11; and (f) a light chain variable region CDR3 comprising the sequence set forth in SEQ ID NO: 12 118. The method of any one of claims 90 to 117, comprising:

119. 119. The method of any one of claims 90 to 118, wherein the anti-LAG-3 antibody comprises a heavy chain variable region and a light chain variable region comprising the sequences set forth in SEQ ID NO:3 and SEQ ID NO:5, respectively.

120. 120. The method of any one of claims 90 to 114 and 116 to 119, wherein the anti-LAG-3 antibody comprises a heavy chain and a light chain comprising the sequences set forth in SEQ ID NO: 1 and SEQ ID NO: 2, respectively.

121. 120. The method of any one of claims 90 to 114 and 116 to 119, wherein the anti-LAG-3 antibody comprises a heavy chain and a light chain comprising the sequences set forth in SEQ ID NO: 30 and SEQ ID NO: 2, respectively.

122. 122. The method of any one of claims 90 to 121, wherein the anti-CTLA-4 antibody is ipilimumab, tremelimumab, MK-1308, AGEN-1884, or an antigen-binding portion thereof.

123. 123. The method of any one of claims 90 to 122, wherein the anti-CTLA-4 antibody comprises CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 34, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO:

32.

124. The anti-CTLA-4 antibody (a) a heavy chain variable region CDR1 comprising the sequence set forth in SEQ ID NO: 35; (b) a heavy chain variable region CDR2 comprising the sequence set forth in SEQ ID NO: 36; (c) a heavy chain variable region CDR3 comprising the sequence set forth in SEQ ID NO: 37; (d) a light chain variable region CDR1 comprising the sequence set forth in SEQ ID NO: 38; (e) a light chain variable region CDR2 comprising the sequence set forth in SEQ ID NO: 39; and (f) a light chain variable region CDR3 comprising the sequence set forth in SEQ ID NO: 40 124. The method of any one of claims 90 to 123, comprising:

125. 125. The method of any one of claims 90 to 124, wherein the anti-CTLA-4 antibody comprises a heavy chain variable region and a light chain variable region comprising the sequences set forth in SEQ ID NOs: 34 and 32, respectively.

126. 126. The method of any one of claims 90-114 and 116-125, wherein the anti-CTLA-4 antibody comprises the heavy and light chains of ipilimumab.

127. 127. The method of any one of claims 90 to 126, further comprising administering a PD-1 pathway inhibitor.

128. The method of claim 127, wherein the PD-1 pathway inhibitor is an anti-PD-1 antibody and / or an anti-PD-L1 antibody.

129. 129. The method of claim 128, wherein the anti-PD-1 antibody is nivolumab, pembrolizumab, PDR001 (spartalizumab), MEDI-0680, TSR-042, REGN2810 (cemiplimab), JS001, PF-06801591, BGB-A317, BI 754091, INCSHR1210, GLS-010, AM-001, STI-1110, AGEN2034, MGA012, BCD-100, IBI308, SSI-361, or an antigen-binding portion thereof.

130. 130. The method of any one of claims 90 to 129, wherein the anti-LAG-3 antibody and the anti-CTLA-4 antibody are formulated for intravenous administration.

131. 131. The method of any one of claims 90 to 130, wherein the anti-LAG-3 antibody and the anti-CTLA-4 antibody are formulated separately.

132. 131. The method of any one of claims 90 to 130, wherein the anti-LAG-3 antibody and the anti-CTLA-4 antibody are formulated as a single agent.

133. 132. The method of any one of claims 90 to 131, wherein the anti-LAG-3 antibody and the anti-CTLA-4 antibody are co-administered.

134. 132. The method of any one of claims 90 to 131, wherein the anti-LAG-3 antibody and the anti-CTLA-4 antibody are administered separately.

135. 132. The method of any one of claims 90 to 131, wherein the anti-LAG-3 antibody is administered simultaneously with the anti-CTLA-4 antibody.

136. 132. The method of any one of claims 90 to 131, wherein the anti-LAG-3 antibody is administered prior to administration of the anti-CTLA-4 antibody.

137. 132. The method of any one of claims 90 to 131, wherein the anti-LAG-3 antibody is administered after administration of the anti-CTLA-4 antibody.

138. 1. A method of treating unresectable or metastatic melanoma in a human patient, comprising: (a) an anti-LAG-3 antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO:3, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO:5; and (b) an anti-CTLA-4 antibody comprising the CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 34, and the CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO:

32. to said patient, wherein the patient has previously received a PD-1 pathway inhibitor as a treatment for melanoma.

139. 1. A method of treating unresectable or metastatic melanoma in a human patient, comprising: (a) an approximately 360 mg dose of an anti-LAG-3 antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO:3, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO:5; and (b) about a 3 mg / kg dose of anti-CTLA-4 comprising the CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 34, and the CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO:

32. to said patient.

140. 1. A method of treating unresectable or metastatic melanoma in a human patient, comprising: (a) an approximately 720 mg dose of an anti-LAG-3 antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO:3, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO:5; and (b) about a 3 mg / kg dose of anti-CTLA-4 comprising the CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 34, and the CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO:

32. to said patient.

141. 1. A method of treating unresectable or metastatic melanoma in a human patient, comprising: (a) an approximately 1080 mg dose of an anti-LAG-3 antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO:3, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO:5; and (b) about a 3 mg / kg dose of anti-CTLA-4 comprising the CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 34, and the CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO:

32. to said patient.

142. 1. A method of treating unresectable or metastatic melanoma in a human patient, comprising: (a) an approximately 1200 mg dose of an anti-LAG-3 antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO:3, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO:5; and (b) about a 3 mg / kg dose of anti-CTLA-4 comprising the CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 34, and the CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO:

32. to said patient.

143. 1. A method of treating unresectable or metastatic melanoma in a human patient, comprising: (a) an approximately 360 mg dose of an anti-LAG-3 antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO:3, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO:5; and (b) about a 3 mg / kg dose of an anti-CTLA-4 antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 34, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO:

32. to said patient, wherein the patient has previously received a PD-1 pathway inhibitor as a treatment for melanoma.

144. 1. A method of treating unresectable or metastatic melanoma in a human patient, comprising: (a) an approximately 720 mg dose of an anti-LAG-3 antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO:3, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO:5; and (b) about a 3 mg / kg dose of an anti-CTLA-4 antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 34, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO:

32. to said patient, wherein the patient has previously received a PD-1 pathway inhibitor as a treatment for melanoma.

145. 1. A method of treating unresectable or metastatic melanoma in a human patient, comprising: (a) an approximately 1080 mg dose of an anti-LAG-3 antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO:3, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO:5; and (b) about a 3 mg / kg dose of an anti-CTLA-4 antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 34, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO:

32. to said patient, wherein the patient has previously received a PD-1 pathway inhibitor as a treatment for melanoma.

146. 1. A method of treating unresectable or metastatic melanoma in a human patient, comprising: (a) an approximately 1200 mg dose of an anti-LAG-3 antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO:3, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO:5; and (b) about a 3 mg / kg dose of an anti-CTLA-4 antibody comprising CDR1, CDR2, and CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 34, and CDR1, CDR2, and CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO:

32. to said patient, wherein the patient has previously received a PD-1 pathway inhibitor as a treatment for melanoma.

147. 147. The method of any one of claims 138 to 146, wherein the anti-LAG-3 antibody and the anti-CTLA-4 antibody are administered about once every week, about once every two weeks, about once every three weeks, about once every four weeks, about once every five weeks, about once every six weeks, about once every seven weeks, about once every eight weeks, about once every nine weeks, about once every ten weeks, about once every eleven weeks, or about once every twelve weeks.

148. 148. The method of any one of claims 138 to 147, wherein the anti-LAG-3 antibody and the anti-CTLA-4 antibody are administered about once every three weeks.

149. 149. The method of any one of claims 138 to 148, wherein the patient has histologically confirmed unresectable stage III or stage IV melanoma.

150. 150. The method of any one of claims 138 to 149, wherein the patient has an Eastern Cooperative Oncology Group (ECOG) performance status of 0 or 1.

151. 151. The method of any one of claims 138 to 150, wherein one or more immune cells in the tumor tissue from the patient express LAG-3.

152. 152. The method of claim 151, wherein at least about 1%, at least about 3%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or about 100% of the immune cells express LAG-3.

153. 153. The method of claim 151 or 152, wherein at least about 1% of immune cells express LAG-3.

154. 154. The method of any one of claims 151 to 153, wherein the immune cells are tumor-infiltrating lymphocytes.

155. Tumor-infiltrating lymphocytes are CD8 ＋ The method of claim 154, wherein the cell is a cell.

156. 156. The method of claim 154 or 155, wherein greater than about 1% of the patient's tumor-infiltrating lymphocyte cells express LAG-3.

157. 157. The method of any one of claims 138 to 156, wherein one or more tumor cells in the tumor tissue from the patient express PD-L1.

158. 158. The method of claim 157, wherein at least about 1%, at least about 3%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or about 100% of the tumor cells express PD-L1.

159. The method of claim 157 or 158, wherein at least about 1% of the tumor cells express PD-L1.

160. 160. The method of any one of claims 157 to 159, wherein greater than 1% of the patient's tumor cells express PD-L1.

161. 161. The method of any one of claims 138 to 160, wherein the patient's tumor cells comprise a BRAF V600 mutation.

162. 1. A method of treating unresectable or metastatic melanoma in a human patient, comprising: (a) about 360 mg, about 720 mg, about 1080 mg, or about 1200 mg doses; (i) a heavy chain variable region CDR1 comprising the sequence set forth in SEQ ID NO: 7; (ii) a heavy chain variable region CDR2 comprising the sequence set forth in SEQ ID NO:8; (iii) a heavy chain variable region CDR3 comprising the sequence set forth in SEQ ID NO:9; (iv) a light chain variable region CDR1 comprising the sequence set forth in SEQ ID NO: 10; (v) a light chain variable region CDR2 comprising the sequence set forth in SEQ ID NO: 11; and (vi) a light chain variable region CDR3 comprising the sequence set forth in SEQ ID NO: 12 an anti-LAG-3 antibody comprising (b) a dose of about 3 mg / kg; (i) a heavy chain variable region CDR1 comprising the sequence set forth in SEQ ID NO: 35; (ii) a heavy chain variable region CDR2 comprising the sequence set forth in SEQ ID NO: 36; (iii) a heavy chain variable region CDR3 comprising the sequence set forth in SEQ ID NO: 37; (iv) a light chain variable region CDR1 comprising the sequence set forth in SEQ ID NO: 38; (v) a light chain variable region CDR2 comprising the sequence set forth in SEQ ID NO: 39; and (vi) a light chain variable region CDR3 comprising the sequence set forth in SEQ ID NO: 40 administering to said patient an anti-CTLA-4 antibody comprising:

163. 1. A method of treating unresectable or metastatic melanoma in a human patient, comprising: (a) about 360 mg, about 720 mg, about 1080 mg, or about 1200 mg doses; (i) a heavy chain variable region CDR1 comprising the sequence set forth in SEQ ID NO: 7; (ii) a heavy chain variable region CDR2 comprising the sequence set forth in SEQ ID NO:8; (iii) a heavy chain variable region CDR3 comprising the sequence set forth in SEQ ID NO:9; (iv) a light chain variable region CDR1 comprising the sequence set forth in SEQ ID NO: 10; (v) a light chain variable region CDR2 comprising the sequence set forth in SEQ ID NO: 11; and (vi) a light chain variable region CDR3 comprising the sequence set forth in SEQ ID NO: 12; an anti-LAG-3 antibody comprising (b) a dose of about 3 mg / kg; (i) a heavy chain variable region CDR1 comprising the sequence set forth in SEQ ID NO: 35; (ii) a heavy chain variable region CDR2 comprising the sequence set forth in SEQ ID NO: 36; (iii) a heavy chain variable region CDR3 comprising the sequence set forth in SEQ ID NO: 37; (iv) a light chain variable region CDR1 comprising the sequence set forth in SEQ ID NO: 38; (v) a light chain variable region CDR2 comprising the sequence set forth in SEQ ID NO: 39; and (vi) a light chain variable region CDR3 comprising the sequence set forth in SEQ ID NO: 40 administering to the patient an anti-CTLA-4 antibody comprising wherein the patient has previously received a PD-1 pathway inhibitor as a treatment for melanoma.

164. 1. A method of treating unresectable or metastatic melanoma in a human patient, comprising: (a) about a 360 mg, about 720 mg, about 1080 mg, or about 1200 mg dose of an anti-LAG-3 antibody comprising a heavy chain variable region and a light chain variable region comprising the sequences set forth in SEQ ID NOs: 3 and 5, respectively; and (b) about a 3 mg / kg dose of an anti-CTLA-4 antibody comprising a heavy chain variable region and a light chain variable region comprising the sequences set forth in SEQ ID NOs: 34 and 32, respectively. administering to said patient an effective amount of each of:

165. 1. A method of treating unresectable or metastatic melanoma in a human patient, comprising: (a) about a 360 mg, about 720 mg, about 1080 mg, or about 1200 mg dose of an anti-LAG-3 antibody comprising a heavy chain variable region and a light chain variable region comprising the sequences set forth in SEQ ID NOs: 3 and 5, respectively; and (b) about a 3 mg / kg dose of an anti-CTLA-4 antibody comprising a heavy chain variable region and a light chain variable region comprising the sequences set forth in SEQ ID NOs: 34 and 32, respectively. to said patient, wherein the patient has previously received a PD-1 pathway inhibitor as a treatment for melanoma.

166. 166. The method of any one of claims 162 to 165, wherein the anti-LAG-3 antibody and the anti-CTLA-4 antibody are administered about once every week, about once every two weeks, about once every three weeks, about once every four weeks, about once every five weeks, about once every six weeks, about once every seven weeks, about once every eight weeks, about once every nine weeks, about once every ten weeks, about once every eleven weeks, or about once every twelve weeks.

167. 167. The method of any one of claims 162 to 166, wherein the anti-LAG-3 antibody and the anti-CTLA-4 antibody are administered about once every three weeks.

168. The method of any one of claims 90 to 167, wherein the anti-LAG-3 antibody is liratolimab.

169. 169. The method of any one of claims 90 to 168, wherein the anti-CTLA-4 antibody is ipilimumab.

170. 1. A method of treating unresectable or metastatic melanoma in a human patient, comprising: (a) a heavy chain variable region CDR1 comprising the sequence set forth in SEQ ID NO:7; (b) a heavy chain variable region CDR2 comprising the sequence set forth in SEQ ID NO:8; (c) a heavy chain variable region CDR3 comprising the sequence set forth in SEQ ID NO:9; (d) a light chain variable region CDR1 comprising the sequence set forth in SEQ ID NO: 10; (e) a light chain variable region CDR2 comprising the sequence set forth in SEQ ID NO: 11; and (f) a light chain variable region CDR3 comprising the sequence set forth in SEQ ID NO: 12 administering to the patient an anti-LAG-3 antibody comprising wherein the patient has previously received a PD-1 pathway inhibitor as a treatment for melanoma; and at least one dose of the anti-LAG-3 antibody is administered at a dose of about 360 mg, about 720 mg, about 1080 mg, or about 1200 mg.

171. 171. The method of any one of claims 90 to 170, wherein the patient is further administered chemotherapy.

172. 172. The method of any one of claims 90 to 171, wherein the patient's tumor cells express fibrinogen-like protein 1 (FGL1).

173. 173. The method of any one of claims 1 to 172, wherein the presence of the BRAF V600E mutation in the tumor specimen is confirmed before initiation of treatment.

174. 174. The method of claim 173, wherein the presence of the BRAF V600E mutation is confirmed using the Cobas® 4800 BRAF V600 Mutation Test.

175. 175. The method of any one of claims 90-126 and 130-174, wherein the patient is not further administered a PD-1 pathway inhibitor.

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