Composition comprising a combination of an anti-LAG-3 antibody, a PD-1 pathway inhibitor, and an immunotherapy agent.

Abstract

This provides an improved method for treating malignant tumors in human patients. [Solution] A method comprising administering a therapeutically effective dose of a LAG-3 inhibitor; a PD-1 pathway inhibitor; and an immunotherapy agent.

Description

[Technical Field]

[0001] Cross-reference of related applications This application claims priority under U.S. Provisional Application 62 / 512,618 filed May 30, 2017, and U.S. Provisional Application 62 / 513,812 filed June 1, 2017, which are incorporated herein by reference as a whole.

[0002] Field of Invention The present invention provides a method for treating malignant tumors (e.g., advanced solid tumors) with a pharmaceutical composition comprising a combination of an anti-LAG-3 antibody, a PD-1 pathway inhibitor, and an immunotherapy agent. [Background technology]

[0003] Background of the Invention Human cancers possess numerous genetic and epigenetic alterations that produce neoantigens that can be recognized by the immune system (Sjoblom et al., Science 314(5797):268-274 (2006)). The adaptive immune system, composed of T and B lymphocytes, has potent anti-cancer capabilities with broad capabilities and sophisticated specificity to respond to diverse tumor antigens. Furthermore, the immune system exhibits considerable flexibility and memory components. The successful utilization of all the characteristics of the adaptive immune system makes immunotherapy unique among all cancer treatment modalities.

[0004] Until recently, cancer immunotherapy has largely focused on attempts to enhance the anti-tumor immune response through adoptive transfer of activated effector cells, immunization against appropriate antigens, or the provision of nonspecific immunostimulatory factors such as cytokines. However, intensive efforts over the past decade to develop specific immune checkpoint pathway inhibitors have begun to offer novel immunotherapeutic approaches for cancer treatment, including the development of antibodies such as ipilimumab (Yervoy®) (Hodi et al., N Engl J Med 363:711-723 (2010)), which binds to and inhibits CTLA-4 for the treatment of patients with advanced melanoma, and antibodies such as nivolumab and pembrolizumab (formerly lambrolizumab; USAN Council Statement, (2013)), which specifically bind to the programmed death-1 (PD-1) receptor and block the inhibitory PD-1 / PD-1 ligand pathway (Topalian et al., N Engl J Med 366:2443-54 (2012a); Topalian et al., Curr Opin Immunol 24:207-12 (2012b); Topalian et al., J Clin Oncol 32(10):1020-30 (2014); Hamid et al., N Engl J Med 369:134-144 (2013); Hamid and Carvajal, Expert Opin Biol Ther 13(6):847-61 (2013); and McDermott and Atkins, Cancer Med 2(5):662-73 (2013).

[0005] In any case, the immune tolerance observed in tumor development and recurrence is thought to be mediated not simply by LAG-3, but by the co-expression of various T cell negative regulatory receptors. Chronic viral infection models (Blackburn et al., Nat. Immunol 10:29-37 (2009), Grosso et al., J. Clin. Invest. 117:3383-3392 (2007) and Lyford-Pike et al., Cancer Res. 73(6):1733-41 (2013)), knockout mice (Woo et al., Cancer Res. 72:917-927 (2012); Okazaki et al., J. Exp Med. 208:395-407 (2011), and Bettini et al., J. Immunol. 187:3493-3498 (2011)), tumor recurrence models (Goding et al., J. Immunol. 190(9):4899-4909) Data from (2013) and, to a more limited extent, from human cancer patients (Matsuzaki et al., Proc. Natl. Acad. Sci., USA. 107:7875-7880 (2010) and Gandhi MK, et al., Blood. 108:2280-2289 (2006)) support a model in which T cells, continuously exposed to antigens, are gradually inactivated through a process called “exhaustion.” Exhausted T cells are characterized by the expression of T cell negative regulatory receptors, primarily CTLA-4, PD-1, and LAG-3, whose function is to limit the cell’s ability to proliferate, produce cytokines, kill target cells, and / or increase Treg activity. Thus, combination therapies including anti-PD-1 and anti-LAG-3 antibodies have shown promising results in certain types of cancer (US Publication 2016 / 0222116A1).

[0006] Lymphocyte-activating gene-3 (LAG-3; CD223) is a type I transmembrane protein expressed on the cell surface of activated CD4+ and CD8+ T cells, as well as subsets of NK and dendritic cells (Triebel et al., J. Exp. Med. 171:1393-1405 (1990); Workman et al., J. Immunol. 182(4):1885-91 (2009)). LAG-3 is closely related to CD4, a co-receptor for T helper cell activation. Both molecules possess four extracellular Ig-like domains and require binding to their ligand, major histocompatibility complex (MHC) class II, for functional activity. In contrast to CD4, LAG-3 is expressed only on the cell surface of activated T cells, and its cleavage from the cell surface halts LAG-3 signaling. LAG-3 has also been found as a soluble protein, but it does not bind to MHC class II, and its function is unknown.

[0007] LAG-3 has been reported to play an important role in promoting regulatory T cell (Treg) activity and negatively regulating T cell activation and proliferation (Workman et al., J. Immunol. 174:688-695 (2005)). Expression of LAG-3 with increased native and induced Tregs is necessary for its maximal inhibitory function (Camisaschi et al., J. Immunol. 184:6545-6551 (2010) and Huang et al., Immunity. 21:503-513 (2004)). Furthermore, ectopic expression of LAG-3 in CD4+ effector T cells reduces their proliferative capacity and gives them regulatory ability over third-party T cells (Huang et al., Immunity. 21:503-513 (2004)). Recent studies have shown that high LAG-3 expression in exhausted lymphocytic choriomeningitis virus (LCMV)-specific CD8+ T cells is involved in their unresponsiveness and limits the CD8+ T cell antitumor response (Blackburn et al., Nat. Immunol. 10:29-37 (2009) and Grosso et al., J. Clin. Invest. 117:3383-3392 (2007)). In fact, LAG-3 maintains tolerance to autologous and tumor antigens through direct action on CD8+ T cells in two mouse models (Grosso et al., J. Clin. Invest. 117:3383-3392 (2007)).

[0008] Programmed cell death 1 (PD-1) is a cell surface signaling receptor that plays a crucial role in regulating T cell activation and tolerance (Keir et al., Annu Rev Immunol 26:677-704 (2008)). It is a type I transmembrane protein and, along with BTLA, CTLA-4, ICOS, and CD28, constitutes the CD28 family of T cell costimulatory receptors. PD-1 is primarily expressed in activated T cells, B cells, and myeloid cells (Dong et al., Nat Med. 5:1365-1369 (1999)). It is also expressed in natural killer (NK) cells (Terme et al., Cancer Res 71:5393-5399 (2011)). When PD-1 binds to its ligands, PD-L1 and PD-L2, phosphorylation of tyrosine residues occurs in the proximal intracellular immune receptor tyrosine inhibitory domain, followed by the recruitment of phosphatase SHP-2, ultimately leading to downregulation of T cell activation. One important role of PD-1 is to limit T cell activity in peripheral tissues during inflammatory responses to infection, thereby limiting the development of autoimmunity (Pardoll Nat Rev Cancer 12:252-264 (2012)). Evidence for this negative regulatory role comes from the discovery that PD-1-deficient mice develop lupus-like autoimmune diseases, including arthritis and nephritis, along with cardiomyopathy (Nishimura H, et al., Immunity, 1999; 11:141-151; and Nishimura H, et al., Science, 2001; 291:319-322). In tumor settings, this results in the development of immune resistance within the microenvironment. PD-1 is highly expressed in tumor-infiltrating lymphocytes, and its ligand is upregulated on the cell surface of various tumors (Dong H, et al., Nat Med 2002; 8:793-800). Multiple mouse cancer models have shown that ligand binding to PD-1 leads to immune evasion. Furthermore, blocking this interaction results in antitumor activity (Topalian SL, et al. NEJM 2012; 366(26):2443-2454; Hamid O, et al., NEJM 2013; 369:134-144).Furthermore, preclinical models have shown that inhibition of PD-1 / PD-L1 interaction is involved in potent antitumor activity (U.S. Patents 8,008,449 and 7,943,743). [Overview of the project] [Problems that the invention aims to solve]

[0009] The prognosis for patients with certain malignant tumors (e.g., metastatic or refractory solid tumors) is extremely poor (Rosenberg SA, et al., Cancer immunotherapy in Cancer: Principles & Practice of Oncology (Eds DeVita VT, Lawrence TS and Rosenberg SA) 2011; 332-344 (Lippincott Williams & Wilkins, Philadelphia Pa.)). Despite advances in multi-faceted treatments, the increase in overall survival in this patient population is limited. Therefore, it is the object of the present invention to provide an improved method (e.g., a composition comprising a combination of an anti-PD-1 antibody, an anti-LAG-3 antibody, and an immunotherapy agent) for the treatment of subjects with such tumors (e.g., advanced, refractory solid tumors). [Means for solving the problem]

[0010] Summary of the Invention The present invention provides a method for treating a subject having a malignant tumor, comprising administering to the subject a therapeutically effective dose of a combination of (a) a LAG-3 inhibitor, (b) a PD-1 pathway inhibitor, and (c) an immunotherapy agent.

[0011] In one embodiment, the LAG-3 inhibitor is an anti-LAG-3 antibody or its antigen-binding fragment. In another embodiment, the anti-LAG-3 antibody is a bispecific antibody. In yet another embodiment, the anti-LAG-3 antibody or its antigen-binding fragment comprises (a) a heavy chain variable region CDR1 containing the sequence shown in SEQ ID NO: 7; (b) a heavy chain variable region CDR2 containing the sequence shown in SEQ ID NO: 8; (c) a heavy chain variable region CDR3 containing the sequence shown in SEQ ID NO: 9; (d) a light chain variable region CDR1 containing the sequence shown in SEQ ID NO: 10; (e) a light chain variable region CDR2 containing the sequence shown in SEQ ID NO: 11; and (f) a light chain variable region CDR3 containing the sequence shown in SEQ ID NO: 12. In one embodiment, the anti-LAG-3 antibody or its antigen-binding fragment comprises a heavy chain and a light chain variable region containing the sequences shown in SEQ ID NOs: 3 and 5, respectively. In one embodiment, the anti-LAG-3 antibody is BMS 986016, MK-4280 (28G-10), REGN3767, GSK2831781, IMP731 (H5L7BW), BAP050, IMP-701 (LAG-5250), IMP321, TSR-033, LAG525, BI 754111, or FS-118.

[0012] In one embodiment, the LAG-3 inhibitor is a soluble LAG-3 polypeptide. In another embodiment, the soluble LAG-3 polypeptide is a fusion polypeptide. In yet another embodiment, the soluble LAG-3 polypeptide comprises a ligand-binding fragment of the LAG-3 extracellular domain. In one embodiment, the ligand-binding fragment of the LAG-3 extracellular domain comprises an amino acid sequence having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity with SEQ ID NO: 44. In one embodiment, the soluble LAG-3 polypeptide further comprises an Fc domain.

[0013] In one embodiment, the PD-1 pathway inhibitor is an anti-PD-1 antibody or its antigen-binding fragment. In one embodiment, the anti-PD-1 antibody is pembrolizumab (Keytruda; MK-3475), pidilizumab (CT-011), nivolumab (Opdivo; BMS-936558), PDR001, MEDI0680 (AMP-514), TSR-042, REGN2810, JS001, AMP-224 (GSK-2661380), PF-06801591, BGB-A317, BI 754091, or SHR-1210.

[0014] In one embodiment, the PD-1 pathway inhibitor is an anti-PD-L1 antibody or its antigen-binding fragment. In one embodiment, the anti-PD-L1 antibody is atezolizumab (Tecentriq; RG7446; MPDL3280A; RO5541267), durvalumab (MEDI4736), BMS-936559, avelumab (Bavencio), LY3300054, CX-072 (Proclaim-CX-072), FAZ053, KN035, or MDX-1105.

[0015] In one embodiment, the PD-1 pathway inhibitor is a small molecule drug. In one embodiment, the PD-1 pathway inhibitor is CA-170. In another embodiment, the PD-1 pathway inhibitor is a cell-based therapy. In one embodiment, the cell-based therapy is a MiHA-loaded PD-L1 / L2 expression-suppressing dendritic cell vaccine. In another embodiment, the cell-based therapy is an anti-programmed cell death protein 1 antibody expressing pluripotent killer T lymphocytes, autologous PD-1-targeted chimeric switch receptor-modified T lymphocytes, or PD-1 knockout autologous T lymphocytes.

[0016] In one embodiment, the PD-1 pathway inhibitor is an anti-PD-L2 antibody or its antigen-binding fragment. In another embodiment, the anti-PD-L2 antibody is rHIgM12B7.

[0017] In one embodiment, the PD-1 pathway inhibitor is a soluble PD-1 polypeptide. In one embodiment, the soluble PD-1 polypeptide is a fusion polypeptide. In one embodiment, the soluble PD-1 polypeptide comprises a ligand-binding fragment of the PD-1 extracellular domain. In another embodiment, the soluble PD-1 polypeptide comprises a ligand-binding fragment of the PD-1 extracellular domain. In one embodiment, the ligand-binding fragment of the PD-1 extracellular domain comprises amino acids having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity with SEQ ID NO: 29. In another embodiment, the soluble PD-1 polypeptide further comprises an Fc domain.

[0018] In one embodiment, the immunotherapy agent is a modulator of CTLA-4 activity, CD28 activity, CD80 activity, CD86 activity, 4-1BB activity, OX40 activity, KIR activity, Tim-3 activity, CD27 activity, CD40 activity, GITR activity, TIGIT activity, CD20 activity, CD96 activity, IDO1 activity, STING activity, GARP activity, A2aR activity, CEACAM1 activity, CEA activity, CD47 activity, PVRIG activity, TDO activity, VISTA activity, cytokines, chemokines, interferons, interleukins, lymphokines, members of the tumor necrosis factor (TNF) family, or immunostimulatory oligonucleotides.

[0019] In certain embodiments, the immunotherapeutic agent is an immune checkpoint inhibitor. In certain embodiments, the immune checkpoint inhibitor is a CTLA-4 antagonist, a CD80 antagonist, a CD86 antagonist, a Tim-3 antagonist, a TIGIT antagonist, a CD20 antagonist, a CD96 antagonist, an IDO1 antagonist, a STING antagonist, a GARP antagonist, a CD40 antagonist, an A2aR antagonist, a CEACAM1 (CD66a) antagonist, a CEA antagonist, a CD47 antagonist, a PVRIG antagonist, a TDO antagonist, a VISTA antagonist or a KIR antagonist.

[0020] In certain embodiments, the immune checkpoint inhibitor is a CTLA-4 antagonist. In certain embodiments, the CTLA-4 antagonist is an anti-CTLA-4 antibody or an antigen-binding fragment thereof. In certain embodiments, the anti-CTLA-4 antibody is ipilimumab (Yervoy), tremelimumab (ticilimumab; CP-675,206), AGEN-1884 or ATOR-1015.

[0021] In certain embodiments, the CTLA-4 antagonist is a soluble CTLA-4 polypeptide. In certain embodiments, the soluble CTLA-4 polypeptide is abatacept (Orencia), belatacept (Nulojix), RG2077 or RG-1046. In other embodiments, the CTLA-4 antagonist is a cell-based therapy. In certain embodiments, the CTLA-4 antagonist is an anti-CTLA4 mAb RNA / GITRL RNA transfected autologous dendritic cell vaccine or an anti-CTLA-4 mAb RNA transfected autologous dendritic cell vaccine.

[0022] In certain embodiments, the immune checkpoint inhibitor is a KIR antagonist. In certain embodiments, the KIR antagonist is an anti-KIR antibody or an antigen-binding fragment thereof. In certain embodiments, the anti-KIR antibody is lirilumab (1-7F9, BMS-986015, IPH 2101) or IPH4102.

[0023] In certain embodiments, the immune checkpoint inhibitor is a TIGIT antagonist. In certain embodiments, the TIGIT antagonist is an anti-TIGIT antibody or an antigen-binding fragment thereof. In certain embodiments, the anti-TIGIT antibody is BMS-986207, AB 154, COM902 (CGEN-15137) or OMP-313M32.

[0024] In certain embodiments, the immune checkpoint inhibitor is a Tim-3 antagonist. In certain embodiments, the Tim-3 antagonist is an anti-Tim-3 antibody or an antigen-binding fragment thereof. In certain embodiments, the anti-Tim-3 antibody is TSR-022 or LY3321367.

[0025] In certain embodiments, the immune checkpoint inhibitor is an IDO1 antagonist. In other embodiments, the IDO1 antagonist is indoximod (NLG8189; 1-methyl- D -TRP), epacadostat (INCB-024360, INCB-24360), KHK2455, PF-06840003, navoximod (RG6078, GDC-0919, NLG919), BMS-986205 (F001287) or a pyrrolidine-2,5-dione derivative.

[0026] In one embodiment, the immune checkpoint inhibitor is a STING antagonist. In one embodiment, the STING antagonist is a 2' or 3'-mono-fluorosubstituted cyclic dinucleotide; a 2'-3'-di-fluorosubstituted mixed bond 2',5'-3',5' cyclic dinucleotide; a 2'-fluorosubstituted, bis-3',5' cyclic dinucleotide; a 2',2''-diF-Rp,Rp,bis-3',5' cyclic dinucleotide; or a fluorinated cyclic dinucleotide.

[0027] In one embodiment, the immune checkpoint inhibitor is a CD20 antagonist. In one embodiment, the CD20 antagonist is an anti-CD20 antibody or its antigen-binding fragment. In one embodiment, the anti-CD20 antibody is rituximab (Rituxan; IDEC-102; IDEC-C2B8), ABP 798, ofatumumab, or obinutuzumab.

[0028] In one embodiment, the immune checkpoint inhibitor is a CD80 antagonist. In another embodiment, the CD80 antagonist is an anti-CD80 antibody or its antigen-binding fragment. In another embodiment, the anti-CD80 antibody is galiximab or AV 1142742.

[0029] In one embodiment, the immune checkpoint inhibitor is a GARP antagonist. In another embodiment, the GARP antagonist is an anti-GARP antibody or its antigen-binding fragment. In another embodiment, the anti-GARP antibody is ARGX-115.

[0030] In one embodiment, the immune checkpoint inhibitor is a CD40 antagonist. In one embodiment, the CD40 antagonist is an anti-CD40 antibody or its antigen-binding fragment. In one embodiment, the anti-CD40 antibody is BMS3h-56, lucatumumab (HCD122 and CHIR-12.12), CHIR-5.9, or dasetuzumab (huS2C6, PRO 64553, RG 3636, SGN 14, SGN-40). In another embodiment, the CD40 antagonist is a soluble CD40 ligand (CD40-L). In one embodiment, the soluble CD40 ligand is a fusion polypeptide. In one embodiment, the soluble CD40 ligand is CD40-L / FC2 or monomeric CD40-L.

[0031] In one embodiment, the immune checkpoint inhibitor is an A2aR antagonist. In one embodiment, the A2aR antagonist is a small molecule. In one embodiment, the A2aR antagonist is CPI-444, PBF-509, istradefylline (KW-6002), preradianant (SCH420814), tozadenant (SYN115), bipardenant (BIIB014), HTL-1071, ST1535, SCH412348, SCH442416, SCH58261, ZM241385, or AZD4635.

[0032] In one embodiment, the immune checkpoint inhibitor is a CEACAM1 antagonist. In another embodiment, the CEACAM1 antagonist is an anti-CEACAM1 antibody or its antigen-binding fragment. In another embodiment, the anti-CEACAM1 antibody is CM-24 (MK-6018).

[0033] In one embodiment, the immune checkpoint inhibitor is a CEA antagonist. In one embodiment, the CEA antagonist is an anti-CEA antibody or its antigen-binding fragment. In one embodiment, the anti-CEA antibody is cergutuzumabu amnaleukin (RG7813, RO-6895882) or RG7802 (RO6958688).

[0034] In one embodiment, the immune checkpoint inhibitor is a CD47 antagonist. In one embodiment, the CD47 antagonist is an anti-CD47 antibody or its antigen-binding fragment. In one embodiment, the anti-CD47 antibody is HuF9-G4, CC-90002, TTI-621, ALX148, NI-1701, NI-1801, SRF231, or Effi-DEM.

[0035] In one embodiment, the immune checkpoint inhibitor is a PVRIG antagonist. In one embodiment, the PVRIG antagonist is an anti-PVRIG antibody or its antigen-binding fragment. In one embodiment, the anti-PVRIG antibody is COM701 (CGEN-15029).

[0036] In one embodiment, the immune checkpoint inhibitor is a TDO antagonist. In one embodiment, the TDO antagonist is a 4-(indole-3-yl)-pyrazole derivative, a 3-indole-substituted derivative, or a 3-(indole-3-yl)-pyridine derivative. In another embodiment, the immune checkpoint inhibitor is a dual IDO and TDO antagonist. In one embodiment, the dual IDO and TDO antagonist is a small molecule.

[0037] In one embodiment, the immune checkpoint inhibitor is a VISTA antagonist. In one embodiment, the VISTA antagonist is CA-170 or JNJ-61610588.

[0038] In one embodiment, the immunotherapy agent is an immune checkpoint enhancer or stimulator. In one embodiment, the immune checkpoint enhancer or stimulator is a CD28 agonist, a 4-1BB agonist, an OX40 agonist, a CD27 agonist, a CD80 agonist, a CD86 agonist, a CD40 agonist, an ICOS agonist, a CD70 agonist, or a GITR agonist.

[0039] In one embodiment, the immune checkpoint enhancer or stimulator is an OX40 agonist. In one embodiment, the OX40 agonist is an anti-OX40 antibody or its antigen-binding fragment. In one embodiment, the anti-OX40 antibody is tavorixizumab (MEDI-0562), pogalizumab (MOXR0916, RG7888), GSK3174998, ATOR-1015, MEDI-6383, MEDI-6469, BMS 986178, PF-04518600, or RG7888 (MOXR0916). In another embodiment, the OX40 agonist is a cell-based therapy. In one embodiment, the OX40 agonist is GINAKIT cells (iC9-GD2-CD28-OX40 expressing T lymphocytes).

[0040] In one embodiment, the immune checkpoint enhancer or stimulator is a CD40 agonist. In one embodiment, the CD40 agonist is an anti-CD40 antibody or its antigen-binding fragment. In one embodiment, the anti-CD40 antibody is ADC-1013 (JNJ-64457107), RG7876 (RO-7009789), HuCD40-M2, APX005M (EPI-0050), or Chi Lob 7 / 4. In another embodiment, the CD40 agonist is a soluble CD40 ligand (CD40-L). In one embodiment, the soluble CD40 ligand is a fusion polypeptide. In one embodiment, the soluble CD40 ligand is trimer CD40-L (AVREND®).

[0041] In one embodiment, the immune checkpoint enhancer or stimulator is a GITR agonist. In one embodiment, the GITR agonist is an anti-GITR antibody or its antigen-binding fragment. In one embodiment, the anti-GITR antibody is BMS-986156, TRX518, GWN323, INCAGN01876, or MEDI1873. In one embodiment, the GITR agonist is a soluble GITR ligand (GITRL). In one embodiment, the soluble GITR ligand is a fusion polypeptide. In another embodiment, the GITR agonist is a cell-based therapy. In one embodiment, the cell-based therapy is an anti-CTLA4 mAb RNA / GITRL RNA-transfected autogenous dendritic cell vaccine or a GITRL RNA-transfected autogenous dendritic cell vaccine.

[0042] In one embodiment, the immune checkpoint enhancer or stimulator is a 4-1BB agonist. In one embodiment, the 4-1BB agonist is an anti-4-1BB antibody or its antigen-binding fragment. In one embodiment, the anti-4-1BB antibody is urelumab or PF-05082566.

[0043] In one embodiment, the immune checkpoint enhancer or stimulator is a CD80 agonist or a CD86 agonist. In one embodiment, the CD80 agonist or CD86 agonist is a soluble CD80 or CD86 ligand (CTLA-4). In one embodiment, the soluble CD80 or CD86 ligand is a fusion polypeptide. In one embodiment, the CD80 or CD86 ligand is CTLA4-Ig (CTLA4-IgG4m, RG2077 or RG1046) or abatacept (Orencia, BMS-188667). In another embodiment, the CD80 agonist or CD86 agonist is a cell-based therapy. In one embodiment, the cell-based therapy is MGN1601 (allogeneic renal cell carcinoma vaccine).

[0044] In one embodiment, the immune checkpoint enhancer or stimulator is a CD28 agonist. In one embodiment, the CD28 agonist is an anti-CD28 antibody or its antigen-binding fragment. In one embodiment, the anti-CD28 antibody is TGN1412.

[0045] In one embodiment, the CD28 agonist is a cell-based therapy. In one embodiment, the cell-based therapy is JCAR015 (anti-CD19-CD28-zeta modified CAR CD3+ T lymphocytes); CD28CAR / CD137CAR expressing T lymphocytes; allogeneic CD4+ memory Th1-like T cells / microparticle-bound anti-CD3 / anti-CD28; anti-CD19 / CD28 / CD3 zeta CAR gamma retroviral vector-transduced autologous T lymphocytes KTE-C19; anti-CEA IgCD28TCR-transduced autologous T lymphocytes; anti-EGFRvIII CAR-transduced allogeneic T lymphocytes; autologous CD123CAR-CD28-CD3 zeta-EGFRt-expressing T lymphocytes; autologous CD171-specific CAR-CD28 zeta-4-1-BB-EGFRt-expressing T lymphocytes; autologous CD19CAR-CD28-CD3 zeta-EGFRt-expressing Tcm-enriched T cells; autologous PD-1-targeted chimeric switch receptor-modified T lymphocytes (chimeric with CD28); CD19CAR-CD28-CD3 zeta-EGFRt-expressing T These include cm-enriched T lymphocytes; CD19CAR-CD28-CD3 zeta-EGFRt-expressing Tn / mem-enriched T lymphocytes; CD19CAR-CD28 zeta-4-1BB-expressing allogeneic T lymphocytes; CD19CAR-CD3 zeta-4-1BB-CD28-expressing autologous T lymphocytes; CD28CAR / CD137CAR-expressing T lymphocytes; CD3 / CD28 costimulated vaccine-stimulated autologous T lymphocytes; or iC9-GD2-CD28-OX40-expressing T lymphocytes.

[0046] In one embodiment, the immune checkpoint enhancer or stimulator is a CD27 agonist. In one embodiment, the CD27 agonist is an anti-CD27 antibody or its antigen-binding fragment. In one embodiment, the anti-CD27 antibody is varylumab (CDX-1127).

[0047] In one embodiment, the immune checkpoint enhancer or stimulator is a CD70 agonist. In one embodiment, the CD70 agonist is an anti-CD70 antibody or its antigen-binding fragment. In one embodiment, the anti-CD70 antibody is ARGX-110.

[0048] In one embodiment, the immune checkpoint enhancer or stimulator is an ICOS agonist. In one embodiment, the ICOS agonist is an anti-ICOS antibody or its antigen-binding fragment. In one embodiment, the anti-ICOS antibody is BMS986226, MEDI-570, GSK3359609, or JTX-2011. In another embodiment, the ICOS agonist is a soluble ICOS ligand. In one embodiment, the soluble ICOS ligand is a fusion polypeptide. In one embodiment, the soluble ICOS ligand is AMG 750.

[0049] In one embodiment, the immunotherapy agent is an anti-CD73 antibody or its antigen-binding fragment. In one embodiment, the anti-CD73 antibody is MEDI9447.

[0050] In one embodiment, the immunotherapy agent is a TLR9 agonist. In one embodiment, the TLR9 agonist is agatrimod sodium.

[0051] In one embodiment, the immunotherapy agent is a cytokine. In one embodiment, the cytokine is a chemokine, interferon, interleukin, lymphokine, or a member of the tumor necrosis factor family. In one embodiment, the cytokine is IL-2, IL-15, or interferon-gamma.

[0052] In one embodiment, the immunotherapy agent is a TGF-β antagonist. In one embodiment, the TGF-β antagonist is fresolimmab (GC-1008); NIS793; IMC-TR1 (LY3022859); ISTH0036; Travedersen (AP 12009); recombinant transforming growth factor-beta-2; autologous HPV-16 / 18 E6 / E7 specific TGF-beta resistant T lymphocytes; or TGF-beta resistant LMP specific cytotoxic T lymphocytes.

[0053] In one embodiment, the immunotherapy agent is an iNOS antagonist. In one embodiment, the iNOS antagonist is N-acetyl-cysteine ​​(NAC), aminoguanidine, L-nitroarginine methyl ester, or S,S-1,4-phenylene-bis(1,2-ethanediyl)bis-isothiourea.

[0054] In one embodiment, the immunotherapy agent is an SHP-1 antagonist.

[0055] In one embodiment, the immunotherapy agent is a CSF1R (colony-stimulating factor 1 receptor) antagonist. In one embodiment, the CSF1R antagonist is an anti-CSF1R antibody or its antigen-binding fragment. In one embodiment, the anti-CSF1R antibody is emuctuzumab.

[0056] In one embodiment, the immunotherapy agent is a TNF family member agonist. In one embodiment, the TNF family member agonist is ATOR 1016, ABBV-621, or adalimumab.

[0057] In one embodiment, the immunotherapy agent is aldesleukin, tocilizumab, or MEDI5083.

[0058] In one embodiment, the immunotherapy agent is a CD160(NK1) agonist. In one embodiment, the CD160(NK1) agonist is an anti-CD160 antibody or its antigen-binding fragment. In one embodiment, the anti-CD160 antibody is BY55.

[0059] In one embodiment, the LAG-3 inhibitor, PD-1 pathway inhibitor, and immunotherapy agent are formulated for intravenous administration. In another embodiment, the LAG-3 inhibitor, PD-1 pathway inhibitor, and immunotherapy agent are formulated together. In yet another embodiment, the LAG-3 inhibitor, PD-1 pathway inhibitor, and immunotherapy agent are formulated separately.

[0060] In one embodiment, malignant tumors include liver cancer, bone cancer, pancreatic cancer, skin cancer, oral cancer, head and neck cancer, breast cancer, lung cancer including small cell and non-small cell lung cancer, cutaneous or intraocular malignant melanoma, kidney cancer, uterine cancer, ovarian cancer, colorectal cancer, colon cancer, rectal cancer, anal cancer, gastric cancer, testicular cancer, uterine cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, non-Hodgkin lymphoma, esophageal cancer, small intestine cancer, endocrine cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, childhood cancer, lymphocytic lymphoma, bladder cancer, kidney or ureteral cancer, renal pelvis cancer, central nervous system (CNS) neoplasms, primary CNS lymphoma, tumor angiogenesis, spinal axis tumors, brainstem gliomas, pituitary adenomas, and Kaposi's positivity. Selected from the group consisting of sarcomas, epidermoid carcinomas, squamous cell carcinomas, environment-induced cancers including those induced by asbestos, such as multiple myeloma, B-cell lymphoma, Hodgkin lymphoma / primary mediastinal B-cell lymphoma, non-Hodgkin lymphoma, acute myeloid lymphoma, chronic myeloid leukemia, chronic lymphoid leukemia, follicular lymphoma, generalized large B-cell lymphoma, Burkitt lymphoma, immunoblastic large cell lymphoma, precursor B-lymphoblastic lymphoma, mantle cell lymphoma, acute lymphoblastic leukemia, mycosis fungoides, anaplastic large cell lymphoma, T-cell lymphoma and precursor T-lymphoblastic lymphoma, and any combination thereof.

[0061] In one embodiment, the malignant tumor is non-small cell lung cancer (NSCLC), virus-associated cancer-associated tumor, or gastric adenocarcinoma. In another embodiment, the malignant tumor is melanoma, gastric cancer, gastroesophageal junction cancer, non-small cell lung cancer, bladder cancer, head and neck squamous cell carcinoma, or renal cell carcinoma. In yet another embodiment, the tumor is lung cancer, melanoma, head and neck squamous cell carcinoma, kidney cancer, gastric cancer, or hepatocellular carcinoma.

[0062] In one embodiment, an anti-LAG-3 antibody or its antigen-binding fragment and an immunotherapy agent are administered as first-line treatment. In another embodiment, a LAG-3 inhibitor, a PD-1 pathway inhibitor, and an immunotherapy agent are administered as second-line treatment. In one embodiment, the malignant tumor is refractory to first-line treatment.

[0063] In one embodiment, the method for treating a subject having the above-mentioned malignant tumor further includes the administration of at least one additional therapeutic agent. In one embodiment, the at least one additional therapeutic agent is a chemotherapeutic agent.

[0064] Other features and benefits of the present invention are evident from the following detailed description and examples, which should not be construed as limiting. [Modes for carrying out the invention]

[0065] Detailed description of the invention I. Terminology The term "patient" as used herein includes all patients with cancer (e.g., melanoma). The terms "subject" and "patient" are used interchangeably here.

[0066] As used herein, “administer” means the physical delivery of a composition comprising a therapeutic agent (e.g., a combination of an anti-PD-1 antibody, an anti-LAG-3 antibody, and an additive immunotherapy agent) to a subject using any of the various methods and delivery systems known to those skilled in the art. Routes of administration include intravenous, intramuscular, subcutaneous, intraperitoneal, spinal, or other non-enteral administration routes, such as by injection or infusion. As used herein, “non-enteral administration” means administration methods other than enteral and topical administration, usually by injection, and includes, but is not limited to, intravenous, intramuscular, intra-arterial, intrathecal, intralymphatic, intrafocal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subepidermal, intra-articular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal injections and infusions, as well as in vivo electroporation. Other non-enteral routes include topical, epithelial, or mucosal administration routes, such as intranasal, vaginal, rectal, sublingual, or topical. The administration may also be carried out, for example, once, multiple times, and / or over a long period of time.

[0067] As used herein, “effective treatment” refers to a treatment that produces a beneficial effect, such as improvement of at least one symptom of a disease or disorder. Beneficial effects may take the form of improvement beyond baseline, i.e., improvement beyond the measurements or observations made before the initiation of treatment with this method. Beneficial effects may also take the form of cessation, slowing, delaying, or stabilization of the harmful progression of solid tumor markers. Effective treatment may refer to the alleviation of at least one symptom of a solid tumor. Such effective treatments may, for example, reduce patient pain, reduce the size and / or number of lesions, reduce or prevent tumor metastasis, and / or slow tumor growth.

[0068] The term “effective dose” refers to the amount of a drug that provides a desired biological, therapeutic, and / or prophylactic outcome. The outcome may be a reduction, improvement, mitigation, decrease, delay, and / or mitigation of the signs, symptoms, or causes of a disease, or any other desirable modification of the biological system. With respect to solid tumors, an effective dose includes an amount sufficient to shrink the tumor and / or reduce the rate of tumor growth (e.g., tumor growth inhibition) or to inhibit or delay other unwanted cell proliferation. In some embodiments, an effective dose is sufficient to delay tumor progression. In some embodiments, an effective dose may be administered in one or more doses, and the effective dose may be sufficient to prevent or delay tumor recurrence. An effective dose of a drug or composition may (i) reduce the number of cancer cells; (ii) reduce tumor size; (iii) inhibit, delay, slow, or halt to some extent cancer cell infiltration into peripheral organs; (iv) inhibit tumor metastasis (i.e., slow or halt to some extent); (v) inhibit tumor growth; (vi) prevent or delay tumor development and / or recurrence; and / or (vii) alleviate to some extent one or more cancer-related symptoms. In one example, “effective dose” is the amount of combined anti-LAG-3 antibody and anti-PD-1 antibody that has been clinically proven to have an effect on a significant reduction in cancer or delaying cancer progression, such as in advanced solid tumors. The terms “constant dose,” “uniform dose,” and “uniform-constant dose” as used herein are interchangeable and refer to the dose administered to a patient regardless of the patient’s body weight or body surface area (BSA). Constant or uniform doses are therefore provided as absolute doses of the drug (e.g., anti-LAG-3 antibody and / or anti-PD-1 antibody), rather than mg / kg doses.

[0069] As used herein, the term “immunotherapy” refers to the treatment of a subject who has, is at risk of developing, or has a relapse of a disease, by inducing, enhancing, suppressing, or otherwise modifying the immune response. The “treatment” or “therapy” of a subject refers to any type of intervention or procedure or administration of an activator (e.g., a composition including a combination of anti-PD-1 antibodies, anti-LAG-3 antibodies, and additive immunotherapy agents) performed on the subject with the aim of restoring, reducing, improving, inhibiting, slowing, or preventing the onset, progression, advancement, worsening, or relapse of symptoms, complications, or conditions, or biochemical signs associated with the disease.

[0070] The terms “cell-based therapy,” “cell therapy,” “cell treatment,” or “cytotherapy” as used herein refer to the transplantation or delivery of cellular material to a patient for the purpose of treating a disease or disorder (e.g., malignant tumor). Cellular material may be cell fragments or intact cells, or living cells (e.g., T lymphocytes, dendritic cells, or stem cells).

[0071] The use of the term "fixed dose" in relation to the compositions of the present invention means that two or more different antibodies in a single composition are present in the composition in a specific (fixed) ratio to one another. In some embodiments, the fixed dose is based on the weight of the antibodies (e.g., mg). In some embodiments, the fixed dose is based on the concentration of the antibodies (e.g., mg / ml). In some embodiments, 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 1:160, about 1 :180, approximately 1:200, approximately 200:1, approximately 180:1, approximately 160:1, approximately 140:1, approximately 120:1, approximately 100:1, approximately 90:1, approximately 80:1, approximately 70:1, approximately 60:1, approximately 50:1, approximately 40:1, approximately 30:1, approximately 20:1, approximately 15:1, approximately 10:1, approximately 9:1, approximately 8:1, approximately 7:1, approximately 6:1, approximately 5:1, approximately 4:1, approximately 3:1, or approximately 2:1. This represents mg of primary antibody to mg of secondary antibody. For example, a 3:1 ratio of primary to secondary antibody may mean that the vial contains approximately 240 mg of primary antibody and 80 mg of secondary antibody, or approximately 3 mg / ml of primary antibody and 1 mg / ml of secondary antibody.

[0072] The use of the term "uniform dose" in relation to the compositions of the present invention means a dose administered to a patient regardless of the patient's body weight or body surface area (BSA). The uniform dose is therefore provided as an absolute dose of the drug (e.g., anti-LAG-3 antibody and / or anti-PD-1 antibody), rather than as a mg / kg dose. For example, a 60kg person and a 100kg person receive the same dose of the composition (e.g., 240mg of anti-PD-1 antibody and 80mg of anti-LAG-3 antibody in a single fixed-dose formulation vial containing both 240mg of anti-PD-1 antibody and 80mg of anti-LAG-3 antibody (or two fixed-dose formulation vials containing 120mg of anti-PD-1 antibody and 40mg of anti-LAG-3 antibody, etc.)).

[0073] The term "weight-based dose" used here means that the dose administered to a patient is calculated based on the patient's weight. For example, if a patient weighing 60 kg requires a combination of 3 mg / kg anti-LAG-3 antibody and 3 mg / kg anti-PD-1 antibody, the appropriate amounts of anti-LAG-3 antibody (i.e., 180 mg) and anti-PD-1 antibody (i.e., 180 mg) can be immediately drawn from a 1:1 ratio fixed-dose formulation of anti-LAG-3 antibody and anti-PD-1 antibody.

[0074] An antibody (Ab) is a glycoprotein immunoglobulin or its antigen-binding moiety that specifically binds to an antigen and contains at least two heavy (H) chains and two light (L) chains interconnected by disulfide bonds. Each heavy chain contains a heavy chain variable region (here, V H It consists of a light chain variable region (abbreviated as CH) and a heavy chain constant region (abbreviated as CH). In some antibodies, for example, naturally occurring IgG antibodies, the heavy chain constant region consists of a hinge and three domains, CH1, CH2, and CH3. In some antibodies, for example, naturally occurring IgG antibodies, each light chain consists of a light chain variable region (abbreviated as V) L It consists of a (abbreviated as) and a light chain steady region. The light chain steady region consists of one domain (abbreviated as CL here). V H and V L The domain can be further subdivided into a more conserved domain called the framework domain (FR) and a hyper-variable domain called the complementarity-determining domain (CDR). H and V L It consists of three CDRs and four FRs arranged in the following order from the amino terminus to the carboxyl terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable regions of the heavy and light chains contain binding domains that interact with the antigen. The constant region of the antibody may mediate the binding of immunoglobulin to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component of the classical complement system (C1q). The heavy chain may or may not have a C-terminal lysine. Unless otherwise specified, the amino acids of the variable region are numbered using the Kabat numbering system, and the amino acids of the constant region are numbered using the EU system.

[0075] Immunoglobulins may originate from any of the known isotypes, including IgA, secretory IgA, IgD, IgE, IgG, and IgM. IgG isotypes are divided into subclasses in some species: IgG1, IgG2, IgG3, and IgG4 in humans, and IgG1, IgG2a, IgG2b, and IgG3 in mice. “Isotype” refers to an antibody class or subclass (e.g., IgM or IgG1) encoded by a heavy chain constant region gene. The term “antibody” includes, by example, monoclonal and polyclonal antibodies; chimeric and humanized antibodies; human or non-human antibodies; fully synthetic antibodies; and single-chain antibodies. Non-human antibodies may be humanized by recombinant methods to reduce their immunogenicity in humans. Unless otherwise specified and indicated by the context, the term “antibody” includes monospecific, bispecific, or polyspecific antibodies and single-chain antibodies. In one embodiment, the antibody is a bispecific antibody. In another embodiment, the antibody is a monospecific antibody.

[0076] The "IgG antibody" used herein has the structure of a naturally occurring IgG antibody, that is, it has the same number of heavy and light chains and disulfide bonds as a naturally occurring IgG antibody of the same subclass. For example, an anti-ICOS IgG1, IgG2, IgG3, or IgG4 antibody consists of two heavy chains (HC) and two light chains (LC), where these two heavy and light chains are linked by the same number and positions of disulfide crosslinks as in naturally occurring IgG1, IgG2, IgG3, and IgG4 antibodies, respectively (unless the antibody has undergone a mutation that modifies the disulfide bonds).

[0077] An "isolated antibody" is an antibody that is substantially free of other antibodies with different antigen specificity (for example, an isolated antibody that specifically binds to PD-1 is substantially free of antibodies that specifically bind to antigens other than PD-1), however, an isolated antibody that specifically binds to PD-1 may cross-react with other antigens, such as PD-1 molecules from different species. Furthermore, an isolated antibody may be substantially free of other cellular material and / or chemical substances.

[0078] Antibodies can be modified antibodies (e.g., by mutation, deletion, substitution, or conjugation to a non-antibody portion). For example, an antibody may contain one or more variant amino acids (compared to naturally occurring antibodies) that alter the properties of the antibody (e.g., functional properties). Numerous such alterations are known in the field, for example, affecting the half-life in a patient, effector function, and / or the immune response to the antibody. The term antibody also includes artificial polypeptide constructs that contain at least one antibody-derived antigen-binding site.

[0079] The term “monoclonal antibody” (“mAb”) refers to a preparation of an antibody molecule that does not exist in nature and has a single molecular composition; that is, an antibody molecule whose primary sequence is essentially identical and which exhibits a single binding specificity and affinity to a particular epitope. MAbs can be produced by hybridoma, recombinant, transgenic, or other techniques known to those skilled in the art.

[0080] A “human” antibody (HuMAb) refers to an antibody having a variable region in which both the framework and CDR region are derived from human germline immunoglobulin sequences. Furthermore, if the antibody includes a constant region, the constant region is also derived from a human germline immunoglobulin sequence. The human antibodies of this invention may contain amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced in vitro by random or site-directed mutagenesis or in vivo by somatic mutation). However, the term “human antibody” as used herein is not intended to include antibodies in which germline-derived CDR sequences from other mammalian species, such as mice, have been transplanted into a human framework sequence. The terms “human” antibody and “fully human” antibody are used synonymously.

[0081] A "humanized antibody" is an antibody in which some, most, or all of the amino acids outside the CDR domain of a non-human antibody are replaced with corresponding amino acids derived from human immunoglobulin. In some embodiments of the humanized form of an antibody, some, most, or all of the amino acids outside the CDR domain are replaced with amino acids from human immunoglobulin, while some, most, or all of the amino acids within the CDR domain remain unchanged. Small additions, deletions, insertions, substitutions, or modifications of amino acids are acceptable as long as they do not inhibit the antibody's ability to bind to a particular antigen. A "humanized" antibody retains antigen specificity similar to that of the original antibody.

[0082] A "chimeric antibody" refers to an antibody in which the variable region originates from one species and the constant region originates from another species, such as an antibody in which the variable region originates from a mouse antibody and the constant region originates from a human antibody.

[0083] An "anti-antigen" antibody is an antibody that specifically binds to a particular antigen. For example, an anti-PD-1 antibody specifically binds to PD-1, and an anti-CTLA-4 antibody specifically binds to CTLA-4.

[0084] The "antigen-binding portion" (also called the "antigen-binding fragment") of an antibody refers to one or more fragments of the antibody that retain the ability of the entire antibody to specifically bind to the antibody. It has been shown that the antigen-binding function of an antibody can be performed by a fragment or a portion of a full-length antibody. Examples of binding fragments that fall within the scope of the term "antigen-binding portion" or "antigen-binding fragment" for an antibody, such as the anti-ICOS antibody described herein, are: (1) Fab fragments (fragments cleaved by papain) or similar monovalent fragments consisting of VL, VH, LC, and CH1 domains; (2) F(ab')2 fragment (pepsin-cleaved fragment) or similar divalent fragment containing two Fab fragments linked by disulfide crosslinking in the hinge region; (3) Fd fragments consisting of VH and CH1 domains; (4) Fv fragment consisting of the VL and VH domains of a single arm of the antibody, (5) A single-domain antibody (dAb) fragment consisting of a VH domain (Ward et al., (1989) Nature 341:544-46); (6) A dual-domain antibody consisting of two VH domains linked by a hinge (dual-affinity retargeting antibody (DART)); (7) Dual variable domain immunoglobulin; (8) Isolation complementarity determination region (CDR); and (9) A combination of two or more isolated CDRs, which may be linked by a synthetic linker if desired. Furthermore, the two domains of the Fv fragment, VL and VH, are encoded by separate genes, but may be linked by a synthetic linker that allows them to be produced as a single protein chain in which the VL and VH regions pair to form a monovalent molecule using a recombinant method (known as single-stranded Fv (scFv); see, e.g., Bird et al. (1988) Science 242:423-426; and Huston et al. (1988) Proc. Natl. Acad. Sci. USA 85:5879-5883). Such single-stranded antibodies are also intended to be included in the terms “antigen-binding portion” or “antigen-binding fragment” of an antibody. These antibody fragments are obtained using conventional techniques known to those skilled in the art, and the fragments are screened for usefulness in the same manner as intact antibodies. The antigen-binding portion can be produced by recombinant DNA techniques or by enzymatic or chemical cleavage of intact immunoglobulins.

[0085] The terms "LAG-3," "LAG3," or "lymphocyte-activating gene-3" refer to lymphocyte-activating gene-3. The term LAG-3 as used herein includes human LAG-3 (hLAG-3), variants, isoforms, and species homologs of hLAG-3, and analogs having at least one common epitope with hLAG-3. The term LAG-3 as used herein includes variants, isoforms, homologs, orthologs, and paralogs. For example, an antibody specific to the human LAG-3 protein may, in some cases, cross-react with LAG-3 proteins from non-human species. In other embodiments, an antibody specific to the human LAG-3 protein may be perfectly specific to the human LAG-3 protein and may not exhibit species or other type cross-reactivity, or may cross-react with LAG-3 from some other species but not all other species (e.g., cross-reacting with monkey LAG-3 but not mouse LAG-3). The term "human LAG-3" refers to human LAG-3 sequences, such as the complete amino acid sequence of human LAG-3 with GenBank Accession No. NP_002277 (Sequence ID 13). The term "mouse LAG-3" refers to mouse LAG-3 sequences, such as the complete amino acid sequence of mouse LAG-3 with Genbank Accession No. NP_032505. LAG-3 is also known in this field, for example, as CD223. Human LAG-3 sequences may differ from human LAG-3 of Genbank Accession No. NP_002277, for example, by having conserved mutations or mutations in non-conserved regions, and LAG-3 has substantially the same biological function as human LAG-3 of Genbank Accession No. NP_002277 (Sequence ID 44). For example, the biological function of human LAG-3 may be having an epitope in the extracellular domain of LAG-3 that is specifically bound by the antibody of the present invention, or the biological function of human LAG-3 may be binding to MHC class II molecules.

[0086] A particular human LAG-3 sequence is generally at least 90% identical in amino acid sequence to the human LAG-3 of Genbank Accession No. NP_002277 and contains amino acid residues that identify the sequence as human when compared to LAG-3 amino acid sequences of other species (e.g., mouse). In some cases, human LAG-3 may be at least 95% or at least 96%, 97%, 98%, or 99% identical in amino acid sequence to the LAG-3 of Genbank Accession No. NP_002277. In some embodiments, a human LAG-3 sequence exhibits no more than 10 amino acid differences from the LAG-3 sequence of Genbank Accession No. NP_002277. In some embodiments, human LAG-3 may exhibit no more than 5 amino acid differences from the LAG-3 sequence of Genbank Accession No. NP_002277, or no more than 4, 3, 2, or 1. Percentage identity may be determined as described herein.

[0087] The terms “programmed death 1,” “programmed cell death 1,” “protein PD-1,” “PD-1,” “PD1,” “PDCD1,” “hPD-1,” and “hPD-I” used herein are interchangeable and include variants, isoforms, species homologs of human PD-1, and analogs having at least one common epitope with PD-1. The complete PD-1 sequence can be found under GenBank Accession Nos. U64863 (SEQ ID NO. 29) and AAC51773.1 (SEQ ID NO. 45).

[0088] Programmed death 1 (PD-1) is an inhibitory member of the CD28 family of receptors, which also includes CD28, CTLA-4, ICOS, and BTLA. PD-1 is expressed in activated B cells, T cells, and myeloid cells (Agata et al., supra; Okazaki et al. (2002) Curr. Opin. Immunol. 14: 391779-82; Bennett et al. (2003) J Immunol 170:711-8). The initial members of this family, CD28 and ICOS, were discovered due to their functional effects of enhancing T cell proliferation after the addition of monoclonal antibodies (Hutloff et al. Nature (1999); 397:263-266; Hansen et al. Immunogenics (1980); 10:247-260). PD-1 was discovered through differential expression screening in apoptotic cells (Ishida et al. EMBO J (1992); 11:3887-95). Other members of this family, CTLA-4 and BTLA, were discovered through differential expression screening in cytotoxic T lymphocytes and TH1 cells, respectively. CD28, ICOS, and CTLA-4 all possess unpaired cysteine ​​residues that enable homodimerization. In contrast, PD-1 lacks the unpaired cysteine ​​residues characteristic of other CD28 family members, suggesting that it exists as a monomer.

[0089] The PD-1 gene is a 55 kDa type I transmembrane protein that is part of the Ig gene superfamily (Agata et al. (1996) Int Immunol 8:765-72). PD-1 contains a membrane-proximal immunoreceptor tyrosine inhibitory motif (ITIM) and a membrane-distal tyrosine-based switch motif (ITSM) (Thomas, ML (1995) J Exp Med 181:1953-6; Vivier, E and Daeron, M (1997) Immunol Today 18:286-91). PD-1 is structurally similar to CTLA-4 but lacks the MYPPPY motif crucial for B7-1 and B7-2 binding. Two ligands for PD-1, PD-L1 and PD-L2, have been identified and shown to downregulate T cell activation by binding to PD-1 (Freeman et al. (2000) J Exp Med 192:1027-34; Latchman et al. (2001) Nat Immunol 2:261-8; Carter et al. (2002) Eur J Immunol 32:634-43). Both PD-L1 and PD-L2 are B7 homologs that bind to PD-1 but not to other CD28 family members. PD-L1 is abundant in a variety of human cancers (Dong et al. (2002) Nat. Med. 8:787-9). The interaction between PD-1 and PD-L1 leads to a decrease in tumor-infiltrating lymphocytes, reduced T-cell receptor-mediated proliferation, and immune evasion by cancer cells (Dong et al. (2003) J. Mol. Med. 81:281-7; Blank et al. (2005) Cancer Immunol. Immunother. 54:307-314; Konishi et al. (2004) Clin. Cancer Res. 10:5094-100).Immunosuppression can be reversed by inhibiting the local interaction between PD-1 and PD-L1, and the effect is additive when the interaction between PD-1 and PD-L2 is similarly blocked (Iwai et al. (2002) Proc. Nat'l. Acad. Sci. USA 99:12293-7; Brown et al. (2003) J. Immunol. 170:1257-66).

[0090] Consistent with PD-1 being an inhibitory member of the CD28 family, PD-1-deficient animals develop a variety of autoimmune phenotypes, including autoimmune cardiomyopathy and lupus-like syndrome with arthritis and nephritis (Nishimura et al. (1999) Immunity 11:141-51; Nishimura et al. (2001) Science 291:319-22). Furthermore, PD-1 has also been found to play a role in autoimmune encephalomyelitis, systemic lupus erythematosus, graft-versus-host disease (GVHD), type 1 diabetes mellitus, and rheumatoid arthritis (Salama et al. (2003) J Exp Med 198:71-78; Prokunina and Alarcon-Riquelme (2004) Hum Mol Genet 13:R143; Nielsen et al. (2004) Lupus 13:510). In mouse B-cell tumor lines, PD-1 ITSM is BCR-mediated Ca 2+ It has been shown to be essential for blocking tyrosine phosphorylation of flow and downstream effector molecules (Okazaki et al. (2001) PNAS 98:13866-71).

[0091] Programmed death ligand-1 (PD-L1) is one of two cell surface glycoprotein ligands for PD-1 (the other being PD-L2), which downregulates T cell activation and cytokine secretion by binding to PD-1. The term "PD-L1" used herein includes human PD-L1 (hPD-L1), variants, isoforms, and species homologs of hPD-L1, as well as five analogs that share at least one common epitope with hPD-L1. The complete hPD-L1 sequence can be found under GenBank Accession No. Q9NZQ7.

[0092] The terms "programmed death ligand-2" and "PD-L2" used herein include human PD-L2 (hPD-L2), variants, isoforms, and species homologs of hPD-L2, as well as analogs that share at least one common epitope with hPD-L2. The complete hPD-L2 sequence can be found under GenBank Accession No. Q9BQ51.

[0093] "Cancer" refers to a diverse group of diseases characterized by the uncontrolled proliferation of abnormal cells in the body. Uncontrolled cell division and proliferation lead to the formation of malignant tumors, which can invade neighboring tissues and metastasize to distal parts of the body via the lymphatic system or bloodstream. "Cancer" or "cancer tissue" may include tumors.

[0094] As used herein, the term "tumor" refers to any mass of tissue resulting from excessive cell growth or proliferation, whether benign (non-cancerous) or malignant (cancerous), including precancerous lesions.

[0095] The term "and / or," as used herein, is to be interpreted as a specific disclosure of each of the two identified properties or components, with or without the other. Thus, the term "and / or," as used herein in terms such as "A and / or B," includes "A and B," "A or B," "A" (alone) and "B" (alone). Similarly, the term "and / or," as used in terms 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).

[0096] Where an aspect is described herein using the term “including,” it is understood that equivalent aspects are also provided, except those described using the terms “consisting of” and / or “essentially consisting of.”

[0097] The terms “approximately” or “essentially” mean a value or composition within an acceptable margin of error for a particular value or composition, as determined by those skilled in the art, which is partly due to how the value or composition was measured or determined, i.e., the limits of the measuring system. For example, “approximately” or “essentially” may mean within one or more standard deviations according to the conventions of the art. Alternatively, “approximately” or “essentially” may mean a range of up to 10% or 20% (i.e., ±10% or ±20%). For example, approximately 3 mg may include any number between 2.7 mg and 3.3 mg (for 10%) or between 2.4 mg and 3.6 mg (for 20%). Furthermore, particularly with respect to biological systems or processes, the term may mean up to one order of magnitude 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 “approximately” or “essentially” should be assumed to be within an acceptable margin of error for that particular value or composition.

[0098] Any concentration ranges, percentage ranges, ratio ranges, or integer ranges used herein are understood to include any integer values ​​within the quoted range, and, where appropriate, fractions thereof (e.g., 1 / 10 and 1 / 100 of an integer), unless otherwise specified.

[0099] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art to which this disclosure belongs. For example, *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 many general dictionaries of the terms used herein.

[0100] Units, prefixes, and symbols are given in the form recognized by the International System of Units (SI). Numerical ranges include the numerical values ​​that define those ranges. The titles provided herein are not limited to the various forms of disclosure that may be obtained by referring to the specification as a whole. Accordingly, the terms defined immediately below are more fully defined by referring to this specification as a whole.

[0101] Various aspects of the present invention will be described in further detail in the following subsections.

[0102] IIa. Anti-LAG-3 antibody An anti-human LAG-3 antibody (or a VH / VL domain derived thereof) suitable for use in the present invention can be produced using methods well known in the art. Alternatively, an anti-LAG-3 antibody recognized in the art can be used. For example, an anti-human LAG-3 antibody described in US2011 / 0150892A1, which is incorporated herein by reference, and referred to as monoclonal antibody 25F7 (also known as "25F7" and "LAG3.1") can be used. Other anti-LAG-3 antibodies available for use and recognized in this field include IMP731(H5L7BW) as described in US2011 / 007023, MK-4280(28G-10) as described in WO2016028672, REGN3767 as described in Journal for ImmunoTherapy of Cancer, (2016) Vol. 4, Supp. Supplement 1 Abstract Number: P195, BAP050, IMP-701(LAG-525), IMP321(Eftilagimod Alpha)), Sym022, TSR-033, MGD013, BI754111, FS118, AVA-017, and GSK2831781 as described in WO2017 / 019894. Examples of anti-LAG-3 antibodies useful in the present invention include: WO2016 / 028672, WO2017 / 106129, WO2017 / 062888, WO2009 / 044273, WO2018 / 069500, WO2016 / 126858, WO2014 / 179664, WO2016 / 200782, WO2015 / 200119, WO2017 / 019846, WO2017 / 198741, WO2017 / 220555, WO20 These can be found in 17 / 220569, WO2018 / 071500, WO2017 / 015560, WO2017 / 025498, WO2017 / 087589, WO2017 / 087901, WO2018 / 083087, WO2017 / 149143, WO2017 / 219995, US2017 / 0260271, WO2017 / 086367, WO / 2017 / 086419, WO2018 / 034227, and WO2014 / 140180. The contents of each of these cited documents are incorporated herein by reference.

[0103] Regarding binding to LAG-3, antibodies that compete with any of the control antibodies recognized in this field may also be used.

[0104] Exemplary anti-LAG-3 antibodies are BMS-986016 or its antigen-binding fragments and variants, which include heavy and light chains containing the sequences shown in SEQ ID NOs. 1 and 2, respectively, as described in U.S. Patent No. 9,505,839, as incorporated herein by reference.

[0105] In other embodiments, the antibody has heavy and light chain CDRs or variable regions of BMS-986016. Thus, in one embodiment, the antibody includes CDR1, CDR2, and CDR3 domains of the VH region of BMS-986016 having the sequence shown in SEQ ID NO: 3, and CDR1, CDR2, and CDR3 domains of the VL region of BMS-986016 having the sequence shown in SEQ ID NO: 5. In another embodiment, the antibody includes CDR1, CDR2, and CDR3 domains containing the sequences shown in SEQ ID NOs: 7, 8, and 9, respectively, and CDR1, CDR2, and CDR3 domains containing the sequences shown in SEQ ID NOs: 10, 11, and 12, respectively. In another embodiment, the antibody includes VH and / or VL regions containing the amino acid sequences shown in SEQ ID NO: 3 and / or SEQ ID NO: 5, respectively. In another embodiment, the antibody includes heavy chain variable (VH) and / or light chain variable (VL) regions encoded by the nucleic acid sequences shown in SEQ ID NO: 4 and / or SEQ ID NO: 6, respectively. In other embodiments, the antibody binds to the same epitope of LAG-3 that competes for binding with the antibody described above. In other embodiments, the antibody binds to an epitope of human LAG-3 comprising the amino acid sequence PGHPLAPG (SEQ ID NO: 14). In other embodiments, the antibody binds to an epitope of human LAG-3 comprising the amino acid sequence HPAAPSSW (SEQ ID NO: 15) or PAAPSSWG (SEQ ID NO: 16).

[0106] In other embodiments, the antibody has at least about 90% variable region amino acid sequence identity with the above antibody (e.g., at least about 90%, 95% or 99% variable region identity with SEQ ID NO: 3 or SEQ ID NO: 5).

[0107] In certain embodiments, the anti-LAG-3 antibody is a bispecific antibody. In certain embodiments, the anti-LAG-3 antibody is a bispecific antibody that binds to both PD-1 and LAG-3.

[0108] Ib. Anti-PD-1 antibody

[0109] Human monoclonal antibodies (HuMAbs) that specifically bind to PD-1 with high affinity are disclosed in U.S. Patents 8,008,449 and 8,779,105. Other anti-PD-1 mAbs are described, for example, in U.S. Patents 6,808,710, 7,488,802, 8,168,757 and 8,354,509 and PCT Publication WO2012 / 145493. Each of the anti-PD-1 HuMAbs disclosed in U.S. Patent 8,008,449 has been shown to exhibit one or more of the following characteristics: (a) binds to human PD-1 with a K -7 of 1×10 D or less determined by surface plasmon resonance using a Biacore biosensor system; (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 monkey 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 Ab response; and (j) inhibits tumor cell proliferation in vivo. Anti-PD-1 antibodies useful in the present invention include mAbs that specifically bind to human PD-1 and exhibit at least one, preferably at least five, of the above characteristics.

[0110] In certain embodiments, the anti-PD-1 antibody is nivolumab. Nivolumab (Opdivo(登録商標) Also known as ;BMS-936558; formerly named 5C4, BMS-936558, MDX-1106 or ONO-4538), is a fully human IgG4(S228P) PD-1 immune checkpoint inhibitor antibody that selectively inhibits interaction with PD-1 ligands (PD-L1 and PD-L2), thereby blocking the downregulation of antitumor T cell function (US Patent 8,008,449; Wang et al., 2014 Cancer Immunol Res. 2(9):846-56). In other embodiments, the anti-PD-1 antibody or its fragments cross-compete with nivolumab. In other embodiments, the anti-PD-1 antibody or its fragments bind to the same epitope as nivolumab. In some embodiments, the anti-PD-1 antibody has the same CDR as nivolumab.

[0111] In other embodiments, the anti-PD-1 antibody is pembrolizumab. Pembrolizumab is a humanized monoclonal IgG4 (S228P) antibody against the human cell surface receptor PD-1 (programmed death-1 or programmed cell death-1). Pembrolizumab is described, for example, in U.S. Patents 8,354,509 and 8,900,587.

[0112] Anti-human PD-1 antibodies (or VH and / or VL domains derived therefrom) suitable for use in the present invention can be produced using methods well known in the art. Alternatively, anti-PD-1 antibodies recognized in the art can be used. For example, monoclonal antibodies 5C4 (hereinafter referred to as nivolumab or BMS-936558), 17D8, 2D3, 4H1, 4A11, 7D3, and 5F4 described in WO2006 / 121168, the teachings of which are incorporated herein by reference, can be used. Other known PD-1 antibodies include lambrolizumab (MK-3475) described in WO2008 / 156712 and AMP-514 described in WO2012 / 145493, the teachings of which are incorporated herein by reference. Further known anti-PD-1 antibodies and other PD-1 inhibitors include those described in WO2009 / 014708, WO03 / 099196, WO2009 / 114335 and WO2011 / 161699, the teachings of which are incorporated herein by reference. In one embodiment, the anti-PD-1 antibody is REGN2810. In another embodiment, the anti-PD-1 antibody is PDR001. Another known anti-PD-1 antibody is pidilizumab (CT-011). Antibodies that compete with any of these antibodies, or their antigen-binding fragments or inhibitors of binding to PD-1, may also be used.

[0113] Other anti-PD-1 monoclonal antibodies include, for example, US Patents 6,808,710, 7,488,802, 8,168,757 and 8,354,509, US Publication 2016 / 0272708 and PCT Publications WO2012 / 145493, WO2008 / 156712, WO2015 / 112900, WO2012 / 145493, WO2015 / 112800, WO2014 / 206107, WO2015 / 35606, WO2015 / 085847, WO2014 / 179664, WO2017 / 020291, WO2017 / 020858, and WO2016 / 19736. 7, WO2017 / 024515, WO2017 / 025051, WO2017 / 123557, WO2016 / 106159, WO2014 / 194302, WO2017 / 040790, WO2017 / 133540, WO2017 / 132827, WO2017 / 024465, WO2017 / 025016, WO2017 / 106061, WO2017 / 19846, WO2017 / 024465, WO2017 / 025016, WO2017 / 132825 and WO2017 / 133540, each of which is incorporated herein by reference.

[0114] In one embodiment, the anti-PD-1 antibody is 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; see WO2015 / 112900), MEDI-0680 (AstraZeneca; also known as AMP-514; see WO2012 / 145493), semiprimab (Regeneron; also known as REGN-2810; see WO2015 / 112800), JS001 (TAIZHOU JUNSHI PHARMA; Si-Yang Liu et al., J. Hematol. Oncol. See 10:136 (2017), BGB-A317 (Beigene; see WO2015 / 35606 and US2015 / 0079109), INCSHR1210 (Jiangsu Hengrui Medicine; also known as SHR-1210; see Si-Yang Liu et al., J. HematoWO2015 / 085847; Si-Yang Liu et al., J. Hematol. Oncol. 10:136 (2017)l. Oncol. 10:136 (2017)), TSR-042 (Tesaro Biopharmaceutical; also known as ANB011; see WO2014 / 179664), GLS-010 (Wuxi / Harbin Gloria Pharmaceuticals; also known as WBP3055; see Si-Yang Liu et al., J. Hematol. Oncol.Selected from the group consisting of 10:136 (see 2017), AM-0001 (Armo), STI-1110 (Sorrento Therapeutics; see WO2014 / 194302), AGEN2034 (Agenus; see WO2017 / 040790), MGA012 (Macrogenics; see WO2017 / 19846), and IBI308 (Innovent; see WO2017 / 024465, WO2017 / 025016, WO2017 / 132825, and WO2017 / 133540), these references are incorporated herein by citation.

[0115] In other embodiments, the anti-PD-1 antibody or its antigen-binding fragment cross-competes with pembrolizumab. In one embodiment, the anti-PD-1 antibody or its antigen-binding fragment binds to the same epitope as pembrolizumab. In one embodiment, the anti-PD-1 antibody or its antigen-binding fragment has the same CDR as pembrolizumab. In another embodiment, the anti-PD-1 antibody is pembrolizumab. Pembrolizumab ("Keytruda") (登録商標) Pembrolizumab (also known as lambrolizumab and MK-3475) is a humanized monoclonal IgG4 antibody against the human cell surface receptor PD-1 (programmed death-1 or programmed cell death-1). Pembrolizumab is described, for example, in U.S. Patents 8,354,509 and 8,900,587; see also http: / / www.cancer.gov / drugdictionary?cdrid=695789 (last accessed: May 25, 2017). Pembrolizumab is approved by the FDA for the treatment of relapsed or refractory melanoma.

[0116] In other embodiments, the anti-PD-1 antibody or its antigen-binding fragment cross-competes with MEDI0608. In yet another embodiment, the anti-PD-1 antibody or its antigen-binding fragment binds to the same epitope as MEDI0608. In one embodiment, the anti-PD-1 antibody has the same CDR as MEDI0608. In another embodiment, the anti-PD-1 antibody is MEDI0608 (formerly AMP-514), which is a monoclonal antibody. MEDI0608 is described, for example, in U.S. Patent 8,609,089 or http: / / www.cancer.gov / drugdictionary?cdrid=756047 (last accessed May 25, 2017).

[0117] In other embodiments, the anti-PD-1 antibody or its antigen-binding fragment cross-competes with BGB-A317. In some embodiments, the anti-PD-1 antibody or its antigen-binding fragment binds to the same epitope as BGB-A317. In some embodiments, the anti-PD-1 antibody or its antigen-binding fragment has the same CDR as BGB-A317. In some embodiments, the anti-PD-1 antibody or its antigen-binding fragment is BGB-A317, a humanized monoclonal antibody. BGB-A317 is described in U.S. Publication 2015 / 0079109.

[0118] Anti-PD-1 antibodies useful for the disclosed compositions include isolated antibodies that specifically bind to human PD-1 and cross-compete with nivolumab for binding to human PD-1 (see, e.g., U.S. Patents 8,008,449 and 8,779,105; International Publication WO2013 / 173223). 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, sterically preventing the binding of the other cross-competing antibody to that particular epitope region. These cross-competing antibodies are expected to have functional properties very similar to nivolumab due to their binding to the same epitope region of PD-1. Cross-competing antibodies can be readily identified based on their ability to cross-compete with nivolumab in standard PD-1 binding assays such as Biacore analysis, ELISA assay, or flow cytometry (see, e.g., International Publication WO2013 / 173223).

[0119] The anti-PD-1 antibodies useful in the disclosed method also include isolated antibodies that specifically bind to human PD-1 and compete for binding to human PD-1 with any of the anti-PD-1 antibodies disclosed herein, for example, nivolumab (see, e.g., U.S. Patents 8,008,449 and 8,779,105; WO2013 / 173223), which are incorporated herein by reference. In some embodiments, the anti-PD-1 antibody binds to the same epitope as any of the anti-PD-1 antibodies described herein, for example, nivolumab. The ability of antibodies to cross-compete for binding to an antigen indicates that these monoclonal antibodies bind to the same epitope region of the antigen, sterically preventing the other cross-competitive antibody from binding to that particular epitope region. These cross-competitive antibodies are expected to have functional properties very similar to a control antibody, for example, nivolumab, due to their binding to the same epitope region of PD-1. Cross-competitive antibodies can be readily identified based on their ability to cross-compete with nivolumab in standard PD-1 binding assays such as Biacore analysis, ELISA assay, or flow cytometry (see, e.g., WO2013 / 173223).

[0120] In one embodiment, an antibody or its antigen-binding fragment that cross-competes with nivolumab for binding to human PD-1 or binds to the same epitope region of human PD-1 is an mAb. For administration to human subjects, these cross-competing antibodies may be chimeric antibodies or humanized or human antibodies. Such chimeric, humanized or human mAbs may be manufactured and isolated by methods well known in the art.

[0121] The anti-PD-1 antibody useful in the composition of the present invention also includes the antigen-binding moiety of the antibody. It has been well demonstrated that the antigen-binding function of an antibody can be carried out by fragments of a full-length antibody. Examples of binding fragments that fall within the “antigen-binding moiety” of an antibody include (i) the Fab fragment, which is a monovalent fragment consisting of VL, VH, CL, and CH1 domains; (ii) the F(ab')2 fragment, which is a bivalent fragment containing two Fab fragments linked by disulfide crosslinking at a hinge region; (iii) the Fd fragment, which consists of VH and CH1 domains; and (iv) the Fv fragment, which consists of the VL and VH domains of a single arm of the antibody.

[0122] An anti-PD-1 antibody suitable for use in the disclosed composition is an antibody that binds to PD-1 with high specificity and affinity, blocks the binding of PD-L1 and / or PD-L2, and inhibits the immunosuppressive effect of the PD-1 signaling pathway. In one embodiment, the anti-PD-1 antibody or its antigen-binding moiety cross-competes with nivolumab for binding to human PD-1. In another embodiment, the anti-PD-1 antibody or its antigen-binding moiety is a chimeric, humanized, or human monoclonal antibody or a part thereof. In one embodiment, the antibody is a humanized antibody. In another embodiment, the antibody is a human antibody. Antibodies of the IgG1, IgG2, IgG3, or IgG4 isotype may be used.

[0123] In one embodiment, the anti-PD-1 antibody or its antigen-binding fragment comprises a heavy chain constant region of a human IgG1 or IgG4 isotype. In another embodiment, the sequence of the IgG4 heavy chain constant region of the anti-PD-1 antibody or its antigen-binding fragment comprises an S228P mutation that replaces a serine residue in the hinge region with a proline residue normally found in the corresponding position of the IgG1 isotype antibody. This mutation present in nivolumab prevents Fab arm exchange with endogenous IgG4 antibody while maintaining low affinity to the associated activated Fc receptor with wild-type IgG4 antibody (Wang et al., 2014). In yet another embodiment, the antibody comprises a light chain constant region which is a human kappa or lambda constant region. In yet another embodiment, the anti-PD-1 antibody or its antigen-binding fragment is an mAb or its antigen-binding moiety. In any embodiment of any of the therapeutic methods described herein, comprising administration of an anti-PD-1 antibody, the anti-PD-1 antibody is nivolumab. In yet another embodiment, the anti-PD-1 antibody is pembrolizumab. In other embodiments, the anti-PD-1 antibody is selected from the human antibodies 17D8, 2D3, 4H1, 4A11, 7D3, and 5F4 described in U.S. Patent 8,008,449. In yet another embodiment, the anti-PD-1 antibody is MEDI0608 (formerly AMP-514), AMP-224, or pidilizumab (CT-011).

[0124] In one embodiment, the anti-PD-1 antibody is a bispecific antibody. In one embodiment, the anti-PD-1 antibody is a bispecific antibody that binds to both PD-1 and LAG-3.

[0125] IIc. Anti-PD-L1 antibody Anti-human PD-L1 antibodies (or VH and / or VL domains derived therefrom) suitable for use in the present invention can be produced using methods well known in the art. Examples of anti-PD-L1 antibodies useful in the methods of the present invention include the antibody disclosed in U.S. Patent 9,580,507, which is incorporated herein by reference. The anti-PD-L1 human monoclonal antibody disclosed in U.S. Patent 9,580,507 has been shown to exhibit one or more of the following features: (a) Determined by surface plasmon resonance using a Biacore biosensor system, it has a 1 × 10⁻¹⁶ ion to human PD-L1. -7 (b) binds with a KD of M or less; (c) increases T cell proliferation in a mixed lymphocyte reaction (MLR) assay; (d) increases interferon-γ production in an MLR assay; (e) stimulates an antibody response; and (f) reverses the effect of T regulatory cells on T cell effector cells and / or dendritic cells. Anti-PD-L1 antibodies useful in the present invention include monoclonal antibodies that specifically bind to human PD-L1 and exhibit at least one, preferably at least five, of the above features.

[0126] Approved anti-PD-L1 antibodies may be used. For example, the human anti-PD-L1 antibody disclosed in U.S. Patent 7,943,743, the contents of which are incorporated herein by reference, may be used. Such anti-PD-L1 antibodies include 3G10, 12A4 (also known as BMS-936559), 10A5, 5F8, 10H10, 1B12, 7H1, 11E6, 12B7, and 13G4. Other art-approved anti-PD-L1 antibodies that may be used include those described in, for example, U.S. Patents 7,635,757 and 8,217,149, U.S. Publication 2009 / 0317368 and PCT Publications WO2011 / 066389 and WO2012 / 145493, the teachings of which are incorporated herein by reference. Other examples of anti-PD-L1 antibodies include atezolizumab (Tecentriq; RG7446), durvalumab (Imfinzi; MEDI4736), or avelumab (Bavencio). Antibodies that compete with any of these art-recognized antibodies, or inhibitors of their antigen-binding fragments or binding to PD-L1, may also be used.

[0127] In one embodiment, the anti-PD-L1 antibody is BMS-936559 (formerly 12A4 or MDX-1105) (see, e.g., U.S. Patent 7,943,743; WO2013 / 173223). In other embodiments, the anti-PD-L1 antibody is MPDL3280A (also known as RG7446 and atezolizumab) (see, e.g., Herbst et al. 2013 J Clin Oncol 31(suppl):3000; U.S. Patent 8,217,149), MEDI4736 (Khleif, 2013, In: Proceedings from the European Cancer Congress 2013; September 27-October 1, 2013; Amsterdam, The Netherlands. Abstract 802), or MSB0010718C (also known as avelumab; see US2014 / 0341917). In one embodiment, the antibody that cross-competes with the control PD-L1 antibody for binding to human PD-L1, or that binds to the same epitope region of human PD-L1, is an mAb. For administration to human subjects, these cross-competing antibodies may be chimeric antibodies, or they may be humanized or human antibodies. Such chimeric, humanized, or human mAbs can be manufactured and isolated by methods well known in the art. In one embodiment, the anti-PD-L1 antibody is BMS-936559 (also known as 12A4, MDX-1105; see, for example, U.S. Patents 7,943,743 and WO2013 / 173223), atezolizumab (Roche; Tecentriq®; also known as MPDL3280A, RG7446; see US8,217,149; also see Herbst et al. (2013) J Clin Oncol 31(suppl):3000), durvalumab (AstraZeneca; Imfinzi) TM, also known as MEDI-4736; see WO2011 / 066389), avelumab (Pfizer; Bavencio®, also known as 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), and CK-301 (Checkpoint Therapeutics; Gorelik et al., AACR:Abtract 4606 (Apr Selected from the group consisting of (see 2016).

[0128] In one embodiment, the PD-L1 antibody is atezolizumab (Tecentriq®). Atezolizumab is a fully humanized IgG1 monoclonal anti-PD-L1 antibody.

[0129] In one embodiment, the PD-L1 antibody is durvalumab (Imfinzi). TM Durvalumab is a human IgG1 kappa monoclonal anti-PD-L1 antibody.

[0130] In one embodiment, the PD-L1 antibody is avelumab (Bavencio®). Avelumab is a human IgG1 lambda monoclonal anti-PD-L1 antibody.

[0131] In another embodiment, the anti-PD-L1 monoclonal antibody is selected from the group consisting of 28-8, 28-1, 28-12, 29-8, 5H1, and any combination thereof.

[0132] The anti-PD-L1 antibodies useful in the disclosed method include isolated antibodies that specifically bind to human PD-L1 and cross-compete with any of the anti-PD-L1 antibodies disclosed herein, e.g., atezolizumab, durvalumab, and / or avelumab, for binding to human PD-L1. In one embodiment, 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. 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, sterically preventing the other cross-competing antibody from binding to that particular epitope region. These cross-competing antibodies are expected to have functional properties very similar to a control antibody, e.g., atezolizumab and / or avelumab, due to their binding to the same epitope region of PD-L1. Cross-competitive antibodies can be readily identified based on their ability to cross-compete with atezolizumab and / or avelumab in standard PD-L1 binding assays such as Biacore analysis, ELISA assay, or flow cytometry (see, e.g., WO2013 / 173223).

[0133] In one embodiment, an antibody that cross-competes with atezolizumab, durvalumab, and / or avelumab for binding to human PD-L1, or that binds to the same epitope region of a human PD-L1 antibody, is a monoclonal antibody. For administration to human subjects, these cross-competing antibodies are chimeric antibodies, modified antibodies, or humanized or human antibodies. Such chimeric, modified, humanized, or human monoclonal antibodies can be manufactured and isolated by methods well known in the art.

[0134] The anti-PD-L1 antibody useful in the disclosed method also includes the antigen-binding portion of the antibody. It has been well demonstrated that the antigen-binding function of the antibody can be performed by a fragment of a full-length antibody.

[0135] An anti-PD-L1 antibody suitable for use in the disclosed method or composition is an antibody that binds to PD-L1 with high specificity and affinity, blocks PD-1 binding, and inhibits the immunosuppressive effect of the PD-1 signaling pathway. In any of the compositions or methods disclosed herein, the anti-PD-L1 "antibody" comprises an antigen-binding moiety or fragment that binds to PD-L1 and exhibits functional properties similar to the whole antibody in receptor binding inhibition and immune system upregulation. In some embodiments, the anti-PD-L1 antibody or its antigen-binding moiety cross-competes with atezolizumab, durvalumab, and / or avelumab for binding to human PD-L1.

[0136] IId. Immunotherapy agents The "immunotherapy agents" and "cancer immunotherapy drugs" used in this invention include any agents, compounds, or biological agents capable of modulating the host immune system. Immunotherapy agents may be immune checkpoint inhibitors, immune checkpoint enhancers, or immune checkpoint stimulants. The immunotherapy agents described herein may be used in combination with one or more further immunotherapy agents (e.g., anti-PD-1 antibodies and anti-LAG-3 antibodies).

[0137] In one embodiment, the immunotherapeutic agent is an immune checkpoint inhibitor. Non-limiting examples of such immunotherapeutic agents include: (i) CTLA-4 (CD152) antagonists (e.g., Yervoy® (ipilimumab) (US Patent 6,984,720); tremelimumab (formerly tisilimumab and CP-675,206); AGEN-1884; and ATOR-1015 (anti-OX40 and anti-CTLA-4 bispecific antibody) disclosed in US Patent 6,984,720, which binds specifically to CTLA-4 with high affinity) human monoclonal antibodies. Other anti-CTLA-4 monoclonal antibodies are disclosed, for example, in U.S. Patents 5,977,318, 6,051,227, 6,682,736 and 7,034,121 and International Publications WO2012 / 122444, WO2007 / 113648, WO2016 / 196237 and WO2000 / 037504. (ii) TIM-3 (HAVCR2) antagonists (e.g., TSR-022 and LY3321367); (iii) TIGIT (T cell immune receptor having Ig and ITIM domains) antagonists (e.g., BMS-986207, OMP-313M32, COM902 (CGEN-15137), and AB154); (iv) IDO1 (indoleamine-2,3-dioxygenase 1) antagonists (e.g., indoximod (NLG8189, 1-methyl-D-TRP), epacadostat (INCB-024360), KHK2455, PF-06840003 (PCT Publication WO2016 / 181348A1), pyrrolizidine-2,5-dione derivatives (PCT Publication WO2015 / 173764A1), napoximod (RG6078, GDC-0919, NLG919), and BMS-986205 (F001287)); (v) KIR (killer cell immunoglobulin-like receptor) antagonists (e.g., lirirumab (I-7F9, BMS-980615, or IPH2101) and IPH4102 (anti-KIR3DL2 monoclonal antibody); (vi) TDO (tryptophan 2,3-dioxygenase) antagonists (e.g., 4-(indole-3-yl)-pyrazole derivatives (US Patent 9,126,984B2 and US Publication 2016 / 0263087A1); 3-indole-substituted derivatives (PCT Publications WO2015140717A1, WO2017025868A1, WO2016147144A1), 3-(indole-3-yl)-pyridine derivatives (US Publication 20150225367A1 and PCT Publication WO2015121812A1); (vii) Dual IDO / TDO antagonists (e.g., small molecule dual IDO / TDO inhibitors disclosed in PCT publications WO2015150097A1, WO2015082499A2, WO2016026772A1, WO2016071283A1, WO2016071293A2 and WO2017007700A1); (viii) CD40 antagonists (e.g., Lineage BMS3h-56 (US Patent 9,475,879), lucatumumab (HCD122 and CHIR-12.12), CHIR-5.9, and dasetuzumab (huS2C6, PRO 64553, RG 3636, SGN 14, SGN-40)); (ix) Adenosine A2a receptor (A2aR) antagonists (e.g., CPI-444, PBF-509, istradefylline (KW-6002), preradianant (SCH420814), tozadenant (SYN115), bipardenant (BIIB014), HTL-1071, ST1535, SCH412348, SCH442416, SCH58261, ZM241385, and AZD4635 (small molecule A2aR inhibitors)); (x) VISTA (V-domain immunoglobulin (Ig)-containing inhibitor of T cell activation) antagonist (e.g., CA-170 (anti-PD-L1 / L2 and anti-VISTA small molecule) and JNJ-61610588); (xi)CEACAM1(CD66a) Antagonist (e.g., CM-24(MK-6018)); (xii) CEA (carcinoembryonic antigen) antagonists (e.g., cerguzumab amnaleukin (RG7813, RO-6895882), RG7802 (RO6958688)); (xiii) CD47 antagonists (e.g., HuF9-G4, CC-90002, TTI-621, ALX148, NI-1701, NI-1801, SRF231 and Effi-DEM); (xiv) PVRIG (CD122R-containing poliovirus receptor-associated immunoglobulin domain) antagonist (e.g., COM701); (xv) GARP (repeated dominant glycoprotein A) antagonist (e.g., ARGX-115); (xvi) STING (IFN gene stimulator) agonists (e.g., 2' or 3'-mono-fluorosubstituted or 2'3'-di-fluorosubstituted mixed bonded 2',5'-3',5' cyclic dinucleotide (PCT Publication WO2017 / 075477A1); 2'-fluorosubstituted, bis-3',5' cyclic dinucleotide and 2',2''-diF-Rp,Rp,bis-3',5' cyclic dinucleotide (PCT Publication WO2016 / 145102A1); and fluorinated cyclic dinucleotide (PCT Publication WO2016 / 096174A1); (xvii) CD20 agonists (e.g., Rituxan® and ABP 798); (xviii) CD80 antagonists (e.g., galiximab (IDEC-114) and AV 1142742 (RhuDex)); (xix)CD86 Antagonist; and (xx) CD96 Antagonist Includes.

[0138] In other embodiments, the immunotherapeutic agent is an immune checkpoint stimulator or enhancer. Non-limiting examples of such immunotherapeutic agents include: (i) CD28 agonists (e.g., TGN1412 (anti-CD28 antibody) and JCAR015 (anti-CD19-CD28-zeta modified chimeric antigen receptor)); (ii) CD80 or CD86 agonists (e.g., CTLA4-Ig fusion constructs (CTLA-4-IgG4m, RG2077 or RG1046); Orencia® (avatacept or BMS-188667); and MGN1601); (iii) ICOS or ICOS-ligand agonists (e.g., BMS986226, MEDI-570, GSK3359609, JTX-2011 and AMG 570); (iv) 4-1BB(CD137) agonists (e.g., urelumab and PF-05082566); (v) OX40 (CD134 or TNFRS4) agonists (e.g., tavorixizumab (MEDI-0562); pogalizumab (MOXR0916, RG7888); GSK3174998; ATOR-1015 (anti-OX40 and anti-CTLA-4 bispecific antibody); MEDI-6383; MEDI-6469; BMS 986178; PF-04518600; and GINAKIT cells (iC9-GD2-CD28-OX40 expressing T lymphocytes)); (vi) CD27 agonists (e.g., valrirumab (CDX-1127)); (vii) CD40 agonists (e.g., ADC-1013 (JNJ-64457107), RG7876 (RO-7009789), HuCD40-M2; APX005M (EPI-0050) (US Patent 9,556,278); Chi Lob7 / 4 (IgG1 chimeric agonist CD40 monoclonal antibody)); (viii) CD70 agonists (e.g., ARGX-110); and (ix) GITR agonists (e.g., BMS-986156, TRX518, GWN323, INCAGN01876 and MEDI1873) Includes.

[0139] In one embodiment, the immunotherapy agent is a cytokine, e.g., chemokines, interferons (e.g., interferon-gamma), interleukins (e.g., aldesleukin (a recombinant analog of IL-2 with immunomodulatory and antineoplastic activity), tocilizumab (an anti-IL-6 receptor antibody)); lymphokines or members of the tumor necrosis factor (TNF) family (e.g., ATOR-1016, ABBV-621, and adalimumab). Other examples of immunotherapies include CSF1R (colony-stimulating factor 1 receptor, CD115) antagonists (e.g., emactuzumab); Toll-like receptor 9 (TLR9) agonists (e.g., agatrimod sodium); CD160 (NK1) agonists (e.g., BY55); CD73 antagonists (5'-nucleotidase or ecto-5'-nucleotidase) (e.g., MEDI9447); iNOS (inducible NO synthase, NOS2) antagonists (e.g., N-acetylcysteine ​​(NAC), aminoguanidine, L-nitroarginine methyl ester, S,S-1,4-phenylene-bis(1,2-ethanediyl)bis-isothiourea); and SHP-1 (Src homology 2-domain-containing protein tyrosine phosphatase 1) antagonists (Watson et al., Biochem Soc Trans) See 44(2): 356-362 (2016).

[0140] III. Pharmaceutical Compositions Pharmaceutical compositions suitable for administration to human patients are generally formulated to be suitable for reconstitution in liquid carriers, for example, in liquid solutions or suspensions for non-enteral or intravenous administration.

[0141] Generally, such compositions typically contain a pharmaceutically acceptable carrier. As used herein, “pharmaceutically acceptable” means approved by a government regulatory authority or listed in the United States Pharmacopeia or other generally recognized pharmacopoeias for use in animals, particularly humans. The term “carrier” refers to any diluent, adjuvant, additive, or medium administered with the compound. Such pharmaceutical carriers may be sterile liquids such as water and oils (including, for example, petroleum, animal, plant, or synthetic sources such as peanut oil, soybean oil, mineral oil, sesame oil, and glycerol polyethylene glycol lysine oleate). Water or saline solutions and aqueous dextrose and glycerol solutions may be used as carriers, particularly for injectable solutions (including, for example, anti-PD-1 antibodies, anti-LAG-3 antibodies, and / or other immunotherapeutic agents). Non-enteral liquid compositions may be formulated for administration by injection or continuous infusion. Routes of administration by injection or infusion include intravenous, intraperitoneal, intramuscular, intrathecal, and subcutaneous. In one embodiment, a composition comprising an anti-PD-1 antibody, an anti-LAG-3 antibody, and an immunotherapy agent is administered intravenously (for example, in separate formulations or as a single formulation).

[0142] IV. Patient population Provided herein is a medical method for treating solid tumors (e.g., advanced, refractory solid tumors) in human patients using a combination of an anti-LAG-3 antibody, a PD-1 pathway inhibitor, and an additive immunotherapy agent.

[0143] Examples of cancers that can be treated using the method of the present invention include liver cancer, bone cancer, pancreatic cancer, skin cancer, head and neck cancer, oral cancer, breast cancer, lung cancer including small cell and non-small cell lung cancer, cutaneous or intraocular malignant melanoma, kidney cancer, uterine cancer, ovarian cancer, colorectal cancer, colon cancer, rectal cancer, anal cancer, gastric cancer, testicular cancer, uterine cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, non-Hodgkin lymphoma, esophageal cancer, small intestine cancer, endocrine cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, pediatric solid tumors, lymphocytic lymphoma, bladder cancer, kidney or ureteral cancer, renal pelvis cancer, central nervous system (CNS) neoplasms, primary CNS lymphoma, tumor angiogenesis, spinal axis tumors, brainstem gliomas, and more. This invention includes hematological malignancies and any combination thereof, including pituitary adenoma, Kaposi's sarcoma, epidermoid carcinoma, squamous cell carcinoma, and environment-induced cancers, such as those induced by asbestos, including multiple myeloma, B-cell lymphoma, Hodgkin lymphoma / primary mediastinal B-cell lymphoma, non-Hodgkin lymphoma, acute myeloid lymphoma, chronic myeloid leukemia, chronic lymphoid leukemia, follicular lymphoma, generalized large B-cell lymphoma, Burkitt lymphoma, immunoblastic large cell lymphoma, precursor B-lymphoblastic lymphoma, mantle cell lymphoma, acute lymphoblastic leukemia, mycosis fungoides, anaplastic large cell lymphoma, T-cell lymphoma, and precursor T-lymphoblastic lymphoma. The invention is also applicable to the treatment of metastatic cancer.

[0144] In one embodiment, the human patient has non-small cell lung cancer (NSCLC), a virus-associated cancer (e.g., human papillomavirus (HPV)-associated tumor), or gastric adenocarcinoma. In a particular embodiment, the HPV-associated tumor is HPV+ head and neck cancer (HNC). In another particular embodiment, the gastric adenocarcinoma is associated with Epstein-Barr virus (EBV) infection.

[0145] Patients may be tested or selected for one or more of the above clinical characteristics before, during, or after the procedure.

[0146] V. Combination Therapy The combination therapies provided herein involve the administration of anti-LAG-3 antibodies, PD-1 pathway inhibitors, and other immunotherapeutic agents that block inhibitory immune receptors (e.g., receptors that inhibit / neutralize activity such as cytotoxic activity by binding to native ligands) to treat subjects with malignant tumors (e.g., advanced, refractory solid tumors).

[0147] In one embodiment, the present invention provides an anti-LAG-3 antibody, an anti-PD-1 antibody, and other immunotherapeutic agents in a combination therapy administered according to a defined clinical dosing schedule for treating subjects with malignant tumors (e.g., advanced, refractory solid tumors). In a particular embodiment, the anti-LAG-3 antibody is BMS-986016. In another embodiment, the anti-PD-1 antibody is BMS-936558. In another embodiment, the dosing schedule is adjusted to provide an optimal desired response (e.g., an effective response).

[0148] The adjunct or combination administration (co-administration) used herein includes the simultaneous administration of multiple compounds in the same or different formulations, or the separate administration of multiple compounds (e.g., sequential administration). Therefore, anti-LAG-3 antibodies, PD-1 pathway inhibitors, and immunotherapeutic agents may be administered simultaneously in a single formulation. Alternatively, anti-LAG-3 antibodies, PD-1 pathway inhibitors, and immunotherapeutic agents may be formulated for separate administration and administered simultaneously, sequentially (e.g., one antibody administered within approximately 30 minutes before the administration of the second antibody), and in any order.

[0149] For example, an anti-PD-1 antibody may be administered first, followed (for example, immediately afterward) by an anti-LAG-3 antibody and / or an immunotherapy agent. In one embodiment, a PD-1 pathway inhibitor is administered before the administration of the anti-LAG-3 antibody and / or immunotherapy agent. In another embodiment, the PD-1 pathway inhibitor is administered after the administration of the anti-LAG-3 antibody and / or immunotherapy agent. In yet another embodiment, the anti-LAG-3 antibody, PD-1 pathway inhibitor, and immunotherapy agent are administered simultaneously. Such simultaneous or sequential administration preferably ensures that all three components are present in the treated patient at the same time.

[0150] VI. Treatment Protocol Appropriate treatment protocols for malignant tumors in human patients include, for example, (a) An anti-LAG-3 antibody comprising CDR1, CDR2, and CDR3 domains of the heavy chain variable region having the sequence shown in SEQ ID NO: 3 and CDR1, CDR2, and CDR3 domains of the light chain variable region having the sequence shown in SEQ ID NO: 5, (b) an anti-PD-1 antibody comprising the CDR1, CDR2, and CDR3 domains of the heavy chain variable region having SEQ ID NO: 19 and the CDR1, CDR2, and CDR3 domains of the light chain variable region having SEQ ID NO: 21, (c) Immunotherapy agents This includes administering each effective dose to the patient, Herein, the method comprises at least one administration cycle, the cycle having a duration of 8 weeks, and for each of at least one cycle, an anti-LAG-3 antibody is administered at least four times in uniform doses of approximately 1 mg, 3 mg, 10 mg, 20 mg, 50 mg, 80 mg, 100 mg, 130 mg, 150 mg, 180 mg, 200 mg, 240 mg, or 280 mg; an anti-PD-1 antibody is administered at least four times in uniform doses of approximately 50 mg, 80 mg, 100 mg, 130 mg, 150 mg, 180 mg, 200 mg, 240 mg, or 280 mg; and an immunotherapy agent is administered at least four times in uniform doses. In another embodiment, an anti-LAG-3 antibody is administered four times at doses of 0.01 mg / kg, 0.03 mg / kg, 0.25 mg / kg, 0.1 mg / kg, 0.3 mg / kg, 1 mg / kg, or 3 mg / kg, 5 mg / kg, 8 mg / kg, or 10 mg / kg body weight, an anti-PD-1 antibody is administered four times at doses of 0.1 mg / kg, 0.3 mg / kg, 1 mg / kg, 3 mg / kg, 5 mg / kg, 8 mg / kg, or 10 mg / kg body weight, and an immunotherapy agent is administered four times at a uniform dose.

[0151] In one embodiment, the dose of the anti-LAG-3 antibody, PD-1 pathway inhibitor, and / or immunotherapy agent is calculated per body weight, for example, mg / kg body weight. In another embodiment, the dose of the anti-LAG-3 antibody, PD-1 pathway inhibitor, and / or immunotherapy agent is a uniform-constant dose. In another embodiment, the dose of the anti-LAG-3 antibody, PD-1 pathway inhibitor, and / or immunotherapy agent changes over time. For example, the anti-LAG-3 antibody, PD-1 antibody, and / or immunotherapy agent may be administered at a high dose initially and then reduced over time. In another embodiment, the anti-LAG-3 antibody, PD-1 pathway inhibitor, and / or immunotherapy agent may be administered at a low dose initially and then increased over time.

[0152] In other embodiments, the amount of anti-LAG-3 antibody, PD-1 pathway inhibitor, and / or immunotherapy agent administered is constant with each dose. In other embodiments, the amount of antibody and / or immunotherapy agent administered varies with each dose. For example, the maintenance (or continuation) dose of antibody and / or immunotherapy agent may be higher than or equal to the initial loading dose. In other embodiments, the maintenance dose of antibody and / or immunotherapy agent may be lower than or equal to the loading dose.

[0153] In other embodiments, the anti-LAG-3 antibody, PD-1 pathway inhibitor, and / or immunotherapy agent are formulated for intravenous administration. In one embodiment, the anti-PD-1 antibody is administered on days 1, 15, 29, and 43 of each cycle. In another embodiment, the anti-LAG-3 antibody is administered on days 1, 15, 29, and 43 of each cycle. In one embodiment, the therapeutic agent is administered on days 1, 15, 29, and 43 of each cycle.

[0154] In other embodiments, anti-LAG-3 antibodies, PD-1 pathway inhibitors, and / or immunotherapies are administered once weekly, once every two or three weeks, once a month, or until clinical benefit is observed or until a full response, confirmed disease progression, or uncontrollable toxicity occurs.

[0155] In other embodiments, the administration cycle is 8 weeks and may be repeated as needed. In other embodiments, the treatment consists of up to 12 cycles.

[0156] In another embodiment, four doses of the PD-1 pathway inhibitor are administered per 8-week cycle. In yet another embodiment, four doses of the PD-1 pathway inhibitor are administered per 8-week cycle. In one embodiment, four doses of the therapeutic agent are administered per 8-week cycle.

[0157] In other embodiments, PD-1 pathway inhibitors, anti-LAG-3 antibodies, and / or immunotherapeutic agents are administered as first-line treatment (e.g., initial or first treatment). In other embodiments, PD-1 pathway inhibitors, anti-LAG-3 antibodies, and / or immunotherapeutic agents are administered as second-line treatment (e.g., after initial or first treatment, including after relapse and / or when initial treatment fails).

[0158] In other embodiments, the present invention relates to any of the above embodiments in which the anti-PD-1 antibody is replaced or combined with an anti-PD-L1 or anti-PD-L2 antibody.

[0159] VII. Outcomes Patients treated with the methods disclosed herein preferably experience improvement in at least one sign of cancer. In one embodiment, improvement is measured by a reduction in the amount and / or size of a measurable tumor lesion. In another embodiment, the lesion may be measured by chest X-ray or CT or MRI film. In yet another embodiment, cytological or histological diagnosis may be used to assess the response to treatment.

[0160] In one embodiment, the treated patient exhibits a complete response (CR), partial response (PR), disease stability (SD), immune-related complete disease (irCR), immune-related partial response (irPR), or immune-related disease stability (irSD). In another embodiment, the treated patient experiences tumor reduction and / or a decrease in growth rate, i.e., tumor growth suppression. In yet another embodiment, unwanted cell proliferation is reduced or inhibited. In yet another embodiment, one or more of the following may occur: the number of cancer cells may decrease; the tumor size may decrease; cancer cell invasion into peripheral organs may be inhibited, delayed, slowed or stopped; tumor metastasis may be slowed or inhibited; tumor growth may be inhibited; tumor recurrence may be prevented or delayed; one or more cancer-related symptoms may be alleviated to some extent.

[0161] In other embodiments, the administration of effective doses of an anti-LAG-3 antibody, a PD-1 pathway inhibitor, and an immunotherapy agent by the method provided herein produces at least one therapeutic effect selected from the group consisting of tumor size reduction, reduction in the number of metastatic lesions appearing over time, complete remission, partial remission, or disease stabilization. In yet another embodiment, the treatment method produces a better, equivalent clinical benefit rate (CBR=CR+PR+SD≧6months) than that achieved by the anti-LAG-3 antibody, PD-1 pathway inhibitor, or immunotherapy agent alone. In other embodiments, the improvement in the clinical benefit rate is approximately 20%, 30%, 40%, 50%, 60%, 70%, 80%, or more compared to the anti-LAG-3 antibody, PD-1 pathway inhibitor, or immunotherapy agent alone.

[0162] VIII. Kits and Unit Dosage Forms Also provided herein is a kit comprising a pharmaceutical composition containing an effective dose suitable for use in the aforementioned method, comprising an anti-LAG-3 antibody (e.g., BMS-986016), a PD-1 pathway inhibitor (e.g., BMS-936558), an immunotherapy agent, and a pharmaceutically acceptable carrier. The kit may optionally include instructions, for example, an administration schedule, enabling a practitioner (e.g., a physician, nurse, or patient) to administer the composition contained therein to a patient with cancer (e.g., a solid tumor). The kit may also include a syringe.

[0163] Optionally, the kit includes multiple packages of single-dose pharmaceutical compositions, each containing an effective amount of anti-LAG-3 antibody, PD-1 pathway inhibitor, and / or immunotherapy agent for single-dose administration according to the method provided above. Apparatus or devices necessary for administering the pharmaceutical compositions may also be included in the kit. For example, the kit may provide one or more pre-filled syringes containing an effective amount of anti-LAG-3 antibody, anti-PD-1 antibody, and / or immunotherapy agent.

[0164] In one embodiment, the present invention provides a kit for treating malignant tumors in human patients, the kit being, for example: (a) A dose of an anti-LAG-3 antibody comprising the CDR1, CDR2, and CDR3 domains of the heavy chain variable region having the sequence shown in SEQ ID NO: 3 and the CDR1, CDR2, and CDR3 domains of the light chain variable region having the sequence shown in SEQ ID NO: 5; (b) A dose of a PD-1 pathway inhibitor, such as an antibody containing the CDR1, CDR2, and CDR3 domains of the heavy chain variable region having SEQ ID NO: 19 and the CDR1, CDR2, and CDR3 domains of the light chain variable region having SEQ ID NO: 21; (c) One dose of an immunotherapy agent; and (d) Instructions for using anti-LAG-3 antibodies, anti-PD-1 pathway inhibitors, and immunotherapeutic agents in the manner described herein. Includes.

[0165] The following embodiments are for illustrative purposes only, and many variations and equivalents will become apparent to those skilled in the art by reading this disclosure; therefore, they should not be construed as limiting the scope of this disclosure. [Examples]

[0166] Example 1 Treatment of malignant tumors with nivolumab monotherapy versus nivolumab + BMS 986016 (anti-LAG-3 antibody) + immunotherapy. To determine whether overall survival (OS) is improved compared to nivolumab monotherapy, a pharmaceutical composition containing nivolumab, BMS 986016, and an immunotherapy agent will be tested in patients with recurrent metastatic tumors. A formal pairwise comparison of OS will be performed between experimental arms (i.e., nivolumab monotherapy versus nivolumab + BMS 986016 + immunotherapy combination therapy).

[0167] This trial will also compare progression-free survival (PFS) and objective response rate (ORR) of nivolumab monotherapy versus nivolumab monotherapy, based on blinded, independent central review (BICR) assessments, in patients with recurrent metastatic tumors. Differences in PFS and ORR between different arms will be evaluated.

[0168] Other objectives of this trial include: 1) evaluating the overall safety and tolerability of the combination therapy compared to nivolumab monotherapy; 2) analyzing the pharmacokinetics of the combination therapy and the exposure-safety and exposure-efficacy correlations; 3) analyzing the immunogenicity of the combination therapy; 4) analyzing the immune correlations of the combination therapy; 5) evaluating predictive tumor and peripheral biomarkers for clinical response to the combination therapy; and 6) evaluating the overall physical condition in subjects treated with the combination therapy using EQ-5D indices and visualized analog indices.

[0169] method Clinical trial design The trial is an open-label, two-arm, randomized study in adult men and women (18 years or older) with stage IV or recurrent non-small cell lung cancer (NSCLC) that has not been previously treated for advanced disease and is PD-L1 positive or negative.

[0170] The main inclusion criteria are: 1) ECOG Performance status of 1 or higher; 2) patients with histologically confirmed stage IV or recurrent NSCLC (according to squamous or non-squamous histology in the 7th International Society for Lung Cancer classification) who have not received prior systemic anticancer therapy (including EGFR and ALK inhibitors) as first-line treatment for advanced or metastatic disease; and 3) disease measurable by CT or MRI according to RECIST 1.1 criteria.

[0171] The main exclusion criteria are: 1) subjects with known EGFR mutations sensitive to available targeted inhibitor therapy; 2) subjects with known ALK translocations sensitive to available targeted inhibitor therapy; 3) subjects with untreated CNS metastases; 4) subjects diagnosed or suspected of having an active autoimmune disease (inclusion is permitted for subjects with type 1 diabetes, hypothyroidism requiring hormone replacement only, skin disorders not requiring systemic treatment (e.g., vitiligo, psoriasis, or alopecia), or conditions not expected to relapse in the absence of external triggers); and 5) subjects requiring systemic treatment with corticosteroids (equivalent to >10 mg prednisone per day) or other immunosuppressants within 14 days of randomization (inhaled or topical steroids and adrenal replacement steroids equivalent to >10 mg prednisone per day are acceptable in the absence of active autoimmune disease).

[0172] Participants will be randomized in a 1:1:1:1 ratio and stratified by histological examination (squamous vs. non-squamous epithelium) and PD-L1 status. PD-L1 status will be determined by immunohistochemical (IHC) staining of PD-L1 protein in tumor samples provided before randomization. Participants will be considered PD-L1 positive if 5% or more tumor cell membrane staining is observed in at least 100 evaluable tumor cells, or PD-L1 negative if less than 5% tumor cell membrane staining is observed in at least 100 evaluable tumor cells.

[0173] Participants will receive open-label treatment in one of the two clinical trial arms. The administration schedule is shown in Table 1. [Table 1] Footnote: "Clinical trial closure" should be changed to "Clinical trial termination." Clinical trial closure means the termination of the entire clinical trial.

[0174] Tumor assessments will begin at 6 weeks (±7 days) after randomization during the trial period and will be conducted every 6 weeks (±7 days) until 48 weeks. After 48 weeks, tumor assessments will be conducted every 12 weeks (±7 days) until disease progression or discontinuation of treatment, whichever is later. Patients receiving nivolumab or combination therapy who have progressed beyond the RECIST 1.1 criteria for investigator assessment must also continue tumor assessments until discontinuation of treatment. Enrollment will end after approximately 1200 patients have been left untreated. The primary endpoint of the trial is overall survival (OS). The trial period from the start of enrollment to the analysis of the primary OS endpoint is expected to be approximately 48 months.

[0175] Clinical trial arm Nivolumab monotherapy (Arm A) Nivolumab 240 mg will be administered intravenously (IV) over 30 minutes on day 1 of each cycle every two weeks until disease progression, unacceptable toxicity occurs, withdrawal of consent, or discontinuation of the study. Procedures exceeding the initial investigator assessment RECIST 1.1 progression are permitted if the patient has a clinical benefit in investigator assessment and is tolerable to the procedure. After completion of administration, the patient will enter the follow-up phase.

[0176] Nivolumab + BMS 986016 + immunotherapy combination therapy (Arm B) Nivolumab 1 mg / kg will be administered intravenously over 30 minutes in 4 cycles every 3 weeks, in combination with BMS 986016 1 mg / kg and an immunotherapy agent administered intravenously over 30 minutes, until disease progression, unacceptable toxicity, withdrawal of consent, or discontinuation of the study. Procedures exceeding the initial investigator assessment RECIST 1.1 progression are permitted if the patient has a clinical benefit in investigator assessment and is tolerable to the procedure. After completion of administration, the patient will enter the follow-up phase.

[0177] Post-procedure follow-up Post-treatment follow-up begins from the decision to discontinue all treatment for the subject; this includes any continued maintenance therapy. Subjects whose treatment is discontinued for reasons other than disease progression will continue tumor evaluation until progression or any subsequent treatment occurs, whichever comes first (if clinically feasible). Subjects will be tracked for drug-related toxicity until these toxicities resolve, return to baseline, or are deemed irreversible. All adverse events will be reported for at least 100 days after the last dose of the investigational drug. After completion of the first two follow-up visits, subjects will be tracked for survival every three months.

[0178] Sample size Approximately 1200 subjects will be randomized to four treatment groups in a 1:1:1:1 ratio. The final analysis will be conducted after 257 events have occurred in the control group, and these events will be followed up by an open-label, independent statistician assisting the DMC. Assuming a 20% screening failure rate, approximately 1500 subjects are expected to be enrolled for the randomization of 1200 subjects. Assuming a segmental constant enrollment rate (8 subjects / month in months 1-2, 40 subjects / month in months 3-4, 85 subjects / month in months 5-6, 138 subjects / month in months 7-8, and 170 subjects / month from month 8 onward), it will take approximately 48 months to obtain the number of deaths required for the final OS analysis (14 months for natural increase and 34 months for survival follow-up).

[0179] endpoint Overall survival (OS) is the primary endpoint of this trial. If OS superiority is demonstrated in at least one comparison, a gatekeeping study method for key secondary endpoints will be applied to further trial-versus-control comparisons as described in the statistical analysis plan. Key secondary endpoints include progression-free survival (PFS) and overall response rate (ORR) based on BICR assessment.

[0180] Each of the three primary OS analyses will be performed using a two-sided log-rank test stratified by histological examination and PD-L1 status for all randomized subjects, with the Hochberg method for multiplicity. Hazard ratios (HRs) and corresponding two-sided (1-adjusted α)% confidence intervals (CIs) will be estimated using a Cox proportional hazards model, with treatment group as the sole covariate and stratified by the aforementioned factors. OS curves, median OS at 95% CI, and 12-month and 24-month OS rates at 95% CI will be estimated using the Kaplan-Meier method. If OS superiority is demonstrated in at least one comparison, gatekeeping studies of key secondary endpoints will be applied to further trial-versus-control comparisons as described in the statistical analysis plan. Key secondary endpoints will be tested in the following hierarchical order: 1) PFS (based on BICR assessment) analysis will be performed using a two-sided log-rank test stratified by histological examination and PD-L1 status for all randomized subjects to compare each of the three investigational treatments against the control group. HR and corresponding two-sided (1-adjusted α)%CI will be estimated using a Cox proportional hazards model, with treatment group as the sole covariate and stratified by the above factors. PFS curves, median PFS at 95% CI, and 6-month and 12-month PFS rates at 95% CI will be estimated using the Kaplan-Meier method. 2) ORR (based on BICR assessment) analysis will be performed using a two-sided Cochrane-Mantel-Henzel (CMH) test stratified by PD-L1 status and histological examination to compare each of the three investigational treatments against the control group. Related odds ratios and (1-adjusted α)%CI will also be calculated. In addition, ORRs and their corresponding 95% accurate CIs will be calculated for each of the four treatment groups using the Cropper-Pearson method. 3) Pairwise comparisons of OS between trial arms will be performed using a two-sided log-rank test stratified by histological examination and PD-L1 status. HR and the corresponding two-sided (1-adjusted α)%CI will be estimated using a Cox proportional hazards model, stratified by the above factors with the treatment group as the sole covariate.

[0181] analysis The analysis of PD-L1 expression is descriptive. The distribution of PD-L1 expression is analyzed based on the overall population. The potential correlation between PD-L1 expression and efficacy tests (ORR, OS, and PFS) is evaluated. If there are signs of a meaningful correlation, further evaluation of PD-L1 expression as a predictive biomarker will be considered to estimate the interaction effect between PD-L1 expression and treatment.

[0182] The results will show whether combination therapy (nivolumab + BMS 986016 + immunotherapy) improves overall survival (OS) compared to nivolumab monotherapy.

[0183] array Sequence ID 1: Heavy chain amino acid sequence; anti-LAG-3 mAb (BMS-986016) QVQLQQWGAGLLKPSETLSLTCAVYGGSFSDYYWNWIRQPPGKGLEWIGEINHRGSTNSNPSLKSRVTLSLDTSKNQFSLKLRSVTAADTAVYYCAFGYSDYEYNWFDPWG QGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYG PPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKT ISKAKGQPREPQVYTLPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK

[0184] Sequence ID 2: Light chain amino acid sequence; anti-LAG-3 mAb (BMS-986016) EIVLTQSPATLSLSPGERATLSCRASQSISSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPLTFGQGTNLEIK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0185] Sequence ID 3: Heavy chain variable region (VH) amino acid sequence; anti-LAG-3 mAb (BMS-986016) QVQLQQWGAGLLKPSETLSLTCAVYGGSFSDYYWNWIRQPPGKGLEWIGEINHRGSTNSNPSLKSRVTLSLDTSKNQFSLKLRSVTAADTAVYYCAFGYSDYEYNWFDPWGQGTLVTVSS

[0186] Sequence ID 4: Heavy chain variable region (VH) nucleotide sequence; anti-LAG-3 mAb (BMS-986016) caggtgcagctacagcagtggggcgcaggactgttgaagccttcggagaccctgtccctcacctgcgctgtctatggtgggtccttcagtgattactactggaactggatccgccagcccccagggaaggggctggagtggattggggaaatcaatcatcgtggaagcaccaactccaac ccgtccctcaagagtcgagtcaccctatcactagacacgtccaagaaccagttctccctgaagctgaggtctgtgaccgccgcggacacggctgtgtattactgtgcgtttggatatagtgactacgagtacaactggttcgacccctggggccagggaaccctggtcaccgtctcctca

[0187] Sequence ID No. 5 Light chain variable region (VL) amino acid sequence; anti-LAG-3 mAb (BMS-986016) EIVLTQSPATLSLSPGERATLSCRASQSISSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPLTFGQGTNLEIK

[0188] Sequence ID 6: Light chain variable region (VL) nucleotide sequence; anti-LAG-3 mAb (BMS-986016) gaaattgtgttgacacagtctccagccaccctgtctttgtctccaggggaaagagccaccctctcctgcagggccagtcagagtattagcagctacttagcctggtaccaacagaaacctggccaggctcccaggctcctcatctatgatgcatccaaca gggccactggcatcccagccaggttcagtggcagtgggtctgggacagacttcactctcaccatcagcagcctagagcctgaagattttgcagtttattactgtcagcagcgtagcaactggcctctcacttttggccaggggaccaacctggagatcaaa

[0189] Sequence ID 7 Heavy chain CDR1 amino acid sequence; anti-LAG-3 mAb (BMS-986016) DYYWN

[0190] Sequence ID 8: Heavy chain CDR2 amino acid sequence; anti-LAG-3 mAb (BMS-986016) EINHRGSTNSNPSLKS

[0191] Sequence ID 9: Heavy chain CDR3 amino acid sequence; anti-LAG-3 mAb (BMS-986016) GYSDYEYNWFDP

[0192] Sequence ID 10: Light chain CDR1 amino acid sequence; anti-LAG-3 mAb (BMS-986016) RASQSISSYLA

[0193] Sequence ID 11 Light chain CDR2 amino acid sequence; anti-LAG-3 mAb (BMS-986016) DASNRAT

[0194] Sequence ID 12 Light chain CDR3 amino acid sequence; anti-LAG-3 mAb (BMS-986016) QQRSNWPLT

[0195] Sequence ID 13: Human LAG-3 amino acid sequence MWEAQFLGLLFLQPLWVAPVKPLQPGAEVPVVWAQEGAPAQLPCSPTIPLQDLSLLRRAGVTWQHQPDSGPPAAAPGHPLAPGPHPAAPSSWGPRPRRYTVLSVGPGGLRSGRLPLQPRVQLDERGRQRGD FSLWLRPARRADAGEYRAAVHLRDRALSCRLRLRLGQASMTASPPGSLRASDWVILNCSFSRPDRPASVHWFRNRGQGRVPVRESPHHHLAESFLFLPQVSPMDSGPWGCILTYRDGFNVSIMYNLTVLGL EPPTPLTVYAGAGSRVGLPCRLPAGVGTRSFLTAKWTPPGGGPDLLVTGDNGDFTLRLEDVSQAQAGTYTCHIHLQEQQLNATVTLAITVTPKSFGSPGSLGKLLCEVTPVSGQERFVWSSLDTPSQRSF SGPWLEAQEAQLLSQPWQCQLYQGERLLGAAVYFTELSSPGAQRSGRAPGALPAGHLLLFLTLGVLSLLLLVTGAFGFHLWRRQWRPRRFSALEQGIHPPQAQSKIEELEQEPEPEPEPEPEPEPEPEQL

[0196] Sequence ID 14: LAG-3 epitope PGHPLAPG

[0197] Sequence ID 15 LAG-3 epitope HPAAPSTW

[0198] Sequence ID 16: LAG-3 epitope PAAPSSWG

[0199] Sequence ID 17 Heavy chain amino acid sequence; Anti-PD-1 mAb (BMS936558) QVQLVESGGGVVQPGRSLRLDCKASGITFSNSGMHWVRQAPGKGLEWVAVIWYDGSKRYYADSVKGRFTISRDNSKNTLFLQMNSLRAEDTAVYYCATNDDYWGQGTLVT VSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCP PCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTIS KAKGQPREPQVYTLPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK

[0200] Sequence ID 18 Light chain amino acid sequence; Anti-PD-1 mAb (BMS936558) EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQSSNWPRTFGQGTKVEI KRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0201] Sequence ID 19: Heavy chain variable region (VH) amino acid sequence; anti-PD-1 mAb (BMS936558) QVQLVESGGGVVQPGRSLRLDCKASGITFSNSGMHWVRQAPGKGLEWVAVIWYDGSKRYYADSVKGRFTISRDNSKNTLFLQMNSLRAEDTAVYYCATNDDYWGQGTLVTVSS

[0202] Sequence ID 20: Heavy chain variable region (VH) nucleotide sequence; anti-PD-1 mAb (BMS936558) caggtgcagctggtggagtctgggggaggcgtggtccagcctgggaggtccctgagactcgactgtaaagcgtctggaatcaccttcagtaactctggcatgcactgggtccgccaggctccaggcaaggggctggagtgggtggcagttatttggtatgatggaagta aaagatactatgcagactccgtgaagggccgattcaccatctccagagacaattccaagaacacgctgtttctgcaaatgaacagcctgagagccgaggacacggctgtgtattactgtgcgacaaacgacgactactggggccagggaaccctggtcaccgtctcctca

[0203] Sequence ID 21: Light chain variable region (VL) amino acid sequence; anti-PD-1 mAb (BMS936558) EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQSSNWPRTFGQGTKVEIK

[0204] Sequence ID 22: Light chain variable region (VL) nucleotide sequence; anti-PD-1 mAb (BMS936558) gaaattgtgttgacacagtctccagccaccctgtctttgtctccaggggaaagagccaccctctcctgcagggccagtcagagtgttagtagttacttagcctggtaccaacagaaacctggccaggctcccaggctcctcatctatgatgcatccaaca gggccactggcatcccagccaggttcagtggcagtgggtctgggacagacttcactctcaccatcagcagcctagagcctgaagattttgcagtttattactgtcagcagagtagcaactggcctcggacgttcggccaagggaccaaggtggaaatcaaa

[0205] Sequence ID 23: Heavy chain CDR1 amino acid sequence; anti-PD-1 mAb (BMS936558) NSG H&M

[0206] Sequence ID 24: Heavy chain CDR2 amino acid sequence; anti-PD-1 mAb (BMS936558) VIWYDGSKRYYADSVKG

[0207] Sequence ID 25: Heavy chain CDR3 amino acid sequence; anti-PD-1 mAb (BMS936558) NDDY

[0208] Sequence ID 26: Light chain CDR1 amino acid sequence; anti-PD-1 mAb (BMS936558) RASQSVSSYLA

[0209] Sequence ID 27 Light chain CDR2 amino acid sequence; anti-PD-1 mAb (BMS936558) DASNRAT

[0210] Sequence ID 28 Light chain CDR3 amino acid sequence; anti-PD-1 mAb (BMS936558) QQSSNWPRT

[0211] Sequence ID 29: Complete Homo sapiens PD-1 sequence

[0212] Sequence ID 30: Heavy chain nucleotide sequence; anti-LAG-3 mAb (BMS-986016)

[0213] Sequence ID 31: Light chain nucleotide sequence; anti-LAG-3 mAb (BMS-986016) gaaattgtgttgacacagtctccagccaccctgtctttgtctccaggggaaagagccaccctctcctgcagggccagtcagagtattagcagctacttagcctggtaccaacagaaacctggccaggctcccaggctcctcatctatgatgcatccaacag ggccactggcatcccagccaggttcagtggcagtgggtctgggacagacttcactctcaccatcagcagcctagagcctgaagattttgcagtttattactgtcagcagcgtagcaactggcctctcacttttggccaggggaccaacctggagatcaaac gtacggtggctgcaccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggag agtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgttag

[0214] Sequence ID 32 Motif MYPPPY

[0215] Sequence ID 33: Heavy chain amino acid sequence; anti-ICOS mAb (BMS986226) EVQLVESGGGLVKPGGSLRLSCAASGFTFSDYFMHWVRQAPGKGLEWVGVIDTKSFNYATYYSDLVKGRFTISRDDSKNTLYLQMNSLKTEDTAVYYCTATIAVPYYFDYWG QGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCD KTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVEHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIE KTISKAKGQPREPQVYTLPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

[0216] Sequence ID 34 Light chain amino acid sequence; Anti-ICOS mAb (BMS986226) DIQMTQSPSSLSASVGDRVTITCQASQDISNYLSWYQQKPGKAPKLLIYYTNLLAEGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQYYNYRTFGPGTKVDIK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0217] Sequence ID 35: Heavy chain variable region (VH) amino acid sequence; anti-ICOS mAb (BMS986226) EVQLVESGGGLVKPGGSLRLSCAASGFTFSDYFMHWVRQAPGKGLEWVGVIDTKSFNYATYYSDLVKGRFTISRDDSKNTLYLQMNSLKTEDTAVYYCTATIAVPYYFDYWGQGTLVTVSS

[0218] Sequence ID 36: Light chain variable region (VL) amino acid sequence; anti-ICOS mAb (BMS986226) DIQMTQSPSSLSASVGDRVTITCQASQDISNYLSWYQQKPGKAPKLLIYYTNLLAEGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQYYNYRTFGPGTKVDIK

[0219] Sequence ID 37: Heavy chain CDR1 amino acid sequence; anti-ICOS mAb (BMS986226) GFTFSDYFMH

[0220] Sequence ID 38: Heavy chain CDR2 amino acid sequence; anti-ICOS mAb (BMS986226) VIDTKSFNYATYYSDLVKG

[0221] Sequence ID 39: Heavy chain CDR3 amino acid sequence; anti-ICOS mAb (BMS986226) TIAVPYYFDY

[0222] Sequence ID 40: Light chain CDR1 amino acid sequence; anti-ICOS mAb (BMS986226) QASQDISNYLS

[0223] Sequence ID 41: Light chain CDR2 amino acid sequence; anti-ICOS mAb (BMS986226) YTNLLAE

[0224] Sequence ID 42: Light chain CDR3 amino acid sequence; anti-ICOS mAb (BMS986226) QQYYNYRT

[0225] Sequence ID 43: Light chain CDR3 amino acid sequence; anti-ICOS mAb (BMS986226) QQYYNYRT

[0226] Sequence ID 44 Lymphocyte Activation Gene 3 Protein Amino Acid Sequence (Homo sapiens, NP_002277) MWEAQFLGLLFLQPLWVAPVKPLQPGAEVPVVWAQEGAPAQLPCSPTIPLQDLSLLRRAGVTWQHQPDSG PPAAAPGHPLAPGPHPAAPSSWGPRPRRYTVLSVGPGGLRSGRLPLQPRVQLDERGRQRGDFSLWLRPAR RADAGEYRAAVHLRDRALSCRLRLRLGQASMTASPPGSLRASDWVILNCSFSRPDRPASVHWFRNRGQGR VPVRESPHHHLAESFLFLPQVSPMDSGPWGCILTYRDGFNVSIMYNLTVLGLEPPTPLTVYAGAGSRVGL PCRLPAGVGTRSFLTAKWTPPGGGPDLLVTGDNGDFTLRLEDVSQAQAGTYTCHIHLQEQQLNATVTLAI ITVTPKSFGSPGSLGKLLCEVTPVSGQERFVWSSLDTPSQRSFSGPWLEAQEAQLLSQPWQCQLYQGERL LGAAVYFTELSSPGAQRSGRAPGALPAGHLLLFLILGVLSLLLLVTGAFGFHLWRRQWRPRRFSALEQGI HPPQAQSKIEELEQEPEPEPEPEPEPEPEPEPEPEQL

[0227] Sequence ID 45 PD-1 amino acid sequence (Homo sapiens, AAC51773.1) MQIPQAPWPVVWAVLQLGWRPGWFLDSPDRPWNPPTFFPALLVVTEGDNATFTCSFSNTSESFVLNWYRM SPSNQTDKLAAFPEEDRSQPGQDCRFRVTQLPNGRDFHMSVVRARRNDSGTYLCGAISLAPKAQIKESLRA ELRVTERRAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWVLAVICSRAARGTIGARRTGQPLK EDPSAVPVFSVDYGELDFQWREKTPEPPVPCVPEQTEYATIVFPSGMGTSSPARRGSADGPRSAQPLRPE DGHCSWPL

Claims

1. A method for treating malignant tumors in human patients, wherein the therapeutically effective amount LAG-3 inhibitors; PD-1 pathway inhibitors; and Immunotherapy agents A method including administering [a substance].

2. The method according to claim 1, wherein the LAG-3 inhibitor is an anti-LAG-3 antibody or its antigen-binding fragment.

3. The method according to claim 1 or 2, wherein the anti-LAG-3 antibody is a bispecific antibody.

4. The method according to any one of claims 1 to 3, wherein the anti-LAG-3 antibody or its antigen-binding fragment comprises (a) a heavy chain variable region CDR1 containing the sequence shown in SEQ ID NO: 7; (b) a heavy chain variable region CDR2 containing the sequence shown in SEQ ID NO: 8; (c) a heavy chain variable region CDR3 containing the sequence shown in SEQ ID NO: 9; (d) a light chain variable region CDR1 containing the sequence shown in SEQ ID NO: 10; (e) a light chain variable region CDR2 containing the sequence shown in SEQ ID NO: 11; and (f) a light chain variable region CDR3 containing the sequence shown in SEQ ID NO:

12.

5. The method according to any one of claims 1 to 4, wherein the anti-LAG-3 antibody or its antigen-binding fragment comprises a heavy chain and a light chain variable region having sequences shown in SEQ ID NOs. 3 and 5, respectively.

6. The method according to any one of claims 1 to 5, wherein the anti-LAG-3 antibody is BMS 986016, MK-4280 (28G-10), REGN3767, GSK2831781, IMP731 (H5L7BW), BAP050, IMP-701 (LAG-5250), IMP321, TSR-033, LAG525, BI 754111, or FS-118.

7. The method according to claim 1, wherein the LAG-3 inhibitor is a soluble LAG-3 polypeptide.

8. The method according to claim 7, wherein the soluble LAG-3 polypeptide is a fusion polypeptide.

9. The method according to claim 7 or 8, wherein the soluble LAG-3 polypeptide comprises a ligand-binding fragment of the LAG-3 extracellular domain.

10. The method according to claim 9, wherein the ligand-binding fragment of the LAG-3 extracellular domain comprises an amino acid sequence having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity with SEQ ID NO:

44.

11. The method according to any one of claims 7 to 10, wherein the soluble LAG-3 polypeptide further comprises an Fc domain.

12. The method according to any one of claims 1 to 11, wherein the PD-1 pathway inhibitor is an anti-PD-1 antibody or its antigen-binding fragment.

13. The method according to claim 12, wherein the anti-PD-1 antibody is pembrolizumab (Keytruda; MK-3475), pidilizumab (CT-011), nivolumab (Opdivo; BMS-936558), PDR001, MEDI0680 (AMP-514), TSR-042, REGN2810, JS001, AMP-224 (GSK-2661380), PF-06801591, BGB-A317, BI 754091, or SHR-1210.

14. The method according to any one of claims 1 to 11, wherein the PD-1 pathway inhibitor is an anti-PD-L1 antibody or its antigen-binding fragment.

15. The method according to claim 14, wherein the anti-PD-L1 antibody is atezolizumab (Tecentriq; RG7446; MPDL3280A; RO5541267), durvalumab (MEDI4736), BMS-936559, avelumab (Bavencio), LY3300054, CX-072 (Proclaim-CX-072), FAZ053, KN035, or MDX-1105.

16. The method according to any one of claims 1 to 11, wherein the PD-1 pathway inhibitor is a small molecule drug.

17. The method according to claim 16, wherein the PD-1 pathway inhibitor is CA-170.

18. The method according to claim 16, wherein the PD-1 pathway inhibitor is a cell-based therapy.

19. The method according to claim 18, wherein the cell-based therapy is a MiHA-loaded PD-L1 / L2 expression-suppressing dendritic cell vaccine.

20. The method according to claim 19, wherein the cell-based therapy is an anti-programmed cell death protein 1 antibody expressing pluripotent killer T lymphocytes, autologous PD-1 targeted chimeric switch receptor modified T lymphocytes, or PD-1 knockout autologous T lymphocytes.

21. The method according to any one of claims 1 to 11, wherein the PD-1 pathway inhibitor is an anti-PD-L2 antibody or its antigen-binding fragment.

22. The method according to claim 21, wherein the anti-PD-L2 antibody is rHIgM12B7.

23. The method according to any one of claims 1 to 11, wherein the PD-1 pathway inhibitor is a soluble PD-1 polypeptide.

24. The method according to claim 23, wherein the soluble PD-1 polypeptide is a fusion polypeptide.

25. The method according to claim 23 or 24, wherein the soluble PD-1 polypeptide comprises a ligand-binding fragment of the PD-1 extracellular domain.

26. The method according to any one of claims 23 to 25, wherein the soluble PD-1 polypeptide comprises a ligand-binding fragment of the PD-1 extracellular domain.

27. The method according to claim 26, wherein the ligand-binding fragment of the PD-1 extracellular domain comprises an amino acid sequence having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity with SEQ ID NO:

45.

28. The method according to any one of claims 23 to 27, wherein the soluble PD-1 polypeptide further comprises an Fc domain.

29. The immunotherapy agents are modulators of CTLA-4 activity, CD28 activity, CD80 activity, CD86 activity, 4-1BB activity, OX40 activity, KIR activity, Tim-3 activity, CD27 activity, CD40 activity, GITR activity, TIGIT activity, CD20 activity, CD96 activity, and IDO1 activity. The method according to any one of claims 1 to 28, wherein the modulator is a STING activity modulator, a GARP activity modulator, an A2aR activity modulator, a CEACAM1 activity modulator, a CEA activity modulator, a CD47 activity modulator, a PVRIG activity modulator, a TDO activity modulator, a VISTA activity modulator, a cytokine, a chemokine, an interferon, an interleukin, a lymphokine, a member of the tumor necrosis factor (TNF) family, or an immunostimulatory oligonucleotide.

30. The method according to any one of claims 1 to 28, wherein the immunotherapy agent is an immune checkpoint inhibitor.

31. The method according to claim 30, wherein the immune checkpoint inhibitor is a CTLA-4 antagonist, CD80 antagonist, CD86 antagonist, Tim-3 antagonist, TIGIT antagonist, CD20 antagonist, CD96 antagonist, IDO1 antagonist, STING antagonist, GARP antagonist, CD40 antagonist, A2aR antagonist, CEACAM1 (CD66a) antagonist, CEA antagonist, CD47 antagonist, PVRIG antagonist, TDO antagonist, VISTA antagonist, or KIR antagonist.

32. The method according to claim 31, wherein the immune checkpoint inhibitor is a CTLA-4 antagonist.

33. The method according to claim 32, wherein the CTLA-4 antagonist is an anti-CTLA-4 antibody or its antigen-binding fragment.

34. The method according to claim 33, wherein the anti-CTLA-4 antibody is ipilimumab (Yervoy), tremelimumab (tisilimunab; CP-675, 206), AGEN-1884, or ATOR-1015.

35. The method according to claim 32, wherein the CTLA-4 antagonist is a soluble CTLA-4 polypeptide.

36. The method according to claim 35, wherein the soluble CTLA-4 polypeptide is abatacept (Orencia), beratacept (Nulojix), RG2077, or RG1046.

37. The method according to claim 32, wherein the CTLA-4 antagonist is a cell-based therapy.

38. The method according to claim 37, wherein the CTLA-4 antagonist is an anti-CTLA-4 mAb RNA / GITRL RNA transfected autogenous dendritic cell vaccine or an anti-CTLA-4 mAb RNA transfected autogenous dendritic cell vaccine.

39. The method according to claim 31, wherein the immune checkpoint inhibitor is a KIR antagonist.

40. The method according to claim 39, wherein the KIR antagonist is an anti-KIR antibody or its antigen-binding fragment.

41. The method according to claim 40, wherein the anti-KIR antibody is lirilumab (1-7F9, BMS-986015, IPH 2101) or IPH 4102.

42. The method according to claim 31, wherein the immune checkpoint inhibitor is a TIGIT antagonist.

43. The method according to claim 42, wherein the TIGIT antagonist is an anti-TIGIT antibody or its antigen-binding fragment.

44. The method according to claim 43, wherein the anti-TIGIT antibody is BMS-986207, AB 154, COM902 (CGEN-15137), or OMP-313M32.

45. The method according to claim 31, wherein the immune checkpoint inhibitor is a Tim-3 antagonist.

46. The method according to claim 45, wherein the Tim-3 antagonist is an anti-Tim-3 antibody or its antigen-binding fragment.

47. The method according to claim 46, wherein the anti-Tim-3 antibody is TSR-022 or LY3321367.

48. The method according to claim 31, wherein the immune checkpoint inhibitor is an IDO1 antagonist.

49. IDO1 antagonist is indoximod (NLG8189; 1-methyl- Ｄ The method according to claim 48, wherein the derivative is -TRP), epacadostat (INCB-024360, INCB-24360), KHK2455, PF-06840003, napoximod (RG6078, GDC-0919, NLG919), BMS-986205 (F001287), or a pyrrolidine-2,5-dione derivative.

50. The method according to claim 31, wherein the immune checkpoint inhibitor is a STING antagonist.

51. The method according to claim 50, wherein the STING antagonist is a 2' or 3'-mono-fluorosubstituted cyclic dinucleotide; a 2'3'-di-fluorosubstituted mixed bond 2',5'-3',5' cyclic dinucleotide; a 2'-fluorosubstituted, bis-3',5' cyclic dinucleotide; a 2',2''-diF-Rp,Rp,bis-3',5' cyclic dinucleotide; or a fluorinated cyclic dinucleotide.

52. The method according to claim 31, wherein the immune checkpoint inhibitor is a CD20 antagonist.

53. The method according to claim 52, wherein the CD20 antagonist is an anti-CD20 antibody or its antigen-binding fragment.

54. The method according to claim 53, wherein the anti-CD20 antibody is rituximab (rituxan; IDEC-102; IDEC-C2B8), ABP 798, ofatumumab, or obinutuzumab.

55. The method according to claim 31, wherein the immune checkpoint inhibitor is a CD80 antagonist.

56. The method according to claim 55, wherein the CD80 antagonist is an anti-CD80 antibody or its antigen-binding fragment.

57. The method according to claim 56, wherein the anti-CD80 antibody is galiximab or AV 1142742.

58. The method according to claim 31, wherein the immune checkpoint inhibitor is a GARP antagonist.

59. The method according to claim 58, wherein the GARP antagonist is an anti-GARP antibody or its antigen-binding fragment.

60. The method according to claim 59, wherein the anti-GARP antibody is ARGX-115.

61. The method according to claim 31, wherein the immune checkpoint inhibitor is a CD40 antagonist.

62. The method according to claim 61, wherein the CD40 antagonist is an anti-CD40 antibody or its antigen-binding fragment.

63. The method according to claim 62, wherein the anti-CD40 antibody is BMS3h-56, lucatumumab (HCD122 and CHIR-12.12), CHIR-5.9, or dasetuzumab (huS2C6, PRO 64553, RG 3636, SGN 14, SGN-40).

64. The method according to claim 61, wherein the CD40 antagonist is a soluble CD40 ligand (CD40-L).

65. The method according to claim 64, wherein the soluble CD40 ligand is a fusion polypeptide.

66. The method according to claim 64 or 65, wherein the soluble CD40 ligand is CD40-L / FC2 or monomeric CD40-L.

67. The method according to claim 31, wherein the immune checkpoint inhibitor is an A2aR antagonist.

68. The method according to claim 67, wherein the A2aR antagonist is a small molecule.

69. The method according to claim 67 or 68, wherein the A2aR antagonist is CPI-444, PBF-509, istradefylline (KW-6002), preradiant (SCH420814), tozadenant (SYN115), vipadenant (BIIB014), HTL-1071, ST1535, SCH412348, SCH442416, SCH58261, ZM241385, or AZD4635.

70. The method according to claim 31, wherein the immune checkpoint inhibitor is a CEACAM1 antagonist.

71. The method according to claim 70, wherein the CEACAM1 antagonist is an anti-CEACAM1 antibody or its antigen-binding fragment.

72. The method according to claim 71, wherein the anti-CEACAM1 antibody is CM-24 (MK-6018).

73. The method according to claim 31, wherein the immune checkpoint inhibitor is a CEA antagonist.

74. The method according to claim 73, wherein the CEA antagonist is an anti-CEA antibody or its antigen-binding fragment.

75. The method according to claim 74, wherein the anti-CEA antibody is cergutuzumabu amnaleukin (RG7813, RO-6895882) or RG7802 (RO6958688).

76. The method according to claim 31, wherein the immune checkpoint inhibitor is a CD47 antagonist.

77. The method according to claim 76, wherein the CD47 antagonist is an anti-CD47 antibody or its antigen-binding fragment.

78. The method according to claim 77, wherein the anti-CD47 antibody is HuF9-G4, CC-90002, TTI-621, ALX148, NI-1701, NI-1801, SRF231, or Effi-DEM.

79. The method according to claim 31, wherein the immune checkpoint inhibitor is a PVRIG antagonist.

80. The method according to claim 79, wherein the PVRIG antagonist is an anti-PVRIG antibody or its antigen-binding fragment.

81. The method according to claim 80, wherein the anti-PVRIG antibody is COM701 (CGEN-15029).

82. The method according to claim 31, wherein the immune checkpoint inhibitor is a TDO antagonist.

83. The method according to claim 82, wherein the TDO antagonist is a 4-(indole-3-yl)pyrazole derivative, a 3-indole substituted derivative, or a 3-(indole-3-yl)pyridine derivative.

84. The method according to claim 31, wherein the immune checkpoint inhibitor is a dual IDO and TDO antagonist.

85. The method according to claim 84, wherein the dual IDO and TDO antagonists are small molecules.

86. The method according to claim 31, wherein the immune checkpoint inhibitor is a VISTA antagonist.

87. The method according to claim 86, wherein the VISTA antagonist is CA-170 or JNJ-61610588.

88. The method according to any one of claims 1 to 29, wherein the immunotherapy agent is an immune checkpoint enhancer or stimulator.

89. The method according to claim 88, wherein the immune checkpoint enhancer or stimulator is a CD28 agonist, a 4-1BB agonist, an OX40 agonist, a CD27 agonist, a CD80 agonist, a CD86 agonist, a CD40 agonist, an ICOS agonist, a CD70 agonist, or a GITR agonist.

90. The method according to claim 89, wherein the immune checkpoint enhancer or stimulator is an OX40 agonist.

91. The method according to claim 90, wherein the OX40 agonist is an anti-OX40 antibody or its antigen-binding fragment.

92. The method according to claim 91, wherein the anti-OX40 antibody is tavorixizumab (MEDI-0562), pogalizumab (MOXR0916, RG7888), GSK3174998, ATOR-1015, MEDI-6383, MEDI-6469, BMS 986178, PF-04518600, or RG7888 (MOXR0916).

93. The method according to claim 90, wherein the OX40 agonist is a cell-based therapy.

94. The method according to claim 93, wherein the OX40 agonist is GINAKIT cells.

95. The method according to claim 89, wherein the immune checkpoint enhancer or stimulator is a CD40 agonist.

96. The method according to claim 95, wherein the CD40 agonist is an anti-CD40 antibody or its antigen-binding fragment.

97. The method according to claim 96, wherein the anti-CD40 antibody is ADC-1013 (JNJ-64457107), RG7876 (RO-7009789), HuCD40-M2, APX005M (EPI-0050), or Chi Lob 7 / 4.

98. The method according to claim 95, wherein the CD40 agonist is a soluble CD40 ligand (CD40-L).

99. The method according to claim 98, wherein the soluble CD40 ligand is a fusion polypeptide.

100. The method according to claim 98 or 99, wherein the soluble CD40 ligand is the trimer CD40-L(AVREND).

101. The method according to claim 89, wherein the immune checkpoint enhancer or stimulator is a GITR agonist.

102. The method according to claim 101, wherein the GITR agonist is an anti-GITR antibody or its antigen-binding fragment.

103. The method according to claim 102, wherein the anti-GITR antibody is BMS-986156, TRX518, GWN323, INCAGN01876, or MEDI1873.

104. The method according to claim 101, wherein the GITR agonist is a soluble GITR ligand (GITRL).

105. The method according to claim 104, wherein the soluble GITR ligand is a fusion polypeptide.

106. The method according to claim 101, wherein the GITR agonist is a cell-based therapy.

107. The method according to claim 106, wherein the cell-based therapy is an anti-CTLA4 mAb RNA / GITRL RNA-transfected autogenous dendritic cell vaccine or a GITRL RNA-transfected autogenous dendritic cell vaccine.

108. The method according to claim 89, wherein the immune checkpoint enhancer or stimulator is a 4-1BB agonist.

109. The method according to claim 108, wherein the 4-1BB agonist is an anti-4-1BB antibody or its antigen-binding fragment.

110. The method according to claim 109, wherein the anti-4-1BB antibody is urelumab or PF-05082566.

111. The method according to claim 89, wherein the immune checkpoint enhancer or stimulator is a CD80 agonist or a CD86 agonist.

112. The method according to claim 111, wherein the CD80 agonist or CD86 agonist is a soluble CD80 or CD86 ligand (CTLA-4).

113. The method according to claim 112, wherein the soluble CD80 or CD86 ligand is a fusion polypeptide.

114. The method according to claim 112 or 113, wherein the CD80 or CD86 ligand is CTLA4-Ig (CTLA4-IgG4m, RG2077 or RG1046) or abatacept (Orencia, BMS-188667).

115. The method according to claim 111, wherein the CD80 agonist or CD86 agonist is a cell-based therapy.

116. The method according to claim 115, wherein the cell-based therapy is MGN1601.

117. The method according to claim 89, wherein the immune checkpoint enhancer or stimulator is a CD28 agonist.

118. The method according to claim 117, wherein the CD28 agonist is an anti-CD28 antibody or its antigen-binding fragment.

119. The method according to claim 118, wherein the anti-CD28 antibody is TGN1412.

120. The method according to claim 117, wherein the CD28 agonist is a cell-based therapy.

121. Cell-based therapies include JCAR015 (anti-CD19-CD28-zeta modified CAR CD3+ T lymphocytes); CD28CAR / CD137CAR expressing T lymphocytes; allogeneic CD4+ memory Th1-like T cells / microparticle-bound anti-CD3 / anti-CD28; anti-CD19 / CD28 / CD3 zeta CAR gamma retrovirus vector-transduced autologous T lymphocytes KTE-C19; anti-CEA IgCD28TCR-transduced autologous T lymphocytes; and anti-EGFRvIII. CAR-transduced allogeneic T lymphocytes; autologous CD123CAR-CD28-CD3 zeta-EGFRt-expressing T lymphocytes; autologous CD171-specific CAR-CD28 zeta-4-1-BB-EGFRt-expressing T lymphocytes; autologous CD19CAR-CD28-CD3 zeta-EGFRt-expressing Tcm-enriched T cells; autologous PD-1-targeted chimeric switch receptor-modified T lymphocytes (chimeric with CD28); CD19CAR-CD28-CD3 zeta-EGFRt-expressing Tcm-enriched T lymphocytes The method according to claim 120, wherein the T lymphocytes are: P-cells; CD19CAR-CD28-CD3 zeta-EGFRt-expressing Tn / mem-enriched T lymphocytes; CD19CAR-CD28 zeta-4-1BB-expressing allogeneic T lymphocytes; CD19CAR-CD3 zeta-4-1BB-CD28-expressing autologous T lymphocytes; CD28CAR / CD137CAR-expressing T lymphocytes; CD3 / CD28 costimulated vaccine-stimulated autologous T lymphocytes; or iC9-GD2-CD28-OX40-expressing T lymphocytes.

122. The method according to claim 89, wherein the immune checkpoint enhancer or stimulator is a CD27 agonist.

123. The method according to claim 122, wherein the CD27 agonist is an anti-CD27 antibody or its antigen-binding fragment.

124. The method according to claim 123, wherein the anti-CD27 antibody is varylumab (CDX-1127).

125. The method according to claim 89, wherein the immune checkpoint enhancer or stimulator is a CD70 agonist.

126. The method according to claim 125, wherein the CD70 agonist is an anti-CD70 antibody or its antigen-binding fragment.

127. The method according to claim 126, wherein the anti-CD70 antibody is ARGX-110.

128. The method according to claim 89, wherein the immune checkpoint enhancer or stimulator is an ICOS agonist.

129. The method according to claim 128, wherein the ICOS agonist is an anti-ICOS antibody or its antigen-binding fragment.

130. The method according to claim 129, wherein the anti-ICOS antibody is BMS986226, MEDI-570, GSK3359609, or JTX-2011.

131. The method according to claim 128, wherein the ICOS agonist is a soluble ICOS ligand.

132. The method according to claim 131, wherein the soluble ICOS ligand is a fusion polypeptide.

133. The method according to claim 131 or 132, wherein the soluble ICOS ligand is AMG 750.

134. The method according to claim 89, wherein the immunotherapy agent is an anti-CD73 antibody or its antigen-binding fragment.

135. The method according to claim 134, wherein the anti-CD73 antibody is MEDI9447.

136. The method according to any one of claims 1 to 28, wherein the immunotherapy agent is a TLR9 agonist.

137. The method according to claim 136, wherein the TLR9 agonist is agatrimod sodium.

138. The method according to any one of claims 1 to 28, wherein the immunotherapy agent is a cytokine.

139. The method according to claim 138, wherein the cytokine is a member of the chemokine, interferon, interleukin, lymphokine or tumor necrosis factor family.

140. The method according to claim 138 or 139, wherein the cytokine is IL-2, IL-15, or interferon-gamma.

141. The method according to any one of claims 1 to 28, wherein the immunotherapy agent is a TGF-β antagonist.

142. The method according to claim 141, wherein the TGF-β antagonist is fresolimmab (GC-1008), NIS793, IMC-TR1 (LY3022859), ISTH0036, Travedersen (AP 12009), recombinant transforming growth factor-beta-2, autologous HPV-16 / 18 E6 / E7 specific TGF-beta resistant T lymphocytes, or TGF-beta resistant LMP specific cytotoxic T lymphocytes.

143. The method according to any one of claims 1 to 28, wherein the immunotherapy agent is an iNOS antagonist.

144. The method according to claim 143, wherein the iNOS antagonist is N-acetyl-cysteine ​​(NAC), aminoguanidine, L-nitroarginine methyl ester, or S,S-1,4-phenylene-bis(1,2-ethanediyl)bis-isothiourea.

145. The method according to any one of claims 1 to 28, wherein the immunotherapy agent is an SHP-1 antagonist.

146. The method according to any one of claims 1 to 28, wherein the immunotherapy agent is a CSF1R (colony-stimulating factor 1 receptor) antagonist.

147. The method according to claim 146, wherein the CSF1R antagonist is an anti-CSF1R antibody or its antigen-binding fragment.

148. The method according to claim 147, wherein the anti-CSF1R antibody is emactuzumab.

149. The method according to any one of claims 1 to 28, wherein the immunotherapy agent is an agonist of a TNF family member.

150. The method according to claim 149, wherein the TNF family member agonist is ATOR 1016, ABBV-621, or adalimumab.

151. The method according to any one of claims 1 to 28, wherein the immunotherapy agent is aldesleukin, tocilizumab, or MEDI5083.

152. The method according to any one of claims 1 to 28, wherein the immunotherapy agent is a CD160 (NK1) agonist.

153. The method according to claim 152, wherein the CD160 (NK1) agonist is an anti-CD160 antibody or its antigen-binding fragment.

154. The method according to claim 152 or 153, wherein the anti-CD160 antibody is BY55.

155. The method according to any one of claims 1 to 154, wherein the LAG-3 inhibitor, PD-1 pathway inhibitor, and immunotherapy agent are formulated for intravenous administration.

156. The method according to any one of claims 1 to 155, wherein a LAG-3 inhibitor, a PD-1 pathway inhibitor, and an immunotherapy agent are formulated in a single formulation.

157. The method according to any one of claims 1 to 155, wherein the LAG-3 inhibitor, the PD-1 pathway inhibitor, and the immunotherapy agent are formulated separately.

158. Malignant tumors include liver cancer, bone cancer, pancreatic cancer, skin cancer, head and neck cancer, breast cancer, lung cancer, cutaneous or intraocular malignant melanoma, kidney cancer, uterine cancer, ovarian cancer, colorectal cancer, colon cancer, rectal cancer, anal cancer, stomach cancer, testicular cancer, uterine cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, non-Hodgkin lymphoma, esophageal cancer, small intestine cancer, endocrine cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, childhood cancer, lymphocytic lymphoma, bladder cancer, kidney or ureteral cancer, renal pelvis cancer, central nervous system (CNS) neoplasms, primary CNS lymphoma, tumor angiogenesis, spinal axial tumor, brainstem glioma, pituitary adenoma, Kaposi's sarcoma, epidermal carcinoma, squamous cell carcinoma, and asbestos-induced The method according to any one of claims 1 to 157, selected from the group consisting of hematological malignancies including environment-induced cancers, such as multiple myeloma, B-cell lymphoma, Hodgkin lymphoma / primary mediastinal B-cell lymphoma, non-Hodgkin lymphoma, acute myeloid lymphoma, chronic myeloid leukemia, chronic lymphoid leukemia, follicular lymphoma, disseminated large B-cell lymphoma, Burkitt lymphoma, immunoblastic large cell lymphoma, precursor B-lymphoblastic lymphoma, mantle cell lymphoma, acute lymphoblastic leukemia, mycosis fungoides, anaplastic large cell lymphoma, T-cell lymphoma and precursor T-lymphoblastic lymphoma, and any combination thereof.

159. The method according to claim 158, wherein the malignant tumor is non-small cell lung cancer (NSCLC), virus-associated cancer-associated tumor, or gastric adenocarcinoma.

160. The method according to any one of claims 1 to 157, wherein the malignant tumor is melanoma, gastric cancer, gastroesophageal junction cancer, non-small cell lung cancer, bladder cancer, head and neck squamous cell carcinoma, or renal cell carcinoma.

161. The method according to any one of claims 1 to 157, wherein the tumor is lung cancer, melanoma, head and neck squamous cell carcinoma, kidney cancer, gastric cancer, or hepatocellular carcinoma.

162. The method according to any one of claims 1 to 161, wherein an anti-LAG-3 antibody or its antigen-binding fragment and an immunotherapy agent are administered as first-line treatment.

163. The method according to any one of claims 1 to 161, wherein a LAG-3 inhibitor, a PD-1 pathway inhibitor, and an immunotherapy agent are administered as a second-line treatment.

164. The method according to any one of claims 1 to 163, wherein the malignant tumor is refractory to first-line treatment.

165. The method according to any one of claims 1 to 164, further comprising the administration of at least one additional therapeutic agent.

166. The method according to claim 165, wherein at least one additional therapeutic agent is a chemotherapeutic agent.

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