LAG-3 combination therapy for cancer treatment

Abstract

This provides an improved method for treating LAG-3-positive tumors. [Solution] The present invention provides a pharmaceutical product for use in the treatment of tumors in patients with human gastric cancer or gastroesophageal junction cancer, wherein the pharmaceutical product is either a LAG-3 antagonist, particularly an anti-LAG-3 antibody or soluble LAG-3 alone, or used in combination with a PD-1 pathway inhibitor, particularly an anti-PD-1 antibody, and may be combined with one or more chemotherapeutic agents as required.

Description

Technical Field

[0001] Cross-Reference to Related Applications This application claims the benefit of priority based on U.S. Provisional Patent Application No. 62 / 703,690 filed on July 26, 2018 and U.S. Provisional Patent Application No. 62 / 725,336 filed on August 31, 2018, the entire contents of which are incorporated herein by reference.

[0002] Field of the Invention The invention described herein relates to a method for treating malignant tumors in human patients by combining a LAG-3 inhibitor, a PD-1 pathway inhibitor and a chemotherapeutic agent. Background Art

[0003] Background of the Invention Lymphocyte activation gene-3 (LAG-3; CD223) is activated CD4 + and CD8 + is a type I transmembrane protein expressed on the cell surface of T cells, as well as subsets of NK cells and dendritic cells (Triebel F, et al., J. Exp. Med. 1990; 171:1393-1405; Workman C J, et al., J. Immunol. 2009; 182(4):1885-91). LAG-3 is closely related to CD4, a co-receptor for T helper cell activation. Both molecules have four extracellular Ig-like domains and require binding to their ligand, major histocompatibility complex (MHC) class II, for their functional activity. In contrast to CD4, LAG-3 is only expressed on the cell surface of activated T cells, and cleavage from the cell surface stops LAG-3 signaling. Although LAG-3 is also found as a soluble protein, it does not bind to MHC class II, and the function of soluble LAG-3 remains unclear.

[0004] PD-1 is a cell surface signaling receptor that plays a crucial role in regulating T cell activation and resistance (Keir ME, et al., Annu Rev Immunol 2008; 26:677-704). It is a type I transmembrane protein and, along with BTLA, CTLA-4, ICOS, and CD28, is part of the CD28 family of T cell costimulatory receptors. PD-1 is primarily expressed on activated T cells, B cells, and myeloid cells (Dong H, et al., Nat Med. 1999; 5:1365-1369). It is also expressed on natural killer (NK) cells (Terme M, et al., Cancer Res 2011; 71:5393-5399). The binding of PD-1 by its ligands, PD-L1 and PD-L2, leads to phosphorylation of tyrosine residues in the proximal intracellular immune receptor tyrosine inhibitory domain, followed by the recruitment of the phosphorylation enzyme SHP-2, ultimately resulting in downregulation of T cell activation. One of the important roles of PD-1 is to suppress the activity of peripheral tissue T cells during inflammatory responses to infection, thereby suppressing the development of autoimmunity (Pardoll D M., Nat Rev Cancer 2012; 12:252-264). Evidence for this negative regulatory role comes from the finding 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 the tumor environment, this results in the expression of immune resistance within the tumor microenvironment. PD-1 is highly expressed on tumor-infiltrating lymphocytes, and its ligand is upregulated on the cell surface of many different tumors (Dong H, et al., Nat Med 2002; 8:793-800). Multiple mouse cancer models have demonstrated that ligand binding to PD-1 leads to immune evasion.In addition, 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, inhibition of the PD-1 / PD-L1 interaction has been shown to mediate potent antitumor activity in preclinical models (U.S. Patent Nos. 8,008,449 and 7,943,743).

[0005] The object of the present invention is to provide an improved method for treating LAG-3 positive tumors. [Overview of the project]

[0006] Summary of the Invention One aspect of the present invention as described herein relates to a method for inhibiting the growth of malignant tumors in a human patient, the method comprising administering to the patient each of the following effective amounts: (a) a LAG-3 antagonist; (b) a PD-1 pathway inhibitor; and (c) one or more chemotherapeutic agents; wherein the patient's tumor-associated immune cells express LAG-3. Another aspect of the present invention relates to a method for treating cancer in a human patient, the method comprising administering to the patient each of the following effective amounts: (a) a LAG-3 antagonist; (b) a PD-1 pathway inhibitor; and (c) one or more chemotherapeutic agents; wherein the patient's tumor-associated immune cells express LAG-3. One aspect of the present invention relates to a method for inhibiting the growth of malignant tumors in a human patient, the method comprising administering to the patient each of the following effective amounts: (a) a LAG-3 antagonist; (b) a PD-1 pathway inhibitor; and (c) one or more chemotherapeutic agents. Another aspect of the present invention relates to a method for treating cancer in a human patient, the method comprising administering to the patient an effective amount of each of the following: (a) a LAG-3 antagonist; (b) a PD-1 pathway inhibitor; and (c) one or more chemotherapeutic agents.

[0007] In one aspect, malignant tumors include liver cancer, bone cancer, pancreatic cancer, skin cancer, oral cancer, head and neck cancer, breast cancer, lung cancer including small cell lung cancer and non-small cell lung cancer, melanoma of the skin or eye, kidney cancer, uterine cancer, ovarian cancer, and colorectal cancer. Cancer, colon cancer, rectal cancer, anal cancer, gastric cancer, testicular cancer, Fallopian duct cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, non-Hodgkin lymphoma, esophageal cancer, small intestine cancer, endocrine cancer, thyroid cancer, parathyroid cancer, adrenal adenocarcinoma, soft tissue sarcoma, urethral cancer, penile cancer, childhood cancer, lymphocytic lymphoma, bladder cancer, kidney or ureter cancer, renal pelvis cancer, neoplasms of the central nervous system (CNS), primary CNS lymphoma, tumor angiogenesis, spinal axial tumor, brainstem glioma, pituitary adenoma, Kaposi's sarcoma, epidermal carcinoma, squamous cell carcinoma, environmentally induced cancers including those induced by asbestos, and blood cancers including the following. The fluid-type tumors are selected from the group consisting of, for example, multiple myeloma, B-cell lymphoma, Hodgkin lymphoma / primary mediastinal B-cell lymphoma, non-Hodgkin lymphoma, acute myeloid lymphoma, chronic myeloid leukemia, chronic lymphocytic leukemia, follicular lymphoma, diffuse large B-cell lymphoma, Burkitt lymphoma, immunoblastic large B-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. In another embodiment, the malignant tumor is gastric cancer or gastroesophageal junction cancer. In another embodiment, gastric cancer is adenocarcinoma, lymphoma, gastrointestinal stromal tumor or carcinoid tumor. In another aspect, malignant tumors are selected from melanoma, non-small cell lung cancer (NSCLC), human papillomavirus (HPV)-related tumors, bladder cancer, head and neck squamous cell carcinoma, renal cell carcinoma, and gastric adenocarcinoma.

[0008] In one embodiment of the present invention, the LAG-3 antagonist is an anti-LAG-3 antibody. In another embodiment, the anti-LAG-3 antibody is a full-length antibody. In another embodiment, the antibody is a monoclonal antibody, a human antibody, a humanized antibody, a chimeric antibody, or a multispecific antibody. In another embodiment, the multispecific antibody is a biaffinity retargeted antibody (DART), DVD-Ig, or a bispecific antibody. In another embodiment, the antibody is an F(ab')2 fragment, a Fab' fragment, a Fab fragment, an Fv fragment, an scFv fragment, a dsFv fragment, a dAb fragment, or a single-chain linked polypeptide peptide. In another embodiment, the anti-LAG-3 antibody is BMS-986016, IMP731(H5L7BW), MK-4280(28G-10), REGN3767, GSK2831781, humanized BAP050, IMP-701(LAG-525), aLAG3(0414), aLAG3(0416), Sym022, TSR-033, TSR-075, XmAb22841, BI754111, MGD013, AVA-017, P13B02-30, or FS-118. In another embodiment, the LAG-3 antagonist is IMP321. In another embodiment, the anti-LAG-3 antibody comprises CDR1, CDR2, and CDR3 domains of the heavy chain variable region having the sequence described in SEQ ID NO: 3, and CDR1, CDR2, and CDR3 domains of the light chain variable region having the sequence described in SEQ ID NO: 5. In another embodiment, the anti-LAG-3 antibody comprises (a) heavy chain variable region CDR1 having the sequence described in SEQ ID NO: 7; (b) heavy chain variable region CDR2 having the sequence described in SEQ ID NO: 8; (c) heavy chain variable region CDR3 having the sequence described in SEQ ID NO: 9; (d) light chain variable region CDR1 having the sequence described in SEQ ID NO: 10; (e) light chain variable region CDR2 having the sequence described in SEQ ID NO: 11; and (f) light chain variable region CDR3 having the sequence described in SEQ ID NO: 12. In another embodiment, the anti-LAG-3 antibody comprises a heavy chain variable region and a light chain variable region having the sequences described in SEQ ID NO: 3 and 5, respectively. In another embodiment, the anti-LAG-3 antibody comprises a heavy chain and a light chain having the sequences described in SEQ ID NO: 1 and 2, respectively.

[0009] In one embodiment, the PD-1 pathway inhibitor is an anti-PD-1 antibody or an anti-PD-L1 antibody. In another embodiment, the anti-PD-1 antibody is selected from the group consisting of nivolumab, pembrolizumab, pizilizumab, PDR001, MEDI0680, TSR-042, REGN2810, JS001, PF-06801591, BGB-A317, BI754091, and SHR-1210. In another embodiment, the anti-PD-1 antibody comprises the CDR1, CDR2, and CDR3 domains of the heavy chain variable region having the sequence described in SEQ ID NO: 15, and the CDR1, CDR2, and CDR3 domains of the light chain variable region having the sequence described in SEQ ID NO: 17. In another embodiment, the anti-PD-1 antibody comprises (a) a heavy chain variable region CDR1 containing the sequence described in SEQ ID NO: 19; (b) a heavy chain variable region CDR2 containing the sequence described in SEQ ID NO: 20; (c) a heavy chain variable region CDR3 containing the sequence described in SEQ ID NO: 21; (d) a light chain variable region CDR1 containing the sequence described in SEQ ID NO: 22; (e) a light chain variable region CDR2 containing the sequence described in SEQ ID NO: 23; and (f) a light chain variable region CDR3 containing the sequence described in SEQ ID NO: 24. In another embodiment, the anti-PD-1 antibody comprises a heavy chain variable region and a light chain variable region containing the sequences described in SEQ ID NOs: 15 and 17, respectively. In another embodiment, the anti-PD-1 antibody comprises a heavy chain and a light chain containing the sequences described in SEQ ID NOs: 13 and 14, respectively.

[0010] In one embodiment, one or more chemotherapeutic agents are platinum compounds or fluoropyrimidines. In another embodiment, one or more chemotherapeutic agents are oxaliplatin, cisplatin, fluorouracil, capecitabine, tegafur, gimeracil, or oteracil. In another embodiment, one or more chemotherapeutic agents are oxaliplatin and capecitabine (XELOX). In another embodiment, one or more chemotherapeutic agents are oxaliplatin and fluorouracil. In another embodiment, the chemotherapeutic agents further comprise a chemoprotective agent. In another embodiment, the chemoprotective agent is leucovorin. In another embodiment, one or more chemotherapeutic agents comprise oxaliplatin, leucovorin, and fluorouracil (FOLFOX). In another embodiment, one or more chemotherapeutic agents comprise oxaliplatin and tegafur / gimeracil / oteracil potassium (SOX).

[0011] In one embodiment, a fixed dose combination of anti-LAG-3 antibody and anti-PD-1 antibody is administered. In another embodiment, the fixed dose is determined based on the chemotherapeutic agent administered to the subject.

[0012] In one embodiment, the method comprises at least one administration cycle, the cycle being a period of 6 weeks, in which at least one dose of anti-LAG-3 antibody is administered in doses of 120 or 160 mg and two doses of anti-PD-1 antibody are administered in doses of 360 or 480 mg. In another embodiment, 120 mg of anti-LAG-3 antibody, 360 mg of anti-PD-1 antibody, and XELOX are administered. In yet another embodiment, 160 mg of anti-LAG-3 antibody, 480 mg of anti-PD-1 antibody, and FOLFOX are administered. In yet another embodiment, 120 mg of anti-LAG-3 antibody, 360 mg of anti-PD-1 antibody, and SOX are administered.

[0013] In one embodiment, the anti-LAG-3 antibody and the anti-PD-1 antibody are formulated for intravenous administration. In another embodiment, the anti-LAG-3 antibody and the anti-PD-1 antibody are co-formulated. In yet another embodiment, the anti-LAG-3 antibody and the anti-PD-1 antibody are formulated separately.

[0014] The present invention also relates to a method for inhibiting the growth of gastric adenocarcinoma or gastroesophageal junction adenocarcinoma in a human patient, the method comprising administering to the patient an effective dose of each of the following: (a) an anti-LAG-3 antibody comprising the CDR1, CDR2, and CDR3 domains of the heavy chain variable region having the sequence described in SEQ ID NO: 3 and the CDR1, CDR2, and CDR3 domains of the light chain variable region having the sequence described 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 the sequence described in SEQ ID NO: 15 and the CDR1, CDR2, and CDR3 domains of the light chain variable region having the sequence described in SEQ ID NO: 17; and (c) one or more chemotherapeutic agents selected from the group consisting of oxaliplatin / capecitabine (XELOX), oxaliplatin / leucovorin / fluorouracil (FOLFOX), and oxaliplatin / tegafur / gimeracil / oteracil (SOX), wherein the patient's tumor-associated immune cells express LAG-3.

[0015] The present invention also relates to a method for treating gastric cancer or gastroesophageal junction cancer in a human patient, the method comprising administering to the patient an effective amount of: (a) an anti-LAG-3 antibody comprising the CDR1, CDR2, and CDR3 domains of the heavy chain variable region having the sequence described in SEQ ID NO: 3 and the CDR1, CDR2, and CDR3 domains of the light chain variable region having the sequence described 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 the sequence described in SEQ ID NO: 15 and the CDR1, CDR2, and CDR3 domains of the light chain variable region having the sequence described in SEQ ID NO: 17; and (c) one or more chemotherapeutic agents selected from the group consisting of XELOX, FOLFOX, and SOX, wherein the patient's tumor-associated immune cells express LAG-3.

[0016] In one embodiment, LAG-3 expression is assayed by RT-PCR, in situ hybridization, RNase protection, RT-PCR-based assays, immunohistochemistry, enzyme-linked immunosorbent assays, in vivo imaging, or flow cytometry. In another embodiment, LAG-3 expression is assayed by immunohistochemistry.

[0017] The present invention also relates to a method for treating gastric cancer or gastroesophageal junction adenocarcinoma in human patients, the method comprising administering to the patient an effective amount of: (a) an anti-LAG-3 antibody comprising the CDR1, CDR2, and CDR3 domains of the heavy chain variable region having the sequence described in SEQ ID NO: 3 and the CDR1, CDR2, and CDR3 domains of the light chain variable region having the sequence described 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 the sequence described in SEQ ID NO: 15 and the CDR1, CDR2, and CDR3 domains of the light chain variable region having the sequence described in SEQ ID NO: 17; and (c) one or more chemotherapeutic agents. In one embodiment, the method is administered to a patient who has not received prior therapy (for example, as first-line therapy).

[0018] The present invention also relates to a method for treating recurrent, locally advanced, or metastatic gastric cancer or gastroesophageal junction adenocarcinoma in a human patient, the method comprising administering to the patient an effective dose of (a) an anti-LAG-3 antibody comprising CDR1, CDR2, and CDR3 domains of the heavy chain variable region having the sequence described in SEQ ID NO: 3 and CDR1, CDR2, and CDR3 domains of the light chain variable region having the sequence described in SEQ ID NO: 5, and (b) one or more standard therapeutic regimens, wherein the patient's tumor-associated immune cells express LAG-3.

[0019] The present invention also relates to a method for treating recurrent, locally advanced, or metastatic gastric cancer or gastroesophageal junction adenocarcinoma in a human patient, the method comprising administering to the patient an effective dose of: (a) an anti-LAG-3 antibody comprising the CDR1, CDR2, and CDR3 domains of the heavy chain variable region having the sequence described in SEQ ID NO: 3 and the CDR1, CDR2, and CDR3 domains of the light chain variable region having the sequence described 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 the sequence described in SEQ ID NO: 15 and the CDR1, CDR2, and CDR3 domains of the light chain variable region having the sequence described in SEQ ID NO: 17; and (c) one or more standard therapy regimens, wherein the patient's tumor-associated immune cells express LAG-3. One aspect of the present invention relates to a method for treating recurrent, locally advanced, or metastatic gastric cancer or gastroesophageal junction adenocarcinoma in a human patient, the method comprising administering to the patient an effective dose of: (a) an anti-LAG-3 antibody comprising the CDR1, CDR2, and CDR3 domains of the heavy chain variable region having the sequence described in SEQ ID NO: 3 and the CDR1, CDR2, and CDR3 domains of the light chain variable region having the sequence described 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 the sequence described in SEQ ID NO: 15 and the CDR1, CDR2, and CDR3 domains of the light chain variable region having the sequence described in SEQ ID NO: 17; and (c) one or more standard therapy regimens. In one embodiment, the anti-LAG-3 antibody and the anti-PD-1 antibody are administered as a fixed-dose combination. In another embodiment, one or more standard therapeutic regimens include the administration of docetaxel, doxorubicin hydrochloride, 5-fluorouracil, mitomycin C, fluorouracil / leucovorin calcium (FU-LV), docetaxel / cisplatin / fluorouracil (TPF), or capecitabine / irinotecan hydrochloride (XELIRI). In another embodiment, this method is administered to patients who have received prior treatment (for example, as second-line therapy).In one embodiment, one or more standard therapy regimens include the administration of docetaxel, doxorubicin hydrochloride, 5-fluorouracil, mitomycin C, fluorouracil / leucovorin calcium (FU-LV), docetaxel / cisplatin / fluorouracil (TPF), or capecitabine / irinotecan hydrochloride (XELIRI).

[0020] The present invention also relates to a method for treating gastric cancer or gastroesophageal junction cancer in a human patient, the method comprising administering to the patient an effective amount of: (a) an anti-LAG-3 antibody comprising the CDR1, CDR2, and CDR3 domains of the heavy chain variable region having the sequence described in SEQ ID NO: 3 and the CDR1, CDR2, and CDR3 domains of the light chain variable region having the sequence described 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 the sequence described in SEQ ID NO: 15 and the CDR1, CDR2, and CDR3 domains of the light chain variable region having the sequence described in SEQ ID NO: 17; and (c) one or more chemotherapeutic agents selected from the group consisting of XELOX, FOLFOX, and SOX. In some embodiments, the method is administered to a patient who has not received prior treatment (e.g., as first-line therapy). In some embodiments, the patient has not received HER2 inhibitor therapy. In some embodiments, the present invention is administered to a patient who is HER2-negative. In certain embodiments, the patient has not received prior systemic therapy. In certain embodiments, anti-LAG-3 antibodies and anti-PD-1 antibodies are administered as a fixed-dose combination. In some embodiments, gastric cancer or gastroesophageal junction cancer is recurrent, locally advanced, or metastatic gastric cancer or gastroesophageal adenocarcinoma.

[0021] In one embodiment of the present invention, the anti-LAG-3 antibody is lilatrimab. In one embodiment of the present invention, the anti-LAG-3 antibody contains a serine-to-proline mutation at amino acid residue 228. [Modes for carrying out the invention]

[0022] Detailed description of the invention In one aspect, the present invention relates to an improved method for the treatment of malignant tumors in human patients. In particular, the present invention shows that administration of an anti-LAG-3 antibody in combination with an anti-PD-1 antibody and one or more chemotherapeutic agents achieves surprisingly improved treatment outcomes in a patient population having LAG-3-positive malignant tumors compared to a population including patients having both LAG-3-positive and LAG-3-negative tumors. Accordingly, in one aspect, the invention described herein relates to a method of treating a LAG-3-positive malignant tumor (e.g., gastric adenocarcinoma or gastroesophageal junction adenocarcinoma) by administering a combination of a LAG-3 inhibitor (e.g., an anti-LAG-3 antibody), a PD-1 pathway inhibitor (e.g., an anti-PD-1 antibody), and one or more chemotherapeutic agents.

[0023] 1. Definitions To facilitate an understanding of the description herein, several terms are first defined. As used herein, unless explicitly defined otherwise herein, each of the following terms may have the meaning set forth below. Additional definitions are set forth throughout the specification.

[0024] "Antibody" (Ab) includes, but is not limited to, a glycoprotein immunoglobulin that specifically binds to an antigen and comprises at least two heavy chains (H) and two light chains (L) interconnected by disulfide bonds. Each H chain comprises a heavy chain variable region (herein, V H abbreviated herein) and a heavy chain constant region. The heavy chain constant region comprises C H1 , C H2 and C H3 three constant domains. Each light chain comprises a light chain variable region (herein, V L abbreviated herein) and a light chain constant region. The light chain constant region comprises one constant domain C L . V H and V L regions can be further subdivided into hypervariable regions called complementarity determining regions (CDRs), interspersed with more conserved regions called framework regions (FRs). Each V H and V LThe antibody comprises three CDRs and four FRs, arranged from the amino terminus to the carboxyl terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable regions of the heavy and light chains contain binding domains that interact with the antigen. The constant region of the antibody may mediate the binding of immunoglobulins to various cells of the immune system (e.g., effector cells) and to host tissues or factors including the first component (C1q) of the classical complement system. The heavy chain may or may not have a C-terminal lysine. Unless otherwise specified herein, the amino acids in the variable region are numbered using the Kabat numbering system, and the amino acids in the constant region are numbered using the EU system. In one embodiment, the antibody is an intact antibody.

[0025] Immunoglobulins may be derived from any of the generally known isotypes, including but not limited to IgA, secretory IgA, IgG, and IgM. IgG subclasses are also well known to those skilled in the art and include, but are not limited to, human IgG1, IgG2, IgG3, and IgG4. “Isotype” means 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; totally synthetic antibodies; and single-chain antibodies. Non-human antibodies may be humanized by recombinant methods to reduce their immunogenicity in humans. Unless expressly stated and unless the context indicates otherwise, the term “antibody” includes monospecific antibodies, bispecific or multispecific antibodies, and single-chain antibodies. In one embodiment, an antibody is a bispecific antibody. In another embodiment, an antibody is a monospecific antibody. On the one hand, the constant region isotype is IgG4 with a mutation at amino acid residue 228, for example, S228P.

[0026] As used herein, “IgG antibodies” have the structure of natural IgG antibodies, that is, they have the same number of heavy and light chains and disulfide bonds as natural IgG antibodies of the same subclass. For example, anti-LAG-3 IgG1, IgG2, IgG3, or IgG4 antibodies consist of two heavy chains (HC) and two light chains (LC), where the two heavy and light chains have the same number and position of disulfide bonds as those found in natural IgG1, IgG2, IgG3, and IgG4 antibodies, respectively (unless the antibody has mutated to modify the disulfide bonds).

[0027] An “isolated antibody” means an antibody that substantially does not contain other antibodies with different antigen specificities (for example, an isolated antibody that specifically binds to LAG-3 substantially does not contain antibodies that specifically bind to antigens other than LAG-3). However, an isolated antibody that specifically binds to LAG-3 may have cross-reactivity to other antigens, such as LAG-3 molecules from different species. Furthermore, an isolated antibody may substantially not contain other cellular material and / or chemical substances.

[0028] Antibodies may be modified antibodies (e.g., by mutation, deletion, substitution, or binding to a non-antibody portion). For example, an antibody may contain one or more mutant amino acids (compared to a natural antibody) that alter the properties of the antibody (e.g., functional properties). Numerous such mutations are known in the art, for example, that affect the half-life of the antibody in a patient, effector function, and / or immune response. The term antibody also includes artificial polypeptide constructs that contain at least one antibody-derived antigen-binding site.

[0029] The term “monoclonal antibody” (“mAb”) refers to an antibody molecule in a single-molecule composition, i.e., a preparation of an antibody molecule that does not exist in nature, having essentially identical primary sequences and exhibiting a single binding specificity and affinity for a particular epitope. An mAb is an example of an isolated antibody. mAbs can be produced by hybridoma, recombinant, transgenic, or other techniques known to those skilled in the art.

[0030] A “human” antibody (HuMAb) refers to an antibody having a variable region in which both the framework and CDR region are derived from a human germ cell immunoglobulin sequence. Furthermore, if the antibody includes a constant region, the constant region is also derived from a human germ cell immunoglobulin sequence. The human antibodies of the present invention may contain amino acid residues not encoded by a human germ cell immunoglobulin sequence (e.g., mutations introduced in vitro by random or site-directed mutagenesis, or in vivo by somatic mutation). However, as used herein, the term “human antibody” is not intended to include antibodies in which a CDR sequence derived from the germ cells of another mammalian species, such as mouse, has been transplanted into a human framework sequence. The terms “human” antibody and “fully human” antibody are used synonymously.

[0031] A “humanized antibody” refers to 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 immunoglobulins. In one form of antibody humanization, some, most, or all of the amino acids outside the CDR domain are replaced with amino acids derived from human immunoglobulins, while some, most, or all of the amino acids within one or more CDR regions remain unchanged. Small additions, deletions, insertions, substitutions, or modifications of amino acids are acceptable as long as they do not impair the antibody's ability to bind to a particular antigen. “Humanized” antibodies retain similar antigen specificity to the original antibody.

[0032] 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.

[0033] An "anti-antigen" antibody is an antibody that specifically binds to an antigen. For example, an anti-LAG-3 antibody specifically binds to LAG-3.

[0034] The “antigen-binding portion” (also referred to as 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 antigen to which it is bound. It has been shown that the antigen-binding function of an antibody can be performed by fragments or portions of a full-length antibody. Examples of binding fragments encompassed by the term “antigen-binding portion” or “antigen-binding fragment” of an antibody, such as the anti-LAG-3 antibody described herein, include: (1) Fab fragments (fragments cleaved by papain) or similar monovalent fragments consisting of VL, VH, LC, and CH1 domains; (2) F(ab')2 fragment (fragment from pepsin cleavage) or similar divalent fragment containing two Fab fragments linked by a disulfide bridge in the hinge region; (3) Fd fragment consisting of a VH domain and a CH1 domain; (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 bi-single-domain antibody consisting of two VH domains linked by a hinge (dual-affinity re-targeting antibody (DART)); (7) Dual variable domain immunoglobulin; (8) Isolated complementarity-determining regions (CDRs); and (9) A combination of two or more isolated CDRs that can be linked by a synthetic linker if necessary. Furthermore, although the two domains of the Fv fragment, VL and VH, are encoded by separate genes, they can be linked by a synthetic linker using recombination and produced as a single protein chain in which the VL and VH regions pair to form a monovalent molecule (known as single-chain 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-chain antibodies are also intended to be included in the terminology of the “antigen-binding portion” or “antigen-binding fragment” of an antibody. These antibody fragments are obtained using prior art 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 technology or by enzymatic or chemical cleavage of intact immunoglobulin. In some embodiments, the antibody is the antigen-binding fragment.

[0035] The term “LAG-3” refers to the lymphocyte activation gene-3. The term “LAG-3” includes variants, isoforms, homologs, orthologs, and paralogs. For example, an antibody specific to the human LAG-3 protein may, in certain cases, cross-react with LAG-3 proteins from non-human species. In other embodiments, an antibody specific to the human LAG-3 protein is perfectly specific to the human LAG-3 protein and does not exhibit cross-reactivity with species or other types, or it may cross-react with LAG-3 from certain other species, but not with all other species (e.g., it cross-reacts with monkey LAG-3 but not with mouse LAG-3). The term “human LAG-3” refers to the complete amino acid sequence of human LAG-3, e.g., the complete amino acid sequence of human LAG-3 having GenBank accession number NP_002277. The term “mouse LAG-3” means the complete amino acid sequence of mouse LAG-3, for example, mouse LAG-3 having GenBank accession number NP_032505. LAG-3 is also known in the art, for example, as CD223. Human LAG-3 sequences may differ from human LAG-3 of GenBank accession number NP_002277, for example, by having conserved mutations or mutations in non-conserved regions, and LAG-3 may have substantially the same biological function as human LAG-3 of GenBank accession number NP_002277. For example, the biological function of human LAG-3 may be to have an epitope in the extracellular domain of LAG-3 that is specifically bound by the antibodies herein, or to bind to MHC class II molecules.

[0036] A particular human LAG-3 sequence is generally at least 90% identical in amino acid sequence to the human LAG-3 sequence of GenBank accession number NP_002277 and contains amino acid residues that identify the sequence as human when compared to the LAG-3 amino acid sequence of another species (e.g., mouse). In some cases, human LAG-3 may be at least 95%, or even more than 96%, 97%, 98%, or 99%, identical in amino acid sequence to the LAG-3 sequence of GenBank accession number NP_002277. In certain embodiments, a human LAG-3 sequence does not show more than 10 amino acid sequence differences from the LAG-3 sequence of GenBank accession number NP_002277. In certain embodiments, human LAG-3 does not show more than 5 amino acid differences, or more than 4, 3, 2, or 1 amino acid differences from the LAG-3 sequence of GenBank accession number NP_002277. The percentage of identity may be determined as described herein.

[0037] 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, and species homologs of human PD-1, as well as analogs that share at least one common epitope with PD-1. The complete PD-1 sequence can be found in GenBank accession number U64863.

[0038] The protein “programmed death 1 (PD-1)” is an inhibitory member of the CD28 receptor family, which also includes CD28, CTLA-4, ICOS, and BTLA. PD-1 is expressed on 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 early members of the family, CD28 and ICOS, were discovered by 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 by screening for differential expression in apoptotic cells (Ishida et al. EMBO J (1992); 11:3887-95). Other members of the family, CTLA-4 and BTLA, were discovered by screening for differential expression 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 is suggested to exist as a monomer and lacks the unpaired cysteine ​​residues characteristic of other CD28 family members.

[0039] 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). Structurally similar to CTLA-4, PD-1 lacks the MYPPPY motif (SEQ ID NO: 32), which is important 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 upon 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 various human cancers (Dong et al. (2002) Nat. Med. 8:787-9). The interaction between PD-1 and PD-L1 reduces tumor-infiltrating lymphocytes, decreases T-cell receptor-mediated proliferation, and enables 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).Inhibiting the local interaction between PD-1 and PD-L1 can reverse immunosuppression, and if the interaction between PD-1 and PD-L2 is also inhibited, the effect is additive (Iwai et al. (2002) Proc. Nat'l. Acad. Sci. USA 99:12293-7; Brown et al. (2003) J. Immunol. 170:1257-66).

[0040] Consistent with PD-1 being an inhibitory member of the CD28 family, PD-1-deficient animals express 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 been found to be involved 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 transmitted via BCR-mediated Ca 2+ -It has been shown to be essential for inhibiting tyrosine phosphorylation of flux and downstream effector molecules (Okazaki et al. (2001) PNAS 98:13866-71).

[0041] "Programmed death ligand-1 (PD-L1)" is one of two cell surface glycoprotein ligands of PD-1 (the other being PD-L2) that, upon binding to PD-1, downregulate T cell activation and cytokine secretion. The term "PD-L1" as used herein includes human PD-L1 (hPD-L1), variants, isoforms, and species homologs of hPD-L1, as well as analogs having at least one common epitope with hPD-L1. The complete hPD-L1 sequence can be found in GenBank accession number Q9NZQ7.

[0042] As used herein, the terms “programmed death ligand-2” and “PD-L2” 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 in GenBank accession number Q9BQ51.

[0043] As used herein, “patient” includes patients suffering from cancer (e.g., gastric cancer or gastroesophageal cancer). The terms “subject” and “patient” are used interchangeably.

[0044] "Administer" means physically introducing a composition containing a therapeutic agent into a subject using any of the various methods and delivery systems known to those skilled in the art. The routes of administration of the formulations described herein include intravenous, intramuscular, subcutaneous, intraperitoneal, spinal, or other non-enteral routes of administration, such as by injection or infusion. As used herein, "non-enteral administration" means a method of administration other than enteral and local administration, which is usually by injection and includes, but is not limited to, intravenous, intramuscular, intra-arterial, intrathecal, intrathoracic, intrafocal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intra-articular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal injection and infusion, as well as in vivo electroporation. In some embodiments, the formulation is administered by a route other than non-enteral administration, and in some embodiments, it is administered orally. Other non-enteral administration routes include local, epithelial, or mucosal administration routes, such as intranasal, intravaginal, intrarectal, sublingual, or topical administration. Administration may also be performed, for example, once, multiple times, and / or one or more times over a long period of time.

[0045] The term "treatment" or "therapy" in this context refers to any therapeutic intervention or method performed on the subject, or the administration of an active ingredient to the subject, with the aim of restoring, alleviating, improving, inhibiting, or delaying the progression, onset, worsening, or recurrence of symptoms, complications, or disease-related biochemical signs. The Criteria for Evaluation of Treatment Efficacy in Solid Tumors (RECIST) are measures of treatment effectiveness and established criteria that define when a tumor responds, stabilizes, or progresses during treatment. RECIST 1.1 is the current guideline for solid tumor measurements and definitions for objectively evaluating changes in tumor size for use in clinical trials of adult and pediatric cancers.

[0046] As used herein, “effective treatment” means a treatment that produces a beneficial effect, for example, improvement of at least one symptom of a disease or disorder. Beneficial effects may take the form of improvement over baseline, i.e., improvement over measurements or observations made before initiating treatment according to the method. Beneficial effects may also take the form of cessation, slowing, delaying, or stabilization of the adverse progression of markers of solid tumors. Effective treatment may mean relief of at least one symptom of a solid tumor. Such effective treatments may, for example, reduce patient pain, decrease the size and / or number of lesions, reduce or prevent tumor metastasis, and / or delay tumor growth.

[0047] The term “effective dose” means the amount of a drug that gives a desired biological, therapeutic, and / or preventive outcome. The outcome may be a reduction, improvement, mitigation, regression, delay, and / or remission of one or more signs, symptoms, or causes of disease, or any other desired change in a biological system. With respect to solid tumors, an effective dose is an amount sufficient to shrink the tumor and / or reduce the rate of tumor growth (e.g., inhibit tumor growth) or to prevent or delay other undesirable cell growth. In some embodiments, an effective dose is an amount sufficient to prevent or delay tumor development. An effective dose may be administered in one or more doses. An effective dose of a drug or composition is an amount that can (i) reduce the number of cancer cells; (ii) reduce the size of the tumor; (iii) inhibit, delay, slow to some extent, or stop the invasion of cancer cells into peripheral organs; (iv) inhibit tumor metastasis, i.e., can be delayed to some extent or stopped; (v) inhibit tumor growth; (vi) prevent or delay the development and / or recurrence of tumor; and / or (vii) alleviate to some extent one or more symptoms associated with cancer. In one example, “effective dose” is the amount of anti-LAG-3 antibody, anti-PD-1 antibody, and / or chemotherapeutic agent, or a combination thereof, that has been clinically proven to have an effect on a significant reduction in cancer or a delay in the progression of cancer such as solid tumors. The terms “fixed dose,” “constant dose,” and “constant fixed dose” as used herein are interchangeable and refer to the dose administered to a patient regardless of their body weight or body surface area (BSA). Therefore, fixed doses or constant doses are provided as absolute amounts of the drug (e.g., anti-LAG-3 antibody and / or anti-PD-1 antibody), not as mg / kg doses. For example, a 60 kg person and a 100 kg person may accept the same dose of the composition (e.g., 360 mg of anti-PD-1 antibody and 120 mg of anti-LAG-3 antibody in a single fixed-dose formulation vial containing both 360 mg and 120 mg of anti-LAG-3 antibody (or two fixed-dose formulation vials containing 180 mg of anti-PD-1 antibody and 60 mg of anti-LAG-3 antibody)).

[0048] With respect to the compositions of the present invention, the use of the term “fixed-dose combination” 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 antibody (e.g., mg). In certain embodiments, the fixed dose is based on the concentration of the antibody (e.g., mg / ml). In some embodiments, the ratio of the first antibody to the second antibody (mg) 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: The ratios are approximately 160, 1:180, 1:200, 200:1, 180:1, 160:1, 140:1, 120:1, 100:1, 90:1, 80:1, 70:1, 60:1, 50:1, 40:1, 30:1, 20:1, 15:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, or 2:1. For example, a 3:1 ratio of the first antibody to the second antibody may mean that the vial contains approximately 240 mg of the first antibody and 80 mg of the second antibody, or approximately 3 mg / ml of the first antibody and 1 mg / ml of the second antibody.

[0049] As used herein, the term “body weight-based dose” means that the dose administered to a patient is calculated based on the patient’s body weight. For example, when 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 amount of anti-LAG-3 antibody (i.e., 180 mg) and anti-PD-1 antibody (i.e., 180 mg) can be aspirated and withdrawn at once from a fixed-dose combination solution of anti-LAG-3 antibody and anti-PD-1 antibody in a 1:1 ratio.

[0050] As used herein, the term “progression-free survival” can be abbreviated as PFS and refers to the length of time during and after treatment for a solid tumor (i.e., melanoma) in which a patient lives with the disease but does not worsen.

[0051] As used herein, “dosing interval” refers to the time elapsed between multiple administrations of the formulation described herein to a subject. Therefore, the dosing interval can be expressed as a time range.

[0052] As used herein, the term "dosage frequency" means the frequency at which multiple doses of the formulation described herein are administered at a given time. Dosage frequency can be expressed as the number of doses per given time, for example, once a week or once every two weeks.

[0053] The terms “about once a week,” “about once a week,” “about once every two weeks,” or other similar dosing interval terms used herein mean approximate frequency, and “about once a week” or “about once a week” may encompass every 7 ± 2 days, i.e., every 5 to 9 days. Thus, the dosing frequency of “once a week” may be every 5 days, every 6 days, every 7 days, every 8 days, or every 9 days. “About once every two weeks” may include once every 14 ± 3 days, i.e., once every 11 to 17 days. Similar approximations apply, for example, to once every 3 weeks, once every 4 weeks, once every 5 weeks, once every 6 weeks, and once every 12 weeks. In some embodiments, an interval of approximately every six weeks or approximately every twelve weeks means that the first dose may be administered on any day in the first week, and the next dose may be administered on any day in the sixth or twelfth week, respectively. In other embodiments, an interval of approximately every six weeks or approximately every twelve weeks means that the first dose may be administered on a specific day in the first week (e.g., Monday), and the next dose may be administered on the same day in the sixth or twelfth week (e.g., Monday), respectively.

[0054] "Cancer" refers to a broad group of diseases characterized by the uncontrolled proliferation of abnormal cells in the body. Uncontrolled cell division and proliferation can lead to the formation of malignant tumors, which can invade adjacent tissues and metastasize to distant parts of the body via the lymphatic system or bloodstream. "Cancer" or "cancer tissue" includes tumors. "Gastric cancer" and "stomach cancer" are used interchangeably herein. As used herein, "stomach cancer" can originate in any part of the stomach and may spread throughout the stomach or to other organs. It may grow along the stomach wall toward the esophagus or small intestine. It may also spread through the stomach wall to nearby lymph nodes as well as to organs such as the liver, pancreas, and large intestine. It may also metastasize to distant organs such as the lungs, supraclavicular lymph nodes, and ovaries. Various types of stomach cancer include adenocarcinoma, lymphoma, gastrointestinal stromal tumor (GIST), and carcinoid tumor.

[0055] As used herein, the term “tumor” means any mass of tissue resulting from excessive cell proliferation or growth, whether benign (non-cancerous) or malignant (cancerous), including precancerous lesions.

[0056] The term “LAG-3 positive” or “LAG-3 expression positive” in relation to LAG-3 expression refers to the percentage of cells in a biopsy tissue sample, including tumor cells and tumor-infiltrating inflammatory cells, that are recorded as expressing LAG-3. In some embodiments, for LAG-3 expression assayed by immunohistochemistry (IHC), a LAG-3 positive tumor or LAG-3 expression positive tumor means that at least about 0.01%, at least about 0.5%, at least about 1%, at least about 2%, at least about 3%, at least about 4%, at least about 5%, at least about 6%, at least about 7%, at least about 8%, at least about 9%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or 100% of the total cell number express LAG-3. In other embodiments, LAG-3-positive tumors or LAG-3 expression-positive tumors are defined as tumor-associated inflammatory cells (e.g., T cells, CD8) as assayed by immunohistochemistry (IHC) or flow cytometry. + T cells, CD4 + T cells, FOXP3 +At least about 0.01%, at least about 0.01%, at least about 0.5%, at least about 1%, at least about 2%, at least about 3%, at least about 4%, at least about 5%, at least about 6%, at least about 7%, at least about 8%, at least about 9%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or 100% of the total number of cells express LAG-3. A LAG-3-positive tumor or LAG-3 expression-positive tumor may also be expressed herein as a tumor that expresses LAG-3. In some embodiments, a LAG-3-positive tumor or LAG-3 expression-positive tumor means that at least about 0.1% to at least about 20% of the total number of cells express LAG-3. In some embodiments, LAG-3-positive tumors or LAG-3 expression-positive tumors are tumor-associated inflammatory cells (e.g., T cells, CD8). + T cells, CD4 + T cells, FOXP3 + This means that at least approximately 0.1% to at least approximately 20% of the total number of cells express LAG-3. In certain embodiments, a LAG-3-positive tumor or LAG-3 expression-positive tumor means that at least approximately 0.1% to at least approximately 10% of the total number of cells express LAG-3. In certain embodiments, a LAG-3-positive tumor or LAG-3 expression-positive tumor is characterized by tumor-infiltrating inflammatory cells (e.g., T cells, CD8). + T cells, CD4 + T cells, FOXP3 + This means that at least about 0.1% to at least about 10% of the total number of cells express LAG-3. In some embodiments, a LAG-3-positive tumor or LAG-3 expression-positive tumor means that at least about 1% of the total number of cells express LAG-3 on the cell surface. In some embodiments, a LAG-3-positive tumor or LAG-3 expression-positive tumor means that tumor-infiltrating inflammatory cells (e.g., T cells, CD8 cells) express LAG-3. + T cells, CD4 + T cells, FOXP3 +In another embodiment, a LAG-3-positive tumor or LAG-3 expression-positive tumor means that at least about 1% of the total number of cells express LAG-3 on their cell surface. In yet another embodiment, a LAG-3-positive tumor or LAG-3 expression-positive tumor means that at least about 5% of the total number of cells express LAG-3 on their cell surface. + T cells, CD4 + T cells, FOXP3 + This means that at least about 5% of the total number of cells express LAG-3 on the cell surface. In a particular embodiment, a LAG-3-positive tumor or LAG-3 expression-positive tumor means that at least about 1%, or in the range of 1-5%, of the total number of cells express LAG-3 on the cell surface. In a particular embodiment, a LAG-3-positive tumor or LAG-3 expression-positive tumor is tumor-infiltrating inflammatory cells (e.g., T cells, CD8 + T cells, CD4 + T cells, FOXP3 + This means that LAG-3 is expressed on the cell surface in at least about 1% of the total number of cells, or in the range of 1-5%.

[0057] "LAG-3 negative" or "LAG-3 expression negative" refers to the percentage of cells in a biopsy tissue sample that are LAG-3 positive or LAG-3 expression negative, including tumor cells and tumor-infiltrating inflammatory cells.

[0058] The terms “PD-L1 positive” or “PD-L1 expression positive” in relation to cell surface PD-L1 expression refer to the percentage of cells in a biopsy tissue sample, including tumor cells and tumor-infiltrating inflammatory cells, that are recorded as expressing cell surface PD-L1. For cell surface expression assayed by immunohistochemistry (IHC), for example using mAb28-8, a PD-L1 positive tumor or PD-L1 expression positive tumor means that at least about 0.01%, at least about 0.5%, at least about 1%, at least about 2%, at least about 3%, at least about 4%, at least about 5%, at least about 6%, at least about 7%, at least about 8%, at least about 9%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, or at least about 30% of the total cell number express PD-L1. A PD-L1 positive tumor or PD-L1 expression positive tumor may be referred to herein as a tumor expressing PD-L1. In other embodiments, a PD-L1-positive tumor or PD-L1-expression-positive tumor means that at least about 0.1% to at least about 20% of the total cell number express PD-L1. In certain embodiments, a PD-L1-positive tumor or PD-L1-expression-positive tumor means that at least about 0.1% to at least about 10% of the total cell number express PD-L1. In some embodiments, a PD-L1-positive tumor or PD-L1-expression-positive tumor means that at least about 1% of the total cell number express PD-L1 on the cell surface. In other embodiments, a PD-L1-positive tumor or PD-L1-expression-positive tumor means that at least about 5% of the total cell number express PD-L1 on the cell surface. In certain embodiments, a PD-L1-positive tumor or PD-L1-expression-positive tumor means that at least about 1% of the total cell number, or in the range of about 1-5%, express PD-L1 on the cell surface.

[0059] The term "PD-L1 negative" or "PD-L1 expression negative" in relation to PD-L1 expression on the cell surface refers to the percentage of cells in a biopsy tissue sample, including tumor cells and tumor-infiltrating inflammatory cells, that are not PD-L1 positive or PD-L1 expression negative.

[0060] The term “evaluable PD-L1 expression status” refers to the measurable expression level of PD-L1, which is generally 5% or greater, or less than 5%.

[0061] As used herein, the term “tumor mutation burden” (TMB) refers to the number of somatic mutations in the tumor genome and / or the number of somatic mutations per region of the tumor genome. Germline (genetic) variants are excluded when determining the TMB because the immune system is more likely to recognize them as self. Tumor mutation burden (TMB) may also be used interchangeably with “tumor mutation load” (tumor mutational burden, tumor mutational load).

[0062] Tumor genome sequencing (TMB) is a genetic analysis of the tumor genome and can therefore be determined using sequencing methods well known to those skilled in the art. Tumor DNA can be compared with DNA from normal tissue adapted to the patient to rule out germline mutations or polymorphisms.

[0063] In one embodiment, TMB is determined by sequencing tumor DNA using high-throughput sequencing techniques, such as next-generation sequencing (NGS) or NGS-based methods. In one embodiment, NGS-based methods include whole-genome sequencing (WGS), whole-exome sequencing (WES), or FOUNDATIONONE® CDX (商標)The cancer gene panel is selected from comprehensive genomic profiling (CGP), such as the MSK-IMPACT clinical trial. In one embodiment, TMB as used herein means the number of somatic mutations per megabase (Mb) of sequenced DNA. In one embodiment, TMB is determined using the total number of non-synonymous mutations, such as missense mutations (i.e., altering specific amino acids in a protein) and / or nonsense mutations (premature termination of a protein sequence and consequently causing cleavage of the protein sequence), and is determined by normalizing matched tumors with germline samples to exclude hereditary germline variations. In another embodiment, TMB is determined using the total number of missense mutations in the tumor. A sufficient amount of sample is required to determine TMB. In one embodiment, tissue samples (e.g., at least 10 slides) are used for evaluation. In one embodiment, TMB is expressed as NsMs per megabase (NsM / Mb). One megabase represents one million bases.

[0064] The TMB state can be a numerical or relative value within the highest quantile or tertile of the control set, e.g., high, medium, or low.

[0065] As used herein, the term “high TMB” means the number of somatic mutations in the tumor genome that exceeds a large number of somatic mutations that are normal or mean. In one embodiment, the TMB has a score of at least 210, at least 215, at least 220, at least 225, at least 230, at least 235, at least 240, at least 245, at least 250, at least 255, at least 255, at least 260, and at least 265. Having a score of at least 270, at least 275, at least 280, at least 285, at least 290, at least 295, at least 300, at least 305, at least 310, at least 315, at least 320, at least 325, at least 330, at least 335, at least 340, at least 345, at least 350, at least 355, at least 360, at least 365, at least 370, at least 375, at least 380, at least 385, at least 390, at least 395, at least 400, at least 405, at least 410, at least 415, at least 420, at least 425, at least 430, at least 435, at least 440, at least 445, at least 450, at least 455, at least 460, at least 465, at least 470, at least 475, at least 480, at least 485, at least 490, at least 495, or at least 500. In other embodiments, high TMB has a score of at least 221, at least 222, at least 223, at least 224, at least 225, at least 226, at least 227, at least 228, at least 229, at least 230, at least 231, at least 232, at least 233, at least 234, at least 235, at least 236, at least 237, at least 238, at least 239, at least 240, at least 241, at least 242, at least 243, at least 244, at least 245, at least 246, at least 247, at least 248, at least 249, or at least 250; and in specific embodiments, high TMB has a score of at least 243.In another embodiment, “high TMB” refers to the TMB within the highest quantile of the control TMB value. For example, all subjects with evaluable TMB data are grouped according to the fractal distribution of TMB, i.e., subjects are ranked from the highest to the lowest number of gene variations and classified into a defined number of groups. In one embodiment, all subjects with evaluable TMB data are ranked and divided into three groups, and “high TMB” is the highest tertile of the control TMB value. In a particular embodiment, the tertile boundaries are 0 < 100 gene variations; 100 to 243 gene variations; and > 243 gene variations. Once ranked, subjects with evaluable TMB data should be understood to be classifiable into any of the group numbers (e.g., quartile, quintile, etc.). In one embodiment, “high TMB” means TMB with at least approximately 20 mutations / tumor, at least approximately 25 mutations / tumor, at least approximately 30 mutations / tumor, at least approximately 35 mutations / tumor, at least approximately 40 mutations / tumor, at least approximately 45 mutations / tumor, at least approximately 50 mutations / tumor, at least approximately 55 mutations / tumor, at least approximately 60 mutations / tumor, at least approximately 65 mutations / tumor, at least approximately 70 mutations / tumor, at least approximately 75 mutations / tumor, at least approximately 80 mutations / tumor, at least approximately 85 mutations / tumor, at least approximately 90 mutations / tumor, at least approximately 95 mutations / tumor, or at least approximately 100 mutations / tumor. In one embodiment, “high TMB” means TMB with at least approximately 105 mutations / tumor, at least approximately 110 mutations / tumor, at least approximately 115 mutations / tumor, at least approximately 120 mutations / tumor, at least approximately 125 mutations / tumor, at least approximately 130 mutations / tumor, at least approximately 135 mutations / tumor, at least approximately 140 mutations / tumor, at least approximately 145 mutations / tumor, at least approximately 150 mutations / tumor, at least approximately 175 mutations / tumor, or at least approximately 200 mutations / tumor. In a particular embodiment, a tumor with high TMB has at least approximately 100 mutations / tumor.

[0066] “High TMB” is also used in mutation assays, such as FOUNDATIONONE® CDX. (商標)This can mean the number of mutations per megabase of the sequenced tumor genome, as measured by the assay. In one embodiment, high TMB may refer to FOUNDATIONONE® CDX (商標) As measured by the assay, this means at least approximately 9, at least approximately 10, at least approximately 11, at least approximately 12, at least approximately 13, at least approximately 14, at least approximately 15, at least approximately 16, at least approximately 17, at least approximately 18, at least approximately 19, or at least approximately 20 mutations per megabase of genome. In certain embodiments, “High TMB” refers to FOUNDATIONONE® CDX (商標) The assay implies at least 10 mutations per megabase of the sequenced genome.

[0067] As used herein, the term “intermediate TMB” means a number of somatic mutations in the tumor genome that is normal or average, or a number of somatic mutations of that magnitude, and the term “low TMB” means a number of somatic mutations in the tumor genome that is less than normal or average. In certain embodiments, “high TMB” has a score of at least 243, “intermediate TMB” has a score of 100 to 242, and “low TMB” has a score of 100 or less (or 0 to 100). “Intermediate or low TMB” is, for example, FOUNDATIONONE® CDX (商標) This means fewer than 9 mutations per megabase of the sequenced genome, as measured by assays.

[0068] Microsatellite instability (MSI) is a state of genetic hypermutability resulting from impaired DNA mismatch repair (MMR). The presence of MSI is evidence of a phenotype in which MMR is not functioning properly. In most cases, the genetic basis for the instability in MSI tumors is a hereditary germline mutation in one of the five human MMR genes: MSH2, MLH1, MSH6, PMS2, and PMS1. In certain embodiments, subjects treated with tumors have high levels of microsatellite instability (MSI-H) and have at least one mutation in the MSH2, MLH1, MSH6, PMS2, or PMS1 gene. In other embodiments, subjects treated with tumors in the control group do not have microsatellite instability (MSS or MSI stability) and do not have mutations in the MSH2, MLH1, MSH6, PMS2, or PMS1 gene.

[0069] "Immune response" refers to the action of cells of the immune system (e.g., T lymphocytes, B lymphocytes, natural killer (NK) cells, macrophages, eosinophils, mast cells, dendritic cells, or neutrophils) and soluble macromolecules (including antibodies, cytokines, and complement) produced by any of these cells or the liver, which as a result selectively target, bind to, damage, destroy, and / or eliminate from the body of vertebrates in the case of invading pathogens, pathogen-infected cells or tissues, cancerous or other abnormal cells, or, in the case of autoimmune or pathological inflammation, normal human cells or tissues.

[0070] "Tumor-infiltrating inflammatory cells" or "tumor-associated inflammatory cells" are any type of cell that is commonly involved in the inflammatory response in question and infiltrates tumor tissue. Such cells include tumor-infiltrating lymphocytes (TILs), macrophages, monocytes, eosinophils, histiocytes, and dendritic cells.

[0071] The use of alternative words (e.g., “or”) should be understood to mean one, both, or any combination thereof of the alternative words. The indefinite articles “a” or “an” used herein should be understood to mean “one or more” of the listed or enumerated components.

[0072] As used herein, the term “and / or” should be interpreted as meaning that each of the two identified features or components is specifically disclosed, with or without the other. Accordingly, as used herein, the term “and / or” in phrases such as “A and / or B” is intended to include “A and B,” “A or B,” “A” (alone), and “B” (alone). Similarly, as used in phrases such as “A, B and / or C” is intended to include each of the following aspects: 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).

[0073] If a side is described with the term "contains," it is understood that similar sides are also provided, described with the terms "consist of" and / or "essentially composed of."

[0074] The terms “about” or “essentially including” mean a value or composition that lies within the tolerance range of a particular value or composition as determined by those skilled in the art, which may in part depend on how this value or composition is measured or determined, i.e., the limits of the measuring system. For example, “about” or “essentially including” may mean within one or two or more standard deviations for practice in the art. Alternatively, “about” or “essentially including” may mean a range of up to 10% or 20% (i.e., ±10% or ±20%). For example, about 3 mg may include any number between 2.7 mg and 3.3 mg (in the case of 10%) or between 2.4 mg and 3.6 mg (in the case of 20%). Furthermore, particularly with respect to biological systems or processes, these terms may mean up to one order of magnitude higher or up to five times higher. When a particular value or composition is provided herein and in the claims, unless otherwise specified, the meaning of “about” or “essentially including” should be assumed to be within the tolerance range of that particular value or composition.

[0075] As described herein, any concentration range, percentage range, ratio range, or integer range should be understood to include any integer value within the range described, and, where appropriate, fractions thereof (e.g., one-tenth and one-hundredth of an integer), unless otherwise specified.

[0076] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to whom this disclosure relates. For example, the Concise Dictionary of Biomedicine and Molecular Biology, Juo, Pei-Show, 2nd ed., 2002, CRC Press; The Dictionary of Cell and Molecular Biology, 5th ed., 2013, Academic Press; and the Oxford Dictionary of Biochemistry and Molecular Biology, 2006, Oxford University Press provide general dictionaries of many of the terms used herein.

[0077] Units, prefixes, and symbols are expressed in the form approved by the International System of Units (SI) for their base units. Numerical ranges include the digit defining the range. The headings provided herein are not intended to limit the various aspects of the disclosure herein and may be provided for reference to the entire specification. Accordingly, the terms defined immediately below are defined in more detail by referring to the entire specification.

[0078] Various aspects of the present invention are described in more detail in the following subsections.

[0079] 2. The method of the present invention In one respect, the present invention relates to a method for treating LAG-3-positive malignancies (e.g., gastric cancer or gastroesophageal junction cancer) in a target population where such treatment is needed. Combination therapy of a LAG-3 inhibitor (e.g., an anti-LAG-3 antibody), a PD-1 pathway inhibitor (e.g., an anti-PD-1 antibody), and one or more chemotherapeutic agents yields better treatment outcomes (e.g., objective response rate and disease control rate) in a patient population with LAG-3-positive malignancies (e.g., gastric tumors or gastroesophageal junction cancer) than in a general patient population with a mixture of LAG-3-negative and LAG-3-positive malignancies. In one respect, to improve the treatment of malignancies, the present invention identifies patients with LAG-3-positive tumors and provides immunotherapy of a LAG-3 inhibitor (e.g., an anti-LAG-3 antibody), a PD-1 pathway inhibitor (e.g., an anti-PD-1 antibody), and one or more chemotherapeutic agents.

[0080] One aspect of the present invention relates to a method for inhibiting the growth of a malignant tumor in a human patient, comprising administering to the patient an effective amount of each of (a) a LAG-3 antagonist; (b) a PD-1 pathway inhibitor; and (c) one or more chemotherapeutic agents. Another aspect of the present invention relates to a method for treating cancer in a human patient, comprising administering to the patient an effective amount of each of (a) a LAG-3 antagonist; (b) a PD-1 pathway inhibitor; and (c) one or more chemotherapeutic agents. One aspect of the present invention relates to a method for treating recurrent, locally advanced, or metastatic gastric cancer or gastroesophageal junction adenocarcinoma in a human patient, comprising administering to the patient an effective dose of: (a) an anti-LAG-3 antibody comprising the CDR1, CDR2, and CDR3 domains of the heavy chain variable region having the sequence described in SEQ ID NO: 3 and the CDR1, CDR2, and CDR3 domains of the light chain variable region having the sequence described 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 the sequence described in SEQ ID NO: 15 and the CDR1, CDR2, and CDR3 domains of the light chain variable region having the sequence described in SEQ ID NO: 17; and (c) one or more standard therapeutic regimens. In one embodiment, the present invention relates to a method for treating gastric cancer or gastroesophageal junction cancer in a human patient, comprising administering to the patient an effective amount of: (a) an anti-LAG-3 antibody comprising the CDR1, CDR2, and CDR3 domains of the heavy chain variable region having the sequence described in SEQ ID NO: 3 and the CDR1, CDR2, and CDR3 domains of the light chain variable region having the sequence described 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 the sequence described in SEQ ID NO: 15 and the CDR1, CDR2, and CDR3 domains of the light chain variable region having the sequence described in SEQ ID NO: 17; and (c) one or more chemotherapeutic agents selected from the group consisting of XELOX, FOLFOX, and SOX. In one embodiment, the present invention relates to a method for selecting a malignant tumor in a human patient for immunotherapy, comprising (a) determining the level of LAG-3 expression in a tumor sample; and (b) selecting the tumor for immunotherapy if the tumor is a LAG-3 positive tumor.In one embodiment, the present invention includes a method for identifying malignant tumors in human patients as eligible for immunotherapy, comprising (a) determining the level of LAG-3 expression in a tumor sample; and (b) identifying the tumor as eligible for immunotherapy if the tumor is a LAG-3 positive tumor. In one embodiment, the present invention includes a method for identifying malignant tumors in human patients that are likely to respond to immunotherapy, the method comprising (a) determining the level of LAG-3 expression in a tumor sample; and (b) identifying the tumor as potentially responsive to treatment if the tumor is a LAG-3 positive tumor. In one embodiment, the present invention includes a method for identifying malignant tumors in human patients that are likely to respond to immunotherapy, the method comprising (a) determining the level of LAG-3 expression in a tumor sample; and (b) identifying the tumor as potentially responsive to treatment if the tumor is a LAG-3 positive tumor. In one embodiment, the present invention includes a method for classifying malignant tumors in human patients that are likely to respond to immunotherapy, the method comprising (a) determining the level of LAG-3 expression in a tumor sample; and (b) classifying the tumor as likely to respond to immunotherapy if the tumor is a LAG-3 positive tumor. In one embodiment, the immunotherapy comprises contacting the tumor with a therapeutically effective amount of a LAG-3 inhibitor, a PD-1 pathway inhibitor, and one or more chemotherapeutic agents. In one embodiment, the LAG-3 inhibitor is an anti-LAG-3 antibody, and the PD-1 pathway inhibitor is an anti-PD-1 antibody. In a particular embodiment, any of the methods of the present invention further comprises determining PD-L1 expression in a tumor sample.

[0081] In one embodiment, the present invention includes a method for identifying patients with malignant tumors likely to respond to immunotherapy, the method comprising (a) determining the level of LAG-3 expression in a tumor sample; and (b) identifying patients likely to respond to treatment if the tumor is a LAG-3 positive tumor. In one embodiment, the present invention includes a method for selecting patients with malignant tumors for immunotherapy, the method comprising (a) determining the level of LAG-3 expression in a tumor sample; and (b) selecting patients for immunotherapy if the tumor is a LAG-3 positive tumor. In one embodiment, the immunotherapy comprises contacting the tumor with a therapeutically effective amount of a LAG-3 inhibitor, a PD-1 pathway inhibitor, and one or more chemotherapeutic agents. In one embodiment, the LAG-3 inhibitor is an anti-LAG-3 antibody, and the PD-1 pathway inhibitor is an anti-PD-1 antibody. In a particular embodiment, any of the methods of the present invention comprises determining PD-L1 expression in a tumor sample.

[0082] In one embodiment, the present invention comprises a method for treating a malignant tumor in a human patient, comprising administering to the patient an immunotherapy described herein; wherein the patient is expected to respond to treatment with a LAG-3 inhibitor, a PD-1 pathway inhibitor, and a chemotherapeutic agent based on LAG-3 expression or based on LAG-3 and PD-L1 expression in a tumor sample of the patient. In one embodiment, the LAG-3 inhibitor is an anti-LAG-3 antibody, and the PD-1 pathway inhibitor is an anti-PD-1 antibody.

[0083] In one embodiment, the present invention includes a method for treating a tumor in a human patient, comprising: administering the patient the immunotherapy described herein; wherein the patient is expected to respond to treatment with a LAG-3 inhibitor, a PD-1 pathway inhibitor, and a chemotherapeutic agent based on having a high tumor mutational burden (TMB) status. In one embodiment, the present invention includes a method for treating cancer in a human patient, comprising: administering the patient the immunotherapy described herein; wherein the patient is expected to respond to treatment with a LAG-3 inhibitor, a PD-1 pathway inhibitor, and a chemotherapeutic agent based on having a high tumor mutational burden (TMB) status. In one embodiment, the cancer is gastric cancer or gastroesophageal junction cancer.

[0084] In one embodiment, the present invention includes a method for treating a tumor in a human patient, comprising: administering the patient the immunotherapy described herein; wherein the patient is expected to respond to treatment with a LAG-3 inhibitor, a PD-1 pathway inhibitor, and a chemotherapeutic agent based on having a high degree of microsatellite instability (MSI-H). In one embodiment, the present invention includes a method for treating cancer in a human patient, comprising: administering the patient the immunotherapy described herein; wherein the patient is expected to respond to treatment with a LAG-3 inhibitor, a PD-1 pathway inhibitor, and a chemotherapeutic agent based on having a high degree of microsatellite instability (MSI-H). In one embodiment, the cancer is gastric cancer or gastroesophageal junction cancer.

[0085] In one embodiment, the present invention includes a method for treating a malignant tumor in a human patient who needs it, comprising: (a) determining the LAG-3 expression level or the LAG-3 and PD-L1 expression levels in a tumor sample; and (b) administering to the patient a therapeutically effective dose of a LAG-3 inhibitor if the tumor is a LAG-3-positive tumor or a LAG-3-positive PD-L1-positive tumor. In one embodiment, the present invention includes a method for treating a malignant tumor in a human patient who needs it, comprising: (a) identifying the patient as having a LAG-3-positive malignant tumor or a LAG-3-positive PD-L1-positive malignant tumor; and (b) administering to the patient a therapeutically effective dose of a LAG-3 inhibitor, a PD-1 pathway inhibitor and one or more chemotherapeutic agents. In one embodiment, the present invention provides a method for treating a malignant tumor in a human patient requiring such treatment, comprising administering a therapeutically effective dose of a LAG-3 inhibitor to the patient, wherein the patient is identified prior to administration as having a LAG-3 positive malignant tumor or a LAG-3 positive PD-L1 positive malignant tumor. In one embodiment, the LAG-3 inhibitor is an anti-LAG-3 antibody, and the PD-1 pathway inhibitor is an anti-PD-1 antibody.

[0086] In one embodiment, the present invention provides a method for treating a malignant tumor in a human patient who needs it, comprising: (a) determining the LAG-3 expression level or the LAG-3 and PD-L1 expression levels in a tumor sample; and (b) administering to the patient a therapeutically effective dose of a LAG-3 inhibitor, a PD-1 pathway inhibitor, and one or more chemotherapeutic agents if the tumor is a LAG-3-positive tumor or a LAG-3-positive / PD-L1-positive tumor. In one embodiment, the present invention provides a method for treating a malignant tumor in a human patient who needs it, comprising: (a) identifying a patient having a LAG-3-positive malignant tumor or a LAG-3-positive / PD-L1-positive malignant tumor; and (b) administering to the patient a therapeutically effective dose of a LAG-3 inhibitor, a PD-1 pathway inhibitor, and one or more chemotherapeutic agents. In one embodiment, the LAG-3 inhibitor is an anti-LAG-3 antibody, and the PD-1 pathway inhibitor is an anti-PD-1 antibody.

[0087] In one embodiment, the present invention provides a method for treating a malignant tumor in a human patient who requires it, comprising: (a) determining the LAG-3 expression level or the LAG-3 and PD-L1 expression levels in a tumor sample; and (b) administering to the patient a therapeutically effective dose of a LAG-3 inhibitor, a PD-1 pathway inhibitor, and one or more chemotherapeutic agents if the tumor is a LAG-3-positive tumor or a LAG-3-positive PD-L1-positive tumor. In one embodiment, the present invention provides a method for treating a malignant tumor in a human patient who requires it, comprising: (a) identifying a patient having a LAG-3-positive malignant tumor or a LAG-3-positive PD-L1-positive malignant tumor; and (b) administering to the patient a therapeutically effective dose of a LAG-3 inhibitor, a PD-1 pathway inhibitor, and one or more chemotherapeutic agents. In one embodiment, the present invention provides a method for treating a malignant tumor in a human patient in need thereof, comprising administering to the patient a therapeutically effective amount of a LAG-3 inhibitor, a PD-1 pathway inhibitor, and one or more chemotherapeutic agents, wherein the patient has been identified prior to administration as having a LAG-3 positive malignant tumor or a LAG-3 positive PD-L1 positive malignant tumor. In one embodiment, the LAG-3 inhibitor is an anti-LAG-3 antibody, and the PD-1 pathway inhibitor is an anti-PD-1 antibody.

[0088] In another embodiment, the present invention provides a method for treating a malignant tumor in a human patient in need thereof, comprising administering to the patient an immunotherapy described herein, wherein the patient has been identified prior to administration as having a LAG-3 positive malignant tumor or a LAG-3 positive PD-L1 positive malignant tumor. In one embodiment, the immunotherapy comprises administering a therapeutically effective amount of a LAG-3 inhibitor, a PD-1 pathway inhibitor, and one or more chemotherapeutic agents. In one embodiment, the LAG-3 inhibitor is an anti-LAG-3 antibody, and the PD-1 pathway inhibitor is an anti-PD-1 antibody.

[0089] In certain embodiments, the present invention provides a method for extending progression-free survival for 12 months or more in a human patient with a malignant tumor, comprising administering an immunotherapy described herein to the patient, wherein the patient has been identified prior to administration as having a LAG-3 positive malignant tumor or a LAG-3 positive PD-L1 positive malignant tumor and has a progression-free survival of 12 months or more. In some embodiments, the patient's progression-free survival can be extended beyond approximately 13 months, approximately 14 months, approximately 15 months, approximately 16 months, approximately 17 months, approximately 18 months, approximately 2 years, approximately 3 years, approximately 4 years, approximately 5 years, approximately 6 years, approximately 7 years, approximately 8 years, approximately 9 years, or approximately 10 years after administration. In some embodiments, the immunotherapy comprises administering a therapeutically effective dose of a LAG-3 inhibitor, a PD-1 pathway inhibitor, and one or more chemotherapeutic agents. In some embodiments, the LAG-3 inhibitor is an anti-LAG-3 antibody, and the PD-1 pathway inhibitor is an anti-PD-1 antibody.

[0090] In yet another embodiment, the present invention provides a method for reducing tumor size by at least 10% in a human patient having a malignant tumor, comprising administering to the patient an immunotherapy described herein, wherein the patient has been identified prior to administration as having a LAG-3 positive malignant tumor (e.g., gastric cancer or gastroesophageal junction cancer) or a LAG-3 positive PD-L1 positive malignant tumor, and the administration reduces the tumor size by at least about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, or 100% compared to the tumor size prior to administration. In one embodiment, the method comprises identifying the patient prior to administration as having a LAG-3 positive malignant tumor or a LAG-3 positive PD-L1 positive malignant tumor. In one embodiment, the LAG-3 inhibitor is an anti-LAG-3 antibody, and the PD-1 pathway inhibitor is an anti-PD-1 antibody.

[0091] The present invention may also include a method for preventing relapse and / or inducing remission in a patient, comprising administering the immunotherapy described herein to the patient, wherein the patient has been identified prior to administration as having a LAG-3 positive malignancy (e.g., gastric cancer or gastroesophageal junction cancer) or a LAG-3 positive PD-L1 positive malignancy. In one embodiment, the method of the present invention comprises (i) identifying the patient as having a LAG-3 positive malignancy or a LAG-3 positive PD-L1 positive malignancy; and (ii) administering the immunotherapy described herein to the patient. In one embodiment, the immunotherapy comprises administering to the patient a therapeutically effective amount of a LAG-3 inhibitor, a PD-1 pathway inhibitor, and one or more chemotherapeutic agents. In one embodiment, the LAG-3 inhibitor is an anti-LAG-3 antibody, and the PD-1 pathway inhibitor is an anti-PD-1 antibody.

[0092] In certain embodiments, the present invention provides a method for a cancer treatment comprising administering an immunotherapy described herein to a patient, wherein each patient is identified prior to administration as having a LAG-3 positive malignancy (e.g., gastric cancer or gastroesophageal junction cancer) or a LAG-3 positive PD-L1 positive malignancy, and the objective response rate is higher than approximately 55%, approximately 60%, approximately 65%, approximately 70%, or approximately 75%. In some embodiments, the method comprises identifying the patient as having a LAG-3 positive malignancy or a LAG-3 positive PD-L1 positive malignancy prior to administration. In some embodiments, the immunotherapy comprises administering a therapeutically effective dose of a LAG-3 inhibitor, a PD-1 pathway inhibitor, and one or more chemotherapeutic agents. In some embodiments, the LAG-3 inhibitor is an anti-LAG-3 antibody, and the PD-1 pathway inhibitor is an anti-PD-1 antibody.

[0093] In certain embodiments, the present invention provides a method for achieving a disease control rate greater than 55% in a patient population in which each patient has a malignant tumor, in a cancer treatment comprising administering an immunotherapy described herein to a patient, wherein each patient is identified prior to administration as having a LAG-3 positive malignant tumor (e.g., gastric cancer or gastroesophageal junction cancer) or a LAG-3 positive PD-L1 positive malignant tumor, and the disease control rate is greater than about 55%, about 60%, about 65%, about 70%, or about 75%. In some embodiments, the method comprises identifying the patient prior to administration as having a LAG-3 positive malignant tumor or a LAG-3 positive PD-L1 positive malignant tumor. In some embodiments, the immunotherapy comprises administering a therapeutically effective dose of a LAG-3 inhibitor, a PD-1 pathway inhibitor, and one or more chemotherapeutic agents. In some embodiments, the LAG-3 inhibitor is an anti-LAG-3 antibody, and the PD-1 pathway inhibitor is an anti-PD-1 antibody.

[0094] In other embodiments, each patient of this method experiences (i) an extension of progression-free survival of 12 months or longer, (ii) a reduction in tumor size of at least approximately 10%, 20%, 30%, 40%, or 50% compared to the tumor size before administration, or (iii) both. In one embodiment, the patient population may be at least 100 patients with LAG-3 positive malignancies (e.g., gastric cancer or gastroesophageal junction cancer) or LAG-3 positive PD-L1 positive malignancies. In another embodiment, the patient population may be at least approximately 200, 300, 400, 500, 600, 700, 800, 900, or 1000 patients with LAG-3 positive malignancies or LAG-3 positive PD-L1 positive malignancies.

[0095] In a further embodiment, the present invention provides a method for selecting a human patient suitable for combination therapy, comprising: (a) identifying the patient as having a LAG-3 positive malignancy or a LAG-3 positive PD-L1 positive malignancy; and (b) instructing a healthcare provider to administer the immunotherapy described herein to the patient. The method may further comprise administering the immunotherapy described herein. In one embodiment, the immunotherapy comprises administering a therapeutically effective dose of a LAG-3 inhibitor, a PD-1 pathway inhibitor, and one or more chemotherapeutic agents. In one embodiment, the LAG-3 inhibitor is an anti-LAG-3 antibody, and the PD-1 pathway inhibitor is an anti-PD-1 antibody. In one embodiment, the administration treats the malignancy.

[0096] As a result of administering the immunotherapy described herein, the methods of the present invention may treat malignant tumors, reduce tumor size, suppress tumor growth, remove tumors from patients, prevent tumor recurrence, induce remission in patients, or result in any combination thereof. In certain embodiments, the administration of the immunotherapy described herein induces a complete response. In other embodiments, the administration of the immunotherapy described herein induces a partial response. In some embodiments, the immunotherapy comprises administering a therapeutically effective dose of a LAG-3 inhibitor, a PD-1 pathway inhibitor, and one or more chemotherapeutic agents. In some embodiments, the LAG-3 inhibitor is an anti-LAG-3 antibody, and the PD-1 pathway inhibitor is an anti-PD-1 antibody.

[0097] In one embodiment, a LAG-3-positive tumor contains at least about 1%, at least about 2%, at least about 3%, at least about 4%, at least about 5%, at least about 7%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or 100% LAG-3-expressing cells. In one embodiment, the LAG-3-expressing cells include tumor-infiltrating lymphocytes.

[0098] In one embodiment, identification includes determining LAG-3 expression in a malignant tumor.

[0099] In one embodiment, LAG-3 expression is determined by receiving the results of an assay capable of determining LAG-3 expression.

[0100] In certain embodiments, any of the methods of the present invention further include determining PD-L1 expression in a tumor sample.

[0101] In certain embodiments, any of the methods of the present invention further includes identifying a PD-L1-positive malignant tumor before administration. In certain embodiments, any of the methods of the present invention further includes determining PD-L1 expression in the malignant tumor.

[0102] In any particular embodiment of the method of the present invention, the patient is identified as having a PD-L1-positive malignant tumor prior to administration. In any particular embodiment of the method of the present invention, the patient is identified as having a PD-L1-negative malignant tumor prior to administration.

[0103] Methods for determining PD-L1 expression in tumor samples, methods for identifying patients with PD-L1-positive malignancies, and methods for determining PD-L1 expression in malignancies are described in PCT / US2016 / 029878, which is incorporated herein by reference in its entirety.

[0104] Measurement of LAG-3 expression In certain embodiments, identifying patients suitable for LAG-3 inhibitor / PD-1 pathway inhibitor / chemotherapy combination therapy for the present invention involves measuring or evaluating LAG-3 expression in malignant tumor biopsy tissue samples, including tumor cells and tumor-infiltrating inflammatory cells. The terms “LAG-3 expressing tumor,” “LAG-3 expressing tumor,” “LAG-3 positive tumor,” and “LAG-3 expression positive tumor” are used interchangeably herein and encompass tumors containing LAG-3 expressing tumor-infiltrating lymphocytes. Methods for measuring or evaluating LAG-3 expression can be achieved by any applicable method.

[0105] In one embodiment, to evaluate LAG-3 expression, biopsy tissue samples are obtained from patients in need of treatment. In one embodiment, biopsy tissue samples include, but are not limited to, clinically relevant tissue samples such as tumor biopsies, core biopsy tissue samples, fine needle aspirates, or samples of bodily fluids such as blood, plasma, serum, lymph, ascites, cystic fluid, or urine. In one embodiment, the biopsy tissue sample is from a primary tumor. In one embodiment, the biopsy tissue sample is from metastases. In one embodiment, biopsy tissue samples are taken from the subject at multiple time points, for example, before, during, and / or after the procedure. In one embodiment, biopsy tissue samples are taken from different sites of the subject, for example, a sample from a primary tumor and a sample from metastases at distant sites.

[0106] In one embodiment, the biopsy tissue sample is a paraffin-embedded tissue sample. In one embodiment, the biopsy tissue sample is a formalin-fixed paraffin-embedded (FFPE) tissue sample. In one embodiment, the biopsy tissue sample is a fresh tissue (e.g., tumor) sample. In one embodiment, the biopsy tissue sample is a frozen tissue sample. In one embodiment, the biopsy tissue sample is a fresh frozen (FF) tissue (e.g., tumor) sample. In one embodiment, the biopsy tissue sample is cells isolated from body fluid (fluid). In one embodiment, the biopsy tissue sample contains circulating tumor cells (CTCs). In one embodiment, the biopsy tissue sample contains tumor-infiltrating lymphocytes (TILs). In one embodiment, the biopsy tissue sample contains tumor cells and tumor-infiltrating lymphocytes (TILs). In one embodiment, the biopsy tissue sample contains circulating lymphocytes. In one embodiment, the biopsy tissue sample is an archival tissue sample. In one embodiment, the biopsy tissue sample is an archival tissue sample with a known history of diagnosis, treatment, and / or outcome. In one embodiment, the sample is a tissue mass. In another embodiment, the biopsy tissue sample is dispersed cells. In another embodiment, the sample size is approximately 1 x 10⁶ cells. 6 It consists of more than one cell. In one embodiment, the sample size is approximately 1 x 10⁶ cells from about one cell. 5 The sample size is a single cell. In one embodiment, the sample size is approximately 1 to 10,000 cells. In one embodiment, the sample size is approximately 1 to 1,000 cells. In one embodiment, the sample size is approximately 1 to 100 cells. In one embodiment, the sample size is approximately 1 to 10 cells. In one embodiment, the sample size is a single cell.

[0107] In another embodiment, the evaluation of LAG-3 expression can be achieved without obtaining a biopsy tissue sample. In one embodiment, selecting a suitable patient includes (i) optionally providing a biopsy tissue sample obtained from a patient with cancerous tissue, the biopsy tissue sample containing tumor cells and / or tumor-infiltrating inflammatory cells; and (ii) evaluating the percentage of cells in the biopsy tissue sample that express LAG-3 on the cell surface, based on the evaluation that the percentage of cells in the biopsy tissue sample that express LAG-3 on the cell surface is higher than a predetermined threshold level.

[0108] However, in a method that includes measuring LAG-3 expression in a biopsy tissue sample, the step of providing a biopsy tissue sample obtained from a patient should be understood as an optional step. That is, in certain embodiments, the method includes this step, and in other embodiments, this step is not included in the method. Also, in certain embodiments, the “measurement” or “evaluation” step for identifying or determining the number or percentage of cells expressing LAG-3 in a biopsy tissue sample should be understood as being performed by a transformation method that assays LAG-3 expression, for example, by performing a reverse transcriptase polymerase chain reaction (RT-PCR) assay or an IHC assay. In some other embodiments, the transformation step is not included, and LAG-3 expression is evaluated, for example, by reviewing a report of test results from a laboratory. In some embodiments, LAG-3 expression is evaluated by reviewing the results of an immunohistochemical assay from a laboratory. In certain embodiments, the steps of the method, including those up to the evaluation of LAG-3 expression, provide interim results that can be provided to a physician or healthcare provider for use in selecting a suitable candidate combination therapy of a LAG-3 inhibitor, a PD-1 pathway inhibitor, and one or more chemotherapeutic agents. In certain embodiments, the step of providing interim results is performed by a physician or another person performing the procedure under the guidance of a physician. In other embodiments, these steps are performed by an independent laboratory or an independent person, for example, a laboratory technician.

[0109] In certain embodiments of the method of the present invention, the percentage of cells expressing LAG-3 is assessed by performing an assay to detect the presence of LAG-3 RNA. In further embodiments, the presence of LAG-3 RNA is determined by RT-PCR, in situ hybridization, or RNase protection. In one embodiment, the presence of LAG-3 RNA is detected by an RT-PCR-based assay. In one embodiment, scoring the RT-PCR-based assay involves evaluating the level of LAG-3 RNA expression in a biopsy tissue sample by comparing it to a predetermined level.

[0110] In other embodiments, the percentage of cells expressing LAG-3 is assessed by performing an assay to detect the presence of the LAG-3 polypeptide. In further embodiments, the presence of the LAG-3 polypeptide is determined by immunohistochemistry (IHC), enzyme immunosorbent assay (ELISA), in vivo imaging, or flow cytometry. In some embodiments, LAG-3 expression is assayed by IHC. In all other embodiments of these methods, cell surface expression of LAG-3 is assayed, for example, using IHC or in vivo imaging.

[0111] In another embodiment, the percentage of cells expressing LAG-3 in a biopsy tissue sample is assessed by flow cytometry. In one embodiment, the biopsy tissue sample assayed by flow cytometry contains tumor-infiltrating immune cells. In one embodiment, the malignant tumor is a hematological malignant, and the tissue sample assayed by flow cytometry contains peripheral blood cells. In one embodiment, flow cytometry is a multiplex assay. In one embodiment, flow cytometry scoring involves detecting the expression of markers including LAG-3, CD4, CD8, FOXP3, and any combination thereof. In one embodiment, flow cytometry scoring involves assessing the percentage of T cells in a biopsy tissue sample expressing LAG-3. In one embodiment, flow cytometry scoring involves CD8 in a biopsy tissue sample expressing LAG-3. + This includes evaluating the proportion of T cells. In one embodiment, flow cytometry scoring involves CD4 in a biopsy tissue sample expressing LAG-3. + This includes evaluating the proportion of T cells. In one embodiment, flow cytometry scoring involves FOXP3 in biopsy tissue samples expressing LAG-3. + This includes evaluating the proportion of T cells.

[0112] In any particular embodiment of the present invention, the percentage of cells expressing LAG-3 in a biopsy tissue sample is assessed by performing an assay to detect the presence of LAG-3 polypeptides. In some embodiments, the presence of LAG-3 polypeptides is detected by an immunohistochemical assay. In some embodiments, the biopsy tissue sample is a tumor biopsy. In some embodiments, the biopsy tissue sample is a formalin-fixed paraffin-embedded (FFPE) sample.

[0113] In one embodiment, the immunohistochemical assay is a monoplex assay. In another embodiment, the immunohistochemical assay is a multiplex assay. In another embodiment, the multiplex immunohistochemical assay can detect the presence of CD4, CD8, FOXP3, or any combination thereof.

[0114] In one embodiment, the immunohistochemical assay comprises contacting a tumor sample with a 17B4 mouse anti-human LAG-3IgG1 monoclonal antibody. In another embodiment, the immunohistochemical assay comprises contacting a tumor sample with an anti-LAG-3 antibody containing heavy chain and light chain variable regions containing the sequences described in SEQ ID NOs: 3 and 5, respectively. In another embodiment, the immunohistochemical assay comprises contacting a tumor sample with an SP346 rabbit anti-human LAG-3IgG monoclonal antibody. In another embodiment, the immunohistochemical assay comprises contacting a tumor sample with an 11E3 (Novusbio), 874501 (Novusbio), or EPR4392(2)(Abcam) anti-human LAG-3 monoclonal antibody.

[0115] In one embodiment, the immunohistochemical assay is scored at a low magnification. In one embodiment, the low magnification is approximately 20x. In one embodiment, the immunohistochemical assay is scored at a high magnification. In one embodiment, the high magnification is approximately 40x.

[0116] In one embodiment, the immunohistochemical assay is scored by image analysis software. In another embodiment, the immunohistochemical assay is scored by a visual immunoscore of a pathologist. In another embodiment, the immunohistochemical assay is scored manually.

[0117] In one embodiment, scoring of the immunohistochemical assay includes evaluating the percentage of cells in the biopsy tissue sample that express LAG-3. In one embodiment, scoring of the immunohistochemical assay includes evaluating the percentage of immune cells in the biopsy tissue sample that express LAG-3. In one embodiment, scoring of the immunohistochemical assay includes evaluating the percentage of T cells in the biopsy tissue sample that express LAG-3. In one embodiment, scoring of the immunohistochemical assay includes evaluating CD8 in the biopsy tissue sample that express LAG-3. + This includes evaluating the percentage of T cells. In one embodiment, scoring of the immunohistochemical assay involves CD4 in a biopsy tissue sample expressing LAG-3. + This includes evaluating the proportion of T cells. In one embodiment, scoring of the immunohistochemical assay involves FOXP3 in biopsy tissue samples expressing LAG-3. + This includes evaluating the proportion of T cells.

[0118] LAG-3 polypeptide localization includes partial membrane / cytoplasmic localization, dot-like localization, and complete membrane / cytoplasmic localization. In one embodiment, cells with partial membrane / cytoplasmic LAG-3 localization are scored. In one embodiment, cells with dot-like LAG-3 localization are scored. In one embodiment, cells with complete membrane / cytoplasmic LAG-3 localization are scored. In one embodiment, cells with any of the LAG-3 localization patterns are scored.

[0119] In one embodiment, the immunohistochemical assay is a multiplex assay further comprising detecting the expression of MHC class II by tumor cells. In one embodiment, scoring of the immunohistochemical assay includes assessing the percentage of cells in a biopsy tissue sample that express MHC class II. In one embodiment, scoring of the immunohistochemical assay includes assessing the percentage of non-immune cells in a biopsy tissue sample that express MHC class II.

[0120] Imaging technologies provide essential tools in cancer research and treatment. Recent developments in molecular imaging systems, including positron emission tomography (PET), single-photon emission computed tomography (SPECT), fluorescence reflectance imaging (FRI), fluorescence-mediated tomography (FMT), bioluminescence imaging (BLI), laser scanning confocal microscopy (LSCM), and multiphoton microscopy (MPM), foreshadow further applications of these technologies in cancer research. Some of these molecular imaging systems allow clinicians to visualize not only the location of tumors within the body, but also the expression and activity of specific molecules, cellular, and biological processes that influence tumor behavior and / or responsiveness to therapeutic agents (Condeelis and Weissleder, Cold Spring Harb. Perspect. Biol. 2(12):a003848 (2010)). Regarding antibody specificity, the combination of PET sensitivity and resolution allows immunoPET imaging to monitor and assay antigen expression, particularly in tissue samples (McCabe and Wu, Cancer Biother. Radiopharm. 25(3):253-61 (2010); Olafsen et al., Protein Eng. Des. Sel. 23(4):243-9 (2010)). In any particular embodiment of the method of the present invention, LAG-3 expression is assayed by immunoPET imaging. In any particular embodiment of the method of the present invention, the percentage of cells in a biopsy tissue sample expressing LAG-3 is assessed by performing an assay to determine the presence of LAG-3 polypeptide on the cell surface in the biopsy tissue sample. In a particular embodiment, the biopsy tissue sample is an FFPE tissue sample. In other embodiments, the presence of LAG-3 polypeptide is determined by an IHC assay. In further embodiments, the IHC assay is performed using an automated method. In some embodiments, the IHC assay is performed using an anti-LAG-3 mAb that binds to the LAG-3 polypeptide.

[0121] Automated IHC assay of LAG-3 expression In one embodiment of this method, an automated IHC method is used to assay LAG-3 expression in FFPE tissue samples. The present invention provides a method for detecting the presence of human LAG-3 antigen in a biopsy tissue sample or for quantifying the level or percentage of human LAG-3 antigen in a sample expressing human LAG-3 antigen, comprising contacting a test sample and a negative control sample with an antibody that specifically binds to human LAG-3, or a portion thereof, under conditions that allow for the formation of a complex between the antibody and human LAG-3. In a particular embodiment, the test and control tissue samples are FFPE samples. Next, the formation of a complex is detected, where the difference in the formation of the complex between the test sample and the negative control sample indicates the presence of human LAG-3 antigen in the sample. Various methods are used to quantify LAG-3 expression.

[0122] In certain embodiments, an automated IHC method includes operating an automated staining apparatus, which comprises the steps of (a) deparaffinizing and rehydrating embedded tissue sections in an automated staining apparatus; (b) recovering antigens in an automated staining apparatus; (c) setting reagents in the automated staining apparatus; (d) neutralizing endogenous peroxidase in the tissue sample; blocking nonspecific protein binding sites on the slide; incubating the slide with a primary antibody; incubating with a post-primary blocking agent; incubating with a post-primary antibody detection reagent, such as another antibody that may or may not be bound to the detection enzyme; adding a chromogen substrate and allowing it to develop color; and then counterstaining with hematoxylin. In certain embodiments, antigen recovery includes the use of any thermal antigen recovery apparatus.

[0123] In one aspect, to evaluate LAG-3 expression in tumor tissue samples, pathologists examine LAG-3 in each field of view under a microscope. +The number of tumor cells is examined, the percentage of positive cells is estimated mentally, and then the estimated values ​​for each field are averaged to obtain the final percentage value. Different staining intensities are defined as 0 / negative, 1+ / weak, 2+ / intermediate, and 3+ / strong. Generally, percentage values ​​are assigned first to the 0 and 3+ buckets, then to the intermediate 1+ and 2+ intensities. For highly heterogeneous tissues, the sample is divided into sections, each section is scored separately, and then combined into a single set of percentage values. The percentages of negative and positive cells for each staining intensity are determined from each section, and the median is assigned to each section. Final percentage values ​​are assigned to the tissue for each staining intensity category: negative, 1+, 2+, and 3+. The sum of all staining intensities must be 100%.

[0124] In one aspect, staining is also evaluated for tumor-infiltrating inflammatory cells such as macrophages and lymphocytes. Macrophages and lymphocytes are evaluated for LAG-3 staining and recorded only as positive or negative for each cell category for all samples. Staining is also characterized according to the naming of intratumoral / extratumoral immune cells. “Intratumoral” means that immune cells are located on the boundary of the tumor area without being physically inserted within the tumor tissue and / or between tumor cells. “Extratumoral” means that there is no physical association with the tumor, and immune cells are found in peripheral or adjacent tissues associated with connective tissue.

[0125] In certain embodiments of these scoring methods, samples are scored by two or more independent pathologists, and these scores are later combined. In other embodiments, the identification of positive and negative cells is scored using appropriate software.

[0126] The Organization Score (H-score) is used as a more quantitative measure of IHC data. The Organization Score is calculated as follows: Tissue score = [(% tumor x 1 (low intensity)) + (% tumor x 2 (intermediate intensity)) + (% tumor x 3 (high intensity)].

[0127] To determine the tissue score, the pathologist estimates the percentage of stained cells in each intensity category within the sample. Because the expression of many biomarkers is heterogeneous, the tissue score is a more accurate representation of overall expression. The final tissue score ranges from 0 (minimum score, no expression) to 300 (maximum score, maximum expression and inclusive).

[0128] 3. LAG-3 inhibitors In one respect, the present invention is characterized by a method of using LAG-3 inhibitors in the treatment of malignant tumors. LAG-3 inhibitors used herein include, but are not limited to, LAG-3 conjugates and soluble LAG-3 polypeptides. LAG-3 conjugates include antibodies that specifically bind to LAG-3.

[0129] In one embodiment, the LAG-3 inhibitor is a LAG-3 conjugate, such as an anti-LAG-3 antibody. In another embodiment, the LAG-3 inhibitor is a soluble LAG-3 polypeptide, such as a LAG-3-Fc fusion polypeptide that can bind to MHC class II.

[0130] Anti-human LAG-3 antibodies (or VH / VL domains derived therefrom) can be prepared using methods well known in the art. Alternatively, anti-LAG-3 antibodies approved in the art can be used.

[0131] In one embodiment, the anti-LAG-3 antibody is BMS-986016, comprising a heavy chain and a light chain containing the sequences shown in SEQ ID NOs. 1 and 2, respectively, as described in PCT / US13 / 48999 (the contents of which are incorporated herein by reference), or their antigen-binding fragments and variants. In one embodiment, the BMS-986016 antibody does not contain the heavy chain terminal lysine amino acid of SEQ ID NO. 1.

[0132] In other embodiments, the antibody has heavy and light chain CDRs or variable regions of BMS-986016. Thus, in one embodiment, the antibody comprises CDR1, CDR2, and CDR3 domains of the VH region of BMS-986016 having the sequence described in SEQ ID NO: 3, and CDR1, CDR2, and CDR3 domains of the VL region of BMS-986016 having the sequence described in SEQ ID NO: 5. In another embodiment, the antibody comprises CDR1, CDR2, and CDR3 domains containing the sequences described in SEQ ID NOs: 7, 8, and 9, respectively, and CDR1, CDR2, and CDR3 domains containing the sequences described in SEQ ID NOs: 10, 11, and 12, respectively. In another embodiment, the antibody comprises a VH region and / or a VL region containing the amino acid sequences described in SEQ ID NO: 3 and / or SEQ ID NO: 5, respectively. In another embodiment, the antibody comprises a heavy chain variable (VH) region and / or a light chain variable (VL) region encoded by the nucleic acid sequences described in SEQ ID NO: 4 and / or SEQ ID NO: 6, respectively. In another embodiment, the antibody competes for binding to the same LAG-3 epitope as the antibody described above, and / or binds to the same LAG-3 epitope as the antibody described above. In another embodiment, the antibody binds to a human LAG-3 epitope containing the amino acid sequence PGHPLAPG (SEQ ID NO: 14). In another embodiment, the antibody binds to a human LAG-3 epitope containing the amino acid sequence HPAAPSSW (SEQ ID NO: 15) or PAAPSSWG (SEQ ID NO: 16).

[0133] In another embodiment, the antibody has at least about 90% variable region amino acid sequence identity with the antibody described above (for example, at least about 90%, 95%, or 99% variable region identity with SEQ ID NO: 3 or SEQ ID NO: 5).

[0134] In one embodiment, the anti-LAG-3 antibody or its antigen-binding moiety cross-compete with BMS-986016 (lilatrimab) for binding to human LAG-3. In another embodiment, the anti-LAG-3 antibody or its antigen-binding moiety binds to the same epitope as BMS-986016 (lilatrimab). In one embodiment, the anti-LAG-3 antibody or its antigen-binding moiety cross-compete with TSR-033 for binding to human LAG-3. In another embodiment, the anti-LAG-3 antibody or its antigen-binding moiety binds to the same epitope as TSR-033. In one embodiment, the anti-LAG-3 antibody or its antigen-binding moiety cross-compete with TSR-075 for binding to human LAG-3. In another embodiment, the anti-LAG-3 antibody or its antigen-binding moiety binds to the same epitope as TSR-075. In one embodiment, the anti-LAG-3 antibody is a chimeric antibody, a humanized antibody, a human monoclonal antibody, or their antigen-binding moiety. In other embodiments, the anti-LAG-3 antibody or its antigen-binding moiety comprises a heavy chain constant region of a human IgG1 isotype or a human IgG4 isotype. In certain embodiments, the anti-PD-1 antibody or its antigen-binding moiety is BMS-986016 (liratrimab). In some embodiments, the anti-LAG-3 antibody or its antigen-binding moiety is a biosimilar of BMS-986016 (liratrimab). In certain embodiments, the anti-PD-1 antibody or its antigen-binding moiety is TSR-033. In some embodiments, the anti-LAG-3 antibody or its antigen-binding moiety is a biosimilar of TSR-033. In certain embodiments, the anti-PD-1 antibody or its antigen-binding moiety is TSR-075. In some embodiments, the anti-LAG-3 antibody or its antigen-binding moiety is a biosimilar of TSR-075.

[0135] In one embodiment, anti-LAG-3 antibodies known in the art can be used in the therapeutic methods of the present invention. For example, the anti-human LAG-3 antibody described in US2011 / 0150892 A1 (as incorporated herein by reference) and referred to as monoclonal antibody 25F7 (also known as “25F7” and “LAG-3.1”) can be used. Other anti-LAG-3 antibodies known in the art that can be used include IMP731(H5L7BW) described in US2011 / 007023, MK-4280(28G-10) described in WO2016028672, and Journal for ImmunoTherapy of Cancer, (2016) Vol. 4, Supp. Supplement 1 Abstract Number: Examples include REGN3767 listed on P195, BAP050, IMP-701(LAG-525), aLAG3(0414), aLAG3(0416), Sym022, TSR-033, TSR-075, XmAb22841, MGD013, BI754111, FS118, P13B02-30, AVA-017, and GSK2831781, as listed in WO2017 / 019894. These and other anti-LAG-3 antibodies useful in the present invention include, for example, US10,188,730, WO2016 / 028672, WO2017 / 106129, WO2017 / 062888, WO2009 / 044273, WO2018 / 069500, WO2016 / 126858, WO2014 / 179664, WO2016 / 200782, WO2015 / 200119, WO2017 / 019846, WO2017 / 198741, WO2017 / 220555, WO2017 / 220569, WO2018 / 071500, WO It can be found in 2017 / 015560, WO2017 / 025498, WO2017 / 087589, WO2017 / 087901, WO2018 / 083087, WO2017 / 149143, WO2017 / 219995, US2017 / 0260271, WO2017 / 086367, WO2017 / 086419, WO2018 / 034227, WO18 / 185046, WO18 / 185043, WO2018 / 217940, WO19 / 011306, WO2018 / 208868 and WO2014 / 140180.In one embodiment, the LAG-3 inhibitor is IMP321 (eftyragimod alfa). The entire contents of each of these references are incorporated herein by reference.

[0136] Regarding binding to LAG-3, antibodies that compete with any of the antibodies recognized in the art described above can also be used.

[0137] In certain embodiments, anti-LAG-3 antibodies are used to determine LAG-3 expression. In one embodiment, anti-LAG-3 antibodies are selected for their ability to bind to LAG-3 in formalin-fixed paraffin-embedded (FFPE) tissue samples. In other embodiments, anti-LAG-3 antibodies can bind to LAG-3 in frozen tissue. In further embodiments, anti-LAG-3 antibodies can distinguish between membrane-bound, cytoplasmic, and / or soluble forms of LAG-3.

[0138] In one embodiment, an anti-LAG-3 antibody useful for assaying, detecting, and / or quantifying LAG-3 expression according to the method described herein is a 17B4 mouse IgG1 anti-human LAG-3 monoclonal antibody or its antigen-binding fragment. See, for example, J. Matsuzaki, et al.; PNAS 107, 7875 (2010).

[0139] 4. PD-1 pathway inhibitors In one respect, the present invention is characterized by a method of using PD-1 inhibitors in the treatment of malignant tumors. The “PD-1 pathway inhibitors” as used herein include, but are not limited to, PD-1 conjugates, PD-L1 conjugates, and PD-L2 conjugates. PD-1 conjugates include antibodies that specifically bind to PD-1. PD-L1 conjugates and PD-L2 conjugates include antibodies that specifically bind to PD-L1 and / or PD-L2, as well as soluble PD-1 polypeptides that bind to PD-L1 and / or PD-L2.

[0140] In one embodiment, the PD-1 pathway inhibitor is a PD-1 conjugate, such as an anti-PD-1 antibody. In another embodiment, the PD-1 pathway inhibitor is a PD-L1 conjugate, such as an anti-PD-L1 antibody. In another embodiment, the PD-1 pathway inhibitor is a PD-L2 conjugate, such as an anti-PD-L2 antibody. In a further embodiment, the PD-L1 conjugate is a soluble PD-1 polypeptide, such as a PD-1-Fc fusion polypeptide that can bind to PD-L1. In a further embodiment, the PD-L2 conjugate is a soluble PD-1 polypeptide, such as a PD-1-Fc fusion polypeptide that can bind to PD-L2.

[0141] Anti-human PD-1 antibodies (or VH and / or VL domains derived therefrom) suitable for use in the present invention can be prepared 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 entire contents 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, which are incorporated herein by reference. Further known PD-1 antibodies and other PD-1 inhibitors include, for example, those described in WO2009 / 014708, WO03 / 099196, WO2009 / 114335 and WO2011 / 161699 (which are incorporated herein by reference). In one embodiment, the anti-PD-1 antibody is REGN2810. In one embodiment, the anti-PD-1 antibody is PDR001. Another known anti-PD-1 antibody is pizilizumab (CT-011).

[0142] In one embodiment, the anti-PD-1 antibody is nivolumab. Nivolumab (also known as "OPDIVO®"; formerly referred to as 5C4, BMS-936558, MDX-1106, or ONO-4538) is a fully human IgG4(S228P) PD-1 immune checkpoint inhibitor antibody that selectively blocks interaction with PD-1 ligands (PD-L1 and PD-L2), thereby preventing downregulation of antitumor T cell function (U.S. Patent No. 8,008,449; Wang et al., Cancer Immunol Res. 2(9):846-56 (2014)). In another embodiment, the anti-PD-1 antibody or its fragments cross-compete with nivolumab. In yet another embodiment, the anti-PD-1 antibody or its fragments bind to the same epitope as nivolumab. In a particular embodiment, the anti-PD-1 antibody has the same CDR as nivolumab.

[0143] 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 binds to the same epitope as nivolumab. In one embodiment, the anti-PD-1 antibody is a chimeric antibody, a humanized antibody, a human monoclonal antibody, or their antigen-binding moiety. In another embodiment, the anti-PD-1 antibody or its antigen-binding moiety contains a heavy chain constant region of a human IgG1 isotype or a human IgG4 isotype. In a specific embodiment, the anti-PD-1 antibody or its antigen-binding moiety is nivolumab or pembrolizumab. In one embodiment, the anti-PD-1 antibody or its antigen-binding moiety is a biosimilar of nivolumab. In another embodiment, the anti-PD-1 antibody or its antigen-binding moiety is a biosimilar of pembrolizumab.

[0144] In one embodiment, the anti-PD-1 antibody comprises a heavy chain and a light chain containing the sequences shown in SEQ ID NOs. 17 and 18, respectively, or antigen-binding fragments and variants thereof.

[0145] In other embodiments, the antibody comprises the heavy and light chain CDRs or variable regions of nivolumab. Thus, in one embodiment, the antibody comprises the CDR1, CDR2, and CDR3 domains of the VH of nivolumab having the sequence described in SEQ ID NO: 19, and the CDR1, CDR2, and CDR3 domains of the VL of nivolumab having the sequence described in SEQ ID NO: 21. In another embodiment, the antibody comprises the CDR1, CDR2, and CDR3 domains having the sequences described in SEQ ID NOs: 23, 24, and 25, respectively, and the CDR1, CDR2, and CDR3 domains having the sequences described in SEQ ID NOs: 26, 27, and 28, respectively. In yet another embodiment, the antibody comprises the VH region and / or VL region having the amino acid sequence described in SEQ ID NO: 19 and / or SEQ ID NO: 21, respectively. In yet another embodiment, the antibody comprises the heavy chain variable (VH) region and / or light chain variable (VL) region encoded by the nucleic acid sequence described in SEQ ID NO: 20 and / or SEQ ID NO: 22, respectively. In another embodiment, the antibody competes for binding to the same PD-1 epitope as the antibody described above, and / or binds to the same PD-1 epitope as the antibody described above. In another embodiment, the antibody has at least about 90% variable region amino acid sequence identity with the antibody described above (e.g., at least about 90%, 95%, or 99% variable region identity with SEQ ID NO: 19 or SEQ ID NO: 21).

[0146] Human monoclonal antibodies that specifically bind to PD-1 with high affinity are described in U.S. Patents 8,008,449 and 8,779,105 (which are incorporated herein by reference). 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 (which are incorporated herein by reference). In some embodiments, anti-PD-1 antibodies have been demonstrated to exhibit one or more of the following characteristics: (a) 1 × 10⁻¹⁶ ions, as determined by surface plasmon resonance using a Biacore biosensor system. -7 M or less K D(b) binds to human PD-1; (c) substantially does not bind to human CD28, CTLA-4, or ICOS; (d) increases T cell proliferation in mixed lymphocyte reaction (MLR) assays; (e) increases interferon-γ production in MLR assays; (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 antigen-specific memory responses; (i) stimulates antibody responses; and (j) inhibits tumor cell proliferation in vivo. Anti-PD-1 antibodies useful for the present invention include mAbs that specifically bind to human PD-1 and exhibit at least one, at least two, at least three, at least four, or at least five of the above features. Anti-PD-1 antibodies exhibiting one or more of these characteristics are described in U.S. Patents No. 8,008,449, 8,779,105, 6,808,710, 7,488,802, 8,168,757, and 8,354,509, as well as PCT Publication WO2012 / 145493 (these are incorporated herein by reference). In another embodiment, 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 No. 8,354,509 and 8,900,587 (these are incorporated herein by reference).

[0147] In one embodiment, the anti-PD-1 antibody or its fragment cross-competes with pembrolizumab. In one embodiment, the anti-PD-1 antibody or its fragment binds to the same epitope as pembrolizumab. In a particular embodiment, the anti-PD-1 antibody has the same CDR as pembrolizumab. In another embodiment, the anti-PD-1 antibody is pembrolizumab. Pembrolizumab ("KEYTRUDA®," 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. Patent No. 8,354,509 and No. 8,900,587 (as incorporated herein by reference; see also http: / / www.cancer.gov / drugdictionary-cdrid=695789 (last accessed December 14, 2014)).

[0148] In other embodiments, the anti-PD-1 antibody or its fragment cross-compete with MEDI0608. In yet another embodiment, the anti-PD-1 antibody or its fragment binds to the same epitope as MEDI0608. In certain embodiments, the anti-PD-1 antibody has the same CDR as MEDI0608. In other embodiments, the anti-PD-1 antibody is the monoclonal antibody MEDI0608 (formerly AMP-514). MEDI0608 is described, for example, in U.S. Patent No. 8,609,089B2 (as incorporated herein by reference or at http: / / www.cancer.gov / drugdictionary-cdrid=756047 (last accessed December 14, 2014)).

[0149] In certain embodiments, the first antibody is an anti-PD-1 antagonist. An example of an anti-PD-1 antagonist is AMP-224, a B7-DC Fc fusion protein. AMP-224 is described in U.S. Patent Publication 2013 / 0017199 (embodied herein by reference) or in http: / / www.cancer.gov / publications / dictionaries / cancer-drug-cdrid=700595 (last accessed July 8, 2015).

[0150] In other embodiments, the anti-PD-1 antibody or a fragment thereof cross-competes with BGB-A317. In some embodiments, the anti-PD-1 antibody or a fragment thereof binds to the same epitope as BGB-A317. In certain embodiments, the anti-PD-1 antibody has the same CDR as BGB-A317. In certain embodiments, the anti-PD-1 antibody is BGB-A317, which is a humanized monoclonal antibody. BGB-A317 is described in U.S. Patent Publication 2015 / 0079109 (which is incorporated herein by reference).

[0151] In one embodiment, the antibody is pizilizumab (CT-011), which is an antibody previously reported to bind to PD-1 and is thought to bind to a different target. Pizilizumab is described in U.S. Patent No. 8,686,119 B2 or WO 2013 / 014668 A1 (as incorporated herein by reference).

[0152] In certain embodiments, antibodies that compete with nivolumab for binding to human PD-1, or that bind to the same epitope region as human PD-1, are mAbs. For administration to human subjects, these cross-competitive antibodies may be chimeric antibodies, humanized antibodies, or human antibodies. Such chimeric antibodies, humanized antibodies, or human mAbs can be prepared and isolated by methods well known in the art.

[0153] Other anti-PD-1 monoclonal antibodies are, for example, U.S. Patent Nos. 6,808,710, 7,488,802, 8,168,757, and 8,354,509, U.S. Patent Publication 2016 / 0272708, and PCT Publications WO2012 / 145493 and WO2008 / 15671. 2, WO2015 / 112900, WO2012 / 145493, WO2015 / 112800, WO2014 / 206107, WO2015 / 35 606, WO2015 / 085847, WO2014 / 179664, WO2017 / 020291, WO2017 / 020858, WO2016 / These are described in 197367, 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 in its entirety).

[0154] 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; also known as toripalimab; Si-Yang See Liu et al., J. Hematol. Oncol. 10:136 (2017), BGB-A317 (Beigene; see WO2015 / 35606 and US2015 / 0079109), INCSHR1210 (Jiangsu Hengrui Medicine; also known as SHR-1210; WO2015 / 085847; see Si-Yang Liu et al., J. Hematol. 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. The group is selected from the following: 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).The contents of each of these documents are incorporated herein by reference in their entirety.

[0155] Other anti-PD-1 antibodies useful in the compositions described herein include the antigen-binding sites of the above antibodies. It has been demonstrated that the antigen-binding function of an antibody can be performed by fragments of a full-length antibody. Examples of binding fragments encompassed by the term “antigen-binding site” of an antibody include (i)V L , V H , C L and C H1 (ii) A monovalent fragment consisting of domains, the Fab fragment; (ii) A bivalent fragment, the F(ab')2 fragment, containing two Fab fragments linked by disulfide bridges in a hinge region; (iii) V H and C H1 Fd fragment consisting of domains; (iv) V of a single arm of the antibody L and V H An example is an Fv fragment consisting of a domain.

[0156] Other anti-PD-1 antibodies that can be used in the methods of the present invention include isolated antibodies that specifically bind to human PD-1 and cross-compete for binding to any of the anti-PD-1 antibodies described herein, such as nivolumab (e.g., U.S. Patent Nos. 8,008,449 and 8,779,105; see WO2013 / 173223, which are incorporated herein by reference). In some embodiments, an anti-PD-1 antibody binds to the same epitope as any of the anti-PD-1 antibodies described herein (e.g., 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 and sterically interfere with the binding of other cross-competing antibodies to a particular epitope region. These cross-competing antibodies are expected to have functional properties remarkably similar to a control antibody (e.g., 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 assays, or flow cytometry (see, for example, WO2013 / 173223, which are incorporated herein by reference).

[0157] An anti-PD-1 antibody suitable for use in the method of the present invention is an antibody that binds to PD-1 with high specificity and affinity, inhibits the binding of PD-L1 and / or PD-L2, and inhibits the immunosuppressive effect of the PD-1 signaling pathway. In any of the compositions or methods described herein, the anti-PD-1 "antibody" includes an antigen-binding moiety or fragment that binds to the PD-1 receptor, inhibits ligand binding, and exhibits functional properties similar to a complete antibody in upregulating the immune system. In certain embodiments, the anti-PD-1 antibody or its antigen-binding moiety cross-competes with nivolumab for binding to human PD-1. In other embodiments, the anti-PD-1 antibody or its antigen-binding moiety is a chimeric antibody, a humanized antibody, or a human monoclonal antibody or a portion thereof. In certain embodiments, the antibody is a humanized antibody. In other embodiments, the antibody is a human antibody. Antibodies of the IgG1, IgG2, IgG3, or IgG4 isotype can be used.

[0158] In certain embodiments, the anti-PD-1 antibody or its antigen-binding moiety comprises a heavy chain constant region of a human IgG1 or IgG4 isotype. In certain other embodiments, the sequence of the IgG4 heavy chain constant region or its antigen-binding moiety of the anti-PD-1 antibody 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 inhibits Fab arm exchange with endogenous IgG4 antibody while maintaining low affinity for activating the Fc receptor associated with wild-type IgG4 antibody (Wang et al., 2014 Cancer Immunol Res. 2(9):846-56). In yet another embodiment, the antibody comprises a light chain constant region which is a human κ or λ constant region. In other embodiments, the anti-PD-1 antibody or its antigen-binding moiety is an mAb or its antigen-binding moiety. In certain preferred embodiments of any of the therapeutic methods described herein, which involve administration of an anti-PD-1 antibody, the anti-PD-1 antibody is nivolumab. In another preferred embodiment, the anti-PD-1 antibody is pembrolizumab. In another embodiment, the anti-PD-1 antibody is selected from the human antibodies 17D8, 2D3, 4H1, 4A11, 7D3, and 5F4 described in U.S. Patent No. 8,008,449. In yet another embodiment, the anti-PD-1 antibody is MEDI0608 (formerly AMP-514), AMP-224, or BGB-A317.

[0159] 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.

[0160] 5. Anti-PD-L1 antibody In certain embodiments, the present invention encompasses the use of anti-PD-L1 antibodies as PD-1 pathway inhibitors. In one embodiment, anti-PD-L1 antibodies inhibit the binding of PD-L1 receptors, i.e., inhibit the binding of PD-1 to its ligand, PD-L1.

[0161] Anti-human PD-L1 antibodies (or VH and / or VL domains derived therefrom) suitable for use in the present invention can be prepared using methods well known to those skilled in the art. Alternatively, anti-PD-L1 antibodies recognized in the art can be used. For example, the human anti-PD-L1 antibody described in U.S. Patent No. 7,943,743 can be used. Such anti-PD-L1 antibodies include 3G10, 12A4 (also known as BMS-936559), 10A5, 5F8, 10H10, 1B12, 7H1, 11E6, 12B7, and 13G4. In one embodiment, the anti-PD-L1 antibody is atezolizumab (Tecentriq or RG7446) (e.g., Herbst et al. (2013) J Clin Oncol 31(suppl):3000. Abstract; see US Patent No. 8,217,149), durvalumab (Imfinzi or MEDI4736) (Khleif (2013) In: Proceedings from the European Cancer Congress 2013; September 27-October 1, 2013; Amsterdam, The Netherlands. Abstract 802), or avelumab (Bavencio). Other art-approved and usable anti-PD-L1 antibodies include, for example, those described in U.S. Patent Nos. 7,635,757 and 8,217,149, U.S. Patent Application Publication No. 2009 / 0317368, and PCT Publications WO2011 / 066389 and WO2012 / 145493 (which are incorporated herein by reference). Antibodies that compete with these art-approved antibodies or inhibitors for binding to PD-L1 can also be used. An example of an anti-PD-L1 antibody useful in the method of the present invention is the antibody described in U.S. Patent No. 9,580,507 (which is incorporated herein by reference). The anti-PD-L1 human monoclonal antibody described in U.S. Patent No. 9,580,507 has been shown to exhibit one or more of the following features: (a) 1 x 10⁻¹⁶ as measured by surface plasmon resonance using a Biacore biosensor system -7 M or less KD (b) binds to human PD-L1; (c) increases T cell proliferation in mixed lymphocyte reaction (MLR) assays; (d) increases interferon-γ production in MLR assays; (e) stimulates an antibody response; and (f) reverses the effect of T regulatory cells in T cell effector cells and / or dendritic cells. Anti-PD-L1 antibodies that can be used in the present invention include monoclonal antibodies that specifically bind to human PD-L1 and exhibit at least one, and in some embodiments, at least five of the above features.

[0162] In certain embodiments, the anti-PD-L1 antibody is BMS-936559 (formerly 12A4 or MDX-1105) (see, for example, U.S. Patent No. 7,943,743; WO2013 / 173223) (as incorporated herein by reference). In other embodiments, the anti-PD-L1 antibody is MPDL3280A (also known as RG7446 and atezolizumab) (e.g., Herbst et al. 2013 J Clin Oncol 31(suppl):3000; see U.S. Patent No. 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 referred to as Avelumab; see U.S. 2014 / 0341917) (as incorporated herein by reference). In certain embodiments, an antibody that cross-competes for binding to human PD-1 or binds to the same human PD-L1 epitope region as the above-mentioned reference PD-L1 antibody is an mAb. For administration to human subjects, these cross-competitive antibodies may be chimeric antibodies, humanized antibodies, or human antibodies. Such chimeric mAbs, humanized mAbs, or human mAbs can be prepared and isolated by methods well known in the art. In certain embodiments, the anti-PD-L1 antibody may be BMS-936559 (also known as 12A4, MDX-1105; see, e.g., U.S. Patent No. 7,943,743 and WO2013 / 173223), atezolizumab (Roche; TECENTRIQ®; also known as MPDL3280A, RG7446; see U.S. 8,217,149; also see Herbst et al. (2013) J Clin Oncol 31(suppl):3000), or durvalumab (AstraZeneca; IMFINZI (商標)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:Abstract 4606 (Apr Selected from the group consisting of (see 2016).

[0163] In certain embodiments, the PD-L1 antibody is atezolizumab (TECENTRIQ®). Atezolizumab is a fully humanized IgG1 monoclonal anti-PD-L1 antibody.

[0164] In certain embodiments, the PD-L1 antibody is durvalumab (IMFINZI (商標) Durvalumab is a human IgG1 kappa monoclonal anti-PD-L1 antibody.

[0165] In certain embodiments, the PD-L1 antibody is avelumab (BAVENCIO®). Avelumab is a human IgG1 lambda monoclonal anti-PD-L1 antibody.

[0166] 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.

[0167] Other anti-PD-L1 antibodies that can be used in the method of the present invention include isolated antibodies that specifically bind to human PD-L1 and cross-compete with the anti-PD-L1 antibodies described herein, e.g., atezolizumab, durvalumab, and / or avelumab, for binding to human PD-L1. In some embodiments, 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 the antigen indicates that these antibodies bind to the same epitope region of the antigen and sterically interfere with the binding of other cross-competing antibodies to a particular epitope region. These cross-competing antibodies are expected to have functional properties remarkably similar to control antibodies, e.g., atezolizumab and / or avelumab, due to their binding to the same epitope region of PD-L1. Cross-competing 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 assays, or flow cytometry (see, for example, WO2013 / 173223, which is incorporated herein by reference).

[0168] In certain embodiments, antibodies that cross-compete for binding to human PD-L1, or that bind to the same human PD-L1 epitope region as atezolizumab, durvalumab, and / or avelumab, are monoclonal antibodies. For administration to human subjects, these cross-competing antibodies are chimeric antibodies, modified antibodies, or humanized or human antibodies. Such chimeric antibodies, modified humanized antibodies, or human monoclonal antibodies can be prepared and isolated by methods well known in the art.

[0169] Examples of anti-PD-L1 antibodies that can be used in the method of the present invention include the antigen-binding portion of the above-mentioned antibody. It has been demonstrated that the antigen-binding function of an antibody can be performed by a fragment of a full-length antibody.

[0170] An anti-PD-L1 antibody suitable for use in the methods or compositions of the present invention is an antibody that binds to PD-L1 with high specificity and affinity, inhibits PD-1 binding, and inhibits the immunosuppressive effect of the PD-1 signaling pathway. In any of the compositions or methods described herein, the anti-PD-L1 "antibody" may be an antigen-binding moiety or fragment that binds to PD-L1 and exhibits functional properties similar to a full-length antibody in upregulating the immune system by inhibiting receptor binding. In certain 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.

[0171] Examples of anti-PD-L1 antibodies useful in the present invention include V as described herein. H and / or V L Examples include antibodies prepared starting from an antibody having one or more sequence elements, which may possess modified characteristics from the starting antibody. Anti-PD-L1 antibodies may be manipulated by various modifications as described above for the preparation of the modified anti-PD-1 antibody of the present invention.

[0172] 6. Chemotherapy agents The present invention is characterized by the use of one or more chemotherapeutic agents in combination with a LAG-3 inhibitor and a PD-1 pathway inhibitor for the treatment of malignant tumors. In one embodiment, the chemotherapeutic agents are considered standard therapy for the treatment of malignant tumors. The “chemotherapeutic agents” are compounds useful for the treatment of cancer. Examples of chemotherapeutic agents include alkylating agents such as thiotepa and cyclophosphamide (CYTOXAN®); alkyl sulfates such as busulfan, improsulfan, and piposulfan; aziridines such as benzodopa, carbocon, meturedopa, and uredopa; ethyleneimines and methylamelamines including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, and trimethylolmelamine; acetogenins (especially bratacin and bratacinone); delta-9-tetrahydrocannabinol (dronabinol, MARINOL®); beta-lapacon; lapachol; colchicine; betulinic acid; camptothecin (synthetic analogue topotecan (HYCAMTIN®)), CPT-11 (irinotecan, CAMPTOSA R(registered trademark)), containing acetylcamptothecin, scopoletin and 9-aminocamptothecin); bryostatin; calistatin; CC-1065 (including its adzeresin, karzeresin and bizeresin synthetic analogs); podophyllotoxin; podophyllic acid; teniposide; cryptophycin (especially cryptophycin 1 and cryptophycin 8); drastatin; duocalmycin (including synthetic analogs KW-2189 and CB1-TM1); e Leuterobin; Pancratistatin; Sarcodictyin; Spongistatin; Chlorambucil, Chlornaphazine, Chlorophosphamide, Estramustine, Ifosfamide, Mechloretamine, Mechloretamine Oxide Hydrochloride, Melphalan, Novoenvicin, Fenesterine, Prednimustine, Trophosphamide, Uracil Mustard and other nitrogenous mustards;Nitrosoureas such as carmustine, chlorozotosine, fotemustine, lomustine, nimustine, and ranimustine; antibiotics such as engine antibiotics (e.g., calicheamicin, especially calicheamicin γ1I and calicheamicin ωI1 (e.g., Nicolaou et al., Angew. Chem Intl. Ed. Engl., 33: 183-186) (See 1994); CDP323, oral alpha-4 integrin inhibitor; dynemycin including dynemycin A; esperamicin; and neocardinostatin chromophore and related chromoprotein enediin antibiotic chromophore); acrasinomycin, actinomycin, anthramycin, azaserin, bleomycin, kactinomycin, carabicin, carminomycin, cardinophilin, chromomycinis, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin (Adriamycin®, morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin, doxorubicin HCl liposome injection (DOXIL®), doxorubicin liposome TLC) D-99 (MYOCET®), pegylated liposomal doxorubicin (CAELYX®, and deoxidoxorubicin), epirubicin, esorubicin, idarubicin, marcelomycin, mitomycin such as mitomycin C, mycophenolic acid, nogaramycin, oligomycin, peplomycin, porphyromycin, puromycin, queramycin, rhodorubicin, streptonigrin, streptozocin, tubercidine, ubenimex, dinostatin, zorubicin; methotrexate, gemcitabine (GEMZAR®), tegafur (UFTORAL®), capecitabine Antimetabolites such as (XELODA®), epothilon, and 5-fluorouracil (5-FU); folic acid analogs such as denopterin, methotrexate, pteropterin, and trimethrexate;Purine analogs such as fludarabine, 6-mercaptopurine, thiamiprine, and thioguanine; pyrimidine analogs such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, and phloxuridine; androgens such as carsterone, dromostanolone propionate, epithiostanol, mepitiostane, and testactone; anti-adrenal agents such as aminoglutethimide, mitotane, and trilostane; folic acid replenisher such as floric acid; acegraton; aldofamide glycoside; aminolevulinic acid; enyluracil; amsacrine; bestrabucil; bisantren; edatraxate; defofamine (d efofamine; demecoltin; diaziquan; eflornithine; eriptinium acetate; epotilon; etogluside; gallium nitrate; hydroxyurea; lentinan; ronidynin; maytansinoids such as mytansin and anthamitosin; mitogwazone; mitoxantrone; mopidanmol; nitraerine; pentostatin; fenamet; pirarubicin; losoxantrone; 2-ethylhydrazide; procarbazine; PSK (registered trademark) complex polysaccharides (JHS Natural Products, Eugene, Oreg.); Lazoxane; Rhizoxin; Sizofiran; Spirogermanium; Tenuazonic acid; Triadiquan; 2,2',2'-Trichlorotriethylamine; Trichothecene (especially T-2 toxin, verracurin A, roridin A and anguidin); Urethane; Vindesine (ELDISINE®, FILDESIN®); Dacarbazine; Mannomustine; Mitobronitol; Mitractol; Pipobroman; Gacytosine; Arabinoside ("Ara-C"); Thiotepa;Taxoids, e.g., paclitaxel (Taxol®), albumin-modified nanoparticle formulations of paclitaxel (ABRAXANE®), and docetaxel (Taxotere®); chlorambucil; 6-thioguanine; mercaptopurine; methotrexate; platinum-based drugs such as cisplatin, oxaliplatin (e.g., ELOXATIN®), and carboplatin; vinblastine (VELBAN®), vincristine Retinoids such as retinoic acid, including (ONCOVIN®), vindesine (ELDISINE®, FILDESIN®), and vinorelbine (NAVELBINE®), which prevent tubulin polymerization by forming microtubules; vinca; etoposide (VP-16); ifosfamide; mitoxantrone; leucovorin; novantrone; edatrexate; daunomycin; aminopterin; ibandronate; topoisomerase inhibitor RFS2000; difluoromethylornithine (DMFO); bexarotene (TARGRETIN®), retinoic acid and other retinoids; bisphosphonates such as clodronate (e.g., BONEFOS® or OSTAC®), etidronate (DIDROCAL®), NE-58095, zoledronic acid / zoledronate (ZOMET A(registered trademark), alendronate (FOSAMAX(registered trademark)), pamidronic acid (AREDIA(registered trademark)), chydronic acid (SKELID(registered trademark)) or risedronic acid (ACTONEL(registered trademark)); troxacitabine (1,3-dioxolane nucleoside cytosine analog); antisense oligonucleotides, in particular those that inhibit gene expression in signaling pathways associated with abnormal cell proliferation, such as PKCα, Raf, H-Ras, and epidermal growth factor receptor (EGF-R); THERATOP(registered trademark) vaccine and gene therapy vaccines, such as ALLOVECTIN(registered trademark) vaccine, LEUVECTIN(registered trademark) vaccine, and VAXID(registered trademark) vaccine; topoisomerase 1 inhibitors (e.g., LURTOTECAN(registered trademark)); rmRH (e.g., Abarelix(registered trademark));BAY439006 (Sorafenib; Bayer); SU-11248 (Sunitinib, SUTENT®, Pfizer); Perifosine, COX-2 inhibitors (e.g., celecoxib or etoricoxib), proteosome inhibitors (e.g., PS341); Bortezomib (Velcade®); CCI-779; Tipifarnib (R11577); Olafenib, ABT510; Bcl-2 inhibitors such as Oblimersen sodium (GENASENSE®); Pixantrone; EGFR inhibitors (see definition below); Tyrosine kinase inhibitors (see definition below); Serine-threonine kinase inhibitors such as Rapamycin (Sirolimus, RAPAMUNE®); Ronafarnib (SCH Examples include farnesyltransferase inhibitors such as 6636 and SARASAR(trademark); and any pharmaceutically acceptable salt, acid, or derivative of any of the above; and two or more combinations of the above, such as CHOP, an abbreviation for combination therapy of cyclophosphamide, doxorubicin, vincristine, and prednisolone; IFL, an abbreviation for treatment regimens of irinotecan in combination with 5-fluorouracil and leucovorin; XELOX, an abbreviation for treatment regimens of oxaliplatin (ELOXATIN(trademark)) in combination with capecitabine; SOX, an abbreviation for treatment regimens of oxaliplatin (ELOXATIN(trademark)) in combination with tegafur / gimeracit / oteracil potassium; and FOLFOX, an abbreviation for treatment regimens of oxaliplatin (ELOXATIN(trademark)) in combination with 5-FU and leucovorin. In one embodiment, one or more chemotherapeutic agents of the method of the present invention are XELOX, FOLFOX, or SOX.

[0173] Chemotherapy agents as defined herein include “anti-hormone agents” or “endocrine therapy agents” that act to modulate, reduce, block, or inhibit the action of hormones that can promote the growth of cancer.These may be hormones themselves and include, but are not limited to: anti-estrogens with a mixed agonist / antagonist profile, including selective estrogen receptor modulators (SERMs) such as tamoxifen (NOLVADEX®), 4-hydroxytamoxifen, toremifene (FARESTON®), doxifen, droxifen, raloxifene (EVISTA®), trioxyfen, keoxyfen, and SERM3; pure anti-estrogens without agonist properties, such as fulvestrant (FASLODEX®) and EM800 (such drugs may inhibit estrogen receptor (ER) dimerization, inhibit DNA binding, increase ER turnover, and / or suppress ER levels); aromatase inhibitors, including steroidal aromatase inhibitors such as formestane and exemestane (AROMASIN®), and anastrozole. Nonsteroidal aromatase inhibitors such as (ARIMIDEX®), letrozole (Femara®), and aminoglutethimide, as well as volozol (RIVISOR®) and megstrol acetate. Other aromatase inhibitors including MEGASE®, phadrozol and 4(5)-imidazole; leuprolide (LUPRON® and ELIGARD®), goserelin, buserelin and trypterellin and other lutenogenic hormone-releasing hormone agonists; sex steroids including progestins such as megestrol acetate and medroxyprogesterone acetate, estrogens such as diethylstilbestrol and premarin, and androgens / retinoids such as fluoxymesterone, total trans-retionic acid and fenretinide; onapristone; antiprogesterone; estrogen receptor downregulation (ERD); antiandrogens such as flutamide, nilutamide and bicalutamide; and any pharmaceutically acceptable salts, acids or derivatives of any of the above; as well as any combination of two or more of the above.

[0174] 7. Pharmaceutical compositions Pharmaceutical compositions suitable for administration to human patients are typically formulated to be suitable for non-enteral administration, for example, in a liquid carrier, or for reconstitution into a solution or suspension for intravenous administration.

[0175] In general, such compositions typically include a pharmaceutically acceptable carrier. As used herein, “pharmaceutically acceptable” means approved by a regulatory authority for use in animals, particularly humans, or listed in the United States Pharmacopeia or other generally recognized pharmacopoeia equivalents. The term “carrier” means a diluent, adjuvant, excipient, or vehicle on which the compound is administered. Such pharmaceutical carriers may include sterile solutions such as water and oil, e.g., petroleum-derived, animal-derived, plant-derived, or synthetic-derived, such as peanut oil, soybean oil, mineral oil, sesame oil, polyethylene glycol ricinoleate glycerol. Water or saline, dextrose aqueous solutions, and glycerol aqueous solutions can be used as carriers, particularly for injectable solutions (e.g., containing anti-LAG-3 antibodies and / or anti-PD-1 antibodies). Liquid compositions for non-enteral administration 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, the anti-LAG-3 antibody and / or anti-PD-1 antibody are administered intravenously (e.g., in separate formulations or in a co-formulation (same formulation or separate formulations)).

[0176] 8. Patient population The present invention provides clinical methods for treating malignancies (e.g., progressive, refractory solid tumors and hematological malignancies) in human patients using immunotherapies described herein, such as combinations of LAG-3 inhibitors (e.g., anti-LAG-3 antibodies), PD-1 pathway inhibitors (e.g., anti-PD-1 antibodies), and one or more chemotherapeutic agents.

[0177] Examples of malignant tumors that can be treated using the method of the present invention include liver cancer, hepatocellular carcinoma (HCC), bone cancer, pancreatic cancer, skin cancer, oral cancer, head and neck cancer, breast cancer, lung cancer, small cell lung cancer, NSCLC, cutaneous or intraocular malignant melanoma, kidney cancer, ovarian cancer, colorectal cancer, colon cancer, rectal cancer, anal cancer, gastric cancer, gastroesophageal junction cancer, testicular cancer, uterine cancer, fallopian duct cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, squamous cell carcinoma of the head and neck (SCCHN), non-Hodgkin lymphoma, esophageal cancer, small intestine cancer, endocrine cancer, thyroid cancer, parathyroid cancer, adrenal adenocarcinoma, soft tissue sarcoma, urethral cancer, penile cancer, pediatric solid tumors, lymphocytic lymphoma, bladder cancer, kidney cancer, ureteral cancer, renal pelvis cancer, neoplasms of the central nervous system (CNS), primary CNS lymphoma, tumor blood vessels This invention includes cancers such as plagiocephalic tumors, spinal axial tumors, brainstem gliomas, pituitary adenomas, Kaposi's sarcoma, epidermoid carcinoma, squamous cell carcinoma, environmentally induced cancers including those induced by asbestos, hematological malignancies such as multiple myeloma, B-cell lymphoma, Hodgkin lymphoma / primary mediastinal B-cell lymphoma, non-Hodgkin lymphoma, acute myeloid lymphoma, chronic myeloid leukemia, chronic lymphocytic leukemia, follicular lymphoma, diffuse large B-cell lymphoma, Burkitt lymphoma, immunoblastic large B-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, as well as any combination of these cancers. The invention may also be used to treat metastatic cancers.

[0178] In one embodiment, the human patient has a malignant tumor that is resistant to treatment with immune checkpoint inhibitors. In another embodiment, the patient has a malignant tumor that is resistant to treatment with PD-1 inhibitors. In yet another embodiment, the patient has a malignant tumor that is resistant to treatment with anti-PD-1 antibodies. In yet another embodiment, the patient has a malignant tumor that is resistant to treatment with anti-PD-L1 antibodies. In one embodiment, the malignant tumor is gastric cancer or gastroesophageal junction cancer.

[0179] In one embodiment, a human patient has gastric cancer or gastroesophageal junction cancer. In another embodiment, the patient has gastric cancer or gastroesophageal junction cancer that is resistant to treatment by cancer therapy. In one embodiment, cancer therapy may be radiotherapy, surgery, chemotherapy, gene therapy, DNA therapy, viral therapy, RNA therapy, immunotherapy, bone marrow transplantation, nanotherapy, monoclonal antibody therapy, or a combination of the above. The treatment may take the form of adjuvant therapy or neoadjuvant therapy. As used herein, “adjuvant therapy” means cancer therapy performed after primary therapy to reduce the risk of cancer recurrence. Adjuvant therapy may include chemotherapy, radiotherapy, hormone therapy, targeted therapy, or biological therapy. Adjuvant therapy is often performed after primary therapy such as surgery or radiation. Adjuvant therapy performed before the main procedure is called neoadjuvant therapy. This type of adjuvant therapy can also reduce the likelihood of cancer recurrence and is often used to make primary therapy, such as surgery or radiation therapy, more effective in reducing the tumor burden. In another embodiment, the patient has gastric cancer or gastroesophageal junction cancer that is refractory to chemotherapy. In another embodiment, the patient has gastric cancer or gastroesophageal junction cancer that is resistant to immune checkpoint inhibitor treatment. In another embodiment, the patient has gastric cancer or gastroesophageal junction cancer that is resistant to PD-1 inhibitor treatment. In another embodiment, the patient has gastric cancer or gastroesophageal junction cancer that is resistant to anti-PD-1 antibody treatment. In another embodiment, the patient has gastric cancer or gastroesophageal junction cancer that is resistant to anti-PD-L1 antibody treatment.

[0180] Patients may be tested or selected for one or more of the above clinical characteristics before, during, or after the procedure.

[0181] Malignant tumors can be tested to determine LAG-3 expression according to the methods described herein. In one embodiment, the malignant tumor treated according to the methods described herein is a LAG-3 positive tumor. In one embodiment, the malignant tumor is a LAG-3 positive gastric adenocarcinoma or gastroesophageal junction adenocarcinoma.

[0182] In one embodiment, at least about 0.5%, at least about 1%, at least about 2%, at least about 3%, at least about 4%, at least about 5%, at least about 6%, at least about 7%, at least about 8%, at least about 9%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, or at least about 30% of the total number of cells in a malignant tumor express LAG-3. In one embodiment, the percentage of cells expressing LAG-3 is assessed by performing an assay to detect the presence of LAG-3 RNA. In a further embodiment, the presence of LAG-3 RNA is detected by RT-PCR, in situ hybridization, or RNase protection. In one embodiment, the presence of LAG-3 RNA is detected by an assay based on RT-PCR. In another embodiment, the percentage of cells expressing LAG-3 is assessed by performing an assay to detect the presence of LAG-3 polypeptide. In one embodiment, the presence of LAG-3 polypeptide is detected by IHC, ELISA, in vivo imaging, or flow cytometry. In one embodiment, LAG-3 expression is assayed by IHC.

[0183] Malignant tumors can be tested according to the methods described herein to determine LAG-3 and PD-L1 expression. In one embodiment, the malignant tumor tested according to the methods described herein is a LAG-3-positive, PD-L1-positive tumor. In one embodiment, the malignant tumor is a LAG-3-positive, PD-L1-positive gastric adenocarcinoma or gastroesophageal junction adenocarcinoma.

[0184] In some embodiments, the patient is HER2-negative. In some embodiments, the patient has a histologically or cytologically confirmed diagnosis of unresectable, locally advanced, or metastatic gastric cancer or gastroesophageal junction adenocarcinoma. In certain embodiments, the patient has not previously received systemic therapy. In some embodiments, the patient has not been administered HER2 inhibitors. In certain embodiments, the patient does not have known untreated central nervous system metastases. In some embodiments, the patient does not have uncontrolled or severe cardiovascular disease. In some embodiments, the patient has an ECOG performance status score of 0 or 1.

[0185] 9. Immunotherapy In one respect, the immunotherapies described herein involve the administration of a LAG-3 inhibitor (e.g., an anti-LAG-3 antibody), a PD-1 pathway inhibitor (e.g., an anti-PD-1 antibody or anti-PD-L1 antibody), and one or more chemotherapeutic agents to treat subjects with malignant tumors (e.g., progressive, refractory solid tumors or hematological malignancies).

[0186] In one embodiment, the present invention provides anti-LAG-3 antibodies and anti-PD-1 antibodies in combination with chemotherapeutic agents according to a defined clinical dosing regimen for treating subjects with malignant tumors (e.g., progressive, 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 regimen is fixed. In another embodiment, the dosing regimen is adjusted to provide an optimal desired response (e.g., an effective response).

[0187] As described herein, adjunctive or combined administration (simultaneous administration) includes simultaneous administration of compounds in the same or different dosage forms, or separate administration of compounds (e.g., sequential administration). Therefore, for example, anti-LAG-3 antibody and anti-PD-1 antibody may be administered simultaneously in a single formulation. Alternatively, anti-LAG-3 antibody and anti-PD-1 antibody may be formulated for separate administration and administered simultaneously or sequentially (e.g., the first antibody is administered within approximately 30 minutes prior to the administration of the second antibody).

[0188] For example, an anti-PD-1 antibody may be administered first, followed (for example, immediately after) by an anti-LAG-3 antibody, or vice versa. In one embodiment, the anti-PD-1 antibody is administered before the anti-LAG-3 antibody. In another embodiment, the anti-PD-1 antibody is administered after the anti-LAG-3 antibody. In yet another embodiment, the anti-LAG-3 antibody and the anti-PD-1 antibody are administered simultaneously. Such simultaneous or sequential administration preferably results in both antibodies being present simultaneously in the treated patient.

[0189] 10. Treatment Protocol On the one hand, an appropriate treatment protocol for treating malignant tumors in human patients includes administering an effective dose of a LAG3 inhibitor (e.g., an anti-LAG-3 antibody), a PD-1 pathway inhibitor (e.g., an anti-PD-1 antibody), and one or more chemotherapeutic agents.

[0190] In one embodiment, an appropriate treatment protocol for treating a malignant tumor in a human patient includes, for example, administering to the patient each of the following effective doses: (a) Anti-LAG-3 antibodies, such as those containing the CDR1, CDR2, and CDR3 domains of the heavy chain variable region having the sequence described in SEQ ID NO: 3, and the CDR1, CDR2, and CDR3 domains of the light chain variable region having the sequence described in SEQ ID NO: 5, (b) Anti-PD-1 antibodies, such as those containing the CDR1, CDR2, and CDR3 domains of the heavy chain variable region having the sequence described in SEQ ID NO: 19, and the CDR1, CDR2, and CDR3 domains of the light chain variable region having the sequence described in SEQ ID NO: 21, and (c) One or more chemotherapeutic agents, Herein, the method comprises at least one administration cycle, the cycle being for a period of 6 weeks, and for at least one cycle, at least two doses of anti-LAG-3 antibody are administered in doses of approximately 1 mg, 3 mg, 10 mg, 20 mg, 50 mg, 80 mg, 100 mg, 120 mg, 130 mg, 150 mg, 160 mg, 180 mg, 200 mg, 240 mg, or 280 mg, and at least two doses of anti-PD-1 antibody are administered in doses of approximately 50 mg, 80 mg, 100 mg, 130 mg, 150 mg, 180 mg, 200 mg, 240 mg, 280 mg, 320 mg, 360 mg, 400 mg, 440 mg, or 480 mg. In certain embodiments, the anti-LAG-3 antibody is administered in doses of approximately 320 mg, 360 mg, 400 mg, 440 mg, 480 mg, 520 mg, 560 mg, 600 mg, 640 mg, 680 mg, 720 mg, 760 mg, 800 mg, 840 mg, 880 mg, 920 mg, 960 mg, or 1000 mg. In certain embodiments, the anti-LAG-3 antibody is administered in doses of approximately 1040 mg, 1080 mg, 1120 mg, 1160 mg, 1200 mg, 1240 mg, 1280 mg, 1320 mg, 1360 mg, 1400 mg, 1440 mg, 1480 mg, 1520 mg, 1560 mg, 1600 mg, 1640 mg, 1680 mg, 1720 mg, 1760 mg, 1800 mg, 1840 mg, 1880 mg, 1920 mg, 1960 mg, or 2000 mg. In certain embodiments, the anti-LAG-3 antibody is administered in a dose of approximately 480 mg. In another embodiment, the four doses of anti-LAG-3 antibody are administered at doses of 0.01 mg, 0.03 mg, 0.25 mg, 0.1 mg, 0.3 mg, 1 mg, 3 mg, 5 mg, 8 mg, or 10 mg per kg of body weight, and the four doses of anti-PD-1 antibody are administered at doses of 0.1 mg, 0.3 mg, 1 mg, 3 mg, 5 mg, 8 mg, or 10 mg per kg of body weight.

[0191] In certain embodiments, the anti-LAG-3 antibody is administered at a dose of approximately 300 mg to 500 mg once every three weeks. In some embodiments, the anti-LAG-3 antibody is administered at a dose of approximately 400 mg once every three weeks. In some embodiments, the anti-LAG-3 antibody is administered at a dose of approximately 700 mg to 900 mg once every four weeks.

[0192] In a further embodiment, one or more chemotherapeutic agents are administered. In one embodiment, at least one chemotherapeutic agent is administered intravenously. In one embodiment, at least one chemotherapeutic agent is administered orally.

[0193] In one embodiment, one or more chemotherapeutic agents are administered using a body surface-based administration method.

[0194] In one embodiment, the anti-LAG-3 antibody and the anti-PD-1 antibody are administered in the following doses: (a) 120 mg of anti-LAG-3 antibody and 360 mg of anti-PD-1 antibody; or (b) 160 mg of anti-LAG-3 antibody and 480 mg of anti-PD-1 antibody.

[0195] In one aspect, the tumor is gastric cancer or esophagogastric junction cancer.

[0196] In another embodiment, the amount of anti-LAG-3 and / or anti-PD-1 antibody administered is constant for each dose. In another embodiment, the amount of antibody administered varies with each dose. For example, the maintenance (or subsequent) dose of the antibody may be higher or the same as the initial loading dose. In another embodiment, the maintenance dose of the antibody may be lower or the same as the loading dose.

[0197] In another embodiment, the anti-LAG-3 antibody and the anti-PD-1 antibody are formulated for intravenous administration. In one embodiment, the anti-LAG-3 antibody and the anti-PD-1 antibody are administered on day 1 and day 22 of each cycle. In one embodiment, one or more chemotherapeutic agents are administered on day 1 and day 22 of each cycle. In one embodiment, one or more chemotherapeutic agents are administered at least once daily.

[0198] In another embodiment, the administration cycle is 6 weeks and can be repeated as needed. In yet another embodiment, the treatment consists of up to 12 cycles.

[0199] In one embodiment, the anti-LAG-3 antibody and anti-PD-1 antibody are administered in the following doses: (a) 120 mg of anti-LAG-3 antibody and 360 mg of anti-PD-1 antibody on days 1 and 22 of each treatment cycle every 6 weeks; and oxaliplatin and capecitabine (XELOX) are administered. In a further embodiment, oxaliplatin 130 mg / m² 2 It is administered every 6 weeks on days 1 and 22 of each treatment cycle, with capecitabine 1000 mg / m². 2 It is administered twice daily every six weeks, from day 1 to day 14 and from day 22 to day 35 of each treatment cycle.

[0200] In one embodiment, the anti-LAG-3 antibody and anti-PD-1 antibody are administered in the following doses: (a) 160 mg of anti-LAG-3 antibody and 480 mg of anti-PD-1 antibody, along with oxaliplatin, leucovorin, and fluorouracil (FOLFOX), are administered on days 1 and 29 of odd-numbered cycles (e.g., cycles 1, 3, and 5) and on day 15 of even-numbered cycles (e.g., cycles 2, 4, and 6). In a further embodiment, oxaliplatin 85 mg / m² 2 , leucovorin 400mg / m² 2 , and fluorouracil 400 mg / m² 2 It is administered every 6 weeks on days 1, 15, and 29 of each treatment cycle, with fluorouracil 1200 mg / m². 2It is administered every six weeks on days 1 and 2, 15 and 16, 29 and 30 of each treatment cycle.

[0201] In one embodiment, the anti-LAG-3 antibody and anti-PD-1 antibody are administered in the following doses: (a) 120 mg of anti-LAG-3 antibody and 360 mg of anti-PD-1 antibody on days 1 and 22 of each treatment cycle every 6 weeks; along with oxaliplatin and tegafur / gimeracil / oteracil (Oral S-1) (SOX). In a further embodiment, oxaliplatin 130 mg / m² 2 The first treatment is administered every six weeks on days 1 and 22 of each treatment cycle, and Oral S-1 is administered twice daily every six weeks on days 1 through 14 and days 22 through 35 of each treatment cycle. In a further embodiment, the S-1 dose is calculated by body surface area (BSA, mg / m2 / dose): BSA < 1.25 m² 2 40 mg / dose; ≥1.25 and <1.5 mg 2 , 50mg / dose; ≧1.5m 2 , 60mg / dose.

[0202] In another embodiment, the anti-LAG-3 antibody, anti-PD-1 antibody, and chemotherapeutic agents are administered as first-line therapy (e.g., initiation or initial treatment). In another embodiment, the anti-LAG-3 antibody, anti-PD-1 antibody, and chemotherapeutic agents are administered as second-line therapy (e.g., after relapse and / or after failure of first-line treatment, including in the event of failure of first-line treatment).

[0203] In one embodiment, the anti-LAG-3 antibody is BMS-986016 and the anti-PD-1 antibody is nivolumab. In one embodiment, the anti-LAG-3 antibody is MK-4280 and the anti-PD-1 antibody is pembrolizumab. In one embodiment, the anti-LAG-3 antibody is REGN3767 and the anti-PD-1 antibody is REGN2810. In one embodiment, the anti-LAG-3 antibody is LAG525 and the anti-PD-1 antibody is PDR001.

[0204] In one embodiment, the patient is administered an effective dose of (a) an anti-LAG-3 antibody comprising the CDR1, CDR2, and CDR3 domains of the heavy chain variable region having the sequence described in SEQ ID NO: 3, and the CDR1, CDR2, and CDR3 domains of the light chain variable region having the sequence described 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 the sequence described in SEQ ID NO: 15, and the CDR1, CDR2, and CDR3 domains of the light chain variable region having the sequence described in SEQ ID NO: 17, and (c) one or more chemotherapeutic agents selected from the group consisting of XELOX, FOLFOX, and SOX. In one embodiment, the method is administered to a patient who has not received prior treatment (e.g., as first-line therapy). In one embodiment, the prior treatment is a HER2 inhibitor. In a specific embodiment, the anti-LAG-3 antibody and the anti-PD-1 antibody are administered as a fixed-dose combination. In one embodiment, the patient has recurrent, locally advanced, or metastatic gastric or gastroesophageal adenocarcinoma.

[0205] In another aspect, the present invention is characterized by any of the above embodiments, wherein the anti-PD-1 antibody is substituted with an anti-PD-L1 antibody or an anti-PD-L2 antibody, or is combined with an anti-PD-L1 antibody.

[0206] 11.Results Patients treated according to the methods described 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 can be measured by chest X-ray or CT or MRI film. In yet another embodiment, cytology or histology can be used to assess the response to treatment.

[0207] In one embodiment, the treated patient exhibits a complete response (CR), a partial response (PR), stable disease (SD), an immune-related complete response (irCR), an immune-related partial response (irPR), or an immune-related stable disease (irSD). In another embodiment, the treated patient experiences tumor reduction and / or a reduction in growth rate, i.e., suppression of tumor growth. In yet another embodiment, undesirable cell growth is mitigated or inhibited. In yet another embodiment, one or more of the following may occur: the number of cancer cells may be reduced; the tumor size may be reduced; the invasion of cancer cells into peripheral organs may be inhibited, delayed, stunted, or stopped; tumor metastasis may be delayed or inhibited; tumor growth may be suppressed; tumor recurrence may be prevented or delayed; and one or more cancer-related symptoms may be alleviated to some extent.

[0208] In other embodiments, administration of an effective dose of an anti-LAG-3 antibody, an anti-PD-1 antibody, and one or more chemotherapeutic agents according to any of the methods of the present invention results in at least one therapeutic effect selected from the group consisting of a reduction in tumor size, a reduction in the number of metastatic lesions appearing over time, complete remission, partial remission, or stabilization of the disease.

[0209] In yet another embodiment, the treatment method yields a better clinical benefit rate (CBR=CR+PR+SD≧6months) than that achieved by a treatment method that includes (i) the step of determining the LAG-3 expression level in a tumor sample before treatment, (ii) the step of selecting LAG-3-positive tumors for treatment, (iii) the step of treating tumors identified as LAG-3-positive before treatment, or (iv) any combination thereof. In other embodiments, the improvement in clinical benefit rate is approximately 20%, approximately 30%, approximately 40%, approximately 50%, approximately 60%, approximately 70%, approximately 80%, or more compared to a treatment method that includes (i) the step of determining the LAG-3 expression level in a tumor sample before treatment, (ii) the step of selecting LAG-3-positive tumors for treatment, (iii) the step of treating tumors identified as LAG-3-positive before treatment, or (iv) any combination thereof.

[0210] In yet another embodiment, the treatment method yields an objective response rate (ORR=CR+PR) of at least approximately 15%, at least approximately 20%, at least approximately 25%, at least approximately 30%, at least approximately 40%, at least approximately 50%, at least approximately 60%, at least approximately 70%, at least approximately 80%, at least approximately 90%, or approximately 100%. In one embodiment, the treatment method yields an objective response rate of at least approximately 15%, where the malignant tumor is a LAG-3 positive melanoma resistant to treatment with anti-PD-1 antibody or anti-PD-L1 antibody. In one embodiment, the median duration of response is ≥3 months, ≥6 months, ≥12 months, or ≥18 months. In one embodiment, the median duration of response is ≥6 months. In one embodiment, the proportion of patients with a response duration of 6 months or longer is at least approximately 30%, at least approximately 40%, at least approximately 50%, at least approximately 60%, at least approximately 70%, at least approximately 80%, at least approximately 90%, at least approximately 95%, at least approximately 99%, or 100%.

[0211] In other embodiments, the treatment method yields a superior objective response rate (ORR=CR+PR) compared to treatment methods that do not include (i) the step of determining the LAG-3 expression level in a tumor sample before treatment, (ii) the step of selecting LAG-3-positive tumors for treatment, (iii) the step of treating tumors identified as LAG-3-positive before treatment, or (iv) any combination thereof. In other embodiments, the improvement in objective response rate is approximately 20%, 30%, 40%, 50%, 60%, 70%, 80%, or more compared to treatment methods that do not include (i) the step of determining the LAG-3 expression level in a tumor sample before treatment, (ii) the step of selecting LAG-3-positive tumors for treatment, (iii) the step of treating tumors identified as LAG-3-positive before treatment, or (iv) any combination thereof. In some embodiments, the median duration of response is ≥3 months, ≥6 months, ≥12 months, or ≥18 months. In one embodiment, the median duration of response is 6 months or more.

[0212] In further embodiments, the treatment method yields disease control rates (DCR=CR+PR+SD) of at least approximately 20%, at least approximately 30%, at least approximately 40%, at least approximately 50%, at least approximately 60%, at least approximately 70%, at least approximately 80%, at least approximately 90%, at least approximately 95%, at least approximately 99%, or at least approximately 100%. In one embodiment, the treatment method yields a disease control rate of at least approximately 70%, where the malignant tumor is LAG-3 positive melanoma resistant to treatment with anti-PD-1 antibody or anti-PD-L1 antibody. In one embodiment, the median duration of response is ≥3 months, ≥6 months, ≥12 months, or ≥18 months. In one embodiment, the median duration of response is ≥6 months. In one embodiment, the proportion of patients with a response duration of 6 months or longer is at least approximately 30%, at least approximately 40%, at least approximately 50%, at least approximately 60%, at least approximately 70%, at least approximately 80%, at least approximately 90%, at least approximately 95%, at least approximately 99%, or 100%.

[0213] In yet another embodiment, the treatment method yields a better disease control rate (DCR=CR+PR+SD) than that achieved by a treatment method that does not include (i) the step of determining the LAG-3 expression level in a tumor sample before treatment, (ii) the step of selecting LAG-3-positive tumors for treatment, (iii) the step of treating tumors identified as LAG-3-positive before treatment, or (iv) any combination thereof. In another embodiment, the improvement in disease control rate is approximately 20%, approximately 30%, approximately 40%, approximately 50%, approximately 60%, approximately 70%, approximately 80%, or more compared to a treatment method that does not include (i) the step of determining the LAG-3 expression level in a tumor sample before treatment, (ii) the step of selecting LAG-3-positive tumors for treatment, (iii) the step of treating tumors identified as LAG-3-positive before treatment, or (iv) any combination thereof. In one embodiment, the median duration of response is ≥3 months, ≥6 months, ≥12 months, or ≥18 months. In one embodiment, the median duration of response is 6 months or more.

[0214] 12. Kit and Unit Dosage Forms Diagnostic kits comprising an anti-LAG-3 antibody for assaying LAG-3 expression as a biomarker for screening patients for immunotherapy or for predicting the effectiveness of immunotherapy are also within the scope of the present invention. The kit generally includes a label and instructions for use indicating the intended use of the kit contents. The term “label” includes any documented or recorded material that is attached to or included with the kit or otherwise incorporated into the kit. In certain embodiments of the diagnostic kit, a first anti-LAG-3 antibody for assaying, detecting and / or quantifying LAG-3 expression is packaged together with at least one therapeutic antibody (e.g., a second anti-LAG-3 antibody and an anti-PD-1 antibody) for the treatment of LAG-3-positive tumors. In some embodiments, the kit further comprises an anti-PD-L1 antibody for assaying, detecting and / or quantifying PD-L1 expression as a biomarker for predicting the effectiveness of immunotherapy. In one embodiment, immunotherapy involves administering to a patient a therapeutically effective dose of a LAG-3 inhibitor (e.g., an anti-LAG-3 antibody), a PD-1 pathway inhibitor (e.g., an anti-PD1 antibody), and one or more chemotherapeutic agents.

[0215] In certain embodiments, the diagnostic kit includes an anti-human LAG-3 monoclonal antibody for assaying, detecting, and / or quantifying LAG-3 expression. See, for example, J. Matsuzaki, et al.; PNAS 107, 7875 (2010).

[0216] The present invention also provides a therapeutic kit comprising a pharmaceutical composition containing an anti-LAG-3 antibody, e.g., BMS-986016, an anti-PD-1 antibody, e.g., nivolumab, and one or more chemotherapeutic agents, in a therapeutically effective amount suitable for use in the method described above. In certain embodiments of the therapeutic kit, the anti-LAG-3 antibody is packaged together with the anti-PD-1 antibody in unit dose form. The kit may also include instructions, for example, an administration schedule, to enable a practitioner (e.g., a physician, nurse, or patient) to administer the composition contained therein to a patient having cancer (e.g., a solid tumor), if required. The kit may also include a syringe.

[0217] In essence, a diagnostic and / or therapeutic kit comprises multiple packages of single-dose pharmaceutical compositions, each containing an effective amount of anti-LAG-3 antibody or anti-PD-1 antibody for single administration according to the method described above. The kit may also include any apparatus or equipment necessary for administering the pharmaceutical composition(s). For example, the kit may provide one or more pre-filled syringes containing a certain amount of anti-LAG-3 antibody or anti-PD-1 antibody.

[0218] In one embodiment, the present invention provides a kit for treating a patient suffering from a malignant tumor, the kit is (a) A dose of an anti-LAG-3 antibody, for example, an antibody comprising the CDR1, CDR2, and CDR3 domains of the heavy chain variable region having the sequence described in SEQ ID NO: 3, and the CDR1, CDR2, and CDR3 domains of the light chain variable region having the sequence described in SEQ ID NO: 5; (b) A dose of an anti-PD-1 antibody, for example, an antibody comprising the CDR1, CDR2, and CDR3 domains of the heavy chain variable region having the sequence described in SEQ ID NO: 19, and the CDR1, CDR2, and CDR3 domains of the light chain variable region having the sequence described in SEQ ID NO: 21; (c) One or more chemotherapeutic agents; and, (d) Instructions for using an anti-LAG-3 antibody, an anti-PD-1 antibody, and one or more chemotherapeutic agents in the methods described herein. Includes.

[0219] In one aspect, the malignant tumor is gastric cancer or esophagogastric junction cancer.

[0220] The present invention is further illustrated by the following embodiments, which should not be construed as further limiting. The contents of all references cited herein are expressly incorporated herein by reference. [Examples]

[0221] Examples Example 1 Efficacy of anti-lymphocyte activating gene-3 antibody (anti-LAG-3; BMS-986016) in combination with nivolumab and chemotherapy agents in patients with advanced or metastatic gastric cancer or gastroesophageal junction cancer. The purpose of this clinical trial is to evaluate the combination therapy of BMS-986016 (liratrimab), nivolumab, and chemotherapy agents in the treatment of gastric cancer or gastroesophageal junction cancer.

[0222] Patients are selected based on the following eligibility criteria: (1) having advanced or metastatic gastric cancer or gastroesophageal junction cancer; (2) no prior systemic treatment; (3) a PS of 0-1 as determined by the Eastern Cooperative Oncology Group; (4) HER2-negative; and (5) a LAG-3-positive tumor as determined by IHC. During the treatment period, patients will receive BMS-986016 and nivolumab in combination with a selected chemotherapy agent (XELOX, FOLFOX, or SOX).

[0223] Patients assigned to XELOX will receive the following: • Administer lilatrimab 120 mg / nivolumab 360 mg intravenously over 60 minutes (IV) on day 1 and day 22 of each treatment cycle every 6 weeks. Oxaliplatin 130 mg / m² 2 This is administered intravenously (IV) on day 1 and day 22 of each treatment cycle every 6 weeks, and • Capecitabine 1000 mg / m³ 2 This is administered orally twice daily on days 1 through 14 and 22 through 35 of each treatment cycle every six weeks.

[0224] Patients assigned to FOLFOX will receive the following: • Administer lilatrimab 160 mg / nivolumab 480 mg intravenously over 60 minutes (IV) on days 1 and 29 of odd-numbered cycles (e.g., cycles 1, 3, and 5) and on day 15 of even-numbered cycles (e.g., cycles 2, 4, and 6). Oxaliplatin 85 mg / m² 2 , leucovorin 400mg / m² 2 , and fluorouracil 400 mg / m² 2 This is administered intravenously every 6 weeks on days 1, 15, and 29 of each treatment cycle, along with fluorouracil 1200 mg / m². 2 This is administered intravenously over 24 hours (or according to local criteria) every 6 weeks on days 1 and 2, 15 and 16, and 29 and 30 of each treatment cycle.

[0225] Patients assigned to SOX will receive the following: • Administer lilatrimab 120 mg / nivolumab 360 mg intravenously over 60 minutes on day 1 and day 22 of each treatment cycle every 6 weeks. • Oxaliplatin 130 mg / m² 2 This is administered intravenously on day 1 and day 22 of each treatment cycle every 6 weeks, and Oral S-1 (tegafur / gimeracil / oteracil) is administered intravenously twice daily every six weeks, from day 1 to 14 and from day 22 to 35 of each treatment cycle. The S-1 dose is calculated according to body surface area (BSA, mg / m²). 2 / dose):BSA<1.25m 2 40 mg / dose; ≥1.25 and <1.5 mg 2 , 50mg / dose; ≧1.5m 2 , 60mg / dose.

[0226] array Sequence ID 1: Heavy chain amino acid sequence; anti-LAG-3mAb (BMS-986016) QVQLQQWGAGLLKPSETLSLTCAVYGGSFSDYYWNWIRQPPGKGLEWIGEINHRGSTNSNPSLKSRVTLSLDTSKNQFSLKLRSVTAADTAVYYCAFGYSDYEYNWFDPWG QGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYG PPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKT ISKAKGQPREPQVYTLPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK Sequence ID 2: Light chain amino acid sequence; anti-LAG-3mAb (BMS-986016) EIVLTQSPATLSLSPGERATLSCRASQSISSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPLTFGQGTNLEIK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC Sequence ID 3: Heavy chain variable region (VH) amino acid sequence; anti-LAG-3mAb (BMS-986016) QVQLQQWGAGLLKPSETLSLTCAVYGGSFSDYYWNWIRQPPGKGLEWIGEINHRGSTNSNPSLKSRVTLSLDTSKNQFSLKLRSVTAADTAVYYCAFGYSDYEYNWFDPWGQGTLVTVSS Sequence ID 4: Heavy chain variable region (VH) nucleotide sequence; anti-LAG-3mAb (BMS-986016) caggtgcagctacagcagtggggcgcaggactgttgaagccttcggagaccctgtccctcacctgcgctgtctatggtgggtccttcagtgattactactggaactggatccgccagcccccagggaaggggctggagtggattggggaaatcaatcatcgtggaagcaccaactccaac ccgtccctcaagagtcgagtcaccctatcactagacacgtccaagaaccagttctccctgaagctgaggtctgtgaccgccgcggacacggctgtgtattactgtgcgtttggatatagtgactacgagtacaactggttcgacccctggggccagggaaccctggtcaccgtctcctca Sequence ID 5: Light chain variable region (VL) amino acid sequence; anti-LAG-3mAb (BMS-986016) EIVLTQSPATLSLSPGERATLSCRASQSISSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPLTFGQGTNLEIK Sequence ID 6: Light chain variable region (VL) nucleotide sequence; anti-LAG-3mAb (BMS-986016) gaaattgtgttgacacagtctccagccaccctgtctttgtctccaggggaaagagccaccctctcctgcagggccagtcagagtattagcagctacttagcctggtaccaacagaaacctggccaggctcccaggctcctcatctatgatgcatccaaca gggccactggcatcccagccaggttcagtggcagtgggtctgggacagacttcactctcaccatcagcagcctagagcctgaagattttgcagtttattactgtcagcagcgtagcaactggcctctcacttttggccaggggaccaacctggagatcaaa Sequence ID 7 Heavy chain CDR1 amino acid sequence; anti-LAG-3mAb (BMS-986016) DYYWN Sequence ID 8 Heavy chain CDR2 amino acid sequence; anti-LAG-3mAb (BMS-986016) EINHRGSTNSNPSLKS Sequence ID 9: Heavy chain CDR3 amino acid sequence; anti-LAG-3mAb (BMS-986016) GYSDYEYNWFDP Sequence ID 10 Light chain CDR1 amino acid sequence; anti-LAG-3mAb (BMS-986016) RASQSISSYLA Sequence ID 11 Light chain CDR2 amino acid sequence; anti-LAG-3mAb (BMS-986016) DASNRAT Sequence ID 12 Light chain CDR3 amino acid sequence; anti-LAG-3mAb (BMS-986016) QQRSNWPLT Sequence ID 13: Heavy chain amino acid sequence; anti-PD-1 mAb (BMS936558) QVQLVESGGGVVQPGRSLRLDCKASGITFSNSGMHWVRQAPGKGLEWVAVIWYDGSKRYYADSVKGRFTISRDNSKNTLFLQMNSLRAEDTAVYYCATNDDYWGQGTLVT VSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCP PCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTIS KAKGQPREPQVYTLPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK Sequence ID 14 Light chain amino acid sequence; anti-PD-1 mAb (BMS936558) EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQSSNWPRTFGQGTKVEI KRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC Sequence ID No. 15: Heavy chain variable region (VH) amino acid sequence; anti-PD-1 mAb (BMS936558) QVQLVESGGGVVQPGRSLRLDCKASGITFSNSGMHWVRQAPGKGLEWVAVIWYDGSKRYYADSVKGRFTISRDNSKNTLFLQMNSLRAEDTAVYYCATNDDYWGQGTLVTVSS

[0227] Sequence ID 16: Heavy chain variable region (VH) nucleotide sequence; anti-PD-1 mAb (BMS936558) caggtgcagctggtggagtctgggggaggcgtggtccagcctgggaggtccctgagactcgactgtaaagcgtctggaatcaccttcagtaactctggcatgcactgggtccgccaggctccaggcaaggggctggagtgggtggcagttatttggtatgatggaagta aaagatactatgcagactccgtgaagggccgattcaccatctccagagacaattccaagaacacgctgtttctgcaaatgaacagcctgagagccgaggacacggctgtgtattactgtgcgacaaacgacgactactggggccagggaaccctggtcaccgtctcctca Sequence ID No. 17 Light chain variable region (VL) amino acid sequence; anti-PD-1 mAb (BMS936558) EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQSSNWPRTFGQGTKVEIK Sequence ID 18: Light chain variable region (VL) nucleotide sequence; anti-PD-1 mAb (BMS936558) gaaattgtgttgacacagtctccagccaccctgtctttgtctccaggggaaagagccaccctctcctgcagggccagtcagagtgttagtagttacttagcctggtaccaacagaaacctggccaggctcccaggctcctcatctatgatgcatccaaca gggccactggcatcccagccaggttcagtggcagtgggtctgggacagacttcactctcaccatcagcagcctagagcctgaagattttgcagtttattactgtcagcagagtagcaactggcctcggacgttcggccaagggaccaaggtggaaatcaaa Sequence ID 19: Heavy chain CDR1 amino acid sequence; anti-PD-1 mAb (BMS936558) NSG H&M Sequence ID No. 20 Heavy chain CDR2 amino acid sequence; anti-PD-1 mAb (BMS936558) VIWYDGSKRYYADSVKG Sequence ID 21: Heavy chain CDR3 amino acid sequence; anti-PD-1 mAb (BMS936558) NDDY Sequence ID 22 Light chain CDR1 amino acid sequence; anti-PD-1 mAb (BMS936558) RASQSVSSYLA Sequence ID 23 Light chain CDR2 amino acid sequence; anti-PD-1 mAb (BMS936558) DASNRAT Sequence ID No. 24 Light chain CDR3 amino acid sequence; anti-PD-1 mAb (BMS936558) QQSSNWPRT Sequence ID 25 Heavy chain nucleotide sequence; anti-LAG-3mAb (BMS-986016) SEQ ID NO: 26 Light chain nucleotide sequence; anti-LAG-3 mAb (BMS-986016) gaaattgtgttgacacagtctccagccaccctgtctttgtctccaggggaaagagccaccctctcctgcagggccagtcagagtattagcagctacttagcctggtaccaacagaaacctggccaggctcccaggctcctcatctatgatgcatccaacagggccactggcatcccagccaggttcagtggcagtgggtctgggacagacttcactctcaccatcagcagcctagagcctgaagattttgcagtttattactgtcagcagcgtagcaactggcctctcacttttggccaggggaccaacctggagatcaaacgtacggtggctgcaccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgttag

Claims

1. A method for inhibiting the growth of malignant tumors in human patients, (a) LAG-3 Antagonist; (b) PD-1 pathway inhibitors; and (c) One or more chemotherapeutic agents This includes administering an effective amount of each to the patient, Herein, the tumor-associated immune cells of the patient express LAG-3.

2. A method for treating cancer in human patients, (a) LAG-3 Antagonist; (b) PD-1 pathway inhibitors; and (c) One or more chemotherapeutic agents This includes administering an effective amount of each to the patient, Herein, the tumor-associated immune cells of the patient express LAG-3.

3. A method for inhibiting the growth of malignant tumors in human patients, (a) LAG-3 Antagonist; (b) PD-1 pathway inhibitors; and (c) One or more chemotherapeutic agents A method comprising administering an effective amount of each to the patient.

4. A method for treating cancer in human patients, (a) LAG-3 Antagonist; (b) PD-1 pathway inhibitors; and (c) One or more chemotherapeutic agents A method comprising administering an effective amount of each to the patient.

5. 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, Fallopian duct cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, non-Hodgkin lymphoma, esophageal cancer, small intestine cancer, endocrine cancer, thyroid cancer, parathyroid cancer, adrenal adenocarcinoma, soft tissue sarcoma, urethral cancer, penile cancer, childhood cancer, lymphocytic lymphoma, bladder cancer, kidney or ureteral cancer, renal pelvis cancer, central nervous system neoplasms (CNS), primary CNS lymphoma, tumor angiogenesis, spinal axial tumors, brainstem gliomas, pituitary adenomas, and capillary tumors. Di sarcoma, epidermoid carcinoma, squamous cell carcinoma, environmentally induced cancers including those induced by asbestos, hematological malignancies including, for example, multiple myeloma, B-cell lymphoma, Hodgkin lymphoma / primary mediastinal B-cell lymphoma, non-Hodgkin lymphoma, acute myeloid lymphoma, chronic myeloid leukemia, chronic lymphocytic leukemia, follicular lymphoma, diffuse large B-cell lymphoma, Burkitt lymphoma, immunoblastic large B-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. A method according to any one of claims 1 to 4, selected from the group consisting of the following.

6. The method according to any one of claims 1 to 5, wherein the malignant tumor is gastric cancer or esophagogastric junction cancer.

7. The method according to claim 6, wherein the gastric cancer is an adenocarcinoma, lymphoma, gastrointestinal stromal tumor, or carcinoid tumor.

8. The method according to any one of claims 1 to 5, wherein the malignant tumor is selected from melanoma, non-small cell lung cancer (NSCLC), human papillomavirus (HPV)-related tumor, bladder cancer, head and neck squamous cell carcinoma, renal cell carcinoma, and gastric adenocarcinoma.

9. The method according to any one of claims 1 to 8, wherein the LAG-3 antagonist is an anti-LAG-3 antibody.

10. The method according to claim 9, wherein the anti-LAG-3 antibody is a full-length antibody.

11. The method according to claim 10, wherein the antibody is a monoclonal antibody, a human antibody, a humanized antibody, a chimeric antibody, or a multispecific antibody.

12. The method according to claim 11, wherein the multispecific antibody is a biaffinity retargeting antibody (DART), DVD-Ig, or a bispecific antibody.

13. The antibody is F(ab') ２ The method according to claim 9, wherein the fragment is a Fab' fragment, a Fab fragment, an Fv fragment, an scFv fragment, a dsFv fragment, a dAb fragment, or a single-chain linked polypeptide peptide.

14. The method according to claim 9, wherein the anti-LAG-3 antibody is BMS-986016, IMP731 (H5L7BW), MK-4280 (28G-10), REGN3767, GSK2831781, humanized BAP050, IMP-701 (LAG-5250), aLAG3 (0414), aLAG3 (0416), Sym022, TSR-033, TSR-075, XmAb22841, BI754111, MGD013, AVA-017, P13B02-30, or FS-118.

15. The method according to any one of claims 1 to 8, wherein the LAG-3 antagonist is IMP321.

16. The method according to any one of claims 9 to 13, wherein the anti-LAG-3 antibody comprises CDR1, CDR2, and CDR3 domains of the heavy chain variable region having the sequence described in SEQ ID NO: 3, and CDR1, CDR2, and CDR3 domains of the light chain variable region having the sequence described in SEQ ID NO:

5.

17. Anti-LAG-3 antibody, (a) Heavy chain variable region CDR1 containing the sequence described in Sequence ID No. 7; (b) Heavy chain variable region CDR2 containing the sequence described in Sequence ID No. 8; (c) Heavy chain variable region CDR3 containing the sequence described in Sequence ID No. 9; (d) Light chain variable region CDR1 containing the sequence described in Sequence ID No. 10; (e) Light chain variable region CDR2 containing the sequence described in Sequence ID No. 11; and (f) Light chain variable region CDR3 containing the sequence described in Sequence ID No. 12 The method according to any one of claims 1 to 14, including the method described in any one of claims 1 to 14.

18. The method according to any one of claims 1 to 11, wherein the anti-LAG-3 antibody comprises a heavy chain variable region and a light chain variable region, each containing the sequences described in SEQ ID NOs: 3 and 5, respectively.

19. The method according to any one of claims 1 to 11, wherein the anti-LAG-3 antibody comprises a heavy chain and a light chain having sequences described in SEQ ID NOs: 1 and 2, respectively.

20. The method according to any one of claims 1 to 19, wherein the PD-1 pathway inhibitor is an anti-PD-1 antibody or an anti-PD-L1 antibody.

21. The method according to claim 20, wherein the anti-PD-1 antibody is selected from the group consisting of nivolumab, pembrolizumab, pizilizumab, PDR001, MEDI0680, TSR-042, REGN2810, JS001, PF-06801591, BGB-A317, BI754091, and SHR-1210.

22. The method according to any one of claims 1 to 21, wherein the anti-PD-1 antibody comprises CDR1, CDR2, and CDR3 domains of the heavy chain variable region having the sequence described in SEQ ID NO: 15, and CDR1, CDR2, and CDR3 domains of the light chain variable region having the sequence described in SEQ ID NO:

17.

23. Anti-PD-1 antibodies, (a) Heavy chain variable region CDR1 containing the sequence described in Sequence ID No. 19; (b) Heavy chain variable region CDR2 containing the sequence described in Sequence ID No. 20; (c) Heavy chain variable region CDR3 containing the sequence described in Sequence ID No. 21; (d) Light chain variable region CDR1 containing the sequence described in Sequence ID No. 22; (e) Light chain variable region CDR2 containing the sequence described in Sequence ID No. 23; and (f) Light chain variable region CDR3 containing the sequence described in Sequence ID No. 24 The method according to any one of claims 1 to 21, including the method described in any one of claims 1 to 21.

24. The method according to any one of claims 1 to 21, wherein the anti-PD-1 antibody comprises a heavy chain variable region and a light chain variable region, each comprising the sequences described in SEQ ID NOs. 15 and 17, respectively.

25. The method according to any one of claims 1 to 21, wherein the anti-PD-1 antibody comprises a heavy chain and a light chain having sequences described in SEQ ID NOs: 13 and 14, respectively.

26. The method according to any one of claims 1 to 25, wherein one or more chemotherapeutic agents are platinum compounds or fluoropyrimidines.

27. The method according to any one of claims 1 to 26, wherein one or more chemotherapeutic agents are oxaliplatin, cisplatin, fluorouracil, capecitabine, tegafur, gimeracil, or oteracil.

28. The method according to claim 27, wherein one or more chemotherapeutic agents are oxaliplatin and capecitabine (XELOX).

29. The method according to claim 27, wherein one or more chemotherapeutic agents are oxaliplatin and fluorouracil.

30. The method according to claim 27, wherein the chemotherapeutic agent further comprises a chemoprotective agent.

31. The method according to claim 30, wherein the chemical protective agent is leucovorin.

32. The method according to claim 31, wherein one or more chemotherapeutic agents include oxaliplatin, leucovorin, and fluorouracil (FOLFOX).

33. The method according to claim 27, wherein one or more chemotherapeutic agents are oxaliplatin and tegafur / gimeracil / oteracil potassium (SOX).

34. The method according to any one of claims 1 to 33, wherein a combination of anti-LAG-3 antibody and anti-PD-1 antibody in a fixed dose is administered.

35. The method according to claim 34, wherein the fixed dose is determined based on the chemotherapeutic agent administered to the subject.

36. The method according to any one of claims 1 to 35, wherein the method comprises at least one administration cycle, the cycle having a period of 6 weeks, and for at least one cycle, two doses of anti-LAG-3 antibody are administered at doses of 120 or 160 mg, and two doses of anti-PD-1 antibody are administered at doses of 360 or 480 mg.

37. The method according to any one of claims 1 to 36, wherein 120 mg of anti-LAG-3 antibody, 360 mg of anti-PD-1 antibody, and XELOX are administered.

38. The method according to any one of claims 1 to 36, wherein 160 mg of anti-LAG-3 antibody, 480 mg of anti-PD-1 antibody, and FOLFOX are administered.

39. The method according to any one of claims 1 to 36, wherein 120 mg of anti-LAG-3 antibody, 360 mg of anti-PD-1 antibody, and SOX are administered.

40. The method according to any one of claims 1 to 39, wherein the anti-LAG-3 antibody and the anti-PD-1 antibody are formulated for intravenous administration.

41. The method according to any one of claims 1 to 40, wherein an anti-LAG-3 antibody and an anti-PD-1 antibody are co-formulated.

42. The method according to any one of claims 1 to 40, wherein the anti-LAG-3 antibody and the anti-PD-1 antibody are formulated separately.

43. A method for inhibiting the proliferation of gastric adenocarcinoma or gastroesophageal junction adenocarcinoma in a human patient, wherein the patient (a) An anti-LAG-3 antibody comprising the CDR1, CDR2, and CDR3 domains of the heavy chain variable region having the sequence described in Sequence ID No. 3, and the CDR1, CDR2, and CDR3 domains of the light chain variable region having the sequence described in Sequence ID No. 5, (b) An anti-PD-1 antibody comprising the CDR1, CDR2, and CDR3 domains of the heavy chain variable region having the sequence described in Sequence ID No. 15 and the CDR1, CDR2, and CDR3 domains of the light chain variable region having the sequence described in Sequence ID No. 17, and (c) One or more chemotherapeutic agents selected from the group consisting of oxaliplatin / capecitabine (XELOX), oxaliplatin / leucovorin / fluorouracil (FOLFOX), and oxaliplatin / tegafur / gimeracil / oteracil (SOX). A method comprising administering an effective amount of each of the following, wherein the tumor-associated immune cells of the patient express LAG-3.

44. A method for treating gastric cancer or esophagogastric junction cancer in a human patient, wherein the patient (a) An anti-LAG-3 antibody comprising the CDR1, CDR2, and CDR3 domains of the heavy chain variable region having the sequence described in Sequence ID No. 3, and the CDR1, CDR2, and CDR3 domains of the light chain variable region having the sequence described in Sequence ID No. 5, (b) An anti-PD-1 antibody comprising the CDR1, CDR2, and CDR3 domains of the heavy chain variable region having the sequence described in Sequence ID No. 15 and the CDR1, CDR2, and CDR3 domains of the light chain variable region having the sequence described in Sequence ID No. 17, and (c) One or more chemotherapeutic agents selected from the group consisting of XELOX, FOLFOX, and SOX A method comprising administering an effective dose of [a substance].

45. The method according to claim 44, wherein the patient's tumor-associated immune cells express LAG-3.

46. The method according to claim 44 or 45, administered to a patient who has not received prior treatment (for example, as first-line therapy).

47. The method according to claim 46, wherein the prior treatment is the administration of a HER2 inhibitor.

48. The method according to any one of claims 44 to 47, administered to a patient who is HER2-negative.

49. The method according to any one of claims 44 to 48, wherein the patient has not received prior systemic therapy.

50. The method according to any one of claims 44 to 49, wherein an anti-LAG-3 antibody and an anti-PD-1 antibody are administered as a fixed dose combination.

51. The method according to any one of claims 44 to 50, wherein the gastric cancer or gastroesophageal junction cancer is recurrent, locally advanced, or metastatic gastric cancer or gastroesophageal adenocarcinoma.

52. The method according to any one of claims 1 to 51, wherein LAG-3 expression is assayed by RT-PCR, in situ hybridization, RNase protection, RT-PCR-based assay, immunohistochemistry, enzyme-linked immunoadsorption assay, in vivo imaging, or flow cytometry.

53. The method according to claim 52, wherein LAG-3 expression is assayed by immunohistochemistry.

54. A method for treating gastric cancer or gastroesophageal junction adenocarcinoma in a human patient, wherein the patient (a) An anti-LAG-3 antibody comprising the CDR1, CDR2, and CDR3 domains of the heavy chain variable region having the sequence described in Sequence ID No. 3, and the CDR1, CDR2, and CDR3 domains of the light chain variable region having the sequence described in Sequence ID No. 5, (b) An anti-PD-1 antibody comprising the CDR1, CDR2, and CDR3 domains of the heavy chain variable region having the sequence described in Sequence ID No. 15 and the CDR1, CDR2, and CDR3 domains of the light chain variable region having the sequence described in Sequence ID No. 17, and (c) One or more chemotherapeutic agents A method comprising administering an effective dose of [a substance].

55. The method according to claim 54, administered to a patient who has not received prior treatment (for example, as first-line therapy).

56. A method for treating recurrent, locally advanced, or metastatic gastric cancer or gastroesophageal junction adenocarcinoma in a human patient, wherein the patient (a) an anti-LAG-3 antibody comprising the CDR1, CDR2, and CDR3 domains of the heavy chain variable region having the sequence described in Sequence ID No. 3 and the CDR1, CDR2, and CDR3 domains of the light chain variable region having the sequence described in Sequence ID No. 5, and (b) One or more standard treatment regimens, A method comprising administering an effective dose thereof, wherein the patient's tumor-associated immune cells express LAG-3.

57. A method for treating recurrent, locally advanced, or metastatic gastric cancer or gastroesophageal junction adenocarcinoma in a human patient, wherein the patient (a) An anti-LAG-3 antibody comprising the CDR1, CDR2, and CDR3 domains of the heavy chain variable region having the sequence described in Sequence ID No. 3, and the CDR1, CDR2, and CDR3 domains of the light chain variable region having the sequence described in Sequence ID No. 5, (b) An anti-PD-1 antibody comprising the CDR1, CDR2, and CDR3 domains of the heavy chain variable region having the sequence described in Sequence ID No. 15 and the CDR1, CDR2, and CDR3 domains of the light chain variable region having the sequence described in Sequence ID No. 17, and (c) One or more standard therapy regimens, A method comprising administering an effective dose thereof, wherein the patient's tumor-associated immune cells express LAG-3.

58. A method for treating recurrent, locally advanced, or metastatic gastric cancer or gastroesophageal junction adenocarcinoma in a human patient, wherein the patient (a) An anti-LAG-3 antibody comprising the CDR1, CDR2, and CDR3 domains of the heavy chain variable region having the sequence described in Sequence ID No. 3, and the CDR1, CDR2, and CDR3 domains of the light chain variable region having the sequence described in Sequence ID No. 5, (b) An anti-PD-1 antibody comprising the CDR1, CDR2, and CDR3 domains of the heavy chain variable region having the sequence described in Sequence ID No. 15 and the CDR1, CDR2, and CDR3 domains of the light chain variable region having the sequence described in Sequence ID No. 17, and (c) One or more standard therapy regimens A method comprising administering an effective dose of [a substance].

59. The method according to claim 57 or 58, wherein an anti-LAG-3 antibody and an anti-PD-1 antibody are administered as a fixed dose combination.

60. The method according to any one of claims 57 to 59, wherein one or more standard therapy regimens include the administration of docetaxel, doxorubicin hydrochloride, 5-fluorouracil, mitomycin C, fluorouracil / leucovorin calcium (FU-LV), docetaxel / cisplatin / fluorouracil (TPF), or capecitabine / irinotecan hydrochloride (XELIRI).

61. The method according to any one of claims 57 to 60, administered to a patient who has received prior treatment (for example, as a second-line therapy).

62. The method according to any one of claims 1 to 61, wherein the anti-LAG-3 antibody is lilatrimab.

63. The method according to any one of claims 1 to 62, wherein the anti-LAG-3 antibody comprises a mutation from serine to proline at amino acid residue 228.