Classifications

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  • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
  • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
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  • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
  • A61K39/39533—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals
  • A61K39/3955—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals against proteinaceous materials, e.g. enzymes, hormones, lymphokines
• • A—HUMAN NECESSITIES
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  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P35/00—Antineoplastic agents
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  • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
  • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
  • C07K16/2803—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
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  • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
  • C07K16/2803—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
  • C07K16/2818—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily against CD28 or CD152
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  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
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  • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
  • C07K16/2803—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
  • C07K16/2827—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily against B7 molecules, e.g. CD80, CD86
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
  • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
  • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
  • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
  • G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
  • G01N33/57407—Specifically defined cancers
  • G01N33/5743—Specifically defined cancers of skin, e.g. melanoma
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
  • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
  • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
  • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
  • G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
  • G01N33/57484—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
  • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
  • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
  • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
  • G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K39/00—Medicinal preparations containing antigens or antibodies
  • A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K39/00—Medicinal preparations containing antigens or antibodies
  • A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
  • A61K2039/507—Comprising a combination of two or more separate antibodies
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K2317/00—Immunoglobulins specific features
  • C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
  • C07K2317/21—Immunoglobulins specific features characterized by taxonomic origin from primates, e.g. man
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K2317/00—Immunoglobulins specific features
  • C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
  • C07K2317/31—Immunoglobulins specific features characterized by aspects of specificity or valency multispecific
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K2317/00—Immunoglobulins specific features
  • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
  • C07K2317/76—Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N2800/00—Detection or diagnosis of diseases
  • G01N2800/52—Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N2800/00—Detection or diagnosis of diseases
  • G01N2800/54—Determining the risk of relapse

Definitions

• the invention disclosed herein relates to methods of treating a LAG-3 positive malignant tumor in a human patient with a PD-1 pathway inhibitor, a combination of a PD1 pathway inhibitor and an immune checkpoint inhibitor, a combination of a LAG-3 inhibitor and a PD-1 pathway inhibitor, or an anti-CTLA-4 antibody.
• Lymphocyte activation gene-3 (LAG-3; CD223) is a type I transmembrane protein that is expressed on the cell surface of activated CD4+ and CD8+ T cells and subsets of NK 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, which is a co-receptor for T helper cell activation. Both molecules have 4 extracellular Ig-like domains and require binding to their ligand, major histocompatibility complex (MHC) class II, for their functional activity.
• MHC major histocompatibility complex
• LAG-3 is only expressed on the cell surface of activated T cells and its cleavage from the cell surface terminates LAG-3 signaling.
• LAG-3 can also be found as a soluble protein but it does not bind to MHC class II and the function of soluble LAG-3 is unknown.
• LAG-3 plays an important role in promoting regulatory T cell (Treg) activity and in negatively regulating T cell activation and proliferation (Workman C J, et al., J. Immunol. 2005; 174:688-695). Both natural and induced Treg express increased LAG-3, which is required for their maximal suppressive function (Camisaschi C, et al., J. Immunol. 2010; 184:6545-6551 and Huang C T, et al., Immunity. 2004; 21 :503-513).
• LAG-3 maintained tolerance to self and tumor antigens via direct effects on CD8+ T cells in 2 murine models (Grosso J F, et al., J. Clin. Invest. 2007; 117:3383-3392).
• Exhausted T cells are characterized by the expression of T cell negative regulatory receptors, predominantly Cytotoxic T-Lymphocyte Antigen-4 (CTLA-4), Programmed Cell Death 1 (PD-1), and LAG-3, whose action is to limit the cell's ability to proliferate, produce cytokines, and kill target cells and/or to increase Treg activity.
• CTL-4 Cytotoxic T-Lymphocyte Antigen-4
• PD-1 Programmed Cell Death 1
• LAG-3 LAG-3
• PD-1 is a cell surface signaling receptor that plays a critical role in the regulation of T cell activation and tolerance (Keir M E, et al., Annu Rev Immunol 2008; 26:677-704). It is a type I transmembrane protein and together with BTLA, CTLA-4, ICOS and CD28, comprise the CD28 family of T cell co-stimulatory 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).
• NK natural killer
• PD-1 One important role of PD-1 is to limit the activity of T cells in peripheral tissues at the time of an inflammatory response to infection, thus limiting the development of autoimmunity (Pardoll D M., Nat Rev Cancer 2012; 12:252-264).
• 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).
• the consequence is the development of immune resistance within the tumor microenvironment.
• PD-1 is highly expressed on tumor- infiltrating lymphocytes, and its ligands are up-regulated on the cell surface of many different tumors (Dong H, et al., Nat Med 2002; 8:793-800).
• One aspect of the invention disclosed herein relates to a method of selecting a malignant tumor in a human patient for treating with a PD-1 pathway inhibitor, a LAG-3 inhibitor, a combination of a PD1 pathway inhibitor and an immune checkpoint inhibitor, or a combination of a LAG-3 inhibitor and a PD-1 pathway inhibitor.
• the method comprises detecting LAG-3 expression in the tumor.
• the method comprises detecting LAG-3 expression and PD-L1 expression in the tumor.
• methods of treating LAG-3 positive tumors in a human patient comprising administering a LAG-3 inhibitor and a PD-1 pathway inhibitor.
• One aspect of the invention disclosed herein relates to a method of selecting a malignant tumor in a human patient for immunotherapy, comprising: determining the level of LAG-3 expression in a tumor sample; and selecting the tumor for immunotherapy if the tumor is a LAG-3 positive tumor.
• Another aspect of the invention disclosed herein relates to a method of identifying a malignant tumor in a human patient as eligible for immunotherapy, comprising: determining the level of LAG-3 expression in a tumor sample; and identifying the tumor as eligible for immunotherapy if the tumor is a LAG-3 positive tumor.
• Another aspect of the invention disclosed herein relates to a method of identifying a malignant tumor in a human patient that is likely to be responsive to an immunotherapy, the method comprising: determining the level of LAG-3 expression in a tumor sample; and identifying the tumor as likely to be responsive to treatment if the tumor is a LAG-3 positive tumor.
• Another aspect of the invention disclosed herein relates to a method of classifying a malignant tumor in a human patient as likely to be responsive to an immunotherapy, the method comprising: determining the level of LAG-3 expression in a tumor sample; and classifying the tumor as likely to be responsive to immunotherapy if the tumor is a LAG-3 positive tumor.
• a method disclosed herein further comprises determining the level of PD-L1 expression in the tumor sample.
• the immunotherapy comprises contacting the tumor with a therapeutically effective amount of a LAG-3 inhibitor and a PD-1 pathway inhibitor.
• the immunotherapy comprises contacting the tumor with a therapeutically effective amount of a LAG-3 inhibitor.
• the immunotherapy comprises contacting the tumor with a therapeutically effective amount of a PD-1 pathway inhibitor.
• the immunotherapy comprises contacting the tumor with a therapeutically effective amount of a PD-1 pathway inhibitor and an immune checkpoint inhibitor.
• a method disclosed herein comprises contacting the tumor with a therapeutically effective amount of a LAG-3 inhibitor and a PD-1 pathway inhibitor. In some embodiments, a method disclosed herein comprises contacting the tumor with a therapeutically effective amount of a LAG-3 inhibitor. In some embodiments, a method disclosed herein comprises contacting the tumor with a therapeutically effective amount of a PD-1 pathway inhibitor. In some embodiments, a method disclosed herein comprises contacting the tumor with a therapeutically effective amount of a PD-1 pathway inhibitor and an immune checkpoint inhibitor. In some embodiments, a method disclosed herein comprises administering to the patient a therapeutically effective amount of a LAG-3 inhibitor and a PD-1 pathway inhibitor.
• a method disclosed herein comprises administering to the patient a therapeutically effective amount of a LAG-3 inhibitor. In some embodiments, a method disclosed herein comprises administering to the patient a therapeutically effective amount of a PD-1 pathway inhibitor. In some embodiments, a method disclosed herein comprises administering to the patient a therapeutically effective amount of a PD-1 pathway inhibitor and an immune checkpoint inhibitor.
• Another aspect of the invention disclosed herein relates to a method of identifying a patient with a malignant tumor who is likely to respond to an immunotherapy, the method comprising: determining the level of LAG-3 expression in a tumor sample; and identifying the patient who is likely to respond to treatment if the tumor is a LAG-3 positive tumor.
• Another aspect of the invention disclosed herein relates to a method of selecting a patient with a malignant tumor for immunotherapy, the method comprising: determining the level of LAG-3 expression in a tumor sample; and selecting the patient for immunotherapy if the tumor is a LAG-3 positive tumor.
• a method disclosed herein further comprises determining the level of PD-L1 expression in the tumor sample.
• a method disclosed herein comprises administering to the patient a therapeutically effective amount of a LAG-3 inhibitor and a PD-1 pathway inhibitor. In some embodiments, a method disclosed herein comprises administering to the patient a therapeutically effective amount of a LAG-3 inhibitor. In some embodiments, a method disclosed herein comprises administering to the patient a therapeutically effective amount of a PD-1 pathway inhibitor. In some embodiments, a method disclosed herein comprises administering to the patient a therapeutically effective amount of a PD-1 pathway inhibitor and an immune checkpoint inhibitor.
• Another aspect of the invention disclosed herein relates to a method of treating a malignant tumor in a human patient, comprising: administering to the patient a therapeutically effective amount of a LAG-3 inhibitor and a PD-1 pathway inhibitor; wherein the patient is predicted to respond to treatment with the LAG-3 inhibitor and PD-1 pathway inhibitor based upon LAG-3 expression in a sample of the patient's tumor.
• Another aspect of the invention disclosed herein relates to a method of treating a malignant tumor in a human patient, comprising: administering to the patient a therapeutically effective amount of a LAG-3 inhibitor; wherein the patient is predicted to respond to treatment with the LAG-3 inhibitor based upon LAG-3 expression in a sample of the patient's tumor.
• Another aspect of the invention disclosed herein relates to a method of treating a malignant tumor in a human patient, comprising: administering to the patient a therapeutically effective amount of a PD-1 pathway inhibitor; wherein the patient is predicted to respond to treatment with the PD-1 pathway inhibitor based upon LAG-3 expression in a sample of the patient's tumor.
• Another aspect of the invention disclosed herein relates to a method of treating a malignant tumor in a human patient, comprising: administering to the patient a therapeutically effective amount of a PD-1 pathway inhibitor and an immune checkpoint inhibitor; wherein the patient is predicted to respond to treatment with the PD-1 pathway inhibitor and an immune checkpoint inhibitor based upon LAG-3 expression in a sample of the patient's tumor.
• the patient is predicted to respond to the treatment based upon LAG-3 and PD-L1 expression in a sample of the patient's tumor.
• Another aspect of the invention disclosed herein relates to a method of treating a malignant tumor in a human patient in need thereof, comprising: determining the level of LAG-3 expression in a tumor sample; and administering to the patient a therapeutically effective amount of a LAG-3 inhibitor and a PD-1 pathway inhibitor if the tumor is a LAG-3 positive tumor.
• Another aspect of the invention disclosed herein relates to a method of treating a malignant tumor in a human patient in need thereof, comprising: determining the level of LAG-3 expression in a tumor sample; and administering to the patient a therapeutically effective amount of a LAG- 3 inhibitor if the tumor is a LAG-3 positive tumor.
• Another aspect of the invention disclosed herein relates to a method of treating a malignant tumor in a human patient in need thereof, comprising: determining the level of LAG-3 expression in a tumor sample; and administering to the patient a therapeutically effective amount of a PD-1 pathway inhibitor if the tumor is a LAG- 3 positive tumor.
• Another aspect of the invention disclosed herein relates to a method of treating a malignant tumor in a human patient in need thereof, comprising: determining the level of LAG- 3 expression in a tumor sample; and administering to the patient a therapeutically effective amount of a PD-1 pathway inhibitor and an immune checkpoint inhibitor if the tumor is a LAG-3 positive tumor.
• a method disclosed herein further comprises determining the level of PD-L1 expression in the tumor sample.
• Another aspect of the invention disclosed herein relates to 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 and a PD-1 pathway inhibitor, wherein the patient is identified as having a LAG-3 positive malignant tumor prior to the administration.
• Another aspect of the invention disclosed herein relates to 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, wherein the patient is identified as having a LAG-3 positive malignant tumor prior to the administration.
• Another aspect of the invention disclosed herein relates to 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 PD-1 pathway inhibitor, wherein the patient is identified as having a LAG-3 positive malignant tumor prior to the administration.
• Another aspect of the invention disclosed herein relates to 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 PD-1 pathway inhibitor and an immune checkpoint inhibitor, wherein the patient is identified as having a LAG-3 positive malignant tumor prior to the administration.
• the patient is identified as having a LAG-3 positive PD-L1 positive malignant tumor prior to the administration.
• the patient is identified as having a LAG-3 positive PD-L1 negative malignant tumor prior to the administration.
• Another aspect of the invention disclosed herein relates to a method for treating a malignant tumor in a human patient in need thereof, comprising: identifying the patient as having a LAG-3 positive malignant tumor; and administering to the patient a therapeutically effective amount of a LAG-3 inhibitor and a PD-1 pathway inhibitor.
• Another aspect of the invention disclosed herein relates to a method for treating a malignant tumor in a human patient in need thereof, comprising: identifying the patient as having a LAG-3 positive malignant tumor; and administering to the patient a therapeutically effective amount of a LAG-3 inhibitor.
• Another aspect of the invention disclosed herein relates to a method for treating a malignant tumor in a human patient in need thereof, comprising: identifying the patient as having a LAG-3 positive malignant tumor; and administering to the patient a therapeutically effective amount of a PD-1 pathway inhibitor.
• Another aspect of the invention disclosed herein relates to a method for treating a malignant tumor in a human patient in need thereof, comprising: identifying the patient as having a LAG-3 positive malignant tumor; and administering to the patient a therapeutically effective amount of a PD-1 pathway inhibitor and an immune checkpoint inhibitor.
• a method disclosed herein further comprises identifying the patient as having a LAG-3 positive PD-Ll positive malignant tumor.
• a method disclosed herein further comprises identifying the patient as having a LAG-3 positive PD-Ll negative malignant tumor.
• Another aspect of the invention disclosed herein relates to a method for extending a progression-free survival period for over 12 months in a human patient afflicted with a malignant tumor comprising administering to the patient a LAG-3 inhibitor and a PD-1 pathway inhibitor, wherein the patient is identified as having a LAG-3 positive malignant tumor prior to the administration and wherein the patient demonstrates progression-free survival for over 12 months.
• Another aspect of the invention disclosed herein relates to a method for extending a progression-free survival period for over 12 months in a human patient afflicted with a malignant tumor comprising administering to the patient a LAG-3 inhibitor, wherein the patient is identified as having a LAG-3 positive malignant tumor prior to the administration and wherein the patient demonstrates progression-free survival for over 12 months.
• Another aspect of the invention disclosed herein relates to a method for extending a progression-free survival period for over 12 months in a human patient afflicted with a malignant tumor comprising administering to the patient a PD-1 pathway inhibitor, wherein the patient is identified as having a LAG-3 positive malignant tumor prior to the administration and wherein the patient demonstrates progression- free survival for over 12 months.
• Another aspect of the invention disclosed herein relates to a method for extending a progression-free survival period for over 12 months in a human patient afflicted with a malignant tumor comprising administering to the patient a PD-1 pathway inhibitor and an immune checkpoint inhibitor, wherein the patient is identified as having a LAG-
• the patient is identified as having a LAG-3 positive PD-L1 positive malignant tumor prior to the administration. In some embodiments, the patient is identified as having a LAG-3 positive PD-L1 negative malignant tumor prior to the administration. In some embodiments, the progression-free survival of the patient is extended after the administration for over about 13 months, about 14 months, about 15 months, about 16 months, about 17 months, about 18 months, about 2 years, about 3 years, about
• the progression-free survival of the patient is extended for over 14 months.
• Another aspect of the invention disclosed herein relates to a method for reducing a tumor size at least by 10% in a human patient afflicted with a malignant tumor comprising administering to the patient a therapeutically effective amount of a LAG-3 inhibitor and a PD-1 pathway inhibitor, wherein the patient is identified as having a LAG-3 positive malignant tumor prior to the administration and wherein the administration reduces the tumor size 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 the administration.
• Another aspect of the invention disclosed herein relates to a method for reducing a tumor size at least by 10%> in a human patient afflicted with a malignant tumor comprising administering to the patient a therapeutically effective amount of a LAG-3 inhibitor, wherein the patient is identified as having a LAG-3 positive malignant tumor prior to the administration and wherein the administration reduces the tumor size 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 the administration.
• Another aspect of the invention disclosed herein relates to a method for reducing a tumor size at least by 10% in a human patient afflicted with a malignant tumor comprising administering to the patient a therapeutically effective amount of a PD-1 pathway inhibitor, wherein the patient is identified as having a LAG-3 positive malignant tumor prior to the administration and wherein the administration reduces the tumor size 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 the administration.
• Another aspect of the invention disclosed herein relates to a method for reducing a tumor size at least by 10% in a human patient afflicted with a malignant tumor comprising administering to the patient a therapeutically effective amount of a PD-1 pathway inhibitor and an immune checkpoint inhibitor, wherein the patient is identified as having a LAG-3 positive malignant tumor prior to the administration and wherein the administration reduces the tumor size 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 the administration.
• the patient is identified as having a LAG-3 positive PD-Ll positive malignant tumor prior to the administration.
• the patient is identified as having a LAG-3 positive PD-Ll negative malignant tumor prior to the administration.
• a method disclosed herein further comprises identifying the patient as having a LAG-3 positive malignant tumor prior to the administration.
• a method disclosed herein further comprises identifying the patient as having a LAG-3 positive PD-Ll positive malignant tumor prior to the administration.
• a method disclosed herein further comprises identifying the patient as having a LAG-3 positive PD-Ll negative malignant tumor prior to the administration.
• the patient experiences (i) extended progression-free survival for over 12 months, (ii) tumor size reduction at least about 10%, about 20%, about 30%, about 40%, or about 50% compared to the tumor size prior to the administration, or (iii) both.
• Another aspect of the invention disclosed herein relates to a method for increasing an objective response rate to a cancer treatment to be higher than 50% in a human patient population, each of whom is afflicted with a malignant tumor, to a cancer treatment comprising administering to the patient a therapeutically effective amount of a LAG-3 inhibitor and a PD-1 pathway inhibitor, wherein each patient is identified as having a LAG-3 positive malignant tumor prior to the administration and wherein the objective response rate is higher than 55%, 60%, 65%, 70%), or 75%).
• Another aspect of the invention disclosed herein relates to a method for increasing an objective response rate to a cancer treatment to be higher than 50% in a human patient population, each of whom is afflicted with a malignant tumor, to a cancer treatment comprising administering to the patient a therapeutically effective amount of a LAG-3 inhibitor, wherein each patient is identified as having a LAG-3 positive malignant tumor prior to the administration and wherein the objective response rate is higher than 55%, 60%, 65%, 70%, or 75%.
• Another aspect of the invention disclosed herein relates to a method for increasing an objective response rate to a cancer treatment to be higher than 50% in a human patient population, each of whom is afflicted with a malignant tumor, to a cancer treatment comprising administering to the patient a therapeutically effective amount of a PD-1 pathway inhibitor, wherein each patient is identified as having a LAG-3 positive malignant tumor prior to the administration and wherein the objective response rate is higher than 55%, 60%, 65%, 70%, or 75%.
• Another aspect of the invention disclosed herein relates to a method for increasing an objective response rate to a cancer treatment to be higher than 50% in a human patient population, each of whom is afflicted with a malignant tumor, to a cancer treatment comprising administering to the patient a therapeutically effective amount of a PD-1 pathway inhibitor and an immune checkpoint inhibitor, wherein each patient is identified as having a LAG-3 positive malignant tumor prior to the administration and wherein the objective response rate is higher than 55%, 60%, 65%, 70%, or 75%).
• each patient is identified as having a LAG-3 positive PD-L1 positive malignant tumor prior to the administration.
• each patient is identified as having a LAG-3 positive PD-L1 negative malignant tumor prior to the administration.
• Another aspect of the invention disclosed herein relates to a method for increasing a disease control rate to be higher than 50% in a human patient population, each of whom is afflicted with a malignant tumor, to a cancer treatment comprising administering to the patient a therapeutically effective amount of a LAG-3 inhibitor and a PD-1 pathway inhibitor, wherein each patient is identified as having a LAG-3 positive malignant tumor prior to the administration and wherein the objective response rate is higher than 55%, 60%, 65%, 70%, or 75%.
• Another aspect of the invention disclosed herein relates to a method for increasing a disease control rate to be higher than 50% in a human patient population, each of whom is afflicted with a malignant tumor, to a cancer treatment comprising administering to the patient a therapeutically effective amount of a LAG-3 inhibitor, wherein each patient is identified as having a LAG-3 positive malignant tumor prior to the administration and wherein the objective response rate is higher than 55%), 60%), 65%), 70%), or 75%.
• Another aspect of the invention disclosed herein relates to a method for increasing a disease control rate to be higher than 50% in a human patient population, each of whom is afflicted with a malignant tumor, to a cancer treatment comprising administering to the patient a therapeutically effective amount of a PD-1 pathway inhibitor, wherein each patient is identified as having a LAG-3 positive malignant tumor prior to the administration and wherein the objective response rate is higher than 55%, 60%, 65%, 70%, or 75%).
• Another aspect of the invention disclosed herein relates to a method for increasing a disease control rate to be higher than 50% in a human patient population, each of whom is afflicted with a malignant tumor, to a cancer treatment comprising administering to the patient a therapeutically effective amount of a PD-1 pathway inhibitor and an immune checkpoint inhibitor, wherein each patient is identified as having a LAG-3 positive malignant tumor prior to the administration and wherein the objective response rate is higher than 55%, 60%, 65%, 70%, or 75%.
• each patient is identified as having a LAG-3 positive PD-L1 positive malignant tumor prior to the administration.
• each patient is identified as having a LAG-3 positive PD-L1 negative malignant tumor prior to the administration.
• the median duration of response is > 3 month, > 6 month, > 12 month, or > 18 month.
• a method disclosed herein further comprises identifying each patient of the patient population as having a LAG-3 positive malignant tumor prior to the administration. In some embodiments, a method disclosed herein further comprises identifying each patient of the patient population as having a LAG-3 positive PD-L1 positive malignant tumor prior to the administration. In some embodiments, a method disclosed herein further comprises identifying each patient of the patient population as having a LAG-3 positive PD-L1 negative malignant tumor prior to the administration.
• each patient of the patient population is further characterized by (i) extended progression-free survival for over 12 months, (ii) tumor size reduction at least about 10%, about 20%, about 30%, about 40%, or about 50%) compared to the tumor size prior to the administration, or (iii) both.
• the patient population comprises at least about 100, 200, 300, 400, 500, 600, 700, 800, 900, or 1000 patients having a LAG-3 positive malignant tumor.
• Another aspect of the invention disclosed herein relates to a method for selecting a human patient suitable for a combination therapy comprising: identifying a patient as having a LAG-3 positive malignant tumor; and instructing a healthcare provider to administer to the patient a therapeutically effective amount of a LAG-3 inhibitor and a PD-1 pathway inhibitor.
• Another aspect of the invention disclosed herein relates to a method for selecting a human patient suitable for a combination therapy comprising: identifying a patient as having a LAG-3 positive malignant tumor; and instructing a healthcare provider to administer to the patient a therapeutically effective amount of a LAG-3 inhibitor.
• Another aspect of the invention disclosed herein relates to a method for selecting a human patient suitable for a combination therapy comprising: identifying a patient as having a LAG-3 positive malignant tumor; and instructing a healthcare provider to administer to the patient a therapeutically effective amount of a PD-1 pathway inhibitor.
• Another aspect of the invention disclosed herein relates to a method for selecting a human patient suitable for a combination therapy comprising: identifying a patient as having a LAG-3 positive malignant tumor; and instructing a healthcare provider to administer to the patient a therapeutically effective amount of a PD-1 pathway inhibitor and an immune checkpoint inhibitor.
• a method disclosed herein further comprises identifying the patient as having a LAG-3 positive PD-Ll positive malignant tumor.
• a method disclosed herein further comprises identifying the patient as having a LAG-3 positive PD-Ll negative malignant tumor.
• the administration treats the malignant tumor.
• identifying the patient as having a LAG-3 positive malignant tumor comprises determining LAG-3 expression in the malignant tumor. In some embodiments, identifying the patient as having a LAG-3 positive PD-Ll positive malignant tumor comprises determining PD-Ll expression in the malignant tumor. In some embodiments, identifying the patient as having a LAG-3 positive PD-Ll negative malignant tumor comprises determining PD-Ll expression in the malignant tumor. In some embodiments, LAG-3 expression is determined by reviewing the results of an assay capable of determining LAG-3 expression. In some embodiments, LAG-3 expression is determined by reviewing the results of an immunohistochemistry assay capable of detecting LAG-3 expression.
• PD- Ll expression is determined by reviewing the results of an assay capable of determining PD-Ll expression. In some embodiments, PD-Ll expression is determined by reviewing the results of an immunohistochemistry assay capable of detecting PD-Ll expression.
• a LAG-3 positive tumor comprises 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% cells expressing LAG-3.
• a LAG-3 positive tumor comprises at least about 1% cells expressing LAG-3.
• a LAG-3 positive tumor comprises at least about 5% cells expressing LAG-3.
• the cells expressing LAG-3 comprise tumor infiltrating lymphocytes.
• the cells expressing LAG-3 are the total number of cells. In other embodiments, the cells express LAG-3 on the cell surface.
• the malignant tumor is selected from the group consisting of a liver cancer, bone cancer, pancreatic cancer, skin cancer, oral cancer, cancer of the head or neck, breast cancer, lung cancer, including small cell and non-small cell lung cancer, cutaneous or intraocular malignant melanoma, renal cancer, uterine cancer, ovarian cancer, colorectal cancer, colon cancer, rectal cancer, cancer of the anal region, stomach cancer, testicular cancer, uterine cancer, carcinoma of the fallopian tubes, carcinoma of the endometrium, carcinoma of the cervix, carcinoma of the vagina, carcinoma of the vulva, non-Hodgkin's lymphoma, cancer of the esophagus, cancer of the small intestine, cancer of the endocrine system, cancer of the thyroid gland, cancer of the parathyroid gland, cancer of the adrenal gland
• the malignant tumor is chosen from melanoma, non-small cell lung cancer (NSCLC), human papilloma virus (HPV)-related tumor, and gastric adenocarcinoma.
• NSCLC non-small cell lung cancer
• HPV human papilloma virus
• gastric adenocarcinoma gastric adenocarcinoma
• the malignant tumor is NSCLC, a virally-related cancer related tumor, or gastric adenocarcinoma.
• the malignant tumor is melanoma, gastric cancer, gastroesophageal junction cancer, non-small cell lung cancer, bladder cancer, head and neck squamous cell carcinoma, or renal cell cancer.
• the malignant tumor is lung cancer, melanoma, squamous cell carcinoma of the head and neck, renal cancer, gastric cancer, or hepatocellular carcinoma.
• the LAG-3 positive malignant tumor is a melanoma tumor comprising about 1% or more cells expressing LAG-3.
• the LAG-3 positive malignant tumor is a gastric cancer tumor comprising about 1% or more cells expressing LAG-3.
• the malignant tumor is refractory to treatment with an immune checkpoint inhibitor. In some embodiments, the malignant tumor is refractory to treatment with an anti-PD-1 antibody. In some embodiments, the malignant tumor is refractory to treatment with an anti-PD-Ll antibody.
• Another aspect of the invention disclosed herein relates to a method for treating melanoma in a human patient, comprising: identifying the patient as having a LAG-3 positive melanoma; and administering to the patient a therapeutically effective amount of a LAG-3 inhibitor and a PD-1 pathway inhibitor.
• identifying the patient as having a LAG-3 positive melanoma comprises determining LAG-3 expression in the melanoma tumor.
• LAG-3 expression is determined by reviewing the results of an assay capable of determining LAG-3 expression.
• LAG-3 expression is determined by an immunohistochemistry assay capable of detecting LAG-3 expression.
• a method disclosed herein further comprises identifying the patient as having a LAG-3 positive PD-Ll positive malignant tumor. In some embodiments, a method disclosed herein further comprises identifying the patient as having a LAG-3 positive PD-Ll negative malignant tumor.
• Another aspect of the invention disclosed herein relates to a method for treating a melanoma in a human patient in need thereof comprising administering to the patient a therapeutically effective amount of a LAG-3 inhibitor and a PD-1 pathway inhibitor, wherein the patient is identified as having a LAG-3 positive melanoma prior to the administration.
• Another aspect of the invention disclosed herein relates to a method for extending a progression-free survival period for over 12 months in a human patient afflicted with a melanoma comprising administering to the patient a LAG-3 inhibitor and a PD-1 pathway inhibitor, wherein the patient is identified as having a LAG-3 positive melanoma prior to the administration and wherein the patient demonstrates progression-free survival for over 12 months.
• the patient is identified as having a LAG-3 positive PD-Ll positive melanoma prior to the administration.
• the patient is identified as having a LAG-3 positive PD-Ll negative melanoma prior to the administration.
• Another aspect of the invention disclosed herein relates to a method for increasing an objective response rate to a cancer treatment to be higher than 15% in a human patient population, each of whom is afflicted with melanoma, comprising administering to the patient a therapeutically effective amount of a LAG-3 inhibitor and a PD-1 pathway inhibitor, wherein each patient is identified as having a LAG-3 positive malignant tumor prior to the administration and wherein the objective response rate is higher than 15%.
• Another aspect of the invention disclosed herein relates to a method for increasing a disease control rate to a cancer treatment to be higher than 70% in a human patient population, each of whom is afflicted with melanoma, comprising administering to the patient a therapeutically effective amount of a LAG-3 inhibitor and a PD-1 pathway inhibitor, wherein each patient is identified as having a LAG-3 positive melanoma prior to the administration and wherein the objective response rate is higher than 70%.
• a method disclosed herein further comprises identifying each patient of the patient population as having a LAG-3 positive melanoma prior to the administration.
• the median duration of response is > 3 month, > 6 month, > 12 month, or > 18 month.
• each patient is identified as having a LAG-3 positive PD-L1 positive melanoma prior to the administration. In some embodiments, each patient is identified as having a LAG-3 positive PD-L1 negative melanoma prior to the administration.
• the melanoma is refractory to treatment with an immune checkpoint inhibitor. In some embodiments, the melanoma is refractory to treatment with an anti- PD-1 antibody or an anti-PD-Ll antibody.
• determining the level of LAG-3 and/or PD-L1 expression comprises providing a test tissue sample obtained from the patient, the test tissue sample comprising tumor cells and/or tumor-infiltrating immune cells.
• the test tissue sample is a tumor biopsy.
• the test tissue sample is a formalin-fixed paraffin embedded (FFPE) sample.
• determining comprises detecting LAG-3 and/or PD-L1 protein or RNA expression in the test tissue sample.
• LAG-3 and/or PD-L1 expression is detected by an assay capable of detecting the level of LAG-3 and/or PD-L1 protein, respectively, in the test tissue sample.
• LAG-3 and/or PD-L1 expression is detected by an immunohistochemistry assay.
• the immunohistochemistry assay is a monoplex assay (assay designed to detect/measure the presence of a single analyte, e.g., antigen/antibody pair).
• the immunohistochemistry assay is a multiplex assay (assay designed to detect/measure multiple analytes, either simultaneously or sequentially).
• the immunohistochemistry assay comprises contacting the tumor sample with the 17B4, SP346, 11E3, 874501, or EPR4392(2) anti-human LAG-3 monoclonal antibody.
• the immunohistochemistry assay comprises contacting the tumor sample with an anti-LAG-3 antibody comprising heavy and light chain variable regions comprising the sequences set forth in SEQ ID NOs:3 and 5, respectively.
• the immunohistochemistry assay uses a black or brown chromogen. In some embodiments, the immunohistochemistry assay uses a red chromogen. In some embodiments, the immunohistochemistry assay uses a blue chromogen. In some embodiments, the immunohistochemistry assay uses a green chromogen. In some embodiments, the immunohistochemistry assay uses a purple chromogen. In certain embodiments, the immunohistochemistry assay uses a yellow chromogen.
• the immunohistochemistry assay is scored at a low magnification (e.g., 4X or 10X). In some embodiments, low magnification is about 20X.
• the immunohistochemistry assay is scored at high magnification.
• high magnification is about 40X, or greater (60X, 100X).
• the immunohistochemistry assay is scored by an image analysis software. In some embodiments, the immunohistochemistry assay is scored manually by a pathologist.
• scoring the immunohistochemistry assay comprises assessing the proportion of cells in the test tissue sample that express LAG-3 and/or assessing the proportion of cells in the test tissue sample that express PD-L1. In some embodiments, scoring the immunohistochemistry assay comprises assessing the proportion of tumor cells in the test tissue sample that express LAG-3 and/or assessing the proportion of tumor cells in the test tissue sample that express PD-L1. In some embodiments, scoring the immunohistochemistry assay comprises assessing the proportion of immune cells in the test tissue sample that express LAG-3 and/or assessing the proportion of immune cells in the test tissue sample that express PD-L1.
• scoring the immunohistochemistry assay comprises assessing the proportion of T cells in the test tissue sample that express LAG-3 and/or assessing the proportion of T cells in the test tissue sample that express PD-L1. In some embodiments, scoring the immunohistochemistry assay comprises assessing the proportion of CD8+ T cells in the test tissue sample that express LAG-3 and/or assessing the proportion of CD8+ T cells in the test tissue sample that express PD-L1. In some embodiments, scoring the immunohistochemistry assay comprises assessing the proportion of CD4+ T cells in the test tissue sample that express LAG-3 and/or assessing the proportion of CD4+ T cells in the test tissue sample that express PD- Ll . In some embodiments, scoring the immunohistochemistry assay comprises assessing the proportion of FOXP3+ T cells in the test tissue sample that express LAG-3 and/or assessing the proportion of FOXP3+ T cells in the test tissue sample that express PD-L1.
• cells with partial membrane/cytoplasmic LAG-3 localization are scored as LAG-3 expressing cells.
• cells with dot-like LAG-3 localization are scored as LAG-3 expressing cells.
• cells with complete membrane/cytoplasmic LAG-3 localization are scored as LAG-3 expressing cells.
• cells with any LAG-3 localization pattern are scored as LAG-3 expressing cells.
• the immunohistochemistry assay is a multiplex assay that further comprises detecting the expression of MHC Class II by the tumor cells. In some embodiments, scoring the immunohistochemistry assay comprises assessing the proportion of cells in the test tissue sample that expresses MHC Class II. In some embodiments, scoring the immunohistochemistry assay comprises assessing the proportion of non-immune cells in the test tissue sample that expresses MHC II.
• LAG-3 and/or PD-L1 protein expression is detected by flow cytometry.
• the test tissue sample obtained from the patient comprises tumor infiltrating immune cells.
• the malignant tumor is a hematological malignancy and the tissue sample comprises circulating lymphocytes.
• the flow cytometry is a multiplex assay.
• the flow cytometry comprises detecting the expression of markers comprising LAG-3, PD-L1, CD4, CD8, FOXP3, MHC Class II and any combination thereof.
• scoring the flow cytometry comprises assessing the proportion of T cells in the test tissue sample that express LAG-3. In some embodiments, scoring the flow cytometry comprises assessing the proportion of CD8+ T cells in the test tissue sample that express LAG-3. In some embodiments, scoring the flow cytometry comprises assessing the proportion of CD4+ T cells in the test tissue sample that express LAG-3. In some embodiments, scoring the flow cytometry comprises assessing the proportion of FOXP3+ T cells in the test tissue sample that express LAG-3.
• LAG-3 and/or PD-L1 expression is detected by an assay capable of detecting the level of LAG-3 and/or PD-L1, respectively, RNA in the tumor sample.
• LAG-3 and/or PD-L1 expression is detected by an RT-PCR based assay.
• scoring the RT-PCR based assay comprises assessing the level of LAG-3 and/or PD-L1 RNA expression in the test tissue sample relative to a predetermined level.
• the LAG-3 inhibitor is an anti-LAG-3 antibody or antigen- binding fragment thereof.
• the anti-LAG-3 antibody is a bispecific antibody.
• the anti-LAG-3 antibody or antigen-binding fragment thereof comprises (a) a heavy chain variable region CDRl comprising the sequence set forth in SEQ ID NO:7; (b) a heavy chain variable region CDR2 comprising the sequence set forth in SEQ ID NO:8; (c) a heavy chain variable region CDR3 comprising the sequence set forth in SEQ ID NO:9; (d) a light chain variable region CDRl comprising the sequence set forth in SEQ ID NO: 10; (e) a light chain variable region CDR2 comprising the sequence set forth in SEQ ID NO: 11; and (f) a light chain variable region CDR3 comprising the sequence set forth in SEQ ID NO: 12.
• the anti-LAG-3 antibody or antigen-binding fragment thereof comprises heavy and light chain variable regions comprising the sequences set forth in SEQ ID NOs:3 and 5, respectively.
• the anti-LAG-3 antibody is MK-4280 (28G-10),
• the PD-1 pathway inhibitor is an anti-PD-1 antibody or antigen-binding fragment thereof.
• the anti-PD-1 antibody or antigen- binding fragment thereof comprises (a) a heavy chain variable region CDRl comprising the sequence set forth in SEQ ID NO:23; (b) a heavy chain variable region CDR2 comprising the sequence set forth in SEQ ID NO:24; (c) a heavy chain variable region CDR3 comprising the sequence set forth in SEQ ID NO:25; (d) a light chain variable region CDRl comprising the sequence set forth in SEQ ID NO:26; (e) a light chain variable region CDR2 comprising the sequence set forth in SEQ ID NO:27; and (f) a light chain variable region CDR3 comprising the sequence set forth in SEQ ID NO:28.
• the anti-PD-1 antibody or antigen-binding fragment thereof comprises heavy and light chain variable regions comprising the sequences set forth in SEQ ID NOs: 19 and 21, respectively.
• the anti-PD-1 antibody or antigen-binding fragment thereof comprises heavy and light chains comprising the sequences set forth in SEQ ID NOs: 17 and 18, respectively.
• the anti-PD-1 antibody is pembrolizumab (KEYTRUDA;
• MK-3475 MK-3475
• pidilizumab CT-011
• nivolumab OPDIVO; BMS-936558
• the PD-1 pathway inhibitor is an anti-PD-Ll antibody or antigen-binding fragment thereof.
• the anti-PD-Ll antibody is atezolizumab (Tecentriq or RG7446), durvalumab (Imfinzi or MEDI4736), avelumab (Bavencio) or BMS-936559.
• the PD-1 pathway inhibitor is an anti-PD-L2 antibody or antigen-binding fragment thereof.
• the immune checkpoint inhibitor is a CTLA-4 antagonist, a
• CD80 antagonist a CD86 antagonist, a Tim-3 antagonist, a TIGIT antagonist, a CD20 antagonist, a CD96 antagonist, a IDOl antagonist, a STING antagonist, a GARP antagonist, a CD40 antagonist, A2aR antagonist, a CEACAMl (CD66a) antagonist, a CEA antagonist, a CD47 antagonist a PVRIG antagonist, a TDO antagonist, a VISTA antagonist, or a KIR antagonist.
• CD80 antagonist a CD86 antagonist, a Tim-3 antagonist, a TIGIT antagonist, a CD20 antagonist, a CD96 antagonist, a IDOl antagonist, a STING antagonist, a GARP antagonist, a CD40 antagonist, A2aR antagonist, a CEACAMl (CD66a) antagonist, a CEA antagonist, a CD47 antagonist a PVRIG antagonist, a TDO antagonist, a VISTA antagonist, or a KIR antagonist.
• the method comprises at least one administration cycle, wherein the cycle is a period of eight weeks, wherein for each of the at least one cycles, four doses of the anti-LAG-3 antibody are administered at a dose of 3, 20, 80, 160, or 240 mg.
• the method comprises at least one administration cycle, wherein the cycle is a period of eight weeks, wherein for each of the at least one cycles, four doses of the anti-PD-1 antibody are administered at a dose of 80 or 240 mg.
• the method comprises at least one administration cycle, wherein the cycle is a period of eight weeks, wherein for each of the at least one cycles, four doses of the anti-PD-Ll antibody are administered at a dose of 3, 20, 80, 160, or 240 mg.
• the method comprises at least one administration cycle, wherein the cycle is a period of eight weeks, wherein for each of the at least one cycles, four doses of the anti-LAG-3 antibody are administered at a dose of 3, 20, 80, 160, or 240 mg and four doses of the anti-PD-1 antibody are administered at a dose of 80 or 240 mg.
• the anti-LAG-3 antibody and anti-PD-1 antibody are administered at the following doses: (a) 3 mg of anti-LAG-3 antibody and 80 mg of anti-PD-1 antibody; (b) 3 mg of anti-LAG-3 antibody and 240 mg of anti-PD-1 antibody; (c) 20 mg of anti- LAG-3 antibody and 240 mg of anti-PD-1 antibody; (d) 80 mg of anti-LAG-3 antibody and 160 mg of anti-PD-1 antibody; (e) 80 mg of anti-LAG-3 antibody and 240 mg of anti-PD-1 antibody; (f) 160 mg of anti-LAG-3 antibody and 240 mg of anti-PD-1 antibody, or (g) 240 mg of anti- LAG-3 antibody and 240 mg of anti-PD-1 antibody.
• the anti-LAG-3 antibody and anti-PD-1 antibody are administered at the dose of 80 mg of anti-LAG-3 antibody and 160 mg of anti-PD-1 antibody.
• the anti-LAG-3 antibody and anti-PD-1 antibody are administered at the dose of 80 mg of anti-LAG-3 antibody and 240 mg of anti-PD-1 antibody.
• the anti-LAG-3 antibody and anti-PD-1 antibody are administered at the dose of 160 mg of anti-LAG-3 antibody and 240 mg of anti-PD-1 antibody.
• the anti-PD-1 and anti-LAG-3 antibodies or antigen- binding fragments thereof are formulated for intravenous administration.
• the anti-PD-1 and anti-LAG-3 antibodies or antigen- binding fragments thereof are formulated together. In some embodiments, the anti-PD-1 and anti- LAG-3 antibodies or antigen-binding fragments thereof are formulated separately.
• the treatment consists of up to 12 cycles.
• anti-PD-1 antibody or antigen-binding fragment thereof is administered on Days 1, 15, 29, and 43 of each cycle.
• anti-LAG-3 antibody or antigen-binding fragment thereof is administered on Days 1, 15, 29, and 43 of each cycle.
• the anti-PD-1 antibody or antigen-binding fragment thereof is administered prior to administration of the anti-LAG-3 antibody or antigen-binding fragment thereof. In some embodiments, the anti-LAG-3 antibody or antigen-binding fragment thereof is administered within about 30 minutes prior to administration of the anti-PD-1 antibody or antigen-binding fragment thereof. In some embodiments, the anti-PD-1 antibody or antigen-bind fragment thereof is administered after administration of the anti-LAG-3 antibody or antigen-binding fragment thereof. In some embodiments, the anti-PD-1 antibody or antigen- binding fragment thereof is administered before administration of the anti-LAG-3 antibody or antigen-binding fragment thereof. In some embodiments, the anti-PD-1 antibody or antigen- binding fragment thereof is administered concurrently with the anti-LAG-3 antibody or antigen- binding fragment thereof.
• the anti-LAG-3 antibody or antigen-binding fragment thereof and PD-1 pathway inhibitor are administered as a first line of treatment. In some embodiments, the anti-LAG-3 antibody or antigen-binding fragment thereof and PD-1 pathway inhibitor are administered as a second line of treatment.
• a method disclosed herein further comprises the administration of at least one additional therapeutic agent.
• the at least one additional therapeutic agent is a chemotherapeutic agent.
• the at least one additional therapeutic agent is an immune checkpoint inhibitor.
• the method produces at least one therapeutic effect chosen from a reduction in size of a tumor, reduction in number of metastatic lesions over time, complete response, partial response, and stable disease.
• administering the anti-LAG-3 antibody or antigen-binding fragment thereof and PD-1 pathway inhibitor activates the patient's T cells. In some embodiments, administering the anti-LAG-3 antibody or antigen-binding fragment thereof and PD-1 pathway inhibitor induces the expression activation markers by the patient's T cells.
• administering the anti-LAG-3 antibody or antigen-binding fragment thereof results in the occupancy of at least about 10%, at least about 20%, 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%, at least about 95%, or about 100% of the LAG-3 receptors on the patient's T cells.
• the T cells are CD8+ T cells.
• the T cells are tumor infiltrating T cells.
• the PD-1 pathway inhibitor comprises an anti-PD-1 antibody or antigen-binding fragment thereof.
• kits for treating a patient afflicted with a malignant tumor comprising: a dosage ranging from about 0.1 to about 10 mg/kg body weight of an anti-LAG-3 antibody or an antigen-binding fragment thereof; a dosage ranging from about 0.1 to about 10 mg/kg body weight of an anti-PD-1 antibody or an antigen-binding fragment thereof; and instructions for using the anti-LAG-3 antibody and anti- PD-1 antibody or the antigen-binding fragments thereof in any of the methods disclosed herein.
• kits for treating a patient afflicted with a malignant tumor comprising: a dosage ranging from about 0.1 to about 10 mg/kg body weight of an anti-PDl antibody or an antigen-binding fragment thereof; a dosage an immune checkpoint inhibitor; and instructions for using the anti-PD-1 antibody or antigen-binding fragment thereof and immune checkpoint inhibitor in any of the methods disclosed herein.
• kits for treating a patient afflicted with a malignant tumor comprising: a dosage ranging from about 0.1 to about 10 mg/kg body weight of an anti-LAG-3 antibody or an antigen-binding fragment thereof; and instructions for using the anti-LAG-3 antibody or the antigen-binding fragment thereof in any of the methods disclosed herein.
• kits for treating a patient afflicted with a malignant tumor comprising: a dosage ranging from 0.1 to 10 mg/kg body weight of an anti-PD-1 antibody or an antigen-binding fragment; and instructions for using the anti-PD-1 antibody or the antigen-binding fragment thereof in any of the methods disclosed herein.
• An aspect of the invention relates to a method of identifying a patient that is refractory to treatment with a PD-1 antagonist, the method comprising determining the level of LAG-3 expression, wherein an increased level of LAG-3 expression following treatment with the PD-1 antagonist, relative to the level of LAG-3 expression prior to treatment with the PD-1 antagonist, indicates that a patient is refractory to PD-1 antagonist therapy.
• Another aspect of the invention relates to a method of identifying a patient that is at risk of becoming refractory to treatment with a PD-1 antagonist, the method comprising determining the level of LAG-3 expression, wherein an increased level of LAG-3 expression following treatment with the PD-1 antagonist, relative to the level of LAG-3 expression prior to treatment with the PD-1 antagonist, indicates that a patient is at risk of becoming refractory to PD-1 antagonist therapy.
• Some aspects of the invention relate to a method of identifying a patient who is likely to respond to a LAG-3 therapy, the method comprising determining the level of LAG-3 expression in the patient, wherein an increased level of LAG-3 expression following treatment with a PD-1 antagonist, relative to the level of LAG-3 expression prior to treatment with the PD-1 antagonist, indicates that a patient is likely to respond to a LAG-3 therapy.
• Certain aspects of the invention relate to a method of selecting a patient for treatment with a LAG-3 therapy, the method comprising determining the level of LAG-3 expression in the patient, wherein an increased level of LAG-3 expression following treatment with a PD-1 antagonist, relative to the level of LAG-3 expression prior to treatment with the PD-1 antagonist, indicates that a patient is likely to respond to a LAG- 3 therapy.
• the PD-1 antagonist is a PD-1 inhibitor.
• the PD-1 antagonist is a PD-1 antibody.
• the LAG-3 therapy is a LAG-3 inhibitor.
• the LAG-3 therapy is an anti-LAG-3 antibody.
• the LAG-3 therapy is a combination therapy.
• the LAG- 3 combination therapy is a combination of an anti-LAG-3 antibody and an anti-PD-1 antibody.
• Figure 2 Frequency distribution of LAG-3+ cells as a ratio of total tumor cells in a sample analyzed with monoplex LAG-3 IHC.
• Figures 3A-B (Figure 3 A) Study design and endpoints. ( Figure 3B) Key eligibility criteria for patients in the melanoma prior IO expansion cohort.
• Figure 4 Baseline demographics and disease characteristics.
• Figure 7 Response by investigator assessment of patients with melanoma who progressed on prior anti-PDl/PD-Ll therapy.
• Figure 9 Depth and duration of response by LAG-3 expression.
• Figure 10 Duration of progression-free survival.
• FIG. 12 LAG-3 expression status of gastric tumor samples. 48% (10/21) of the samples were scored as LAG-3 positive using a 1% cut-off in a monoplex IHC assay.
• Figure 13 Change in target lesion size in gastric cancer patients in response to treatment with a combination of anti-LAG-3 and anti-PD-1 antibody. LAG-3 positive tumors were enriched among the patients that were responsive to the treatment. Tumor response was determined according to RECIST. The patients in this study have not been previously exposed to anti-PD-l/PD-Ll treatment.
• SCCFIN renal carcinoma
• HCC hepatocellular carcinoma
• NSCLC tumor samples as determined by a monoplex IHC assay.
• Figure 15A and B Figure 15 A. Pigmented melanoma sections. Nuclei were counterstained with hematoxylin with or without bleaching.
• Figure 15B Pigmented melanoma
• Figure 20 Updated response by baseline characteristics and LAG-3 expression.
• Figure 21 Updated best change in target lesion size by LAG-3 and PD-L1 expression.
• FIG. 25 LAG-3 patterns of expression by IHC staining of total nucleated cells in a melanoma tumor specimen.
• FIGS 26A-F Association of LAG-3 with immune and inflammatory biomarkers: (A) LAG-3 vs CD8, (B) LAG-3 vs FOXP3, (C) LAG-3 vs CD 163, (D) LAG-3 vs CD68, (E) LAG-3 vs PD-L1, (F) LAG-3 vs MHC II.
• FIG. 27 Ratio of LAG-3 positive tumor infiltrating lymphocytes (TILs) in tumors comprising ⁇ 1% or ⁇ 1% MHC II positive tumor cells.
• TILs tumor infiltrating lymphocytes
• Figures 28A-C Relationship between inflammation clusters and biomarker expression in (A) urothelial cancer, (B) NSCLC, and (C) all tumor types.
• FIGS 29A-C Heterogeneous MHC II tumor cell expression and LAG-3 + TILs.
• A LAG-3 + TIL numbers in MHC II high and MHC II low tumor cell regions in urothelial carcinoma.
• B-C Ratio of LAG- 3 + TIL cells in MHC II high and MHC II low tumor cell regions in urothelial and gastric carcinoma samples.
• Figures 30A and B LAG-3 mRNA levels at screening and at week 2-4 of nivolumab monotherapy.
• the present invention relates to an improved method of treatment for malignant tumors in a human patient.
• the present invention shows that the administration of an anti-LAG-3 antibody in combination with an anti-PD-1 antibody achieves surprisingly improved treatment outcomes in a patient population having a LAG-3 positive malignant tumor compared to a population comprising patients having both LAG-3 positive and LAG-3 negative tumors.
• the invention described herein relates to a method for identifying patents having a LAG-3 positive tumor, e.g., melanoma.
• the invention described herein relates to a method of treating a LAG-3 positive malignant tumor by administering a combination of a LAG-3 inhibitor (e.g., anti-LAG-3 antibody) and a PD-1 pathway inhibitor (e.g., an anti-PD-1 antibody).
• a LAG-3 inhibitor e.g., anti-LAG-3 antibody
• a PD-1 pathway inhibitor e.g., an anti-PD-1 antibody
• the invention described herein relates to a method of treating a
• LAG-3 positive malignant tumor by administering a PD-1 pathway inhibitor (e.g., an anti-PD-1 antibody) or a combination of a PD-1 pathway inhibitor and an immune checkpoint inhibitor.
• a PD-1 pathway inhibitor e.g., an anti-PD-1 antibody
• an immune checkpoint inhibitor e.g., an immune checkpoint inhibitor
• the invention described herein relates to a method of treating a
• LAG-3 positive malignant tumor by administering an anti-CTLA4 antibody.
• an "antibody” shall include, without limitation, a glycoprotein immunoglobulin which binds specifically to an antigen and comprises at least two heavy (H) chains and two light (L) chains interconnected by disulfide bonds, or an antigen-binding portion thereof.
• Each H chain comprises a heavy chain variable region (abbreviated herein as Y H ) and a heavy chain constant region.
• the heavy chain constant region comprises three constant domains, CHI, Cm and Cm-
• Each light chain comprises a light chain variable region (abbreviated herein as VL) and a light chain constant region.
• the light chain constant region is comprises one constant domain, C L .
• the Y H and Y L regions can be further subdivided into regions of hypervariability, termed complementarity determining regions (CDRs), interspersed with regions that are more conserved, termed framework regions (FR).
• CDRs complementarity determining regions
• FR framework regions
• Each Y H and Vz comprises three CDRs and four FRs, arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.
• the variable regions of the heavy and light chains contain a binding domain that interacts with an antigen.
• the constant regions of the antibodies may mediate the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (Clq) of the classical complement system.
• a heavy chain may have the C-terminal lysine or not.
• the amino acids in the variable regions are numbered using the Kabat numbering system and those in the
• An immunoglobulin may derive from any of the commonly known isotypes, including but not limited to IgA, secretory IgA, IgG and IgM.
• IgG subclasses are also well known to those in the art and include but are not limited to human IgGl, IgG2, IgG3 and IgG4.
• immunotype refers to the antibody class or subclass (e.g. , IgM or IgGl) that is encoded by the heavy chain constant region genes.
• antibody includes, by way of example, monoclonal and polyclonal antibodies; chimeric and humanized antibodies; human or nonhuman antibodies; wholly synthetic antibodies; and single chain antibodies.
• a nonhuman antibody may be humanized by recombinant methods to reduce its immunogenicity in man.
• the term "antibody” includes monospecific, bispecific, or multi-specific antibodies, as well as a single chain antibody.
• the antibody is a bispecific antibody.
• the antibody is a monospecific antibody.
• an "IgG antibody” has the structure of a naturally occurring IgG antibody, i.e., it has the same number of heavy and light chains and disulfide bonds as a naturally occurring IgG antibody of the same subclass.
• an anti-ICOS IgGl, IgG2, IgG3 or IgG4 antibody consists of two heavy chains (HCs) and two light chains (LCs), wherein the two heavy chains and light chains are linked by the same number and location of disulfide bridges that occur in naturally occurring IgGl, IgG2, IgG3 and IgG4 antibodies, respectively (unless the antibody has been mutated to modify the disulfide bonds)
• an "isolated antibody” refers to an antibody that is substantially free of other antibodies having different antigenic specificities (e.g., an isolated antibody that binds specifically to PD-1 is substantially free of antibodies that bind specifically to antigens other than PD-1).
• An isolated antibody that binds specifically to PD-1 may, however, have cross-reactivity to other antigens, such as PD-1 molecules from different species.
• an isolated antibody may be substantially free of other cellular material and/or chemicals.
• the antibody may be an antibody that has been altered (e.g., by mutation, deletion, substitution, conjugation to a non-antibody moiety).
• an antibody may include one or more variant amino acids (compared to a naturally occurring antibody) which change a property (e.g., a functional property) of the antibody.
• a property e.g., a functional property
• numerous such alterations are known in the art which affect, e.g., half-life, effector function, and/or immune responses to the antibody in a patient.
• the term antibody also includes artificial polypeptide constructs which comprise at least one antibody-derived antigen binding site.
• mAb monoclonal antibody
• a mAb is an example of an isolated antibody.
• MAbs may be produced by hybridoma, recombinant, transgenic or other techniques known to those skilled in the art.
• a “human” antibody refers to an antibody having variable regions in which both the framework and CDR regions are derived from human germline immunoglobulin sequences. Furthermore, if the antibody contains a constant region, the constant region is also derived from human germline immunoglobulin sequences.
• the human antibodies of the invention may include amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutation in vivo).
• the term "human antibody,” as used herein is not intended to include antibodies in which CDR sequences derived from the germline of another mammalian species, such as a mouse, have been grafted onto human framework sequences.
• a “humanized antibody” refers to an antibody in which some, most or all of the amino acids outside the CDR domains of a non-human antibody are replaced with corresponding amino acids derived from human immunoglobulins. In one embodiment of a humanized form of an antibody, some, most or all of the amino acids outside the CDR domains have been replaced with amino acids from human immunoglobulins, whereas some, most or all amino acids within one or more CDR regions are unchanged. Small additions, deletions, insertions, substitutions or modifications of amino acids are permissible as long as they do not abrogate the ability of the antibody to bind to a particular antigen.
• a “humanized” antibody retains an antigenic specificity similar to that of the original antibody.
• a "chimeric antibody” refers to an antibody in which the variable regions are derived from one species and the constant regions are derived from another species, such as an antibody in which the variable regions are derived from a mouse antibody and the constant regions are derived from a human antibody.
• an "anti-antigen” antibody refers to an antibody that binds specifically to the antigen.
• an anti-PD-1 antibody binds specifically to PD-1 and an anti-LAG-3 antibody binds specifically to LAG-3.
• an "antigen-binding portion" of an antibody refers to one or more fragments of an antibody that retain the ability to bind specifically to the antigen bound by the whole antibody. 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 within the term “antigen-binding portion” or “antigen-binding fragment” of an antibody, e.g., an anti-LAG-3 antibody described herein, include:
• Fab fragment fragment from papain cleavage
• a similar monovalent fragment consisting of the VL, VH, LC and CHI domains
• CDR complementarity determining region
• antibody-binding portion or "antigen-binding fragment” of an antibody.
• antigen-binding portion or "antigen-binding fragment” of an antibody.
• Antigen-binding portions can be produced by
• LAG-3 refers to LAG-3
• LAG3 refers to LAG3
• LAG-3 Lymphocyte Activation Gene-3.
• LAG-3 as used herein includes human LAG-3 (hLAG- 3), variants, isoforms, and species homologs of hLAG-3, and analogs having at least one common epitope with hLAG-3.
• LAG-3 as used herein includes variants, isoforms, homologs, orthologs and paralogs.
• antibodies specific for a human LAG-3 protein may, in certain cases, cross-react with a LAG-3 protein from a species other than human.
• the antibodies specific for a human LAG-3 protein may be completely specific for the human LAG-3 protein and may not exhibit species or other types of cross-reactivity, or may cross-react with LAG-3 from certain other species, but not all other species (e.g., cross-react with monkey LAG-3 but not mouse LAG-3).
• human LAG-3 refers to human sequence LAG-3, such as the complete amino acid sequence of human LAG-3 having GenBank Accession No. NP_002277 (SEQ ID NO: 13).
• mouse LAG-3 refers to mouse sequence LAG-3, such as the complete amino acid sequence of mouse LAG-3 having GenBank Accession No. NP 032505.
• LAG-3 is also known in the art as, for example, CD223.
• the human LAG-3 sequence may differ from human LAG-3 of GenBank Accession No. NP_002277 by having, e.g., conserved mutations or mutations in non-conserved regions and the LAG-3 has substantially the same biological function as the human LAG-3 of GenBank Accession No. NP 002277.
• a biological function of human LAG-3 is having an epitope in the extracellular domain of LAG-3 that is specifically bound by an antibody of the instant disclosure or a biological function of human LAG-3 is binding to MHC Class II molecules.
• a particular human LAG-3 sequence will generally be at least 90% identical in amino acid sequence to human LAG-3 of GenBank Accession No. NP 002277 and contains amino acid residues that identify the amino acid sequence as being human when compared to LAG-3 amino acid sequences of other species (e.g., murine).
• a human LAG-3 can be at least 95%, or even at least 96%, 97%, 98%, or 99% identical in amino acid sequence to LAG-3 of GenBank Accession No. NP 002277.
• a human LAG-3 sequence will display no more than 10 amino acid differences from the LAG-3 sequence of GenBank Accession No. NP 002277.
• the human LAG-3 can display no more than 5, or even no more than 4, 3, 2, or 1 amino acid difference from the LAG-3 sequence of GenBank Accession No. NP 002277. Percent identity can be determined as described herein.
• Protein PD-1 "PD-1,” “PD1,” “PDCD1,” “hPD-1 “ and “hPD-I” are used interchangeably, and include variants, isoforms, species homologs of human PD-1, and analogs having at least one common epitope with PD-1.
• the complete PD-1 sequence can be found under GenBank Accession No. U64863 (SEQ ID NO:29).
• the protein Programmed Death 1 is an inhibitory member of the CD28 family of receptors, that 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 initial members of the family, CD28 and ICOS were discovered by functional effects on augmenting T cell proliferation following the addition of monoclonal antibodies (Hutloff et al. Nature (1999); 397:263-266; Hansen et al.
• PD-1 was discovered through screening for differential expression in apoptotic cells (Ishida et al. EMBO J (1992); 11 :3887- 95).
• the other members of the family, CTLA-4 and BTLA were discovered through screening for differential expression in cytotoxic T lymphocytes and TH1 cells, respectively.
• CD28, ICOS and CTLA-4 all have an unpaired cysteine residue allowing for homodimerization.
• PD-1 is suggested to exist as a monomer, lacking the unpaired cysteine residue characteristic in other CD28 family members.
• 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, M. L. (1995) J Exp Med 181 : 1953-6; Vivier, E and Daeron, M (1997) Immunol Today 18:286-91).
• ITIM immunoreceptor tyrosine inhibitory motif
• ITSM membrane distal tyrosine-based switch motif
• PD-1 lacks the MYPPPY motif (SEQ ID NO: 32) that is critical for B7-1 and B7-2 binding.
• PD-Ll and PD-L2 Two ligands for PD- 1 have been identified, PD-Ll and PD-L2, that have been 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-Ll and PD-L2 are B7 homologs that bind to PD-1, but do not bind to other CD28 family members. PD- Ll is abundant in a variety of human cancers (Dong et al. (2002) Nat. Med. 8:787-9).
• PD-1 deficient animals develop various autoimmune phenotypes, including autoimmune cardiomyopathy and a lupus-like syndrome with arthritis and nephritis (Nishimura et al. (1999) Immunity 11 : 141-51; Nishimura et al. (2001) Science 291 :319-22). Additionally, PD-1 has been found to play a role in autoimmune encephalomyelitis, systemic lupus erythematosus, graft- versus-host disease (GVHD), type I diabetes, and rheumatoid arthritis (Salama et al.
• GVHD graft- versus-host disease
• P-Ll Programmed Death Ligand-1
• PD-L1 includes human PD-L1 (hPD- Ll), variants, isoforms, and species homologs of hPD-Ll, and 5 analogs having at least one common epitope with hPD-Ll .
• the complete hPD-Ll sequence can be found under GenBank Accession No. Q9NZQ7.
• Programmed Death Ligand-2 and "PD-L2” as used herein include human PD-L2 (hPD-L2), variants, isoforms, and species homologs of hPD-L2, and analogs having at least one common epitope with hPD-L2.
• the complete hPD-L2 sequence can be found under GenBank Accession No. Q9BQ51.
• a "patient” as used herein includes any patient who is afflicted with a cancer (e.g., melanoma).
• a cancer e.g., melanoma
• subject and patient are used interchangeably herein.
• administering refers to the physical introduction of a composition comprising a therapeutic agent to a subject, using any of the various methods and delivery systems known to those skilled in the art. Routes of administration for the formulations disclosed herein include intravenous, intramuscular, subcutaneous, intraperitoneal, spinal or other parenteral routes of administration, for example by injection or infusion.
• parenteral administration means modes of administration other than enteral and topical administration, usually by injection, and includes, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intralymphatic, intralesional, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, epidural and intrasternal injection and infusion, as well as in vivo electroporation.
• the formulation is administered via a non-parenteral route, in some embodiments, orally.
• non-parenteral routes include a topical, epidermal or mucosal route of administration, for example, intranasally, vaginally, rectally, sublingually or topically.
• Administering can also be performed, for example, once, a plurality of times, and/or over one or more extended periods.
• Treatment refers to any type of intervention or process performed on, or the administration of an active agent to, the subject with the objective of reversing, alleviating, ameliorating, inhibiting, slowing down or preventing the onset, progression, development, severity or recurrence of a symptom, complication or condition, or biochemical indicia associated with a disease.
• effective treatment refers to treatment producing a beneficial effect, e.g., amelioration of at least one symptom of a disease or disorder.
• a beneficial effect can take the form of an improvement over baseline, i.e., an improvement over a measurement or observation made prior to initiation of therapy according to the method.
• a beneficial effect can also take the form of arresting, slowing, retarding, or stabilizing of a deleterious progression of a marker of solid tumor.
• Effective treatment may refer to alleviation of at least one symptom of a solid tumor. Such effective treatment may, e.g., reduce patient pain, reduce the size and/or number of lesions, may reduce or prevent metastasis of a tumor, and/or may slow tumor growth.
• an effective amount refers to an amount of an agent that provides the desired biological, therapeutic, and/or prophylactic result. That result can be reduction, amelioration, palliation, lessening, delaying, and/or alleviation of one or more of the signs, symptoms, or causes of a disease, or any other desired alteration of a biological system.
• an effective amount comprises an amount sufficient to cause a tumor to shrink and/or to decrease the growth rate of the tumor (such as to suppress tumor growth) or to prevent or delay other unwanted cell proliferation.
• an effective amount is an amount sufficient to delay tumor development.
• an effective amount is an amount sufficient to prevent or delay tumor recurrence.
• an effective amount can be administered in one or more administrations.
• the effective amount of the drug or composition may: (i) reduce the number of cancer cells; (ii) reduce tumor size; (iii) inhibit, retard, slow to some extent and may stop cancer cell infiltration into peripheral organs; (iv) inhibit (i.e., slow to some extent and may stop tumor metastasis; (v) inhibit tumor growth; (vi) prevent or delay occurrence and/or recurrence of tumor; and/or (vii) relieve to some extent one or more of the symptoms associated with the cancer.
• an "effective amount” is the amount of anti-LAG-3 antibody and the amount of anti-PD-1 antibody, in combination, clinically proven to affect a significant decrease in cancer or slowing of progression of cancer, such as an advanced solid tumor.
• the terms "fixed dose”, “flat dose” and “flat-fixed dose” are used interchangeably and refer to a dose that is administered to a patient without regard for the weight or body surface area (BSA) of the patient.
• the fixed or flat dose is therefore not provided as a mg/kg dose, but rather as an absolute amount of the agent (e.g., the anti-LAG-3 antibody and/or anti-PD-1 antibody).
• progression-free survival which can be abbreviated as PFS, as used herein refers to the length of time during and after the treatment of a solid tumor (i.e., melanoma) that a patient lives with the disease but it does not get worse.
• Dosing interval means the amount of time that elapses between multiple doses of a formulation disclosed herein being administered to a subject. Dosing interval can thus be indicated as ranges.
• Dosing frequency refers to the frequency of administering doses of a formulation disclosed herein in a given time. Dosing frequency can be indicated as the number of doses per a given time, e.g., once a week or once in two weeks.
• fixed dose with regard to a composition of the invention means that two or more different antibodies in a single composition are present in the composition in particular (fixed) ratios with each other.
• the fixed dose is based on the weight (e.g., mg) of the antibodies.
• the fixed dose is based on the concentration (e.g., mg/ml) of the antibodies.
• the ratio is at least about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, about 1:30, about 1:40, about 1:50, about 1:60, about 1:70, about 1:80, about 1:90, about 1:100, about 1:120, about 1:140, about 1:160, about 1:180, about 1:200, about 200:1, about 180:1, about 160:1, about 140:1, about 120:1, about 100:1, about 90:1, about 80:1, about 70:1, about 60:1, about 50:1, about 40:1, about 30:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, or about 2:1 mg first antibody to mg second antibody.
• the 3:1 ratio of a first antibody and a second antibody can mean that a vial can contain about 240 mg of the first antibody and 80 mg of
• flat dose with regard to the composition of the invention means a dose that is administered to a patient without regard for the weight or body surface area (BSA) of the patient.
• the flat dose is therefore not provided as a mg/kg dose, but rather as an absolute amount of the agent (e.g., the anti-LAG-3 antibody and/or anti-PD-1 antibody).
• a 60 kg person and a 100 kg person would receive the same dose of the composition (e.g., 240 mg of an anti-PD-1 antibody and 80 mg of an anti- LAG-3 antibody in a single fixed dosing formulation vial containing both 240 mg of an anti-PD-1 antibody and 80 mg of an anti- LAG-3 antibody (or two fixed dosing formulation vials containing 120 mg of an anti-PD-1 antibody and 40 mg of an anti- LAG-3 antibody, etc.)).
• the composition e.g., 240 mg of an anti-PD-1 antibody and 80 mg of an anti- LAG-3 antibody in a single fixed dosing formulation vial containing both 240 mg of an anti-PD-1 antibody and 80 mg of an anti- LAG-3 antibody (or two fixed dosing formulation vials containing 120 mg of an anti-PD-1 antibody and 40 mg of an anti- LAG-3 antibody, etc.
• weight based dose means that a dose that is administered to a patient is calculated based on the weight of the patient. For example, when a patient with 60 kg body weight requires 3 mg/kg of an anti-LAG-3 antibody in combination with 3 mg/kg of an anti-PD-1 antibody, one can draw the appropriate amounts of the anti-LAG-3 antibody (i.e., 180 mg) and the anti-PD-1 antibody (i.e., 180 mg) at once from a 1 : 1 ratio fixed dosing formulation of an anti-LAG3 antibody and an anti-PD-1 antibody.
• the terms "about once a week,” “once about every week,” “once about every two weeks,” or any other similar dosing interval terms as used herein means approximate number, and "about once a week” or “once about every week” can include every seven days ⁇ two days, i.e., every five days to every nine days.
• the dosing frequency of "once a week” thus can be every five days, every six days, every seven days, every eight days, or every nine days.
• "Once about every two weeks” can include every fourteen days ⁇ three days, i.e., every eleven days to every seventeen days. Similar approximations apply, for example, to once about every three weeks, once about every four weeks, once about every five weeks, once about every six weeks and once about every twelve weeks.
• a dosing interval of once about every six weeks or once about every twelve weeks means that the first dose can be administered any day in the first week, and then the next dose can be administered any day in the sixth or twelfth week, respectively.
• a dosing interval of once about every six weeks or once about every twelve weeks means that the first dose is administered on a particular day of the first week (e.g., Monday) and then the next dose is administered on the same day of the sixth or twelfth weeks (i.e., Monday), respectively.
• a “cancer” refers a broad group of various diseases characterized by the uncontrolled growth of abnormal cells in the body. Unregulated cell division and growth results in the formation of malignant tumors that invade neighboring tissues and may also metastasize to distant parts of the body through the lymphatic system or bloodstream.
• a “cancer” or “cancer tissue” can include a tumor.
• tumor refers to any mass of tissue that results from excessive cell growth or proliferation, either benign (non-cancerous) or malignant (cancerous), including pre-cancerous lesions.
• LAG-3 positive or "LAG-3 expression positive,” relating to LAG-3 expression, refers to the proportion of cells in a test tissue sample comprising tumor cells and tumor-infiltrating inflammatory cells above which the tissue sample is scored as expressing LAG-3.
• the 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 number of cells express LAG-3.
• the 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 number of tumor-infiltrating inflammatory cells (e.g., T cells, CD8+ T cells, CD4+ T cells, FOXP3+ cells) express LAG-3.
• tumor-infiltrating inflammatory cells e.g., T cells, CD8+ T cells, CD4+ T cells, FOXP3+ cells
• LAG-3 positive tumor or LAG-3 expression positive tumor can also be expressed herein as tumor expressing LAG-3.
• the 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.
• 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 tumor-infiltrating inflammatory cells (e.g., T cells, CD8+ T cells, CD4+ T cells, FOXP3+ cells) express LAG-3.
• a LAG-3 positive tumor or LAG-3 expression positive tumor means that at least about 0.1% to at least about 10% of the total number of cells express LAG-3.
• a LAG-3 positive tumor or LAG-3 expression positive tumor means that at least about 0.1% to at least about 10% of the total number of tumor-infiltrating inflammatory cells (e.g., T cells, CD8+ T cells, CD4+ T cells, FOXP3+ cells) express LAG-3.
• a LAG-3 positive 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.
• a LAG-3 positive or LAG-3 expression positive tumor means that at least about 1% of the total number of tumor-infiltrating inflammatory cells (e.g., T cells, CD8+ T cells, CD4+ T cells, FOXP3+ cells) express LAG-3 on the cell surface. In other embodiments, a LAG-3 positive or LAG-3 expression positive tumor means that at least about 5%) of the total number of cells express LAG-3 on the cell surface.
• tumor-infiltrating inflammatory cells e.g., T cells, CD8+ T cells, CD4+ T cells, FOXP3+ cells
• a LAG-3 positive or LAG-3 expression positive tumor means that at least about 5% of the total number of tumor-infiltrating inflammatory cells (e.g., T cells, CD8+ T cells, CD4+ T cells, FOXP3+ cells) express LAG-3 on the cell surface.
• LAG-3 positive 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.
• LAG-3 positive or LAG-3 expression positive tumor means that at least about 1%, or in the range of 1- 5% of the total number of tumor-infiltrating inflammatory cells (e.g., T cells, CD8+ T cells, CD4+ T cells, FOXP3+ cells) express LAG-3 on the cell surface.
• tumor-infiltrating inflammatory cells e.g., T cells, CD8+ T cells, CD4+ T cells, FOXP3+ cells
• LAG-3 negative refers to the lack of a detectable amount of LAG-3 expression.
• a LAG-3 negative tumor or LAG-3 expression negative tumor means that less than 0.01% of the total number of cells express a detectable level of LAG-3.
• a LAG-3 negative tumor or LAG-3 expression negative tumor means that less than 0.01% of the total number of tumor-infiltrating inflammatory cells (e.g., T cells, CD8+ T cells, CD4+ T cells, FOXP3+ cells) express a detectable level of LAG-3.
• a LAG-3 negative tumor or LAG-3 expression negative tumor means that less than 1% of the total number of cells express a detectable level of LAG-3.
• a LAG-3 negative tumor or LAG-3 expression negative tumor means that less than 1% of the total number of tumor-infiltrating inflammatory cells (e.g., T cells, CD8+ T cells, CD4+ T cells, FOXP3+ cells) express a detectable level of LAG-3.
• a LAG-3 negative tumor or LAG-3 expression negative tumor means that zero (0) cells express a detectable level of LAG-3.
• a LAG-3 negative or a LAG-3 expression negative tumor is any tumor other than a LAG-3 positive or a LAG-3 expression positive tumor.
• PD-Ll expression refers to the proportion of cells in a test tissue sample comprising tumor cells and tumor- infiltrating inflammatory cells above which the sample is scored as expressing cell surface PD-Ll .
• IHC immunohistochemistry
• the PD-Ll positive tumor or PD-Ll 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 number of cells express PD-Ll .
• the PD-Ll positive tumor or PD-Ll expression positive tumor can also be expressed herein as tumor expressing PD-Ll .
• the PD-Ll positive tumor or PD-Ll expression positive tumor means that at least about 0.1% to at least about 20% of the total number of cells express PD-Ll .
• the PD-Ll positive tumor or PD-Ll expression positive tumor means that at least about 0.1%) to at least about 10%> of the total number of cells express PD-Ll .
• the PD-Ll positive or PD-Ll expression positive tumor means that at least about 1%) of the total number of cells express PD-Ll on the cell surface.
• the PD- Ll positive or PD-Ll expression positive tumor means that at least about 5% of the total number of cells express PD-Ll on the cell surface. In one particular embodiment, PD-Ll positive or PD- Ll expression positive tumor means that at least about 1%, or in the range of 1- 5% of the total number of cells express PD-Ll on the cell surface.
• PD-Ll negative or "PD-Ll expression negative,” relating to cell surface PD-Ll expression, refers to the proportion of cells in a test tissue sample comprising tumor cells and tumor- infiltrating inflammatory cells that are not PD-Ll positive or PD-Ll expression positive.
• an "immune response” refers to the action of a cell of the immune system (for example, T lymphocytes, B lymphocytes, natural killer (NK) cells, macrophages, eosinophils, mast cells, dendritic cells and neutrophils) and soluble macromolecules produced by any of these cells or the liver (including antibodies, cytokines, and complement) that results in selective targeting, binding to, damage to, destruction of, and/or elimination from a vertebrate's body of invading pathogens, cells or tissues infected with pathogens, cancerous or other abnormal cells, or, in cases of autoimmunity or pathological inflammation, normal human cells or tissues.
• a cell of the immune system for example, T lymphocytes, B lymphocytes, natural killer (NK) cells, macrophages, eosinophils, mast cells, dendritic cells and neutrophils
• soluble macromolecules produced by any of these cells or the liver including antibodies, cytokines, and complement
• a "tumor-infiltrating inflammatory cell” is any type of cell that typically participates in an inflammatory response in a subject and which infiltrates tumor tissue. Such cells include tumor-infiltrating lymphocytes (TILs), macrophages, monocytes, eosinophils, histiocytes and dendritic cells.
• TILs tumor-infiltrating lymphocytes
• macrophages macrophages
• monocytes eosinophils
• histiocytes histiocytes and dendritic cells.
• the terms "about” or “comprising essentially of refer to a value or composition that is within an acceptable error range for the particular value or composition as determined by one of ordinary skill in the art, which will depend in part on how the value or composition is measured or determined, i.e., the limitations of the measurement system.
• “about” or “comprising essentially of can mean within 1 or more than 1 standard deviation per the practice in the art.
• “about” or “comprising essentially of can mean a range of up to 10% or 20% ⁇ i.e., ⁇ 10%) or ⁇ 20%).
• about 3mg can include any number between 2.7 mg and 3.3 mg (for 10%) or between 2.4 mg and 3.6 mg (for 20%).
• the terms can mean up to an order of magnitude or up to 5-fold of a value.
• the meaning of "about” or “comprising essentially of should be assumed to be within an acceptable error range for that particular value or composition.
• any concentration range, percentage range, ratio range or integer range is to be understood to include the value of any integer within the recited range and, when appropriate, fractions thereof (such as one-tenth and one-hundredth of an integer), unless otherwise indicated.
• the present invention is directed to a method for treating a LAG-3- positive malignant tumor (e.g., melanoma) in a subject in need thereof.
• a combination therapy of a LAG-3 inhibitor (e.g., anti-LAG-3 antibody) and a PD-1 pathway inhibitor (e.g., anti-PD-1 antibody) results in better therapeutic outcomes (e.g., objective response rate and disease control rate) in a patient population with LAG-3 positive malignant tumors (e.g., melanoma) than in a general patient population having a mix of LAG-3 -negative malignant tumors and LAG-3 - positive malignant tumors.
• the present invention provides identifying a patient as having a LAG-3 -positive tumor and providing an immunotherapy of a LAG-3 inhibitor (e.g., anti-LAG-3 antibody) and a PD-1 pathway inhibitor (e.g., anti-PD-1 antibody).
• a LAG-3 inhibitor e.g., anti-LAG-3 antibody
• a PD-1 pathway inhibitor e.g., anti-PD-1 antibody
• the present invention is directed to identifying a patient as having a LAG-3 -positive tumor and treating the LAG-3 positive tumor by administering a PD-1 pathway inhibitor (e.g., an anti-PD-1 antibody) or a combination of a PD-1 pathway inhibitor and an immune checkpoint inhibitor.
• a PD-1 pathway inhibitor e.g., an anti-PD-1 antibody
• the invention includes a method of identifying a patient as having a LAG-3 -positive tumor and treating the LAG-3 positive tumor by administering an anti-PD-1 antibody.
• the invention includes a method of identifying a patient as having a LAG-3 -positive tumor and treating the LAG-3 positive tumor by administering an anti-PD-Ll antibody.
• the present invention is directed to identifying a patient as having a LAG-3 -positive tumor and treating the LAG-3 positive tumor by administering an anti- CTLA-4 antibody.
• the invention includes a method of 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.
• the invention includes a method of identifying a malignant tumor in a human patient 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.
• the invention includes a method of identifying a malignant tumor in a human patient that is likely to be responsive to a immunotherapy, the method comprising: (a) determining the level of LAG-3 expression in a tumor sample; and (b) identifying the tumor as likely to be responsive to treatment if the tumor is a LAG-3 positive tumor.
• the invention includes a method of identifying a malignant tumor in a human patient that is likely to be responsive to a immunotherapy, the method comprising: (a) determining the level of LAG-3 expression in a tumor sample; and (b) identifying the tumor as likely to be responsive to treatment if the tumor is a LAG-3 positive tumor.
• the invention includes a method of classifying a malignant tumor in a human patient as likely to be responsive to a immunotherapy, the method comprising: (a) determining the level of LAG-3 expression in a tumor sample; and (b) classifying the tumor as likely to be responsive to immunotherapy if the tumor is a LAG-3 positive tumor.
• the immunotherapy comprises contacting the tumor with a therapeutically effective amount of a LAG-3 inhibitor and a PD-1 pathway inhibitor.
• the immunotherapy comprises contacting the tumor with a therapeutically effective amount of a LAG-3 inhibitor.
• the immunotherapy comprises contacting the tumor with a therapeutically effective amount of a PD-1 pathway inhibitor.
• the immunotherapy comprises contacting the tumor with a therapeutically effective amount of an anti-PD-1 antibody. In some embodiments, the immunotherapy comprises contacting the tumor with a therapeutically effective amount of an anti-PD-Ll antibody. In some embodiments, the immunotherapy comprises contacting the tumor with a therapeutically effective amount of an anti-CTLA-4 antibody. In some embodiments, the immunotherapy comprises contacting the tumor with a therapeutically effective amount of a PD-1 pathway inhibitor and an immune checkpoint inhibitor. In some embodiments, the method comprises contacting the tumor with a therapeutically effective amount of a LAG-3 inhibitor and a PD-1 pathway inhibitor. In some embodiments, the method comprises contacting the tumor with a therapeutically effective amount of a LAG-3 inhibitor.
• the method comprises contacting the tumor with a therapeutically effective amount of a PD-1 pathway inhibitor. In some embodiments, the method comprises contacting the tumor with a therapeutically effective amount of an anti-PD-1 antibody. In some embodiments, the method comprises contacting the tumor with a therapeutically effective amount of an anti-PD-Ll antibody. In some embodiments, the method comprises contacting the tumor with a therapeutically effective amount of an anti-CTLA-4 antibody. In some embodiments, the method comprises contacting the tumor with a therapeutically effective amount of a PD-1 pathway inhibitor and an immune checkpoint inhibitor. In some embodiments, the method comprises administering to the patient a therapeutically effective amount of a LAG-3 inhibitor and a PD-1 pathway inhibitor.
• the method comprises administering to the patient a therapeutically effective amount of a LAG-3 inhibitor. In some embodiments, the method comprises administering to the patient a therapeutically effective amount of a PD-1 pathway inhibitor. In some embodiments, the method comprises administering to the patient a therapeutically effective amount of an anti-CTLA-4 antibody. In some embodiments, the method comprises administering to the patient a therapeutically effective amount of a PD-1 pathway inhibitor and an immune checkpoint inhibitor. 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. In certain embodiments, any of the present methods further comprise determining PD- Ll expression in the tumor sample.
• the invention includes a method of identifying a patient with a malignant tumor who is likely to respond to a immunotherapy, the method comprising: (a) determining the level of LAG-3 expression in a tumor sample; and (b) identifying the patient who is likely to respond to treatment if the tumor is a LAG-3 positive tumor.
• the invention includes a method of selecting a patient with a malignant tumor for immunotherapy, the method comprising: (a) determining the level of LAG-3 expression in a tumor sample; and (b) selecting the patient for immunotherapy if the tumor is a LAG-3 positive tumor.
• the immunotherapy comprises contacting the tumor with a therapeutically effective amount of a LAG-3 inhibitor and a PD-1 pathway inhibitor. In some embodiments, the immunotherapy comprises contacting the tumor with a therapeutically effective amount of a LAG-3 inhibitor. In some embodiments, the immunotherapy comprises contacting the tumor with a therapeutically effective amount of a PD-1 pathway inhibitor. In some embodiments, the immunotherapy comprises contacting the tumor with a therapeutically effective amount of an anti-PD-1 antibody. In some embodiments, the immunotherapy comprises contacting the tumor with a therapeutically effective amount of an anti-PD-Ll antibody. In some embodiments, the immunotherapy comprises contacting the tumor with a therapeutically effective amount of an anti-CTLA-4 antibody.
• the immunotherapy comprises contacting the tumor with a therapeutically effective amount of a PD-1 pathway inhibitor and an immune checkpoint inhibitor.
• the method comprises contacting the tumor with a therapeutically effective amount of a LAG-3 inhibitor and a PD-1 pathway inhibitor.
• the method comprises contacting the tumor with a therapeutically effective amount of a LAG-3 inhibitor.
• the method comprises contacting the tumor with a therapeutically effective amount of a PD-1 pathway inhibitor.
• the method comprises contacting the tumor with a therapeutically effective amount of an anti-PD-1 antibody.
• the method comprises contacting the tumor with a therapeutically effective amount of an anti-PD-Ll antibody.
• the method comprises contacting the tumor with a therapeutically effective amount of an anti-CTLA-4 antibody. In some embodiments, the method comprises contacting the tumor with a therapeutically effective amount of a PD-1 pathway inhibitor and an immune checkpoint inhibitor. In some embodiments, the method comprises administering to the patient a therapeutically effective amount of a LAG-3 inhibitor and a PD-1 pathway inhibitor. In some embodiments, the method comprises administering to the patient a therapeutically effective amount of a LAG-3 inhibitor. In some embodiments, the method comprises administering to the patient a therapeutically effective amount of a PD-1 pathway inhibitor. In some embodiments, the method comprises administering to the patient a therapeutically effective amount of an anti-CTLA-4 antibody.
• the method comprises administering to the patient a therapeutically effective amount of a PD-1 pathway inhibitor and an immune checkpoint inhibitor.
• the LAG-3 inhibitor is an anti-LAG-3 antibody and the PD-1 pathway inhibitor is an anti-PD-1 antibody.
• any of the present methods further comprise determining PD-L1 expression in the tumor sample.
• the invention includes a method of treating a malignant tumor in a human patient, comprising: administering to the patient an immunotherapy disclosed herein; wherein the patient is predicted to respond to treatment with the LAG-3 inhibitor and PD-1 pathway inhibitor based upon LAG-3 expression or based upon LAG-3 and PD-L1 expression in a sample of the patient's tumor.
• the LAG-3 inhibitor is an anti-LAG-3 antibody and the PD-1 pathway inhibitor is an anti-PD-1 antibody.
• the invention includes a method of treating a malignant tumor in a human patient in need thereof, comprising: (a) determining the level of LAG-3 expression or the level of LAG-3 and PD-L1 expression in a tumor sample; and (b) administering to the patient a therapeutically effective amount of a LAG-3 inhibitor if the tumor is a LAG-3 positive tumor or a LAG-3 positive PD-Ll positive tumor.
• the invention includes a method for treating a malignant tumor in a human patient in need thereof, comprising: (a) identifying the patient as having a LAG-3 positive malignant tumor or a LAG-3 positive PD-Ll positive malignant tumor; and (b) administering to the patient a therapeutically effective amount of a LAG-3 inhibitor.
• the invention includes 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, wherein the patient is identified as having a LAG-3 positive malignant tumor or a LAG-3 positive PD-Ll positive malignant tumor prior to the administration.
• the LAG-3 positive tumor is a LAG-3 positive PD-Ll negative tumor.
• the LAG-3 positive malignant tumor is a LAG-3 positive PD-Ll negative malignant tumor.
• the LAG-3 inhibitor is an anti-LAG-3 antibody.
• the invention includes a method of treating a malignant tumor in a human patient in need thereof, comprising: (a) determining the level of LAG-3 expression or the level of LAG-3 and PD-Ll expression in a tumor sample; and (b) administering to the patient a therapeutically effective amount of a PD-1 pathway inhibitor if the tumor is a LAG-3 positive tumor or a LAG-3 positive PD-Ll positive tumor.
• the invention includes a method for treating a malignant tumor in a human patient in need thereof, comprising: (a) identifying the patient as having a LAG-3 positive malignant tumor or a LAG-3 positive PD-Ll positive malignant tumor; and (b) administering to the patient a therapeutically effective amount of a PD-1 pathway inhibitor.
• the invention includes 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 PD-1 pathway inhibitor, wherein the patient is identified as having a LAG-3 positive malignant tumor or a LAG-3 positive PD-Ll positive malignant tumor prior to the administration.
• the PD-1 pathway inhibitor is an anti-PD-1 antibody.
• the PD-1 pathway inhibitor is an anti-PD-Ll antibody.
• the LAG-3 positive tumor is a LAG-3 positive PD-Ll negative tumor.
• the LAG-3 positive malignant tumor is a LAG-3 positive PD-Ll negative malignant tumor.
• the invention includes a method of treating a malignant tumor in a human patient in need thereof, comprising: (a) determining the level of LAG-3 expression or the level of LAG-3 and PD-Ll expression in a tumor sample; and (b) administering to the patient a therapeutically effective amount of an anti-CTLA-4 antibody if the tumor is a LAG-3 positive tumor or a LAG-3 positive PD-Ll positive tumor.
• the invention includes a method for treating a malignant tumor in a human patient in need thereof, comprising: (a) identifying the patient as having a LAG-3 positive malignant tumor or a LAG-3 positive PD-Ll positive malignant tumor; and (b) administering to the patient a therapeutically effective amount of an anti-CTLA-4 antibody.
• the invention includes a method for treating a malignant tumor in a human patient in need thereof comprising administering to the patient a therapeutically effective amount of an anti-CTLA-4 antibody, wherein the patient is identified as having a LAG-3 positive malignant tumor or a LAG-3 positive PD-Ll positive malignant tumor prior to the administration.
• the LAG-3 positive tumor is a LAG-3 positive PD-Ll negative tumor.
• the LAG-3 positive malignant tumor is a LAG-3 positive PD-Ll negative malignant tumor.
• the invention includes a method of treating a malignant tumor in a human patient in need thereof, comprising: (a) determining the level of LAG-3 expression or the level of LAG-3 and PD-Ll expression in a tumor sample; and (b) administering to the patient a therapeutically effective amount of a LAG-3 inhibitor and PD-1 pathway inhibitor if the tumor is a LAG-3 positive tumor or a LAG-3 positive PD-Ll positive tumor.
• the invention includes a method for treating a malignant tumor in a human patient in need thereof, comprising: (a) identifying the patient as having a LAG-3 positive malignant tumor or a LAG-3 positive PD-Ll positive malignant tumor; and (b) administering to the patient a therapeutically effective amount of a LAG-3 inhibitor and a PD-1 pathway inhibitor.
• the invention includes 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 and a PD-1 pathway inhibitor, wherein the patient is identified as having a LAG-3 positive malignant tumor or a LAG-3 positive PD-Ll positive malignant tumor prior to the administration.
• the LAG-3 positive tumor is a LAG-3 positive PD-Ll negative tumor.
• the LAG-3 positive malignant tumor is a LAG-3 positive PD-Ll negative malignant tumor.
• the LAG-3 inhibitor is an anti-LAG-3 antibody and the PD-1 pathway inhibitor is an anti -PD-Ll antibody.
• the LAG-3 inhibitor is an anti-LAG-3 antibody and the PD-1 pathway inhibitor is an anti-PD-1 antibody.
• the invention includes a method of treating a malignant tumor in a human patient in need thereof, comprising: (a) determining the level of LAG-3 expression or the level of LAG-3 and PD-Ll expression in a tumor sample; and (b) administering to the patient a therapeutically effective amount of a PDl pathway inhibitor and an immune checkpoint inhibitor if the tumor is a LAG-3 positive tumor or a LAG-3 positive PD-Ll positive tumor.
• the invention includes a method for treating a malignant tumor in a human patient in need thereof, comprising: (a) identifying the patient as having a LAG-3 positive malignant tumor or a LAG-3 positive PD-Ll positive malignant tumor; and (b) administering to the patient a therapeutically effective amount of a PDl pathway inhibitor and an immune checkpoint inhibitor.
• the invention includes 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 PDl pathway inhibitor and an immune checkpoint inhibitor, wherein the patient is identified as having a LAG-3 positive malignant tumor or a LAG-3 positive PD-Ll positive malignant tumor prior to the administration.
• the LAG-3 positive tumor is a LAG-3 positive PD-Ll negative tumor.
• the LAG-3 positive malignant tumor is a LAG-3 positive PD-Ll negative malignant tumor.
• the PD-1 pathway inhibitor is an anti-PD-Ll antibody. In some embodiments, the PD-1 pathway inhibitor is an anti-PD-1 antibody.
• the invention includes a method for treating a malignant tumor in a human patient in need thereof comprising administering to the patient an immunotherapy disclosed herein, wherein the patient is identified as having a LAG-3 positive malignant tumor or a LAG-3 positive PD-Ll positive malignant tumor prior to the administration.
• the LAG-3 positive malignant tumor is a LAG-3 positive PD-Ll negative malignant tumor.
• the immunotherapy comprises administering a therapeutically effective amount of a LAG-3 inhibitor and a PD-1 pathway inhibitor.
• the immunotherapy comprises administering a therapeutically effective amount of a LAG-3 inhibitor.
• the immunotherapy comprises administering a therapeutically effective amount of a PD-1 pathway inhibitor. In some embodiments, the immunotherapy comprises administering a therapeutically effective amount of an anti-CTLA-4 antibody. In some embodiments, the immunotherapy comprises administering a therapeutically effective amount of a PD-1 pathway inhibitor and an immune checkpoint inhibitor.
• the LAG-3 inhibitor is an anti -LAG-3 antibody and the PD-1 pathway inhibitor is an anti-PD-Ll antibody. 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.
• the invention includes method for extending a progression-free survival period for over 12 months in a human patient afflicted with a malignant tumor comprising administering to the patient an immunotherapy disclosed herein, wherein the patient is identified as having a LAG-3 positive malignant tumor or a LAG-3 positive PD-L1 positive malignant tumor prior to the administration and wherein the patient demonstrates progression-free survival for over 12 months.
• the LAG-3 positive malignant tumor is a LAG-3 positive PD-L1 negative malignant tumor.
• the progression-free survival of the patient can be extended, after the administration, for over about 13 months, about 14 months, about 15 months, about 16 months, about 17 months, about 18 months, about 2 years, about 3 years, about 4 years, about 5 years, about 6 years, about 7 years, about 8 years, about 9 years, or about 10 years.
• the immunotherapy comprises administering a therapeutically effective amount of a LAG-3 inhibitor and a PD-1 pathway inhibitor. In some embodiments, the immunotherapy comprises administering a therapeutically effective amount of a LAG-3 inhibitor. In some embodiments, the immunotherapy comprises administering a therapeutically effective amount of a PD-1 pathway inhibitor.
• the immunotherapy comprises administering a therapeutically effective amount of an anti-CTLA-4 antibody. In some embodiments, the immunotherapy comprises administering a therapeutically effective amount of a PD-1 pathway inhibitor and an immune checkpoint inhibitor.
• the LAG-3 inhibitor is an anti-LAG-3 antibody and the PD-1 pathway inhibitor is an anti-PD-1 antibody. In some embodiments, the LAG-3 inhibitor is an anti-LAG-3 antibody and the PD-1 pathway inhibitor is an anti-PD-Ll antibody.
• the invention is includes a method for reducing a tumor size at least by 10% in a human patient afflicted with a malignant tumor comprising administering to the patient an immunotherapy disclosed herein, wherein the patient is identified as having a LAG-3 positive malignant tumor (e.g., melanoma) or a LAG-3 positive PD-L1 positive malignant tumor prior to the administration and wherein the administration reduces the tumor size 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 the administration.
• a LAG-3 positive malignant tumor e.g., melanoma
• PD-L1 positive malignant tumor prior to the administration
• the administration reduces the tumor size 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 the administration.
• the method comprises identifying the patient as having a LAG-3 positive malignant tumor or a LAG-3 positive PD-Ll positive malignant tumor prior to the administration.
• the LAG-3 positive malignant tumor is a LAG-3 positive PD-Ll negative malignant tumor.
• the immunotherapy comprises administering a therapeutically effective amount of a LAG-3 inhibitor and a PD-1 pathway inhibitor.
• the immunotherapy comprises administering a therapeutically effective amount of a LAG-3 inhibitor.
• the immunotherapy comprises administering a therapeutically effective amount of a PD-1 pathway inhibitor.
• the immunotherapy comprises administering a therapeutically effective amount of an anti-PD-1 antibody.
• the immunotherapy comprises administering a therapeutically effective amount of an anti -PD-Ll antibody. In some embodiments, the immunotherapy comprises administering a therapeutically effective amount of an anti-CTLA-4 antibody. In some embodiments, the immunotherapy comprises administering a therapeutically effective amount of a PD-1 pathway inhibitor and an immune checkpoint inhibitor.
• the LAG-3 inhibitor is an anti -LAG-3 antibody and the PD-1 pathway inhibitor is an anti-PD-1 antibody. In some embodiments, the LAG-3 inhibitor is an anti -LAG-3 antibody and the PD-1 pathway inhibitor is an anti -PD-Ll antibody.
• the invention can also include a method of preventing a relapse and/or inducing a remission in a patient comprising administering to the patient an immunotherapy disclosed herein, wherein the patient is identified as having a LAG-3 -positive malignant tumor (e.g., melanoma) or a LAG-3 positive PD-Ll positive malignant tumor prior to the administration.
• the method of the invention comprises (i) identifying a patient as having a LAG-3 -positive malignant tumor or a LAG-3 positive PD-Ll positive malignant tumor; (ii) administering to the patient an immunotherapy disclosed herein.
• the immunotherapy comprises administering a therapeutically effective amount of a LAG-3 inhibitor and a PD-1 pathway inhibitor.
• the LAG-3 positive malignant tumor is a LAG-3 positive PD-Ll negative malignant tumor.
• the immunotherapy comprises administering a therapeutically effective amount of a LAG-3 inhibitor.
• the immunotherapy comprises administering a therapeutically effective amount of a PD-1 pathway inhibitor.
• the immunotherapy comprises administering a therapeutically effective amount of an anti-PD-1 antibody.
• the immunotherapy comprises administering a therapeutically effective amount of an anti-PD-Ll antibody.
• the immunotherapy comprises administering a therapeutically effective amount of an anti-CTLA-4 antibody.
• the immunotherapy comprises administering a therapeutically effective amount of a PD-1 pathway inhibitor and an immune checkpoint inhibitor.
• the LAG-3 inhibitor is an anti-LAG-3 antibody and the PD-1 pathway inhibitor is an anti-PD-1 antibody.
• the LAG-3 inhibitor is an anti-LAG-3 antibody and the PD-1 pathway inhibitor is an anti-PD-Ll antibody.
• the invention includes a method for increasing an objective response rate to be higher than 55% in a patient population, wherein each patient of the patient population is afflicted with a malignant tumor, in a cancer treatment comprising administering to the patient an immunotherapy disclosed herein, wherein each patient is identified as having a LAG-3 positive malignant tumor (e.g., melanoma) or a LAG-3 positive PD-Ll positive malignant tumor prior to the administration and wherein the objective response rate is higher than 55%, 60%, 65%, 70%, or 75%.
• the method comprises identifying the patient as having a LAG-3 positive malignant tumor or a LAG-3 positive PD-Ll positive malignant tumor prior to the administration.
• the LAG-3 positive malignant tumor is a LAG-3 positive PD-Ll negative malignant tumor.
• the immunotherapy comprises administering a therapeutically effective amount of a LAG-3 inhibitor and a PD-1 pathway inhibitor. In some embodiments, the immunotherapy comprises administering a therapeutically effective amount of a LAG-3 inhibitor and an anti-PD-1 antibody. In some embodiments, the immunotherapy comprises administering a therapeutically effective amount of a LAG-3 inhibitor and an anti-PD-Ll antibody. In some embodiments, the immunotherapy comprises administering a therapeutically effective amount of a LAG-3 inhibitor. In some embodiments, the immunotherapy comprises administering a therapeutically effective amount of a PD-1 pathway inhibitor.
• the immunotherapy comprises administering a therapeutically effective amount of an anti-CTLA-4 antibody. In some embodiments, the immunotherapy comprises administering a therapeutically effective amount of a PD-1 pathway inhibitor and an immune checkpoint inhibitor.
• the LAG-3 inhibitor is an anti-LAG-3 antibody and the PD-1 pathway inhibitor is an anti-PD-1 antibody. In some embodiments, the LAG-3 inhibitor is an anti-LAG-3 antibody and the PD-1 pathway inhibitor is an anti-PD-Ll antibody.
• the invention includes a method for increasing a disease control rate to be higher than 55% in a patient population, wherein each patient of the patient population is afflicted with a malignant tumor, in a cancer treatment comprising administering to the patient an immunotherapy disclosed herein, wherein each patient is identified as having a LAG-3 positive malignant tumor (e.g., melanoma) or a LAG-3 positive PD-Ll positive malignant tumor prior to the administration and wherein the disease control rate is higher than 55%, 60%), 65%o, 70%), or 75%.
• the method comprises identifying the patient as having a LAG-3 positive malignant tumor or a LAG-3 positive PD-Ll positive malignant tumor prior to the administration.
• the LAG-3 positive malignant tumor is a LAG-3 positive PD-Ll negative malignant tumor.
• the immunotherapy comprises administering a therapeutically effective amount of a LAG-3 inhibitor and a PD-1 pathway inhibitor. In some embodiments, the immunotherapy comprises administering a therapeutically effective amount of a LAG-3 inhibitor. In some embodiments, the immunotherapy comprises administering a therapeutically effective amount of a PD- 1 pathway inhibitor. In some embodiments, the immunotherapy comprises administering a therapeutically effective amount of an anti-PD- 1 antibody. In some embodiments, the immunotherapy comprises administering a therapeutically effective amount of an anti-PD-Ll antibody.
• the immunotherapy comprises administering a therapeutically effective amount of an anti-CTLA-4 antibody. In some embodiments, the immunotherapy comprises administering a therapeutically effective amount of a PD- 1 pathway inhibitor and an immune checkpoint inhibitor.
• the LAG-3 inhibitor is an anti -LAG-3 antibody and the PD- 1 pathway inhibitor is an anti-PD-1 antibody. In some embodiments, the LAG-3 inhibitor is an anti -LAG-3 antibody and the PD- 1 pathway inhibitor is an anti-PD-Ll antibody.
• each patient in the methods experiences (i) extended progression-free survival for over 12 months, (ii) tumor size reduction at least about 10%>, about 20%), about 30%o, about 40%, or about 50% compared to the tumor size prior to the administration, or (iii) both.
• the patient population can be at least 100 patients having a LAG-3 positive malignant tumor (e.g., melanoma) or a LAG-3 positive PD-Ll positive malignant tumor.
• the patient population can be at least 200, 300, 400, 500, 600, 700, 800, 900, or 1000 patients having a LAG-3 positive malignant tumor or a LAG-3 positive PD-Ll positive malignant tumor.
• the LAG-3 positive malignant tumor is a LAG-3 positive PD-Ll negative malignant tumor.
• the invention provides a method for selecting a human patient suitable for a combination therapy comprising: (a) identifying a patient as having a LAG- 3 positive malignant tumor or a LAG-3 positive PD-Ll positive malignant tumor; and (b) instructing a healthcare provider to administer to the patient an immunotherapy disclosed herein.
• the LAG-3 positive malignant tumor is a LAG-3 positive PD-Ll negative malignant tumor.
• the method can further comprise administering an immunotherapy disclosed herein.
• the immunotherapy comprises administering a therapeutically effective amount of a LAG-3 inhibitor and a PD-1 pathway inhibitor.
• the immunotherapy comprises administering a therapeutically effective amount of a LAG-3 inhibitor.
• the immunotherapy comprises administering a therapeutically effective amount of a PD-1 pathway inhibitor. In some embodiments, the immunotherapy comprises administering to the patient a therapeutically effective amount of an anti-PD-1 antibody. In some embodiments, the immunotherapy comprises administering to the patient a therapeutically effective amount of an anti -PD-Ll antibody. In some embodiments, the immunotherapy comprises administering a therapeutically effective amount of an anti-CTLA-4 antibody. In some embodiments, the immunotherapy comprises administering a therapeutically effective amount of a PD-1 pathway inhibitor and an immune checkpoint inhibitor. 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. In some embodiments, the LAG-3 inhibitor is an anti-LAG-3 antibody and the PD-1 pathway inhibitor is an anti-PD-Ll antibody. In some embodiments, the administration treats the malignant tumor.
• the methods of the invention can treat the malignant tumor, reduce the tumor size, prevent growth of the tumor, eliminate the tumor from the patient, prevent a relapse of a tumor, induce a remission in a patient, or any combination thereof.
• the administration of an immunotherapy disclosed herein induces a complete response.
• the administration of the immunotherapy disclosed herein induces a partial response.
• the immunotherapy comprises administering a therapeutically effective amount of a LAG-3 inhibitor and a PD-1 pathway inhibitor.
• the immunotherapy comprises administering a therapeutically effective amount of a LAG-3 inhibitor.
• the immunotherapy comprises administering a therapeutically effective amount of a PD-1 pathway inhibitor. In some embodiments, the immunotherapy comprises administering to the patient a therapeutically effective amount of an anti-PD-1 antibody. In some embodiments, the immunotherapy comprises administering to the patient a therapeutically effective amount of an anti -PD-Ll antibody. In some embodiments, the immunotherapy comprises administering to the patient a therapeutically effective amount of an anti-CTLA-4 antibody. In some embodiments, the immunotherapy comprises administering a therapeutically effective amount of a PD-1 pathway inhibitor and an immune checkpoint inhibitor. 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. In some embodiments, the LAG-3 inhibitor is an anti-LAG-3 antibody and the PD-1 pathway inhibitor is an anti -PD-Ll antibody.
• the LAG-3 positive tumor comprises 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% cells expressing LAG-3.
• the cells expressing LAG-3 comprise tumor infiltrating lymphocytes.
• the identifying comprises determining LAG-3 expression in a malignant tumor.
• LAG-3 expression is determined by receiving the results of an assay capable of determining LAG-3 expression.
• any of the present methods further comprise determining
• any of the present methods further comprise identifying the patient as having a PD-Ll positive malignant tumor prior to the administration. In certain embodiments, any of the present methods further comprise identifying the patient as having a PD-Ll negative malignant tumor prior to the administration.
• any of the present methods further comprise determining
• the patient is identified as having a PD-Ll positive malignant tumor prior to the administration. In certain embodiments of any of the present methods, the patient is identified as having a PD-Ll negative malignant tumor prior to the administration.
• the methods of the invention include methods of treating a human patient with unresectable or metastatic melanoma in need thereof with a combination of a PD-1 pathway inhibitor and a LAG-3 inhibitor, wherein the patient was previously treated with an anti-PD-1 inhibitor and/or an anti-PD-Ll inhibitor.
• the PD-1 pathway inhibitor is an anti-PD-1 antibody.
• the anti-PD-1 antibody is nivolumab.
• the LAG-3 inhibitor is an anti-LAG-3 antibody.
• the LAG-3 antibody is BMS-986016.
• the melanoma is a LAG-3 expressing tumor.
• the melanoma is a LAG-3 expression tumor, with LAG-3 expression > 1%.
• identifying a patient suitable for a LAG-3 inhibitor/ PD-1 pathway inhibitor combination therapy, a PD-1 pathway inhibitor (e.g., an anti-PD-1 antibody) therapy, or an anti-CTLA-4 antibody therapy for the present methods includes measuring or assessing a LAG-3 expression in a sample, for example, a malignant tumor test tissue sample comprising tumor cells and tumor infiltrating inflammatory cells.
• a malignant tumor test tissue sample comprising tumor cells and tumor infiltrating inflammatory cells.
• the phrases "tumors expressing LAG-3,” “LAG-3 expressing tumor,” “LAG-3 positive tumor,” and “LAG-3 expression positive tumor” are used interchangeably herein and encompass tumors comprising LAG-3 expressing tumor-infiltrating lymphocytes. The meaning of the phrases is provided elsewhere herein.
• the methods of measuring or assessing the LAG-3 expression can be achieved by any methods applicable.
• a test tissue sample is obtained from the patient who is in need of the therapy.
• a test tissue sample includes, but is not limited to, any clinically relevant tissue sample, such as a tumor biopsy, a core biopsy tissue sample, a fine needle aspirate, or a sample of bodily fluid, such as blood, plasma, serum, lymph, ascites fluid, cystic fluid, or urine.
• the test tissue sample is from a primary tumor.
• the test tissue sample is from a metastasis.
• test tissue samples are taken from a subject at multiple time points, for example, before treatment, during treatment, and/or after treatment.
• test tissue samples are taken from different locations in the subject, for example, a sample from a primary tumor and a sample from a metastasis in a distant location.
• the test tissue sample is a paraffin-embedded fixed tissue sample.
• the test tissue sample is a formalin-fixed paraffin embedded (FFPE) tissue sample.
• the test tissue sample is a fresh tissue (e.g., tumor) sample.
• the test tissue sample is a frozen tissue sample.
• the test tissue sample is a fresh frozen (FF) tissue (e.g., tumor) sample.
• the test tissue sample is a cell isolated from a fluid.
• the test tissue sample comprises circulating tumor cells (CTCs). In some embodiments, the test tissue sample comprises tumor-infiltrating lymphocytes (TILs). In some embodiments, the test tissue sample comprises tumor cells and tumor-infiltrating lymphocytes (TILs). In some embodiments, the test tissue sample comprises circulating lymphocytes. In some embodiments, the test tissue sample is an archival tissue sample. In some embodiments, the test tissue sample is an archival tissue sample with known diagnosis, treatment, and/or outcome history. In some embodiments, the sample is a block of tissue. In some embodiments, the test tissue sample is dispersed cells. In some embodiments, the sample size is from about 1 cell to about 1 x 10 6 cells or more.
• the sample size is about 1 cell to about 1 x 10 5 cells. In some embodiments, the sample size is about 1 cell to about 10,000 cells. In some embodiments, the sample size is about 1 cell to about 1,000 cells. In some embodiments, the sample size is about 1 cells to about 100 cells. In some embodiments, the sample size is about 1 cell to about 10 cells. In some embodiments, the sample size is a single cell.
• the assessment of LAG-3 expression can be achieved without obtaining a test tissue sample.
• selecting a suitable patient includes (i) optionally providing a test tissue sample obtained from a patient with cancer of the tissue, the test tissue sample comprising tumor cells and/or tumor-infiltrating inflammatory cells; and (ii) assessing the proportion of cells in the test tissue sample that express LAG-3 on the surface of the cells based on an assessment that the proportion of cells in the test tissue sample that express LAG-3 on the cell surface is higher than a predetermined threshold level.
• the step comprising the provision of a test tissue sample obtained from a patient is 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. It should also be understood that in certain embodiments the "measuring" or “assessing” step to identify, or determine the number or proportion of, cells in the test tissue sample that express LAG-3 is performed by a transformative method of assaying for LAG-3 expression, for example by performing a reverse transcriptase-polymerase chain reaction (RT-PCR) assay or an IHC assay.
• RT-PCR reverse transcriptase-polymerase chain reaction
• no transformative step is involved and LAG-3 expression is assessed by, for example, reviewing a report of test results from a laboratory.
• LAG-3 expression is assessed by reviewing the results of an immunohistochemistry assay from a laboratory.
• the steps of the methods up to, and including, assessing LAG-3 expression provides an intermediate result that may be provided to a physician or other healthcare provider for use in selecting a suitable candidate for the combination therapy of a LAG-3 inhibitor and a PD-1 pathway inhibitor.
• the steps of the methods up to, and including, assessing LAG-3 expression provides an intermediate result that may be provided to a physician or other healthcare provider for use in selecting a suitable candidate for PD-1 pathway inhibitor (e.g., anti-PD-1 antibody) therapy.
• the steps of the methods up to, and including, assessing LAG-3 expression provides an intermediate result that may be provided to a physician or other healthcare provider for use in selecting a suitable candidate for anti-CTLA-4 antibody therapy.
• the steps that provide the intermediate result is performed by a medical practitioner or someone acting under the direction of a medical practitioner. In other embodiments, these steps are performed by an independent laboratory or by an independent person such as a laboratory technician.
• the proportion of cells that express LAG-3 is assessed by performing an assay to detect the presence of LAG-3 RNA.
• the presence of LAG-3 RNA is detected by RT-PCR, in situ hybridization or RNase protection.
• the presence of LAG-3 RNA is detected by an RT- PCR based assay.
• scoring the RT-PCR based assay comprises assessing the level of LAG-3 RNA expression in the test tissue sample relative to a predetermined level.
• the proportion of cells that express LAG-3 is assessed by performing an assay to detect the presence of LAG-3 polypeptide.
• the presence of LAG-3 polypeptide is detected by IHC, enzyme-linked immunosorbent assay (ELISA), in vivo imaging, or flow cytometry.
• ELISA enzyme-linked immunosorbent assay
• LAG-3 expression is assayed by IHC.
• cell surface expression of LAG-3 is assayed using, e.g., IHC or in vivo imaging.
• the biomarker measured is LAG-3, CD4, CD8, FOXP3, CD163
• the biomarker is measured using any detection method disclosed herein.
• the proportion of cells that express LAG-3 in the test tissue sample is assessed by flow cytometry.
• the test tissue sample assayed by flow cytometry comprises tumor infiltrating immune cells.
• the malignant tumor is a hematological malignancy and the tissue sample assayed by flow cytometry comprises peripheral blood cells.
• the flow cytometry is a multiplex assay.
• scoring the flow cytometry comprises detecting the expression of markers comprising LAG-3, CD4, CD8, FOXP3, and any combination thereof.
• LAG-3, CD4, CD8, and FOXP3 are detected as single markers.
• scoring the flow cytometry comprises assessing the proportion of T cells in the test tissue sample that express LAG-3.
• scoring the flow cytometry comprises assessing the proportion of CD8+ T cells in the test tissue sample that express LAG-3.
• scoring the flow cytometry comprises assessing the proportion of CD4+ T cells in the test tissue sample that express LAG-3.
• scoring the flow cytometry comprises assessing the proportion of FOXP3+ T cells in the test tissue sample that express LAG-3.
• scoring the flow cytometry comprises detecting the expression of markers comprising CD163 and/or CD68.
• scoring the flow cytometry comprises assessing the proportion of cells in the test tissue sample that express CD 163 and/or CD68.
• the proportion of cells that express LAG-3 in the test tissue sample is assessed by performing an assay to detect the presence of LAG-3 polypeptide.
• the presence of LAG-3 polypeptide is detected by an immunohistochemistry assay.
• the test tissue sample is a tumor biopsy.
• the test tissue sample is a formalin-fixed paraffin embedded (FFPE) sample.
• the immunohistochemistry assay is a monoplex assay. In some embodiments, the immunohistochemistry assay is a multiplex assay. In some embodiments, the multiplex immunohistochemistry assay is capable of detecting the presence of CD4, CD8, FOXP3, CD 163, CD68, or any combination thereof.
• the immunohistochemistry assay comprises contacting the tumor sample with the 17B4 mouse anti -human LAG-3 IgGl monoclonal antibody. In some embodiments, the immunohistochemistry assay comprises contacting the tumor sample with an anti-LAG-3 antibody comprising heavy and light chain variable regions comprising the sequences set forth in SEQ ID NOs: 3 and 5, respectively. In some embodiments, the immunohistochemistry assay comprises contacting the tumor sample with the SP346 rabbit anti- human LAG-3 IgG monoclonal antibody. In some embodiments, the immunohistochemistry assay comprises contacting the tumor sample with the 1 1E3 (Novusbio), 874501 (Novusbio), or EPR4392(2) (Abeam) anti-human LAG-3 monoclonal antibody.
• Melanin for example, in melanoma tumor samples, can interfere with histological analysis by obscuring histological features, and by interfering with and/or masking staining during immunohistochemistry (IHC). Melanin can be removed by bleaching the samples. See, e.g., Shen & Wu, Appl Immunohistochem Mol Morphol, 23(4): 303-307 (2015); Orchard & Calonje, Am J Dermatopathol, 20(4): 357-61 (1998).
• the immunohistochemistry assay comprises melanin bleaching prior to contacting the sample with an anti-LAG-3 antibody. See, e.g., Figure 15.
• the melanin bleaching comprises contacting the sample with dilute hydrogen peroxide (0.1 to 30% v/v), trichloroisocyanuric acid (TCCA), potassium permanganate/oxalic acid, or other traditional oxidation methods for depigmenting (i.e., removing melanin from) tissue samples.
• dilute hydrogen peroxide 0.1 to 30% v/v
• TCCA trichloroisocyanuric acid
• potassium permanganate/oxalic acid potassium permanganate/oxalic acid
• the immunohistochemistry assay uses a black or brown chromogen. In some embodiments, the immunohistochemistry assay uses a red chromogen. In some embodiments, the immunohistochemistry assay uses a blue chromogen. In some embodiments, the immunohistochemistry assay uses a green chromogen. In some embodiments, the immunohistochemistry assay uses a purple chromogen. In some embodiments, the immunohistochemistry assay uses a yellow chromogen.
• the immunohistochemistry assay is scored at a low magnification. In some embodiments, low magnification is about 20X. In some embodiments, the immunohistochemistry assay is scored at high magnification. In some embodiments, high magnification is about 40X.
• the immunohistochemistry assay is scored by an image analysis software. In some embodiments, the immunohistochemistry assay is scored by pathologist visual immune score. In some embodiments, the immunohistochemistry assay is scored manually. [0210] In some embodiments, scoring the immunohistochemistry assay comprises assessing the proportion of cells in the test tissue sample that express LAG-3. In some embodiments, scoring the immunohistochemistry assay comprises assessing the proportion of immune cells in the test tissue sample that express LAG-3. In some embodiments, scoring the immunohistochemistry assay comprises assessing the proportion of T cells in the test tissue sample that express LAG-3.
• scoring the immunohistochemistry assay comprises assessing the proportion of CD8+ T cells in the test tissue sample that express LAG-3. In some embodiments, scoring the immunohistochemistry assay comprises assessing the proportion of CD4+ T cells in the test tissue sample that express LAG-3. In some embodiments, scoring the immunohistochemistry assay comprises assessing the proportion of FOXP3+ T cells in the test tissue sample that express LAG-3.
• LAG-3 polypeptide localization includes partial membrane/cytoplasmic localization, dot like localization, perinuclear, and complete membrane/cytoplasmic localization. In some embodiments, cells with partial membrane/cytoplasmic LAG-3 localization are scored. In some embodiments, cells with dot-like LAG-3 localization are scored. In some embodiments, cells with complete membrane/cytoplasmic LAG-3 localization are scored. In some embodiments, cells with perinuclear LAG-3 localization are scored. In some embodiments, cells with any LAG-3 localization pattern are scored.
• the immunohistochemistry assay is a multiplex assay that further comprises detecting the expression of MHC Class II by the tumor cells.
• scoring the immunohistochemistry assay comprises assessing the proportion of cells in the test tissue sample that expresses MHC Class II.
• scoring the immunohistochemistry assay comprises assessing the proportion of non-immune cells in the test tissue sample that expresses MHC Class II.
• the distribution of MHC II expressing cells is heterogenous in the tumor sample.
• scoring the immunohistochemistry assay comprises assessing the proportion of cells that expresses MHC Class II in regions of the tumor sample comprising a high density of MHC Class II expressing cells.
• the immunohistochemistry assay is a multiplex assay that further comprises detecting the expression of CD 163 and/or CD68 by tumor infiltrating lymphocytes (TIL).
• scoring the immunohistochemistry assay comprises assessing the proportion of TILs in the test tissue sample that expresses CD163 and/or CD68.
• LAG-3 expression is assayed by immunoPET imaging.
• immunoPET is performed using a zirconium-89 radiolabeled anti-LAG-3 antibody.
• the proportion of cells in a test tissue sample that express LAG-3 is assessed by performing an assay to determine the presence of LAG-3 polypeptide on the surface of cells in the test tissue sample.
• the test tissue sample is a FFPE tissue sample.
• the presence of LAG-3 polypeptide is determined by IHC assay.
• the IHC assay is performed using an automated process.
• the IHC assay is performed using an anti-LAG-3 mAb to bind to the LAG-3 polypeptide.
• an automated IHC method is used to assay the expression of LAG-3 in FFPE tissue specimens.
• This disclosure provides methods for detecting the presence of human LAG-3 antigen in a test tissue sample, or quantifying the level of human LAG-3 antigen or the proportion of cells in the sample that express the antigen, which methods comprise contacting the test sample, and a negative control sample, with a mAb that specifically binds to human LAG-3, under conditions that allow for formation of a complex between the antibody or portion thereof and human LAG-3.
• the test and control tissue samples are FFPE samples. The formation of a complex is then detected, wherein a difference in complex formation between the test sample and the negative control sample is indicative of the presence of human LAG-3 antigen in the sample.
• Various methods are used to quantify LAG-3 expression.
• the automated IHC method comprises: (a) deparaffinizing and rehydrating mounted tissue sections in an autostainer; (b) retrieving antigen in an autostainer; (c) setting up reagents on an autostainer; and (d) running the autostainer to include steps of neutralizing endogenous peroxidase in the tissue specimen; blocking nonspecific protein-binding sites on the slides; incubating the slides with primary Ab; incubating with a postprimary blocking agent; incubating with a postprimary antibody detection agent, such as another antibody that may or may not be conjugated to a detection enzyme; incubating with a polymeric-enzyme detection reagent; adding a chromogen substrate and developing; and counterstaining with hematoxylin.
• the retrieving antigen comprises using any heat based antigen retrieval device.
• a pathologist examines the number of LAG-3+ tumor cells in each field under a microscope and mentally estimates the percentage of cells that are positive, then averages them to come to the final percentage.
• the different staining intensities are defined as 0/negative, l+/weak, 2+/moderate, and 3+/strong.
• percentage values are first assigned to the 0 and 3+ buckets, and then the intermediate 1+ and 2+ intensities are considered.
• the specimen is divided into zones, and each zone is scored separately and then combined into a single set of percentage values.
• the percentages of negative and positive cells for the different staining intensities are determined from each area and a median value is given to each zone. A final percentage value is given to the tissue for each staining intensity category: negative, 1+, 2+, and 3+. The sum of all staining intensities needs to be 100%.
• staining is also assessed in tumor-infiltrating inflammatory cells such as macrophages and lymphocytes. Macrophages and lymphocytes are assessed for LAG-3 staining and only recorded for all samples as being positive or negative for each cell category. Staining is also characterized according to an outside/inside tumor immune cell designation. "Inside” means the immune cell is within the tumor tissue and/or on the boundaries of the tumor region without being physically intercalated among the tumor cells. "Outside” means that there is no physical association with the tumor, the immune cells being found in the periphery associated with connective or any associated adjacent tissue. [0219] In certain embodiments of these scoring methods, the samples are scored by two or more pathologists operating independently, and the scores are subsequently consolidated. In certain other embodiments, the identification of positive and negative cells is scored using appropriate software.
• H-score A histoscore (H-score) is used as a more quantitative measure of the IHC data.
• the histoscore is calculated as follows:
• Histoscore [(% tumor x 1 (low intensity)) + (% tumor x 2 (medium intensity))
• the pathologist estimates the percentage of stained cells in each intensity category within a specimen. Because expression of most biomarkers is heterogeneous the histoscore is a truer representation of the overall expression. The final histoscore range is 0 (minimum score, no expression) to 300 (maximum score, strong and inclusive expression).
• the invention features methods of using a LAG-3 inhibitor in the treatment of malignant tumors.
• LAG-3 inhibitor includes, but is not limited to, LAG-3 binding agents and soluble LAG-3 polypeptides.
• LAG-3 binding agents include antibodies that specifically bind to LAG-3.
• a LAG-3 inhibitor is a LAG-3-binding agent, for example an anti-LAG-3 antibody.
• the LAG-3 inhibitor is a soluble LAG-3 polypeptide, for example, a LAG-3 -Fc fusion polypeptide capable of binding to MHC Class II.
• Anti-human-LAG-3 antibodies (or VH/VL domains derived therefrom) suitable for use in the invention can be generated using methods well known in the art. Alternatively, art recognized anti-LAG-3 antibodies can be used.
• LAG-3 inhibitors include an anti-LAG-3 bispecific antibody. In some embodiments, the anti-LAG-3 antibody binds LAG-3 and PD-1.
• the anti-LAG-3 antibody is BMS-986016 comprising heavy and light chains comprising the sequences shown in SEQ ID NOs: l and 2, respectively, or antigen binding fragments and variants thereof, as described in PCT/US 13/48999.
• the antibody has the heavy and light chain CDRs or variable regions of BMS-986016. Accordingly, in one embodiment, the antibody comprises CDR1, CDR2, and CDR3 domains of the VH region of BMS-986016 having the sequence set forth in SEQ ID NO:3, and CDR1, CDR2 and CDR3 domains of the VL region of BMS-986016 having the sequence set forth in SEQ ID NO:5. In another embodiment, the antibody comprises CDR1, CDR2 and CDR3 domains comprising the sequences set forth in SEQ ID NOs:7, 8, and 9, respectively, and CDR1, CDR2 and CDR3 domains comprising the sequences set forth in SEQ ID NOs: 10, 11, and 12, respectively.
• the antibody comprises VH and/or VL regions comprising the amino acid sequences set forth in SEQ ID NO:3 and/or SEQ ID NO: 5, respectively.
• the antibody comprises heavy chain variable (VH) and/or light chain variable (VL) regions encoded by the nucleic acid sequences set forth in SEQ ID NO:4 and/or SEQ ID NO:6, respectively.
• the antibody competes for binding with and/or binds to the same epitope on LAG-3 as the above-mentioned antibodies.
• the antibody binds an epitope of human LAG-3 comprising the amino acid sequence PGHPLAPG (SEQ ID NO: 14).
• the antibody binds an epitope of human LAG-3 comprising the amino acid sequence HPAAPSSW (SEQ ID NO: 15) or PAAPSSWG (SEQ ID NO: 16).
• the antibody has at least about 90% variable region amino acid sequence identity with the above-mentioned antibodies (e.g., at least about 90%, 95% or 99% variable region identity with SEQ ID NO:3 or SEQ ID NO:5).
• art recognized anti-LAG-3 antibodies can be used in the therapeutic methods of the invention.
• the anti-human LAG-3 antibody described in US2011/0150892 Al and referred to as monoclonal antibody 25F7 (also known as “25F7” and "LAG-3.1) can be used.
• Other art recognized anti-LAG-3 antibodies that can be used include IMP731 (H5L7BW) described in US 2011/007023, MK-4280 (28G-10) described in WO2016028672, REGN3767 described in Journal for ImmunoTherapy of Cancer, (2016) Vol. 4, Supp.
• anti-LAG-3 antibodies useful in the claimed invention can be found in, for example: 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, WO2017/015560, WO2017/025498, WO2017/087589, WO2017/087901, WO2018/083087, WO2017/149143, WO2017/219995, US2017/0260271, WO2017/086367, WO2017/086419, WO2018/034227, and WO2014/140180.
• the LAG- 3 inhibitor is IMP321 (eftilagimod alpha). The contents of each of these references are incorporated by reference
• Antibodies that compete with any of the above-referenced art-recognized antibodies for binding to LAG-3 also can be used.
• an anti-LAG-3 antibody is used to determine LAG-3 expression.
• an anti-LAG-3 antibody is selected for its ability to bind to LAG-3 in formalin-fixed, paraffin-embedded (FFPE) tissue specimens.
• FFPE paraffin-embedded
• an anti-LAG-3 antibody is capable of binding to LAG-3 in frozen tissues.
• an anti-LAG-3 antibody is capable of distinguishing membrane bound, cytoplasmic, and/or soluble forms of LAG-3.
• an anti-LAG-3 antibody useful for assaying, detecting, and/or quantifying LAG-3 expression in accordance with the methods described herein is the 17B4 mouse IgGl anti -human LAG-3 monoclonal antibody, or an antigen binding fragment thereof. See, e.g., J. Matsuzaki, et al.; PNAS 107, 7875 (2010).
• the invention features methods of using a PD-1 inhibitor in the treatment of malignant tumors.
• PD-1 pathway inhibitor includes, but is not limited to, PD-1 binding agents, PD-Ll binding agent and PD-L2 binding agents.
• PD-1 binding agents include antibodies that specifically bind to PD-1.
• PD-Ll and PD-L2 binding agents include antibodies that specifically bind to PD-Ll and/or PD-L2, as well as soluble PD-1 polypeptides that bind to PD-Ll and/or PD-L2.
• PD-1 pathway inhibitor is a PD-1 -binding agent, for example an anti-PD-1 antibody.
• the PD-1 pathway inhibitor is a PD-L1- binding agent, for example, an anti-PD-Ll antibody.
• the PD-1 pathway inhibitor is a PD-L2 -binding agent, for example an anti-PD-L2 antibody.
• the PD-Ll-binding agent is a soluble PD-1 polypeptide, for example, a PD-l-Fc fusion polypeptide capable of binding to PD-Ll .
• the PD-L2 -binding agent is a soluble PD-1 polypeptide, for example, a PD-l-Fc fusion polypeptide capable of binding to PD- L2.
• Anti-human-PD-1 antibodies (or VH and/or VL domains derived therefrom) suitable for use in the invention can be generated using methods well known in the art. Alternatively, art recognized anti-PD-1 antibodies can be used. For example, monoclonal antibodies 5C4 (referred to herein as Nivolumab or BMS-936558), 17D8, 2D3, 4H1, 4A11, 7D3, and 5F4, described in WO 2006/121 168 can be used.
• PD-1 antibodies include lambrolizumab (MK-3475) described in WO 2008/156712, and AMP-514 described in WO 2012/145493. Further known PD-1 antibodies and other PD-1 inhibitors include those described in, for example, WO 2009/014708, WO 03/099196, WO 2009/114335 and WO 2011/161699, which are herein incorporated by reference..
• the anti-PD-1 antibody is REGN2810.
• the anti-PD-1 antibody is PDR001.
• Another known anti-PD-1 antibody is pidilizumab (CT-011).
• the anti-PD-1 antibody is nivolumab.
• Nivolumab also known as "OPDIVO ® "; formerly designated 5C4, BMS-936558, MDX-1106, or ONO-4538
• OPDIVO ® is a fully human IgG4 (S228P) PD-1 immune checkpoint inhibitor antibody that selectively prevents interaction with PD-1 ligands (PD-L1 and PD-L2), thereby blocking the down-regulation of antitumor T-cell functions
• S228P PD-1 immune checkpoint inhibitor antibody that selectively prevents interaction with PD-1 ligands (PD-L1 and PD-L2), thereby blocking the down-regulation of antitumor T-cell functions
• the anti-PD-1 antibody or fragment thereof cross- competes with nivolumab. In other embodiments, the anti-PD-1 antibody or fragment thereof binds to the same epitope as nivolumab. In certain embodiments, the anti-PD-1 antibody has the same CDRs as nivolumab.
• the anti-PD-1 antibody comprises heavy and light chains comprising the sequences shown in SEQ ID NOs: 17 and 18, respectively, or antigen binding fragments and variants thereof.
• the antibody has heavy and light chain CDRs or variable regions of nivolumab. Accordingly, in one embodiment, the antibody comprises CDR1, CDR2, and CDR3 domains of the VH of nivolumab having the sequence set forth in SEQ ID NO: 19, and CDR1, CDR2 and CDR3 domains of the VL of nivolumab having the sequence set forth in SEQ ID NO:21. In another embodiment, the antibody comprises CDR1, CDR2 and CDR3 domains comprising the sequences set forth in SEQ ID NOs:23, 24, and 25, respectively, and CDR1, CDR2 and CDR3 domains comprising the sequences set forth in SEQ ID NOs:26, 27, and 28, respectively.
• the antibody comprises VH and/or VL regions comprising the amino acid sequences set forth in SEQ ID NO: 19 and/or SEQ ID NO: 21, respectively.
• the antibody comprises heavy chain variable (VH) and/or light chain variable (VL) regions encoded by the nucleic acid sequences set forth in SEQ ID NO:20 and/or SEQ ID NO:22, respectively.
• the antibody competes for binding with and/or binds to the same epitope on PD-1 as the above-mentioned antibodies.
• the antibody has at least about 90% variable region amino acid sequence identity with the above-mentioned antibodies (e.g., at least about 90%, 95% or 99% variable region identity with SEQ ID NO: 19 or SEQ ID NO:21).
• Human monoclonal antibodies that bind specifically to PD-1 with high affinity have been disclosed in U.S. Patent Nos. 8,008,449 and 8,779,105.
• Other anti-PD-1 mAbs have been described in, for example, U.S. Patent Nos. 6,808,710, 7,488,802, 8, 168,757 and 8,354,509, and PCT Publication No. WO 2012/145493, which are herein incorporated by reference.
• the anti-PD-1 antibody has been demonstrated to exhibit one or more of the following characteristics: (a) binds to human PD-1 with a K D of 1 x 10 "7 M or less, as determined by surface plasmon resonance using a Biacore biosensor system; (b) does not substantially bind to human CD28, CTLA-4 or ICOS; (c) increases T-cell proliferation in a Mixed Lymphocyte Reaction (MLR) assay; (d) increases interferon- ⁇ production in an MLR assay; (e) increases IL-2 secretion in an MLR assay; (f) binds to human PD-1 and cynomolgus monkey PD-1; (g) inhibits the binding of PD-L1 and/or PD-L2 to PD-1; (h) stimulates antigen- specific memory responses; (i) stimulates antibody responses; and (j) inhibits tumor cell growth in vivo.
• MLR Mixed Lymphocyte Reaction
• Anti-PD-1 antibodies useful for the present invention include mAbs that bind specifically to human PD-1 and exhibit at least one, at least two, at least three, at least four, or at least five of the preceding characteristics. Anti-PD-1 antibodies that exhibit one or more of these characteristics have been disclosed in U.S. Patent Nos. 8,008,449, 8,779, 105, 6,808,710, 7,488,802, 8, 168,757 and 8,354,509, and PCT Publication No. WO 2012/145493, which are herein incorporated by reference. In another embodiment, the anti-PD-1 antibody is pembrolizumab.
• Pembrolizumab is a humanized monoclonal IgG4 (S228P) antibody directed against human cell surface receptor PD-1 (programmed death- 1 or programmed cell death- 1). Pembrolizumab is described, for example, in U.S. Patent Nos. 8,354,509 and 8,900,587, which are herein incorporated by reference.
• the anti-PD-1 antibody or fragment thereof cross-competes with pembrolizumab. In some embodiments, the anti-PD-1 antibody or fragment thereof binds to the same epitope as pembrolizumab. In certain embodiments, the anti-PD-1 antibody has the same CDRs as pembrolizumab. In another embodiment, the anti-PD-1 antibody is pembrolizumab.
• Pembrolizumab also known as "KEYTRUDA ® ", lambrolizumab, and MK- 3475
• Pembrolizumab is a humanized monoclonal IgG4 antibody directed against human cell surface receptor PD-1 (programmed death-1 or programmed cell death-1). Pembrolizumab is described, for example, in U.S.
• Patent Nos. 8,354,509 and 8,900,587; see also http://www.cancer.gov/drugdictionary?cdrid 695789 (last accessed: December 14, 2014).
• Pembrolizumab has been approved by the FDA for the treatment of relapsed or refractory melanoma.
• the first antibody is an anti-PD-1 antagonist.
• the anti-PD-1 antagonist is AMP-224, which is a B7-DC Fc fusion protein.
• the anti-PD-1 antibody or fragment thereof cross-competes with BGB-A317.
• the anti-PD-1 antibody or fragment thereof binds the same epitope as BGB-A317.
• the anti-PD-1 antibody has the same CDRs as BGB-A317.
• the anti-PD-1 antibody is BGB-A317, which is a humanized monoclonal antibody. BGB-A317 is described in U.S. Publ. No. 2015/0079109.
• the antibody is pidilizumab (CT-011), which is an antibody previously reported to bind to PD-1 but which is believed to bind to a different target, pidilizumab is described in US Pat. No. 8,686,119 B2 or WO 2013/014668 Al .
• the antibodies that cross-compete for binding to human are [0244] in certain embodiments.
• these cross-competing antibodies can be chimeric antibodies, or humanized or human antibodies.
• Such chimeric, humanized or human mAbs can be prepared and isolated by methods well known in the art.
• the anti-PD-1 antibody is selected from the group consisting of 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 WO 2015/112900), MEDI-0680 (AstraZeneca; also known as AMP-514; see WO 2012/145493), cemiplimab (Regeneron; also known as REGN-2810; see WO 2015/112800), JSOOl (TAIZHOU JUNSHI PHARMA; see Si-Yang Liu et al., J.
• nivolumab also known as OPDIVO®, 5C4, BMS-936558, MDX-1106, and ONO- 4538
• pembrolizumab Merck;
• Anti-PD-1 antibodies useful for the compositions of the disclosed invention also include antigen-binding portions of the above antibodies. It has been amply demonstrated that the antigen-binding function of an antibody can be performed by fragments of a full-length antibody. Examples of binding fragments encompassed within the term "antigen-binding portion" of an antibody include (i) a Fab fragment, a monovalent fragment consisting of the Y ⁇ , V//, C ⁇ and C HI domains; (ii) a F(ab') 2 fragment, a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; (iii) a Fd fragment consisting of the VH and CHI domains; and (iv) a Fv fragment consisting of the Vz, and YH domains of a single arm of an antibody.
• Anti-PD-1 antibodies usable in the disclosed methods also include isolated antibodies that bind specifically to human PD-1 and cross-compete for binding to human PD-1 with any anti-PD-1 antibody disclosed herein, e.g., nivolumab (see, e.g. , U.S. Patent No. 8,008,449 and 8,779, 105; WO 2013/173223).
• the anti-PD-1 antibody binds the same epitope as any of the anti-PD-1 antibodies described herein, e.g., nivolumab.
• cross-competing antibodies are expected to have functional properties very similar those of the reference antibody, e.g., nivolumab, by virtue of their binding to the same epitope region of PD-1.
• Cross-competing antibodies can be readily identified based on their ability to cross-compete with nivolumab in standard PD-1 binding assays such as Biacore analysis, ELISA assays or flow cytometry (see, e.g., WO 2013/173223).
• Anti-PD-1 antibodies suitable for use in the disclosed methods are antibodies that bind to PD-1 with high specificity and affinity, block the binding of PD-Ll and or PD-L2, and inhibit the immunosuppressive effect of the PD-1 signaling pathway.
• an anti-PD-1 "antibody” includes an antigen-binding portion or fragment that binds to the PD-1 receptor and exhibits the functional properties similar to those of whole antibodies in inhibiting ligand binding and upregulating the immune system.
• the anti-PD-1 antibody or antigen-binding portion thereof cross-competes with nivolumab for binding to human PD-1.
• the anti-PD-1 antibody or antigen- binding portion thereof is a chimeric, humanized or human monoclonal antibody or a portion thereof.
• the antibody is a humanized antibody.
• the antibody is a human antibody.
• Antibodies of an IgGl, IgG2, IgG3 or IgG4 isotype can be used.
• the anti-PD-1 antibody or antigen-binding portion thereof comprises a heavy chain constant region which is of a human IgGl or IgG4 isotype.
• the sequence of the IgG4 heavy chain constant region of the anti-PD-1 antibody or antigen-binding portion thereof contains an S228P mutation which replaces a serine residue in the hinge region with the proline residue normally found at the corresponding position in IgGl isotype antibodies. This mutation, which is present in nivolumab, prevents Fab arm exchange with endogenous IgG4 antibodies, while retaining the low affinity for activating Fc receptors associated with wild-type IgG4 antibodies (Wang et al, 2014 Cancer Immunol Res.
• the antibody comprises a light chain constant region which is a human kappa or lambda constant region.
• the anti-PD-1 antibody or antigen-binding portion thereof is a mAb or an antigen-binding portion thereof.
• the anti-PD-1 antibody is nivolumab.
• the anti-PD-1 antibody is pembrolizumab.
• the anti-PD-1 antibody is chosen from the human antibodies 17D8, 2D3, 4H1, 4A11, 7D3 and 5F4 described in U.S. Patent No. 8,008,449.
• the anti-PD-1 antibody is MEDI0608 (formerly AMP-514), AMP-224, or BGB-A317.
• the anti-PD-1 antibody is a bispecific antibody. In embodiments, the anti-PD-1 antibody is a bispecific antibody that binds both PD-1 and LAG-3.
• the present application encompasses use of an anti-PD-
• the anti-PD-Ll antibody inhibits the binding of PD-L1 receptor, i.e., PD-1 to its ligand PD-L1.
• Anti-human-PD-Ll antibodies (or VH and/or VL domains derived therefrom) suitable for use in the invention can be generated using methods well known in the art.
• art recognized anti-PD-Ll antibodies can be used.
• human anti-PD-Ll antibodies disclosed in U.S. Pat. No. 7,943,743 can be used.
• Such anti-PD-Ll antibodies include 3G10, 12A4 (also referred to as BMS-936559), 10A5, 5F8, 10H10, 1B12, 7H1, 11E6, 12B7, and 13G4.
• the anti-PD-Ll antibody is atezolizumab (Tecentriq or RG7446) (see, e.g., Herbst et al. (2013) J Clin Oncol 31(suppl):3000. Abstract; U.S. 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), avelumab (Bavencio).
• Other art recognized anti-PD-Ll antibodies which can be used include those described in, for example, U.S. Pat. Nos. 7,635,757 and 8,217, 149, U.S.
• Antibodies that compete with any of these art-recognized antibodies or inhibitors for binding to PD-Ll also can be used.
• Examples of anti -PD-Ll antibodies useful in the methods of the present disclosure include the antibodies disclosed in US Patent No. 9,580,507.
• 9,580,507 have been demonstrated to exhibit one or more of the following characteristics: (a) bind to human PD-Ll with a KD of 1 x 10-7 M or less, as determined by surface plasmon resonance using a Biacore biosensor system; (b) increase T-cell proliferation in a Mixed Lymphocyte Reaction (MLR) assay; (c) increase interferon- ⁇ production in an MLR assay; (d) increase IL-2 secretion in an MLR assay; (e) stimulate antibody responses; and (f) reverse the effect of T regulatory cells on T cell effector cells and/or dendritic cells.
• Anti-PD-Ll antibodies usable in the present invention include monoclonal antibodies that bind specifically to human PD-Ll and exhibit at least one, in some embodiments, at least five, of the preceding characteristics.
• the anti-PD-Ll antibody is BMS-936559 (formerly 12A4 or MDX-1105) (see, e.g., U.S. Patent No. 7,943,743; WO 2013/173223).
• the anti-PD-Ll antibody is MPDL3280A (also known as RG7446 and atezolizumab) (see, e.g., Herbst et al. 2013 J Clin Oncol 31(suppl):3000; U.S. Patent No. 8,217, 149), MEDI4736 (Khleif, 2013, In: Proceedings from the European Cancer Congress 2013; September 27-October 1, 2013; Amsterdam, The Netherlands.
• antibodies that cross-compete for binding to human PD- Ll with, or bind to the same epitope region of human PD-Ll as the above-references PD-Ll antibodies are mAbs.
• these cross-competing antibodies can be chimeric antibodies, or can be humanized or human antibodies.
• Such chimeric, humanized or human mAbs can be prepared and isolated by methods well known in the art.
• the anti-PD-Ll antibody is selected from the group consisting of BMS-936559 (also known as 12A4, MDX-1105; see, e.g., U.S.
• the PD-L1 antibody is atezolizumab (TECENTRIQ®).
• Atezolizumab is a fully humanized IgGl monoclonal anti-PD-Ll antibody.
• the PD-L1 antibody is durvalumab (IMFINZITM).
• Durvalumab is a human IgGl kappa monoclonal anti-PD-Ll antibody.
• the PD-L1 antibody is avelumab (BAVENCIO®).
• Avelumab is a human IgGl lambda monoclonal anti-PD-Ll antibody.
• the anti-PD-Ll monoclonal antibody is selected from the group consisting of 28-8, 28-1, 28-12, 29-8, 5H1, and any combination thereof.
• Anti-PD-Ll antibodies usable in the disclosed methods also include isolated antibodies that bind specifically to human PD-L1 and cross-compete for binding to human PD- Ll with any anti-PD-Ll antibody disclosed herein, e.g., atezolizumab, durvalumab, and/or avelumab.
• the anti-PD-Ll antibody binds the same epitope as any of the anti-PD-Ll antibodies described herein, e.g., atezolizumab, durvalumab, and/or avelumab.
• antibodies to cross-compete for binding to an antigen indicates that these antibodies bind to the same epitope region of the antigen and sterically hinder the binding of other cross- competing antibodies to that particular epitope region.
• These cross-competing antibodies are expected to have functional properties very similar those of the reference antibody, e.g., atezolizumab and/or avelumab, by virtue of 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, e.g., WO 2013/173223).
• the antibodies that cross-compete for binding to human are [0260] in certain embodiments.
• PD-L1 with, or bind to the same epitope region of human PD-L1 antibody as, atezolizumab, durvalumab, and/or avelumab are monoclonal antibodies.
• these cross-competing antibodies are chimeric antibodies, engineered antibodies, or humanized or human antibodies.
• Such chimeric, engineered, humanized or human monoclonal antibodies can be prepared and isolated by methods well known in the art.
• Anti-PD-Ll antibodies usable in the methods of the disclosed invention also include antigen-binding portions of the above antibodies. It has been amply demonstrated that the antigen-binding function of an antibody can be performed by fragments of a full-length antibody.
• Anti -PD-Ll antibodies suitable for use in the disclosed methods or compositions are antibodies that bind to PD-Ll with high specificity and affinity, block the binding of PD-1, and inhibit the immunosuppressive effect of the PD-1 signaling pathway.
• an anti-PD-Ll "antibody” includes an antigen-binding portion or fragment that binds to PD-Ll and exhibits the functional properties similar to those of whole antibodies in inhibiting receptor binding and up-regulating the immune system.
• the anti-PD-Ll antibody or antigen-binding portion thereof cross-competes with atezolizumab, durvalumab, and/or avelumab for binding to human PD-Ll .
• Anti-PD-Ll antibodies useful for the invention include antibodies engineered starting from antibodies having one or more of the Y H and/or Y L sequences disclosed herein, which engineered antibodies can have altered properties from the starting antibodies.
• An anti- PD-Ll antibody can be engineered by a variety of modifications as described above for the engineering of modified anti -PD-1 antibodies of the invention.
• the present application encompasses use of an anti-
• the anti-CTLA-4 antibody binds to and inhibits CTLA-4.
• the anti-CTLA-4 antibody is ipilimumab (YERVOY), tremelimumab (ticilimumab; CP-675,206), AGEN-1884, or ATOR-1015.
• the invention features methods of using a PD-1 inhibitor in combination with an immune checkpoint inhibitor in the treatment of malignant tumors. Any art recognized immune checkpoint inhibitor can be used.
• the immune checkpoint inhibitor is a CTLA-4 antagonist, a CD80 antagonist, a CD86 antagonist, a Tim-3 antagonist, a TIGIT antagonist, a CD20 antagonist, a CD96 antagonist, a IDOl antagonist, a STING antagonist, a GARP antagonist, a CD40 antagonist, A2aR antagonist, a CEACAMl (CD66a) antagonist, a CEA antagonist, a CD47 antagonist a PVRIG antagonist, a TDO antagonist, a VISTA antagonist, or a KIR antagonist.
• the immune checkpoint inhibitor is a CTLA-4 antagonist.
• the CTLA-4 antagonist is an anti-CTLA-4 antibody or antigen binding fragment thereof.
• the anti-CTLA-4 antibody is ipilimumab (YERVOY), tremelimumab (ticilimumab; CP-675,206), AGEN-1884, or ATOR-1015.
• the CTLA-4 antagonist is a soluble CTLA-4 polypeptide.
• the soluble CTLA-4 polypeptide is abatacept (Orencia), belatacept (Nulojix), RG2077, or RG-1046.
• the CTLA-4 antagonist is a cell based therapy.
• the CTLA-4 antagonist is an anti-CTLA-4 mAb RNA/GITRL RNA- transfected autologous dendritic cell vaccine or an anti-CTLA-4 mAb RNA-transfected autologous dendritic cell vaccine.
• the immune checkpoint inhibitor is a KIR antagonist.
• the KIR antagonist is an anti-KIR antibody or antigen binding fragment thereof.
• the anti-KIR antibody is linlumab (1-7F9, BMS-986015, IPH 2101) or IPH4102.
• the immune checkpoint inhibitor is TIGIT antagonist.
• the TIGIT antagonist is an anti-TIGIT antibody or antigen binding fragment thereof.
• the anti-TIGIT antibody is BMS-986207, AB 154, COM902 (CGEN-15137), or OMP-313M32.
• the immune checkpoint inhibitor is Tim-3 antagonist.
• the Tim-3 antagonist is an anti-Tim-3 antibody or antigen binding fragment thereof.
• the anti-Tim-3 antibody is TSR-022 or LY3321367.
• the immune checkpoint inhibitor is a IDOl antagonist.
• the IDOl antagonist is indoximod ( LG8189; l-methyl- D -TRP), epacadostat (INCB-024360, INCB-24360), KHK2455, PF-06840003, navoximod (RG6078, GDC-0919, LG919), BMS-986205 (F001287), or pyrrolidine-2,5-dione derivatives.
• the immune checkpoint inhibitor is a STING antagonist.
• the STING antagonist is 2' or 3'-mono-fluoro substituted cyclic-di- nucleotides; 2'3'-di-fluoro substituted mixed linkage 2', 5' - 3 ', 5' cyclic-di-nucleotides; 2'-fluoro substituted, bis-3',5' cyclic-di-nucleotides; 2',2"-diF-Rp,Rp,bis-3',5' cyclic-di-nucleotides; or fluorinated cyclic-di-nucleotides.
• the immune checkpoint inhibitor is CD20 antagonist.
• the CD20 antagonist is an anti-CD20 antibody or antigen binding fragment thereof.
• the anti-CD20 antibody is rituximab (RITUXAN; IDEC-102; IDEC- C2B8), ABP 798, ofatumumab, or obinutuzumab.
• the immune checkpoint inhibitor is CD80 antagonist.
• the CD80 antagonist is an anti-CD80 antibody or antigen binding fragment thereof.
• the anti-CD80 antibody is galiximab or AV 1142742.
• the immune checkpoint inhibitor is a GARP antagonist.
• the GARP antagonist is an anti-GARP antibody or antigen binding fragment thereof.
• the anti-GARP antibody is ARGX-115.
• the immune checkpoint inhibitor is a CD40 antagonist.
• the CD40 antagonist is an anti-CD40 antibody for antigen binding fragment thereof.
• the anti-CD40 antibody is BMS3h-56, lucatumumab (HCD122 and CHIR-12.12), CHIR-5.9, or dacetuzumab (huS2C6, PRO 64553, RG 3636, SGN 14, SGN-40).
• the CD40 antagonist is a soluble CD40 ligand (CD40-L).
• the soluble CD40 ligand is a fusion polypeptide.
• the soluble CD40 ligand is a CD40-L/FC2 or a monomelic CD40-L.
• the immune checkpoint inhibitor is an A2aR antagonist.
• the A2aR antagonist is a small molecule.
• the A2aR antagonist is CPI-444, PBF-509, istradefylline (KW-6002), preladenant (SCH420814), tozadenant (SYN115), vipadenant (BIIB014), HTL-1071, ST1535, SCH412348, SCH442416, SCH58261, ZM241385, or AZD4635.
• the immune checkpoint inhibitor is a CEACAMl antagonist.
• the CEACAMl antagonist is an anti-CEACAMl antibody or antigen binding fragment thereof.
• the anti-CEACAMl antibody is CM-24 (MK- 6018).
• the immune checkpoint inhibitor is a CEA antagonist.
• the CEA antagonist is an anti-CEA antibody or antigen binding fragment thereof.
• the anti-CEA antibody is cergutuzumab amunaleukin (RG7813, RO- 6895882) or RG7802 (R06958688).
• the immune checkpoint inhibitor is a CD47 antagonist.
• the CD47 antagonist is an anti-CD47 antibody or antigen binding fragment thereof.
• the anti-CD47 antibody is HuF9-G4, CC-90002, TTI-621, ALX148, NI-1701, NI-1801, SRF231, or Effi-DEM.
• the immune checkpoint inhibitor is a PVRIG antagonist.
• the PVRIG antagonist is an anti-PVRIG antibody or antigen binding fragment thereof.
• the anti-PVRIG antibody is COM701 (CGEN- 15029).
• the immune checkpoint inhibitor is a TDO antagonist.
• the TDO antagonist is a 4-(indol-3-yl)-pyrazole derivative, a 3-indol substituted derivative, or a 3-(indol-3-yl)-pyridine derivative.
• the immune checkpoint inhibitor is a dual IDO and TDO antagonist.
• the dual IDO and TDO antagonist is a small molecule.
• the immune checkpoint inhibitor is a VISTA antagonist.
• the VISTA antagonist is CA-170 or JNJ-61610588.
• compositions suitable for administration to human patients are typically formulated for parenteral administration, e.g., in a liquid carrier, or suitable for reconstitution into liquid solution or suspension for intravenous administration.
• compositions typically comprise a pharmaceutically acceptable carrier.
• pharmaceutically acceptable means approved by a government regulatory agency or listed in the U.S. Pharmacopeia or another generally recognized pharmacopeia for use in animals, particularly in humans.
• carrier refers to a diluent, adjuvant, excipient, or vehicle with which the compound is administered.
• Such pharmaceutical carriers can be sterile liquids, such as water and oils, including those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil, glycerol polyethylene glycol ricinoleate, and the like.
• Liquid compositions for parenteral administration can be formulated for administration by injection or continuous infusion. Routes of administration by injection or infusion include intravenous, intraperitoneal, intramuscular, intrathecal and subcutaneous.
• the anti-LAG-3 and/or anti-PD-1 antibodies are administered intravenously (e.g., in separate formulations or together (in the same formulation or in separate formulations)).
• a LAG-3 inhibitor e.g., an anti-LAG-3 antibody
• a PD-1 pathway inhibitor e.g., an anti-PD-1 antibody
• an anti-CTLA-4 antibody e.g., an anti-CTLA-4 antibody
• 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
• cancers and/or malignant tumors include liver cancer, hepatocellular carcinoma (HCC), bone cancer, pancreatic cancer, skin cancer, oral cancer, cancer of the head or neck, breast cancer, lung cancer, small cell lung cancer, NSCLC, cutaneous or intraocular malignant melanoma, renal cancer, uterine cancer, ovarian cancer, colorectal cancer, colon cancer, rectal cancer, cancer of the anal region, stomach cancer, testicular cancer, uterine cancer, carcinoma of the fallopian tubes, carcinoma of the endometrium, carcinoma of the cervix, carcinoma of the vagina, carcinoma of the vulva, squamous cell carcinoma of the head and neck (SCCHN), non-Hodgkin's lymphoma, cancer of the esophagus, cancer of the small intestine, cancer of the endocrine system, cancer of the thyroid gland, cancer of the parathyroid gland, cancer of the adrenal gland, sarcoma of soft tissue,
• the present invention is also applicable to treatment of metastatic cancers.
• the cancer is renal cell carcinoma (RCC), gastric/gastoesophogeal junction carcinoma, non-small cell lung carcinoma (NSCLC), melanoma, squamous cell carcinoma of the head and neck (SCCHN), hepatocellular carcinoma, or urothelial carcinoma.
• RCC renal cell carcinoma
• NSCLC non-small cell lung carcinoma
• SCCHN squamous cell carcinoma of the head and neck
• hepatocellular carcinoma or urothelial carcinoma.
• the melanoma is unresectable or metastatic melanoma.
• the patient was previously treated with an anti-PD-1 or an anti-PD-Ll antibody.
• the tumor is a LAG-3 expressing tumor.
• the tumor is a LAG-3 expressing tumor with LAG-3 expression > 1%.
• the human patient suffers from unresectable metastatic melanoma and was previously treated with an anti-PD-1 or anti-PD-Ll metastatic inhibitor.
• the human patient suffers from unresectable metastatic melanoma and was previously treated with an anti-PD-1 or anti-PD-Ll metastatic inhibitor and the tumor is a LAG-3 expressing tumor.
• the human patient suffers from unresectable metastatic melanoma and was previously treated with an anti-PD-1 or anti-PD-Ll metastatic inhibitor and the tumor is a LAG-3 expressing tumor.
• the human patient suffers from unresectable metastatic melanoma and was previously treated with an anti- PD-1 or anti-PD-Ll metastatic inhibitor and the tumor is a LAG-3 expressing tumor with LAG-3 expression > 1%.
• the human patient suffers from a malignant tumor that is refractory to treatment with an immune checkpoint inhibitor. In another embodiment, the patient suffers from a malignant tumor that is refractory to treatment with a PD-1 inhibitor. In another embodiment, the patient suffers from a malignant tumor that is refractory to treatment with an anti-PD-1 antibody. In another embodiment, the patient suffers from a malignant tumor that is refractory to treatment with an anti-PD-Ll antibody. In some embodiments, the malignant tumor is gastric cancer, renal cancer, HCC, SCCHN, or NSCLC.
• the human patient suffers from melanoma.
• the patient suffers from melanoma that is refractory to treatment with an immune checkpoint inhibitor.
• the patient suffers from melanoma that is refractory to treatment with a PD-1 inhibitor.
• the patient suffers from melanoma that is refractory to treatment with an anti-PD-1 antibody.
• the patient suffers from melanoma that is refractory to treatment with an anti-PD-Ll antibody.
• the human patient suffers from melanoma, gastric cancer, renal cancer, HCC, SCCHN, or NSCLC. In one embodiment, the human patient suffers from melanoma.
• the human patient suffers from NSCLC or a virally-related cancer (e.g., a human papilloma virus (HPV)-related tumor) or gastric adenocarcinoma.
• HPV-related tumor is HPV+ head and neck cancer (HNC).
• the gastric adenocarcinoma is associated with Epstein-Barr virus (EBV) infection.
• Patients can be tested or selected for one or more of the above described clinical attributes prior to, during or after treatment.
• the malignant tumors can be tested to determine LAG-3 expression.
• the malignant tumors treated in accordance with the methods disclosed herein are LAG-3 positive tumors.
• the malignant tumor is a LAG-3 positive melanoma.
• the malignant tumor is a LAG-3 positive gastric cancer, renal cancer, HCC, SCCHN, or NSCLC.
• the malignant tumor is melanoma, gastric cancer, renal cancer, HCC, SCCHN, or NSCLC.
• the malignant tumors can be tested to determine LAG-3 and PD-Ll expression.
• the malignant tumors treated in accordance with the methods disclosed herein are LAG-3 positive PD-Ll positive tumors.
• the malignant tumor is a LAG-3 positive PD-Ll positive melanoma.
• the malignant tumor is a LAG-3 positive PD-Ll positive gastric cancer, renal cancer, HCC, SCCHN, or NSCLC.
• the malignant tumors treated in accordance with the methods disclosed herein are LAG-3 positive PD-Ll negative tumors.
• the malignant tumor is a LAG-3 positive PD-Ll negative melanoma.
• the malignant tumor is a LAG-3 positive PD-Ll negative gastric cancer, renal cancer, HCC, SCCHN, or NSCLC. 10.
• immunotherapies provided herein involve administration of a LAG-
• immunotherapies provided herein involve administration of an anti-PD-1 antibody or an anti-PD-Ll antibody to treat subjects having malignant tumors (e.g., advanced refractory solid tumors or hematological malignancies).
• immunotherapies provided herein involve administration of an anti-CTLA-4 antibody to treat subjects having malignant tumors (e.g., advanced refractory solid tumors or hematological malignancies).
• the invention provides an anti-LAG-3 antibody and an anti-
• the anti-LAG-3 antibody is BMS-986016.
• the anti-PD-1 antibody is BMS-936558.
• dosage regimens are adjusted to provide the optimum desired response (e.g., an effective response).
• the invention provides an anti-LAG-3 antibody and an anti-PD-Ll antibody in combination according to a defined clinical dosage regimen, to treat subjects having a malignant tumor (e.g., an advanced refractory solid tumor).
• a malignant tumor e.g., an advanced refractory solid tumor.
• the anti-LAG-3 antibody is BMS-986016.
• the anti-PD-Ll antibody is BMS-936559.
• dosage regimens are adjusted to provide the optimum desired response (e.g., an effective response).
• the invention provides an anti-LAG-3 antibody according to a defined clinical dosage regimen, to treat subjects having a malignant tumor (e.g., an advanced refractory solid tumor).
• a malignant tumor e.g., an advanced refractory solid tumor.
• the anti-LAG-3 antibody is BMS-986016.
• dosage regimens are adjusted to provide the optimum desired response (e.g., an effective response).
• the invention provides an anti-PD-1 antibody according to a defined clinical dosage regimen, to treat subjects having a malignant tumor (e.g., an advanced refractory solid tumor).
• a malignant tumor e.g., an advanced refractory solid tumor
• the anti-PD-1 antibody is BMS-936558.
• dosage regimens are adjusted to provide the optimum desired response (e.g., an effective response).
• the invention provides an anti-PD-Ll antibody according to a defined clinical dosage regimen, to treat subjects having a malignant tumor (e.g., an advanced refractory solid tumor).
• a malignant tumor e.g., an advanced refractory solid tumor
• the anti-PD-Ll antibody is BMS-936559.
• dosage regimens are adjusted to provide the optimum desired response (e.g., an effective response).
• the invention provides an anti-CTLA-4 antibody according to a defined clinical dosage regimen, to treat subjects having a malignant tumor (e.g., an advanced refractory solid tumor).
• a malignant tumor e.g., an advanced refractory solid tumor.
• the anti- CTLA4 antibody is ipilimumab (YERVOY).
• the anti- CTLA4 antibody is tremelimumab (ticilimumab; CP-675,206), AGEN-1884, or ATOR-1015.
• dosage regimens are adjusted to provide the optimum desired response (e.g., an effective response).
• immunotherapies provided herein involve administration of an anti-PD-1 antibody and an immune checkpoint inhibitor to treat subjects having malignant tumors (e.g., advanced refractory solid tumors or hematological malignancies).
• the anti-PD-1 antibody is BMS-936558.
• the immune checkpoint inhibitor is a CTLA-4 antagonist, a CD80 antagonist, a CD86 antagonist, a Tim-3 antagonist, a TIGIT antagonist, a CD20 antagonist, a CD96 antagonist, a IDOl antagonist, a STING antagonist, a GARP antagonist, a CD40 antagonist, A2aR antagonist, a CEACAMl (CD66a) antagonist, a CEA antagonist, a CD47 antagonist a PVRIG antagonist, a TDO antagonist, a VISTA antagonist, or a KIR antagonist.
• immunotherapies provided herein involve administration of an anti-PD-Ll antibody and an immune checkpoint inhibitor to treat subjects having malignant tumors (e.g., advanced refractory solid tumors or hematological malignancies).
• the anti-PD-Ll antibody is BMS-936559.
• the immune checkpoint inhibitor is a CTLA-4 antagonist, a CD80 antagonist, a CD86 antagonist, a Tim-3 antagonist, a TIGIT antagonist, a CD20 antagonist, a CD96 antagonist, a IDOl antagonist, a STING antagonist, a GARP antagonist, a CD40 antagonist, A2aR antagonist, a CEACAMl (CD66a) antagonist, a CEA antagonist, a CD47 antagonist a PVRIG antagonist, a TDO antagonist, a VISTA antagonist, or a KIR antagonist.
• adjunctive or combined administration includes simultaneous administration of the compounds in the same or different dosage form, or separate administration of the compounds (e.g., sequential administration).
• the anti-LAG-3 and anti-PD-1 antibodies can be simultaneously administered in a single formulation.
• the anti-LAG-3 and anti-PD-1 antibodies can be formulated for separate administration and are administered concurrently or sequentially (e.g., one antibody is administered within about 30 minutes prior to administration of the second antibody).
• the anti-PD-1 antibody can be administered first followed by (e.g., immediately followed by) the administration of the anti-LAG-3 antibody, or vice versa.
• the anti-PD-1 antibody is administered prior to administration of the anti-LAG-3 antibody.
• the anti-PD-1 antibody is administered after administration of the anti-LAG-3 antibody.
• the anti-LAG-3 antibody and anti-PD-1 antibody are administered concurrently. Such concurrent or sequential administration preferably results in both antibodies being simultaneously present in treated patients.
• suitable treatment protocols for treating a malignant tumor in a human patient include administering to the patient an effective amount of a LAG3 inhibitor (e.g., an anti-LAG-3 antibody).
• a LAG3 inhibitor e.g., an anti-LAG-3 antibody.
• a suitable treatment protocol for treating a malignant tumor in a human patient include, for example, administering to the patient an effective amount of an anti-LAG-3 antibody, such as one comprising CDR1, CDR2 and CDR3 domains of the heavy chain variable region having the sequence set forth in SEQ ID NO:3, and CDR1, CDR2 and CDR3 domains of the light chain variable region having the sequence set forth in SEQ ID NO:5, wherein the method comprises at least one administration cycle, wherein the cycle is a period of eight weeks, wherein for each of the at least one cycles, at least four doses of the anti-LAG-3 antibody are administered at a flat dose of about 1, 3, 10, 20, 50, 80, 100, 130, 150, 16, 180, 200, 240 or 280 mg. In another embodiment, four doses of the anti-LAG-3 antibody are administered at a dose of 0.01, 0.03, 0.25, 0.1, 0.3, 1 or 3, 5, 8 or 10 mg/kg body weight.
• an anti-LAG-3 antibody such as one comprising CDR1,
• suitable treatment protocols for treating a malignant tumor in a human patient include administering to the patient an effective amount of a PD1 pathway inhibitor (e.g., an anti-PDl antibody).
• a suitable treatment protocol for treating a malignant tumor in a human patient include, for example, administering to the patient an effective amount of an anti-PD-1 antibody, such as one comprising CDRl, CDR2 and CDR3 domains of the heavy chain variable region having the sequence set forth in SEQ ID NO: 19, and CDRl, CDR2 and CDR3 domains of the light chain variable region having the sequence set forth in SEQ ID NO:21, wherein the method comprises at least one administration cycle, wherein the cycle is a period of eight weeks, wherein for each of the at least one cycles, at least four doses of the anti-PD-1 antibody are administered at flat dose of about 50, 80, 100, 130, 150, 180, 200, 240 or 280 mg. In another embodiment, four doses of the anti-PD-1 antibody are administered
• suitable treatment protocols for treating a malignant tumor in a human patient include administering to the patient an effective amount of an anti-CTLA-4 antibody.
• a suitable treatment protocol for treating a malignant tumor in a human patient include, for example, administering to the patient an effective amount of an anti-CTLA-4 antibody, wherein the method comprises at least one administration cycle, wherein the cycle is a period of eight weeks, wherein for each of the at least one cycles, at least four doses of the anti-CTLA-4 antibody are administered at flat dose of about 50, 80, 100, 130, 150, 180, 200, 240 or 280 mg. In another embodiment, four doses of the anti-CTLA-4 antibody are administered at a dose of 0.1, 0.3, 1, 3, 5, 8 or 10 mg/kg body weight.
• suitable treatment protocols for treating a malignant tumor in a human patient include administering to the patient an effective amount of each of a LAG3 inhibitor (e.g., an anti-LAG-3 antibody) and a PD-1 pathway inhibitor (e.g., an anti-PD-1 antibody).
• a LAG3 inhibitor e.g., an anti-LAG-3 antibody
• a PD-1 pathway inhibitor e.g., an anti-PD-1 antibody
• a suitable treatment protocol for treating a malignant tumor in a human patient include, for example, administering to the patient an effective amount of each of:
• an anti-LAG-3 antibody such as one comprising CDRl, CDR2 and CDR3 domains of the heavy chain variable region having the sequence set forth in SEQ ID NO:3, and CDRl, CDR2 and CDR3 domains of the light chain variable region having the sequence set forth in SEQ ID N0 5
• an anti-PD-1 antibody such as one comprising CDR1, CDR2 and CDR3 domains of the heavy chain variable region having the sequence set forth in SEQ ID NO: 19, and CDR1, CDR2 and CDR3 domains of the light chain variable region having the sequence set forth in SEQ ID NO:21,
• the method comprises at least one administration cycle, wherein the cycle is a period of eight weeks, wherein for each of the at least one cycles, at least four doses of the anti-LAG-3 antibody are administered at a flat dose of about 1, 3, 10, 20, 50, 80, 100, 130, 150, 16, 180, 200, 240 or 280 mg and at least four doses of the anti-PD-1 antibody are administered at flat dose of about 50, 80, 100, 130, 150, 180, 200, 240 or 280 mg.
• four doses of the anti-LAG-3 antibody are administered at a dose of 0.01, 0.03, 0.25, 0.1, 0.3, 1 or 3, 5, 8 or 10 mg/kg body weight and four doses of the anti-PD-1 antibody are administered at a dose of 0.1, 0.3, 1, 3, 5, 8 or 10 mg/kg body weight.
• the anti-LAG-3 antibody and anti-PD-1 antibody are administered at the following doses:
• the anti-LAG-3 antibody and anti-PD-1 antibody are administered at a dose of 20 mg of anti-LAG-3 antibody and 80 mg of anti-PD-1 antibody.
• the tumor is lung cancer.
• the anti-LAG-3 antibody and anti-PD-1 antibody are administered at a dose of 20 mg of anti-LAG-3 antibody and 240 mg of anti-PD-1 antibody.
• the anti-LAG-3 antibody and anti-PD-1 antibody are administered at a dose of 80 mg of anti-LAG-3 antibody and 240 mg of anti-PD-1 antibody.
• the tumor is melanoma (e.g., anti-PDl/PD-Ll antibody experienced melanoma or first line melanoma treatment), RCC (e.g., IO naive RCC), NSCLC (e.g., anti-PDl/PD-Ll antibody experienced NSCLC), gastric cancer (e.g., IO naive gastric cancer), HCC (e.g., IO naive HCC), NSCLC (e.g., first line treatment of NSCLC), or SCCHN (e.g., IO naive SCCHN).
• the anti-LAG-3 antibody and anti-PD-1 antibody are administered at a dose of 240 mg of anti-LAG-3 antibody and 240 mg of anti-PD-1 antibody.
• the anti-LAG-3 antibody and anti-PD-1 antibody are administered at a dose of 160 mg of anti-LAG-3 antibody and 240 mg of anti-PD-1 antibody.
• the tumor is melanoma (e.g., anti-PDl/PD-Ll antibody experienced melanoma or first line melanoma treatment), RCC (e.g., IO naive RCC), NSCLC (e.g., anti-PDl/PD-Ll antibody experienced NSCLC), gastric cancer (e.g., IO naive gastric cancer), HCC (e.g., IO naive HCC), NSCLC (e.g., first line treatment of NSCLC), or SCCHN (e.g., IO naive SCCHN).
• RCC e.g., IO naive RCC
• NSCLC e.g., anti-PDl/PD-Ll antibody experienced NSCLC
• gastric cancer e.g., IO n
• the tumor is Hodgkin's lymphoma (e.g., prior IO treated Hodgkin's lymphoma); DLBCL, PD-1/PD-L1 naive Hodgkin's lymphoma, or PD-1/PD-L1 progressed/refractory Hodgkin's lymphoma.
• Hodgkin's lymphoma e.g., prior IO treated Hodgkin's lymphoma
• DLBCL PD-1/PD-L1 naive Hodgkin's lymphoma
• PD-1/PD-L1 progressed/refractory Hodgkin's lymphoma.
• the anti-LAG-3 antibody and anti-PD-1 antibody are administered at the following doses:
• the dose of the anti-LAG-3 and/or anti-PD-1 antibody is calculated per body weight, e.g., mg/kg body weight.
• the dose of the anti-LAG-3 and/or anti-PD-1 antibody is a flat-fixed dose.
• the dose of the anti-LAG-3 and/or anti-PD-1 antibody is varied over time.
• the anti-LAG-3 antibody and/or anti-PD-1 antibody may be initially administered at a high dose and may be lowered over time.
• the anti-LAG-3 antibody and/or anti-PD-1 antibody is initially administered at a low dose and increased over time.
• the amount of the anti-LAG-3 and/or anti-PD-1 antibodies administered is constant for each dose. In another embodiment, the amount of antibody administered varies with each dose. For example, the maintenance (or follow-on) dose of the antibody can be higher or the same as the loading dose which is first administered. In another embodiment, the maintenance dose of the antibody can be lower or the same as the loading dose.
• the anti-LAG-3 and/or anti-PD-1 antibodies are formulated for intravenous administration. In one embodiment, the anti-PD-1 antibody is administered on Days 1, 15, 29, and 43 of each cycle. In another embodiment, the anti-LAG-3 antibody is administered on Days 1, 15, 29, and 43 of each cycle.
• the anti-LAG-3 and/or anti-PD-1 antibodies are administered about once per week, once about every or three two weeks, about once per month or as long as a clinical benefit is observed or until there is a complete response, confirmed progressive disease or unmanageable toxicity.
• a cycle of administration is eight weeks, which can be repeated, as necessary.
• the treatment consists of up to 12 cycles.
• 4 doses of the anti-PD-1 antibody are administered per eight week cycle.
• 4 doses of the anti-LAG-3 antibody are administered per eight week cycle.
• the anti-PD-1 antibody and anti-LAG-3 antibody are administered as a first line of treatment (e.g., the initial or first treatment).
• the anti-PD-1 antibody and anti-LAG-3 antibody are administered as a second line of treatment (e.g., after the initial or first treatment, including after relapse and/or where the first treatment has failed).
• the invention provides a method of treating a human patient with unresectable or metastatic melanoma, comprising: administering to the patient a therapeutically effective amount of a LAG-3 inhibitor and a PD-1 pathway inhibitor; wherein the patient has previously been treated with a PD-1 inhibitor.
• the invention provides a method of treating a human patient with unresectable or metastatic melanoma, comprising: administering to the patient a therapeutically effective amount of a LAG-3 inhibitor and a PD-1 pathway inhibitor; wherein the patient has previously been treated with a PD-L1 inhibitor.
• the invention is directed to a method of treating a human patient with unresectable or metastatic melanoma, comprising: administering to the patient a therapeutically effective amount of a LAG-3 inhibitor and a PD-1 pathway inhibitor; wherein the patient has previously been treated with a PD-1 inhibitor, and wherein the melanoma expresses LAG-3.
• the invention is directed to a method of treating a human patient with unresectable or metastatic melanoma, comprising: administering to the patient a therapeutically effective amount of a LAG-3 inhibitor and a PD-1 pathway inhibitor; wherein the patient has previously been treated with a PD-L1 inhibitor, and wherein the melanoma expresses LAG-3.
• the invention provides a method of treating a human patient with melanoma that progressed while-on or after treatment with a PD-1 pathway inhibitor or a PD-L1 pathway inhibitor, comprising: administering to the patient a therapeutically effective amount of a LAG-3 inhibitor and a PD-1 pathway inhibitor; wherein the patient has previously been treated with an anti-PD-1 inhibitor.
• the invention provides a method of treating a human patient with melanoma that progressed while-on or after treatment with a PD-1 pathway inhibitor or a PD-L1 pathway inhibitor, comprising: administering to the patient a therapeutically effective amount of a LAG-3 inhibitor and a PD-1 pathway inhibitor; wherein the patient has previously been treated with an anti-PD-Ll inhibitor.
• the invention provides a method of treating a human patient with melanoma that progressed while-on or after treatment with a PD-1 pathway inhibitor or a PD-L1 pathway inhibitor, comprising: administering to the patient a therapeutically effective amount of a LAG-3 inhibitor and a PD-1 pathway inhibitor; wherein the patient has previously been treated with an anti-PD-1 inhibitor, and wherein the melanoma expresses LAG-3.
• the invention provides a method of treating a human patient with melanoma that progressed while-on or after treatment with a PD-1 pathway inhibitor or a PD-L1 pathway inhibitor, comprising: administering to the patient a therapeutically effective amount of a LAG-3 inhibitor and a PD-1 pathway inhibitor; wherein the patient has previously been treated with an anti-PD-Ll inhibitor, and wherein the melanoma expresses LAG-3.
• the LAG-3 expression of the melanoma is > 1%.
• the PD-1 pathway inhibitor administered is an anti-PD-1 antibody.
• the PD-1 antibody is nivolumab.
• the LAG-3 inhibitor is a LAG-3 antibody.
• the LAG-3 antibody is BMS-986016.
• the PD-1 pathway inhibitor administered is an anti-PD-Ll antibody.
• 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 (Int'l Publ. No. WO2015/138920 ) and the anti-PD-1 antibody is PDR001.
• the invention features any of the aforementioned embodiments, wherein the anti-PD-1 antibody is replaced by, or combined with, an anti-PD-Ll or anti-PD-L2 antibody.
• the invention features any of the aforementioned embodiments, wherein administering the anti-LAG-3 antibody or antigen-binding fragment thereof and PD-1 pathway inhibitor (e.g., anti-PD-1 antibody) activates the patient's T cells.
• administering the anti-LAG-3 antibody or antigen-binding fragment thereof and PD-1 pathway inhibitor induces the expression of activation markers by the patient's T cells. Expression of activation markers by the patient's T cells can be detected by analyzing a patient sample, for example, peripheral lymphocytes or tumor-infiltrating lymphocytes using flow cytometry.
• the invention features any of the aforementioned embodiments, wherein administering the anti-LAG-3 antibody or antigen-binding fragment thereof results in the occupancy of at least about 10%, at least about 20%, 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%, at least about 95%, or about 100% of the LAG-3 receptors on the patient's T cells.
• the T cells are CD8+ T cells.
• the T cells are tumor infiltrating T cells.
• the invention features any of the aforementioned embodiments, wherein the treatment protocol further comprises the administration of at least one additional therapeutic agent.
• the at least one additional therapeutic agent is a chemotherapeutic agent.
• the at least one additional therapeutic agent is an immune checkpoint inhibitor.
• CR Complete Response
• Partial Response At least a 30% decrease in the sum of the diameters of target (RECIST Vl . l) lesions, taking as reference the baseline sum diameters.
• the baseline sum if that is the smallest on study). In addition to the relative increase of 20%, the sum must also demonstrate an absolute increase of at least 5 mm. (Note: the appearance of one or more new lesions is also considered progression).
• Immune-related Partial At least a 30% decrease in the sum of diameters of target lesions Response (irPR) and all new measurable lesions (i.e., Percentage Change in Tumor (irRECIST) Burden), taking as reference the baseline sum diameters. Note: the appearance of new measurable lesions is factored into the overall Tumor Burden, but does not automatically qualify as progressive disease until the sum of the diameters increases by >20% when compared to nadir.
• irPR target lesions Response
• irRECIST Percentage Change in Tumor
• Tumor Burden ie the sum of diameters Progressive Disease of target lesions, and any new measurable lesions
• irPD the sum of diameters Progressive Disease of target lesions, and any new measurable lesions
• irRECIST reference the smallest sum on study (this includes the baseline sum if that is the smallest on study).
• the sum must also demonstrate an absolute increase of at least 5 mm.
• responses to therapy may include:
• lymph nodes must be Response (irCR) non-pathological in size ( ⁇ 10 mm short axis).
• Progressive Disease constitute progressive disease unless/until Tumor Burden increases (irPD) (irRECIST) by 20%) (ie the sum of the diameters at nadir of target lesions and any new measurable lesions increases by the required amount). Non-target lesions are not considered in the definition of Stable Disease and Partial Response.
• Patients treated according to the methods disclosed herein preferably experience improvement in at least one sign of cancer.
• improvement is measured by a reduction in the quantity and/or size of measurable tumor lesions.
• lesions can be measured on chest x-rays or CT or MRI films.
• cytology or histology can be used to evaluate responsiveness to a therapy.
• the patient treated exhibits a complete response (CR), a partial response (PR), stable disease (SD), immune-related complete disease (irCR), immune- related partial response (irPR), or immune-related stable disease (irSD).
• the patient treated experiences tumor shrinkage and/or decrease in growth rate, i.e., suppression of tumor growth.
• unwanted cell proliferation is reduced or inhibited.
• one or more of the following can occur: the number of cancer cells can be reduced; tumor size can be reduced; cancer cell infiltration into peripheral organs can be inhibited, retarded, slowed, or stopped; tumor metastasis can be slowed or inhibited; tumor growth can be inhibited; recurrence of tumor can be prevented or delayed; one or more of the symptoms associated with cancer can be relieved to some extent.
• administration of effective amounts of the anti-LAG-3 antibody, anti-PD-1 antibody, anti-PD-Ll antibody, anti-CTLA-4 antibody, a combination of the anti-LAG-3 antibody and anti-PD-1 antibody, or a combination of the anti-PD-1 antibody and an immune checkpoint inhibitor according to any of the methods provided herein produces at least one therapeutic effect selected from the group consisting of reduction in size of a tumor, reduction in number of metastatic lesions appearing over time, complete remission, partial remission, or stable disease.
• the improvement of clinical benefit rate is about 20% 20%, 30%>, 40%, 50%, 60%, 70%), 80%) or more compared to a method of treatment that does not comprise a step of (i) determining the level of LAG-3 expression in a tumor sample prior to treatment, (ii) selecting a LAG-3 positive tumor for treatment, (iii) treating a tumor that has been identified as LAG-3 positive prior to treatment, or (iv) any combinations thereof.
• the methods of treatment produce an objective response rate of at least about 15%, wherein the malignant tumor is a LAG-3 positive melanoma that is resistant to treatment with an anti-PD-1 or anti-PD- Llantibody.
• the median duration of response is > 3 month, > 6 month, > 12 month, or > 18 month.
• the median duration of response is > 6 month.
• the frequency of patients with duration of response > 6 month is 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%.
• the improvement of objective response rate is about 20% 20%, 30%, 40%, 50%, 60%), 70%), 80%) or more compared to a method of treatment that does not comprise a step of (i) determining the level of LAG-3 expression in a tumor sample prior to treatment, (ii) selecting a LAG-3 positive tumor for treatment, (iii) treating a tumor that has been identified as LAG-3 positive prior to treatment, or (iv) any combinations thereof.
• the median duration of response is > 3 month, > 6 month, > 12 month, or > 18 month. In one embodiment, the median duration of response is > 6 month.
• the methods of treatment produce a disease control rate of at least about 70%, wherein the malignant tumor is a LAG-3 positive melanoma that is resistant to treatment with an anti-PD-1 or anti-PD-Llantibody.
• the median duration of response is > 3 month, > 6 month, > 12 month, or > 18 month.
• the median duration of response is > 6 month.
• the frequency of patients with duration of response > 6 month is 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%.
• the improvement of disease control rate is about 20% 20%, 30%, 40%, 50%, 60%, 70%), 80%) or more compared to a method of treatment that does not comprise a step of (i) determining the level of LAG-3 expression in a tumor sample prior to treatment, (ii) selecting a LAG-3 positive tumor for treatment, (iii) treating a tumor that has been identified as LAG-3 positive prior to treatment, or (iv) any combinations thereof.
• the median duration of response is > 3 month, > 6 month, > 12 month, or > 18 month. In one embodiment, the median duration of response is > 6 month.
• kits comprising an anti-LAG-3 antibody for assaying LAG-3 expression as a biomarker for screening patients for the immunotherapy or for predicting the efficacy of the immunotherapy.
• Kits typically include a label indicating the intended use of the contents of the kit and instructions for use.
• label includes any writing, or recorded material supplied on or with the kit, or which otherwise accompanies the kit.
• a first anti-LAG-3 antibody for assaying, detecting, and/or quantifying LAG-3 expression is co-packaged with at least one therapeutic antibody (e.g., a second anti-LAG-3 antibody, an anti-PD-1 antibody, an anti-PD-Ll antibody, and/or an anti-CTLA-4 antibody) for the treatment of a LAG-3 positive tumor.
• the kit further comprises an anti-PD-Ll antibody for assaying, detecting, and/or quantifying PD-L1 expression as a biomarker for predicting the efficacy of the immunotherapy.
• the immunotherapy comprises administering to the patient a therapeutically effective amount of a LAG-3 inhibitor (e.g., anti-LAG-3 antibody) and a PD-1 pathway inhibitor (e.g., anti-PDl antibody or anti-PD-Ll antibody).
• the immunotherapy comprises administering to the patient a therapeutically effective amount of a LAG-3 inhibitor (e.g., anti-LAG-3 antibody).
• the immunotherapy comprises administering to the patient a therapeutically effective amount of a PD-1 pathway inhibitor (e.g., anti-PDl antibody or anti-PD-Ll antibody).
• the immunotherapy comprises administering to the patient a therapeutically effective amount of an anti-PDl antibody.
• the immunotherapy comprises administering to the patient a therapeutically effective amount of an anti-CTLA-4 antibody. In one embodiment, the immunotherapy comprises administering to the patient a therapeutically effective amount of a PD-1 pathway inhibitor (e.g., anti-PDl antibody or anti-PD-Ll antibody) and an immune checkpoint inhibitor.
• a PD-1 pathway inhibitor e.g., anti-PDl antibody or anti-PD-Ll antibody
• an immune checkpoint inhibitor e.g., anti-PDl antibody or anti-PD-Ll antibody
• the diagnostic kit comprises an anti-human LAG-3 monoclonal antibody for assaying, detecting, and/or quantifying LAG-3 expression. See, e.g., J. Matsuzaki, et al.; PNAS 107, 7875 (2010).
• kits which include a pharmaceutical composition containing an anti-LAG-3 antibody, such as BMS-986016, and an anti-PD-1 antibody, such as nivolumab, and a pharmaceutically-acceptable carrier, in a therapeutically effective amount adapted for use in the preceding methods.
• an anti-LAG-3 antibody such as BMS-986016
• an anti-PD-1 antibody such as nivolumab
• a pharmaceutically-acceptable carrier in a therapeutically effective amount adapted for use in the preceding methods.
• the anti-LAG-3 antibody is co-packaged with an anti-PD-1 antibody in unit dosage form.
• the kits optionally also can include instructions, e.g., comprising administration schedules, to allow a practitioner (e.g., a physician, nurse, or patient) to administer the composition contained therein to administer the composition to a patient having cancer (e.g., a solid tumor).
• the kit also can include a syringe.
• the diagnostic and/or therapeutic kits include multiple packages of the single-dose pharmaceutical compositions each containing an effective amount of the anti-LAG-3 or anti-PD-1 antibody for a single administration in accordance with the methods provided above. Instruments or devices necessary for administering the pharmaceutical composition(s) also may be included in the kits. For instance, a kit may provide one or more pre-filled syringes containing an amount of the anti-LAG-3 or anti-PD-1 antibody.
• the present invention provides a kit for treating a patient afflicted with a malignant tumor, the kit, for example, comprising:
• a dose of an anti-LAG-3 antibody such as one comprising CDRl, CDR2 and CDR3 domains of the heavy chain variable region having the sequence set forth in SEQ ID NO:3, and CDRl, CDR2 and CDR3 domains of the light chain variable region having the sequence set forth in SEQ ID NO:5;
• an anti-PD-1 antibody such as one comprising CDRl, CDR2 and CDR3 domains of the heavy chain variable region having the sequence set forth in SEQ ID NO: 19, and CDRl, CDR2 and CDR3 domains of the light chain variable region having the sequence set forth in SEQ ID NO:21; and (c) instructions for using the anti-LAG-3 antibody and anti-PD-1 antibody in the methods described herein.
• the present invention provides a kit for treating a patient afflicted with a malignant tumor, the kit, for example, comprising:
• a dose of an anti-LAG-3 antibody such as one comprising CDRl, CDR2 and CDR3 domains of the heavy chain variable region having the sequence set forth in SEQ ID NO:3, and CDRl, CDR2 and CDR3 domains of the light chain variable region having the sequence set forth in SEQ ID NO:5; and
• the present invention provides a kit for treating a patient afflicted with a malignant tumor, the kit, for example, comprising:
• a dose of an anti-PD-1 antibody such as one comprising CDRl, CDR2 and CDR3 domains of the heavy chain variable region having the sequence set forth in SEQ ID NO: 19, and CDRl, CDR2 and CDR3 domains of the light chain variable region having the sequence set forth in SEQ ID NO:21; and
• the present invention provides a kit for treating a patient afflicted with a malignant tumor, the kit, for example, comprising:
• the present invention provides a kit for treating a patient afflicted with a malignant tumor, the kit, for example, comprising:
• the present invention provides a kit for treating a patient afflicted with a malignant tumor, the kit, for example, comprising:
• a dose of an anti-LAG-3 antibody such as one comprising CDRl, CDR2 and CDR3 domains of the heavy chain variable region having the sequence set forth in SEQ ID NO:3, and CDRl, CDR2 and CDR3 domains of the light chain variable region having the sequence set forth in SEQ ID NO:5;
• the malignant tumor is a LAG-3 positive tumor. In some embodiments, the malignant tumor is a LAG-3/PD-L1 positive tumor. In some embodiments, the malignant tumor is a LAG-3 positive/PD-Ll negative tumor.
• the malignant tumor is melanoma.
• lymphocyte activation gene- 3 (LAG3)
• FFPE formalin fixed, paraffin embedded
• Immunohistochemistry refers to the process of localizing proteins or other molecules in cells of a tissue section. Immunohistochemical staining is widely used in the diagnosis of cancer and has recently been used to help predict whether patients are likely to respond to a targeted chemotherapeutic agent. As opposed to many other analytical techniques such as the Western blot or ELISA, IHC retains the spatial localization of protein expression within a tissue specimen. This technique involves using an antibody (primary antibody) to specifically bind a target within the cellular context and then using the bound antibody to deposit a dye in the region of the target.
• an antibody primary antibody
• Test System FFPE validation were performed on remnant, de-identified, or anonymized human samples. Tissues used for sensitivity testing and analysis included 40 bladder urothelial cancer, 41 gastric/GEJ cancer, 41 HNSCC, 41 melanoma, 41 NSCLC, and 43 RCC. The positive and negative control selected for LAG3 IHC was a tonsil tissue. Tonsil tissue contains cellular features that are positive and negative for LAG3.
• Test Articles The LAG3 mouse clone 17B4 antibody was purchased from LS
• a mouse IgG isotype control antibody was purchased from BD Pharmingen (San Jose, CA) and stored at 2-8°C.
• Tissues were incubated with Polymer (Bond Polymer Refine Detection Kit) for 8 minutes followed by 3 rinses in Bond Wash buffer for 2 minutes each and 2 rinses in distilled water. Tissues were incubated with DAB (Bond Polymer Refine Detection Kit) for 10 minutes followed by 4 rinses in distilled water.
• Polymer Bond Polymer Refine Detection Kit
• Red Chromogen Assay Tissues were then incubated with 3% hydrogen peroxide for 5 minutes followed by 3 rinses in Bond Wash Buffer. Tissues were incubated with Protein Block, Serum Free for 5 minutes followed by incubation with the primary antibody or isotype negative control reagent diluted in Bond Primary Antibody Diluent for 30 minutes and 3 rinses in Bond Wash Buffer. Tissues were incubated with Post Primary AP (Catalog# DS9390, Bond Polymer Refine Red Detection Kit, Leica) for 20 minutes followed by 3 rinses in Bond Wash buffer for 2 minutes each.
• Post Primary AP Catalog# DS9390, Bond Polymer Refine Red Detection Kit, Leica
• Tissues were incubated with Polymer AP (Bond Polymer Refine Red Detection Kit) for 30 minutes followed by 3 rinses in Bond Wash buffer for 2 minutes each and 2 rinses in distilled water. Tissues were incubated with Red Refine (Bond Polymer Refine Red Detection Kit) for 10 minutes followed by 4 rinses in distilled water.
• Polymer AP Bond Polymer Refine Red Detection Kit
• Image Analysis Tissues stained with LAG3 (mouse clone 17B4) using DAB chromogen or red chromogen were evaluated by image analysis with a Nuclear v9 algorithm from Aperio The ROI includes the area of tumor tissue with intervening stroma. Areas excluded from analysis include normal tissue, larger stromal areas, necrotic tissue, tar (if possible), and staining artifact.
• a nuclear algorithm was selected because heavy cytoplasmic stains in small cells, such as immune cells, often obscure the hematoxylin in the nucleus.
• the cytoplasmic and membrane algorithm require visualization of hematoxylin in the nucleus to quantify a cell.
• the nuclear algorithm has a featured called "fill holes" that will fill the central portion of a lymphocyte if there is hematoxylin present and record it as one cell.
• Pathologist Visual Immune Score A subset of samples within the dynamic range were also scored by a pathologist during QC of image analysis.
• the purpose of the pathologist visual immune score is to provide a back-up result in the event of an image analysis score that does not produce an accurate result as deemed by a board-certified pathologist.
• Reasons for image analysis failure may include but not limited to: 1) light counterstain; 2) crushed tissue; 3) presence of tar in NSCLC tissues; 4) staining of hemosiderin; or 5) presence of melanin that precludes evaluation.
• the pathologist visual immune score is the percentage of positive immune cells within the annotated region (to mimic the algorithm).
• LAG3 IHC Assay Validation - Sensitivity A sensitivity analysis was performed using the optimized LAG3 (mouse clone 17B4) IHC assay on 247 FFPE human tissues (40 bladder urothelial cancer, 41 gastric/GEJ cancer, 41 HNSCC, 41 melanoma, 41 NSCLC, 43 RCC) tissues to demonstrate the dynamic range of the assay within the 6 indications. All specimens were evaluated by image analysis of 1 ROI (tumor + intervening stroma) and a subset of the tissues (10 each within the 6 indications) were also evaluated by pathologist visual immune score.
• ROI tumor + intervening stroma
• Figure 1 shows anti-LAG-3 staining patterns observed in the tumor samples using monoplex IHC.
• the staining patterns observed included partial membrane/cytoplasmic localization, dot like localization, and complete membrane/cytoplasmic localization.
• Figure 2 shows the frequency distribution of LAG-3 positive cell as a ratio of total tumor cells across various tumors as detected by monoplex LAG-3 IHC.
• Simultaneous blockade of the negative T-cell regulators LAG-3 and PD-1 may function synergistically to restore T-cell activation and enhance antitumor immunity.
• Data from a phase l/2a study of BMS-986016 (fully human IgG4 mAb that targets LAG-3) ⁇ nivolumab (fully human IgG4 mAb that targets PD-1) demonstrated that the combination was well tolerated and showed promising antitumor activity in patients with melanoma who were refractory to or relapsed during prior anti-PD-l/PD-Ll therapy (NCT01968109; Ascierto et al. J Clin Oncol. 2017;35(suppl) [abstract 9520]).
• NCT01968109 Ascierto et al. J Clin Oncol. 2017;35(suppl) [abstract 9520]
• Figure 6 shows the LAG-3 expression status of the first 40 IO experienced melanoma samples. 40% (16/40) of the samples were scored as LAG-3 positive using >1% cutoff in a monoplex IHC assay.
• Figure 10 shows the duration of progression-free survival. Of 48 evaluable patients, 46%> (22/48) of patients remain on treatment without progression at data cutoff.
• Figure 18 shows the updated prior treatment history of the mel prior 10 cohort.
• Figure 19 shows the updated efficacy data for the mel prior IO cohort. ORR was
• LAG-3 expression (> 1%) appeared to enrich for response. Median duration of response was not reached (range, 0.1+-39.3+).
• Figure 20 shows the response by baseline characteristics
• LAG-3 expression observed in the mel prior IO cohort. LAG-3 expression (> 1%) enriched for response irrespective of PD-L1 expression.
• Figures 21 and 22 show the best change in target lesion size by LAG-3 and PD-L1 expression and the depth and duration of response by LAG-3 and PD-L1 expression, respectively, observed in the mel prior IO cohort. Responses were more likely in patients with LAG-3 expression > 1%. PD-L1 expression did not appear to enrich for response.
• Figure 23 shows the duration of progression-free survival. Of 61 evaluable patients, 34% (21/61) of patients had not progressed at data cutoff. Of 33 evaluable LAG-3 > 1% patients, 55% (18/33) of patients had not progressed at data cutoff. Of 20 evaluable LAG-3 ⁇ 1% patients, 5% (1/20) of patients had not progressed at data cutoff.
• LAG-3 is a transmembrane receptor that negatively regulates T-cell activation.
• LAG-3 and other T-cell inhibitory receptors can lead to T-cell exhaustion and is a mechanism of immune escape for tumors.
• Simultaneous blockade of LAG-3 and PD-1 may function synergistically to restore T-cell activation and enhance antitumor immunity.
• BMS-986016 IgG4 mAb targeting LAG-3)
• ⁇ nivolumab IgG4 mAb targeting PD-1
• Efficacy of BMS-986016 + nivolumab across several advanced solid tumor expansion cohorts was evaluated in both all-comer and biomarker- enriched populations.
• Figure 12 shows LAG-3 expression status of immuno-oncology-naive gastric tumor samples. 48% (10/21) of the samples were scored as LAG-3 positive using a >1% cut-off in a monoplex IHC assay.
• Figure 13 shows change in target lesion size in immuno-oncology-naive gastric cancer patients in response to treatment with a combination of anti -LAG-3 and anti-PD-1 antibody.
• LAG-3 positive tumors were enriched among the patients that were responsive to the treatment. Tumor response was determined according to RECIST. The group of patients shown have not been previously exposed to anti-PD-l/PD-Ll treatment.
• Figure 14 shows LAG-3 expression status of immuno-oncology-naive SCCHN, renal carcinoma, HCC, and NSCLC tumor samples as determined by a monoplex IHC assay.
• LAG-3 and programmed death- 1 (PD-1) receptors are overexpressed and co-expressed on tumor-infiltrating lymphocytes (TILs).
• TILs tumor-infiltrating lymphocytes
• LAG-3 and PD-1 overexpression may limit treatment response to anti-PD-1 therapy and lead to tumor progression.
• BMS-986016 is a fully human IgG4 antibody that targets LAG-3, blocking binding to its ligand, major histocompatibility complex class II (MHC II) ( Figure 24).
• MHC II major histocompatibility complex class II
• BMS-986016 combined with nivolumab (anti-PD-1) may restore T-cell activation and tumor response in patients whose disease progressed on anti-PD-1 monotherapy. Ascierto P et al.
• IHC Quantitative Immunohistochemistry
• MHC II and PD-Ll expression by IHC on tumor cells were scored manually. Unsupervised clustering (Ward's method) was performed on the IHC data to identify associations between LAG-3 and other immune biomarkers. To determine MHC 11+ and LAG-3+ colocalization, MHC Il-high (>70% MHC 11+) or MHC II-low ( ⁇ 10% MHC 11+) tumor cell regions were assessed for the number of LAG-3 stained cells (average of three 20 ⁇ fields of view each for positive and negative regions).
• mRNA Analysis In patients with RCC and melanoma, changes in LAG-3 mRNA levels were determined by differential gene expression analyses of Affymetrix (RCC) or RNA- sequencing (melanoma) data from tumor biopsy samples collected at screening and 2-4 weeks post-immunotherapy initiation. [0402] Statistical Analyses Correlations between LAG-3 expression and other immune biomarkers were assessed by Spearman's correlation, r. Mann-Whitney test was conducted to assess statistical differences. Differential gene expression analyses were performed using generalized linear models that included treatment group and time as factors..
• LAG-3 expression was associated with cellular inflammation in the tumor microenvironment, as shown by IHC. MHC II tumor cell expression was frequently observed across the 6 tumor types analyzed; LAG-3 expression in immune cells was enriched in tumors with expression of MHC II in tumor cells. Higher frequency of LAG-3+ TILs was observed in MHC II high/positive tumor regions vs MHC II low/negative tumor regions within individual tumor specimens, raising the possibility that co-localization of LAG-3 and MHC II expression in tumor cells may serve as a mechanism of LAG-3 checkpoint activation in certain tumors.

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