Combination therapy of a PD-1 antagonist and LAG3 antagonist and all-trans retinoic acid or a pharmaceutically acceptable salt thereof for treating patients with cancer

Abstract

The present disclosure describes combination therapies comprising an antagonist of Programmed Death 1 receptor (PD-1), a Lymphocyte-Activation Gene 3 (LAG3) antagonist, and all-trans retinoic acid or a pharmaceutically acceptable salt thereof and the use of the combination therapies for the treatment of cancer.

Description

COMBINATION THERAPY OF A PD-1 ANTAGONIST AND LAG3 ANTAGONIST AND ALL-TRANS RETINOIC ACID OR A PHARMACEUTICALLY ACCEPTABLE SALT THEREOF FOR TREATING PATIENTS WITH CANCERCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] The application claims the benefit of priority to U.S. Provisional Application No. 63 / 609.108, filed December 12, 2023, the contents of each of which are incorporated herein by reference in their entirety.REFERENCE TO SEQUENCE LISTING SUBMITTED ELECTRONICALLY

[0002] The instant application contains a Sequence Listing which has been submitted electronically in XML format and is hereby incorporated by reference in its entirety7. The XML file, created on April 3, 2024, is named 2588 l-WO-PCT SL. XML. and is 33 bytes in size.FIELD OF THE INVENTION

[0003] The present invention relates to combination therapies useful for the treatment of cancer. In particular, the invention relates to a combination therapy that comprises an antagonist of a Programmed Death 1 protein (PD-1), an antagonist of Lymphocyte- Activation Gene 3 (LAG3), and all-trans retinoic acid or a pharmaceutically acceptable salt thereof.BACKGROUND OF THE INVENTION

[0004] PD-1 is recognized as an important molecule in immune regulation and the maintenance of peripheral tolerance. PD-1 is moderately expressed on naive T. B and Natural Killer T (NKT) cells and up-regulated by T / B cell receptor signaling on lymphocytes, monocytes and myeloid cells (1).

[0005] Two known ligands for PD-1, PD-L1 (B7-H1) and PD-L2 (B7-DC), are expressed in human cancers arising in various tissues. In large sample sets of cancers, e.g., ovarian, renal, colorectal, pancreatic, liver cancers and melanoma, it was shown that PD-L1 expression correlated with poor prognosis and reduced overall survival irrespective of subsequent treatment (2-13). Similarly, PD-1 expression on tumor infiltrating lymphocytes was found to mark dysfunctional T cells in breast cancer and melanoma (14-15) and to correlate with poor prognosis in renal cancer (16). Thus, it has been proposed that PD-L1 expressing tumor cells interact with PD-1 expressing T cells to attenuate T cell activation and evasion of immune surveillance, thereby contributing to an impaired immune response against the tumor.

[0006] Several monoclonal antibodies that inhibit the interaction between PD-1 and one or both of its ligands PD-L1 and PD-L2 have been approved for treating cancer. Pembrolizumab is a potent humanized immunoglobulin G4 (IgG4) mAb with high specificity of binding to the programmed cell death 1 (PD 1) receptor, thus inhibiting its interaction with programmed cell death ligand 1 (PD-L1) and programmed cell death ligand 2 (PD-L2). Based on preclinical in vitro data, pembrolizumab has high affinity and potent receptor blocking activity for PD-1. Keytruda® (pembrolizumab) is indicated for the treatment of patients across a number of indications.

[0007] Lymphocyte-Activation Gene 3 (LAG3) is an inhibitory immune modulatory- receptor that regulates effector T cell homeostasis, proliferation, and activation, and has a role in the suppressor activity of regulatory T cells (Tregs). LAG3 is expressed on activated CD8+ and CD4+ T cells, Tregs and the Tri regulatory T-cell population, as well as on natural killer cells and a subset of tolerogenic plasmacytoid dendritic cells. Because of its proposed role on both effector T cells and Tregs. LAG3 is one of several immune checkpoint molecules where simultaneous blockade of both cell populations has the potential to enhance antitumor immunity.

[0008] All trans retinoic acid (ATRA) (tretinoin) is a vitamin A derivative that binds the retinoic acid receptor on myeloid-derived suppressor cells (MDSCs) and differentiates immature monocytes into more mature dendritic cells (17). Tretinoin is the generic name of the trade name drug Vesanoid®. It is available in a 10 mg soft gelatin capsule for oral administration.

[0009] There is a large population of advanced melanoma patients (approximately 60% of all unresectable Stage III or Stage IV melanoma patients who receive PD-1 inhibitors as IL therapy for their advanced disease) who do not respond to therapy with anti-PD-1 agents. Thus, there is a need to improve the overall outcome of patients with IL advanced melanoma. In addition, because of the high-risk of recurrence in Stage III melanoma with bulky resectable disease, and the advantages of neoadjuvant approaches, there is a high need to identity' an acceptable treatment in this setting with an acceptable benefit-risk profile.SUMMARY OF THE INVENTION

[0010] The invention provides a method for treating cancer in an individual comprising administering to the individual a combination therapy that comprises a PD-1 antagonist, a LAG3 antagonist, and all-trans retinoic acid or a pharmaceutically acceptable salt thereof. In one embodiment, the cancer is melanoma. In another embodiment, the cancer is melanoma at Stage III or IV. In one embodiment, the PD-1 antagonist and LAG3 antagonist are co-formulated and co-administered with all-trans retinoic acid. In another embodiment, the PD-1 antagonist, LAG3antagomst. and all-trans retinoic acid are co-admimstered. In one embodiment, the PD-1 antagonist is an anti-PD-1 antibody that blocks the binding of PD-1 to PD-L1 and PD-L2. In another embodiment, the LAG3 antagonist is an anti-LAG3 antibody that blocks the binding of LAG3 to MHC Class II molecules. In one embodiment, all-trans retinoic acid is used.DETAILED DESCRIPTION

[0011] Abbreviations. Throughout the detailed description and examples of the invention the following abbreviations will be used: BOR Best overall responseBID One dose twice dailyBICR Blinded Independent Central RadiologyCBR Clinical Benefit RateCDR Complementarity determining regionCHO Chinese hamster ovaryCR Complete ResponseDCR Disease Control RateDFS Disease free survivalDLT Dose limiting toxicityDOR Duration of ResponseDSDR Durable Stable Disease RateEFS Event Free SurvivalFFPE Formalin-fixed, paraffin-embeddedFR Framework regionIgG Immunoglobulin GIHC Immunohistochemistry' or immunohistochemical irRC Immune related response criteriaIV IntravenousMTD Maximum tolerated doseNCBI National Center for Biotechnology7InformationNCI National Cancer InstituteORR Objective response rateOS Overall survivalPD Progressive diseasePD-1 Programmed Death 1PD-L 1 Programmed Cell Death 1 Ligand 1PD-L2 Programmed Cell Death 1 Ligand 2PFS Progression free survivalPR Partial response mPR Major Pathologic Response pCR Pathological Complete ResponseQ2W One dose every’ two weeksQ3W One dose every three weeksQD One dose per dayRECIST Response Evaluation Criteria in Solid TumorsSD Stable diseaseTPI Toxicity' Probability' IntervalVH Immunoglobulin heavy chain variable regionVK Immunoglobulin kappa light chain variable regionDEFINITIONS

[0012] So that the invention may be more readily understood, certain technical and scientific terms are specifically defined below. Unless specifically defined elsewhere in this document, all other technical and scientific terms used herein have the meaning commonly understood by one of ordinary' skill in the art to which this invention belongs.

[0013] As used herein, including the appended claims, the singular forms of words such as “a,” “an,” and “the,” include their corresponding plural references unless the context clearly dictates otherwise.

[0014] “Favezelimab” is defined in the World Health Organization drug information INN Vol. 35, No. l, 2021. Favezelimab corresponds with the monoclonal antibody that consists of two heavy' chain and two light chain sequences of SEQ ID NO: 23 and SEQ ID NO: 22, respectively. When recombinantly expressed in host cells (e.g.. CHO cells), or during purification / storage, post-translational modifications can occur (e g., C-terminal Lysine clipping in the heavy chain, conversion of the N-terminal glutamine to pyroglutamate or pyroglutamic acid in the heavy chain).

[0015] As used herein, a “Favezelimab variant” means a monoclonal antibody that comprises heavy chain and light chain sequences that are substantially identical to those in favezelimab described herein, except for having three, tw o or one conservative amino acid substitutions in the light chain sequence at positions that are located outside of the light chainCDRs and / or six, five, four, three, two or one conservative amino acid substitutions in the heavy chain sequence that are located outside of the heavy chain CDRs, e g., the variant positions are located in the FR regions or the constant region. In other words, favezelimab and a favezelimab variant comprise identical CDR sequences, but differ from each other due to having a conservative amino acid substitution at no more than three or six other positions in their full length light and heavy chain sequences, respectively. A favezelimab variant is substantially the same as favezelimab with respect to the following properties: binding affinity to human LAG3 and ability to block the binding of human LAG3 to human MHC Class II.

[0016] 'Administration” as it applies to an animal, human, experimental subject, cell, tissue, organ, or biological fluid, refers to contact of an exogenous pharmaceutical, therapeutic, diagnostic agent, or composition to the animal, human, subject, cell, tissue, organ, or biological fluid. Treatment of a cell encompasses contact of a reagent to the cell, as well as contact of a reagent to a fluid, where the fluid is in contact wi th the cell. The term “subject'’ includes any organism, preferably an animal, more preferably a mammal (e.g.. rat. mouse, dog, cat, rabbit) and most preferably a human.

[0017] In general, the basic antibody structural unit comprises a tetramer. Each tetramer includes two identical pairs of polypeptide chains, each pair having one “light’' (about 25 kDa) and one “heavy” chain (about 50-70 kDa). The amino-terminal portion of each chain includes a variable region of about 100 to 110 or more amino acids primarily responsible for antigen recognition. The carboxy-terminal portion of the heavy chain may define a constant region primarily responsible for effector function. Typically, human light chains are classified as kappa and lambda light chains. Furthermore, human heavy chains are typically classified as mu, delta, gamma, alpha, or epsilon, and define the antibody's isotype as IgM. IgD. IgG, IgA. and IgE. respectively. Within light and heavy chains, the variable and constant regions are joined by a “J” region of about 12 or more amino acids, with the heavy chain also including a “D” region of about 10 more amino acids. See generally, Fundamental Immunology Ch. 7 (Paul, W., ed.. 2nd ed. Raven Press, N.Y. (1989).

[0018] The variable regions of each light / heavy chain pair form the antibody binding site. Thus, in general, an intact antibody has two binding sites. Except in bifunctional or bispecific antibodies, the two binding sites are. in general, the same.

[0019] Typically, the variable domains of both the heavy and light chains comprise three hypervariable regions, also called complementarity determining regions (CDRs), which are located within relatively conserved framework regions (FR). The CDRs are usually aligned by the framework regions, supporting binding to a specific epitope. In general, from the N-terminalto C-terminal, both light and heavy chains variable domains comprise FR1. CDR1. FR2. CDR2. FR3, CDR3 and FR4. The assignment of amino acids to each domain is, generally, in accordance with the definitions of Sequences of Proteins of Immunological Interest. Kabat, et al , National Institutes of Health, Bethesda, Md.; 5thed.; NIH Publ. No. 91-3242 (1991); Kabat (1978) Adv. Prot. Chem. 32: 1-75; Kabat. et al.. (1977) J. Biol. Chem. 252:6609-6616; Chothia. et al.. (1987) J Mol. Biol. 196:901-917 or Chothia, etal., (1989) Nature 342:878-883.

[0020] As used herein, unless otherwise indicated, "antibody fragment” or “antigen binding fragment” refers to antigen binding fragments of antibodies, i.e., antibody fragments that retain the ability to bind specifically to the antigen bound by the full-length antibody, e.g., fragments that retain one or more CDR regions. Examples of antibody binding fragments include, but are not limited to, Fab, Fab', F(ab')2, and Fv fragments; diabodies; linear antibodies; single-chain antibody molecules, e.g., sc-Fv; nanobodies and multispecific antibodies formed from antibody fragments.

[0021] An antibody that “specifically binds to” a specified target protein is an antibody that exhibits preferential binding to that target as compared to other proteins, but this specificity does not require absolute binding specificity. An antibody is considered “specific” for its intended target if its binding is determinative of the presence of the target protein in a sample, e.g., without producing undesired results such as false positives. Antibodies, or binding fragments thereof, useful in the present invention, will bind to the target protein with an affinity that is at least two fold greater, preferably at least ten times greater, more preferably at least 20-times greater, and most preferably at least 100-times greater than the affinity with non-target proteins. As used herein, an antibody is said to bind specifically to a polypeptide comprising a given amino acid sequence, e.g.. the amino acid sequence of a mature human PD-1 or human PD-L1 molecule, if it binds to polypeptides comprising that sequence but does not bind to proteins lacking that sequence.

[0022] “Chimeric antibody” refers to an antibody in which a portion of the heavy and / or light chain is identical with or homologous to corresponding sequences in an antibody derived from a particular species (e g., human) or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is identical with or homologous to corresponding sequences in an antibody derived from another species (e.g., mouse) or belonging to another antibody class or subclass, as well as fragments of such antibodies, so long as they exhibit the desired biological activity.

[0023] "Co-administration" as used herein for agents such as the PD-1 antagonist or LAG3 antagonist means that the agents are administered so as to have overlapping therapeutic activities,and not necessarily that the agents are administered simultaneously to the subject. The agents may or may not be in physical combination prior to administration. In an embodiment, the agents are administered to a subject simultaneously or at about the same time. For example, the PD-1 antagonist and LAG3 antagonist may be contained in separate vials, when in liquid solution, maybe mixed into the same intravenous infusion bag or injection device, and administered simultaneously to the patient, along with all-trans retinoic acid. In some embodiments, the agents are administered within the same formulation, the same administration, the same infusion (e.g., intravenous infusion), or are administered hours (e.g., any where from about 1 to 48 hours apart) or days apart (e.g., anywhere from about 1 to 30 days apart). In specific embodiments, the PD-1 antagonist, LAG3 antagonist, and all-trans retinoic acid are administered on the same day.

[0024] “Co-formulated” or “co-formulation” or “coformulation” or “coformulated” as used herein refers to at least two different antagonists, antibodies or antigen binding fragments thereof that are formulated together and stored as a combined product in a single vial or vessel (for example an injection device) rather than being formulated and stored individually and then mixed before administration or separately administered. In one embodiment, the co-formulation contains two different antagonists, antibodies or antigen binding fragments thereof.

[0025] “Human antibody” refers to an antibody that comprises human immunoglobulin protein sequences only. A human antibody may contain murine carbohydrate chains if produced in a mouse, in a mouse cell, or in a hybridoma derived from a mouse cell. Similarly, “mouse antibody” or “rat antibody” refer to an antibody that comprises only mouse or rat immunoglobulin sequences, respectively.

[0026] “Humanized antibody” refers to forms of antibodies that contain sequences from nonhuman (e.g., murine) antibodies as well as human antibodies. Such antibodies contain minimal sequence derived from non-human immunoglobulin. In general, the humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the hypervariable loops correspond to those of a non-human immunoglobulin and all or substantially all of the FR regions are those of a human immunoglobulin sequence. The humanized antibody optionally also will comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. The prefix “hum”, “hu” or “h” is added to antibody clone designations when necessary to distinguish humanized antibodies from parental rodent antibodies. The humanized forms of rodent antibodies will generally comprise the same CDR sequences of the parental rodent antibodies, although certain ammo acid substitutions may be included to increase affinity, increase stability- of the humanized antibody, or for other reasons.

[0027] “Anti-tumor response’7when referring to a cancer patient treated with a therapeutic regimen, such as a combination therapy described herein, means at least one positive therapeutic effect on the cancer cells or tumors within the cancer patient, such as for example, reduced number of cancer cells, reduced tumor size, reduced rate of cancer cell infiltration into peripheral organs, reduced rate of tumor metastasis or tumor growth, or progression free survival. Positive therapeutic effects in cancer can be measured in a number of ways (See, W. A. Weber, J. Null. Med. 5O: 1S-1OS (2009); Eisenhauer et al., supra). In some embodiments, an anti-tumor response to a combination therapy described herein is assessed using RECIST 1.1 criteria, bidimensional irRC or unidimensional irRC. In some embodiments, an anti-tumor response is any of SD, PR. CR, PFS, or DFS.

[0028] “Bidimensional irRC” refers to the set of criteria described in Wolchok JD, et al. Guidelines for the evaluation of immune therapy activity7in solid tumors: immune-related response criteria. Clin Cancer Res. 2009;15(23):7412-7420. These criteria utilize bidimensional tumor measurements of target lesions, which are obtained by multiplying the longest diameter and the longest perpendicular diameter (cm2) of each lesion.

[0029] “Biotherapeutic agent” means a biological molecule, such as an antibody or fusion protein, that blocks ligand / receptor signaling in any biological pathway that supports tumor maintenance and / or growth or suppresses the anti-tumor immune response. Classes of biotherapeutic agents include, but are not limited to, antibodies to PD-1, LAG3, VEGF, EGFR, Her2 / neu, other growth factor receptors, CD20, CD40, CD-40L, CTLA-4, OX-40, 4- IBB, and ICOS.

[0030] “CBR” or “Clinical Benefit Rate” means CR + PR + durable SD.

[0031] “CDR” or “CDRs” as used herein means complementarity determining region(s) in an immunoglobulin variable region, defined using the Kabat numbering system, unless otherwise indicated.

[0032] “Chemotherapeutic agent” is a chemical compound useful in the treatment of cancer. Classes of chemotherapeutic agents include, but are not limited to: alkylating agents, antimetabolites, kinase inhibitors, spindle poison plant alkaloids, cytoxic / antitumor antibiotics, topisomerase inhibitors, photosensitizers, anti-estrogens and selective estrogen receptor modulators (SERMs), anti-progesterones, estrogen receptor down-regulators (ERDs), estrogen receptor antagonists, leutinizing hormone-releasing hormone agonists, anti-androgens, aromatase inhibitors, EGFR inhibitors, VEGF inhibitors, and anti-sense oligonucleotides that inhibit expression of genes implicated in abnormal cell proliferation or tumor growth. Chemotherapeuticagents useful in the treatment methods of the present invention include cytostatic and / or cytotoxic agents.

[0033] “Chothia” as used herein means an antibody numbering system described in Al- Lazikani et al., JMB 273:927-948 (1997).

[0034] "Comprising" or variations such as "comprise", “comprises” or “comprised of?are used throughout the specification and claims in an inclusive sense, i.e., to specify the presence of the stated features but not to preclude the presence or addition of further features that may materially enhance the operation or utility of any of the embodiments of the invention, unless the context requires otherwise due to express language or necessary’ implication.

[0035] “Combination therapy” or “in combination” refers to two or more biotherapeutic and / or chemotherapeutic agents administered as a part of a treatment regimen. The two or more biotherapeutic and / or chemotherapeutic agents can be administered co-formulated, coadministered, or both.

[0036] “In sequence” refers to two or more treatment regimens administered sequentially in any order.

[0037] “Conservatively modified variants” or “conservative substitution” refers to substitutions of amino acids in a protein with other amino acids having similar characteristics (e.g., charge, side-chain size, hydrophobicity / hydrophilicity, backbone conformation and rigidity’, etc.), such that the changes can frequently be made without altering the biological activity or other desired property of the protein, such as antigen affinity and / or specificity. Those of skill in this art recognize that, in general, single amino acid substitutions in non-essential regions of a polypeptide do not substantially alter biological activity (see, e.g., Watson et al. (1987) Molecular Biology of the Gene, The Benjamin / Cummings Pub. Co., p. 224 (4th Ed.)). In addition, substitutions of structurally or functionally similar amino acids are less likely to disrupt biological activity’. Exemplary’ conserv ative substitutions are set forth in Table 1 below.TABLE 1. Exemplary Conservative Amino Acid Substitutions

[0038] “Consists essentially of,"’ and variations such as “consist essentially of’ or “consisting essentially of,"’ as used throughout the specification and claims, indicate the inclusion of any recited elements or group of elements, and the optional inclusion of other elements, of similar or different nature than the recited elements, that do not materially change the basic or novel properties of the specified dosage regimen, method, or composition. As a non-limiting example, a PD-1 antagonist that consists essentially of a recited amino acid sequence may also include one or more amino acids, including substitutions of one or more amino acid residues, which do not materially affect the properties of the binding compound.

[0039] “DCR” or “Disease Control Rate” means CR + PR + SD.

[0040] “Diagnostic anti-PD-L monoclonal antibody” means a mAb that specifically binds to the mature form of the designated PD-L (PD-L1 or PD-L2) that is expressed on the surface of certain mammalian cells. A mature PD-L lacks the presecretory leader sequence, also referred to as leader peptide. The terms “PD-L” and “mature PD-L” are used interchangeably herein, and shall be understood to mean the same molecule unless otherwise indicated or readily apparent from the context.

[0041] As used herein, a diagnostic anti-human PD-L1 mAb or an anti-hPD-Ll mAb refers to a monoclonal antibody that specifically binds to mature human PD-L1. A mature human PD- L1 molecule consists of amino acids 19-290 of the following sequence:MRIFAVFIFMTYWHLLNAFTVTVPKDLYVVEYGSNMTIECKFPVEKQLDLAALIVYWE MEDKNIIQFVHGEEDLKVQHSSYRQRARLLKDQLSLGNAALQITDVKLQDAGVYRCMIS YGGADYKRITVKVNAPYNKINQRILVVDPVTSEHELTCQAEGYPKAEVIWTSSDHQVLS GKTTTTNSKREEKLFNVTSTLRINTTTNEIFYCTFRRLDPEENHTAELVIPELPLAHPPNER THLVILGAILLCLGVALTFIFRLRKGRMMDVKKCGIQDTNSKKQSDTHLEET (SEQ ID NO:32).

[0042] Specific examples of diagnostic anti-human PD-L1 mAbs useful as diagnostic mAbs for immunohistochemistry (IHC) detection of PD-L1 expression in formalin-fixed, paraffin- embedded (FFPE) tumor tissue sections are antibody 20C3 and antibody 22C3, which are described in W02014 / 100079. Another anti-human PD-L1 mAb that has been reported to be useful for IHC detection of PD-L1 expression in FFPE tissue sections (Chen. B.J. et al., Clin Cancer Res 19: 3462-3473 (2013)) is a rabbit anti-human PD-L1 mAb publicly available from Sino Biological, Inc. (Beijing, P.R. China; Catalog number 10084-R015).

[0043] “PD-L 1 or ‘PD-L2-’ expression as used herein means any detectable level of expression of the designated PD-L protein on the cell surface or of the designated PD-L mRNA within a cell or tissue. PD-L protein expression may be detected with a diagnostic PD-L antibody in an IHC assay of a tumor tissue section or by flow cytometry. Alternatively, PD-L protein expression by tumor cells may be detected by PET imaging, using a binding agent (e.g., antibody fragment, affibody and the like) that specifically binds to the desired PD-L target, e.g., PD-L1 or PD-L2. Techniques for detecting and measuring PD-L mRNA expression include RT-PCR, realtime quantitative RT-PCR, RNAseq, and the Nanostring platform (J. Clin. Invest.20I7;127(8):2930-2940).

[0044] Several approaches have been described for quantifying PD-L1 protein expression in IHC assays of tumor tissue sections. See, e.g.. Thompson. R. H., et al., PNAS 101 (49): 17174- 17179 (2004); Thompson, R. H. et al., Cancer Res. 66:3381-3385 (2006); Gadiot, I, et al.. Cancer 117:2192-2201 (2011); Taube, J. M. et al., Sci TranslMedt, 127ra37 (2012); andToplian, S. L. et al., New Eng. J Med. 366 (26): 2443-2454 (2012). See US 20170285037 which describes Hematoxylin and Eosin staining used by the pathologist.

[0045] One approach employs a simple binary end-point of positive or negative for PD-L1 expression, with a positive result defined in terms of the percentage of tumor cells that exhibit histologic evidence of cell-surface membrane staining. A tumor tissue section is counted as positive for PD-L1 expression if it is at least 1% of total tumor cells.

[0046] In another approach, PD-L1 expression in the tumor tissue section is quantified in the tumor cells as well as in infiltrating immune cells, which predominantly comprise lymphocytes. The percentage of tumor cells and infiltrating immune cells that exhibit membrane staining are separately quantified as < 5%, 5 to 9%, and then in 10% increments up to 100%. PD-L1 expression in the immune infiltrate is reported as a semi-quantitative measurement called the adjusted inflammation score (AIS), which is determined by multiplying the percent of membrane staining cells by the intensity of the infiltrate, which is graded as none (0), mild (score of 1, rare lymphocytes), moderate (score of 2. focal infiltration of tumor by lymphohistiocytic aggregates), or severe (score of 3, diffuse infiltration). A tumor tissue section is counted as positive for PD-L1 expression by immune infiltrates if the AIS is > 5.

[0047] The level of PD-L mRNA expression may be compared to the mRNA expression levels of one or more reference genes that are frequently used in quantitative RT-PCR.

[0048] In some embodiments, a level of PD-L1 expression (protein and / or mRNA) by malignant cells and / or by infiltrating immune cells within a tumor is determined to be “overexpressed” or “elevated” based on comparison with the level of PD-L 1 expression (protein and / or mRNA) by an appropriate control. For example, a control PD-L1 protein or mRNA expression level may be the level quantified in nonmalignant cells of the same type or in a section from a matched normal tissue. In some preferred embodiments, PD-L1 expression in a tumor sample is determined to be elevated if PD-L1 protein (and / or PD-L1 mRNA) in the sample is at least 10%, 20%. or 30% greater than in the control.

[0049] “Tumor Proportion Score (TPS)” refers to the percentage of tumor cells expressing PD-L1 on the cell membrane at any intensity (weak, moderate or strong). Linear partial or complete cell membrane staining is interpreted as positive for PD-L1.

[0050] “Mononuclear inflammatory density score (MIDS)” refers to the ratio of the number of PD-L 1 expressing mononuclear inflammatory cells (MIC) infiltrating or adjacent to the tumor (small and large lymphocytes, monocytes, and macrophages within the tumor nests and the adjacent supporting stroma) compared to the total number of tumor cells. The MIDS is recorded at a scale from 0 to 4 with 0=none; l=present, but less than one MIC for every 100 tumor cells(<1 %); 2=at least one MIC for even- 100 tumor cells, but less than one MIC per 10 tumor cells (1-9%); 3=at least one MIC for every 10 tumor cells, but fewer MIC's than tumor cells (10-99%); 4=at least as many MIC's as tumor cells (>100%).

[0051] “Combined positive score (CPS)’' refers to the ratio of the number of PD-L1 positive tumor cells and PD-L1 positive mononuclear inflammatory cells (MIC) within the tumor nests and the adjacent supporting stroma (numerator) compared to the total number of tumor cells (denominator; i.e., the number of PD-L1 positive and PD-L1 negative tumor cells). PD-L1 expression at any intensity is considered positive, i.e., weak (1+), moderate (2+), or strong (3+).

[0052] “PD-L1 expression positive” refers to a Tumor Proportion Score. Mononuclear Inflammatory Density Score or Combined Positive Score of at least 1%; AIS is > 5; or elevated level of PD-L1 expression (protein and / or mRNA) by malignant cells and / or by infiltrating immune cells within a tumor compared to an appropriate control.

[0053] “DSDR” or “Durable Stable Disease Rate” means SD for > 23 weeks.

[0054] “Framework region” or “FR” as used herein means the immunoglobulin variable regions excluding the CDR regions.

[0055] “Kabat” as used herein means an immunoglobulin alignment and numbering system pioneered by Elvin A. Kabat ((1991) Sequences of Proteins of Immunological Interest. 5th Ed. Public Health Service, National Institutes of Health. Bethesda, Md.).

[0056] “LAG3 antagonist” means any chemical compound or biological molecule that blocks binding of LAG3 expressed on an immune cell (T cell, Tregs, or NK cell etc.) to its ligand(s), for example, MHC Class II molecules. Human LAG3 comprises the amino acid sequence: MWEAQFLGLLFLQPLWVAPVKPLQPGAEVPVVWAQEGAPAQLPCSPTIPLQDLSLLRRA GVTWQHQPDSGPPAAAPGHPLAPGPHPAAPSSWGPRPRRYTVLSVGPGGLRSGRLPLQP RVQLDERGRQRGDFSLWLRPARRADAGEYRAAVHLRDRALSCRLRLRLGQASMTASPP GSLRASDWVILNCSFSRPDRPASVHWFRNRGQGRVPVRESPHHHLAESFLFLPQVSPMD SGPWGCILTYRDGFNVSIMYNLTVLGLEPPTPLTVYAGAGSRVGLPCRLPAGVGTRSFLT AKWTPPGGGPDLLVTGDNGDFTLRLEDVSQAQAGTYTCHIHLQEQQLNATVTLAIITVT PKSFGSPGSLGKLLCEVTPVSGQERFVWSSLDTPSQRSFSGPWLEAQEAQLLSQPWQCQL YQGERLLGAAVYFTELSSPGAQRSGRAPGALPAGHLLLFLILGVLSLLLLVTGAFGFHLW RRQWRPRRFSALEQGI HPPQAQSKIE ELEQEPEPEP EPEPEPEPEP EPEQL (SEQ ID NO: 33); see also Uniprot accession no. Pl 8627.

[0057] “Microsatellite instability (MSI)” refers to the form of genomic instability associated with defective DNA mismatch repair in tumors. See Boland et al., Cancer Research 58, 5258-5257, 1998. In one embodiment. MSI analysis can be earned out using the five National Cancer Institute (NCI) recommended microsatellite markers: BAT25 (GenBank accession no. 9834508), BAT26 (GenBank accession no. 9834505), D5S346 (GenBank accession no. 181171), D2S123 (GenBank accession no. 187953), D17S250 (GenBank accession no. 177030). Additional markers for example, BAT40, BAT34C4, TGF-[3-RII and ACTC can be used. Commercially available kits for MSI analysis include, for example, the Promega MSI multiplex PCR assay, FoundationOne® CDx (FICDx) next generation sequencing based in vitro diagnostic device using DNA isolated from formalin-fixed, paraffin-embedded (FFPE) tumor tissue specimens.

[0058] "High frequency microsatellite instability” or “microsatellite instability -high (MSI- H)" refers to if two or more of the five NCI markers indicated above show instability or >30-40% of the total markers demonstrate instability (i.e., have insertion / deletion mutations).

[0059] "Low frequency microsatellite instability ” or “microsatellite instability -low (MSI-L)" refers to if one of the five NCI markers indicated above show instability or <30-40% of the total markers exhibit instability (i.e., have insertion / deletion mutations).

[0060] "Non-MSI-H colorectal cancer" as used herein refers to microsatellite stable (MSS) and low frequency MSI (MSI-L) colorectal cancer.

[0061] "Microsatellite Stable (MSS)" refers to if none of the five NCI markers indicated above show instability (i.e., have insertion / deletion mutations)

[0062] Proficient mismatch repair (pMMR) colorectal cancer" refers to normal expression of MMR proteins (MLH1, PMS2, MSH2, and MSH6) in a CRC tumor specimen by IHC. Commercially available kits for MMR analysis include the Ventana MMR IHC assay.

[0063] "Mismatch repair deficient (dMMR) colorectal cancer" refers to low expression of one or more MMR protein(s) (MLH 1. PMS2. MSH2. and MSH6) in a CRC tumor specimen by IHC.

[0064] “Monoclonal antibody” or “mAb” or “Mab”, as used herein, refers to a population of substantially homogeneous antibodies, i.e., the antibody molecules comprising the population are identical in amino acid sequence except for possible naturally occurring mutations that may be present in minor amounts. In contrast, conventional (polyclonal) antibody preparations typically include a multitude of different antibodies having different amino acid sequences in their variable domains, particularly their CDRs, which are often specific for different epitopes. The modifier “monoclonal” indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies and is not to be construed as requiring production of the antibody by any particular method. For example, the monoclonal antibodies to be used in accordance with the present invention may be made by the hybridoma method first described byKohler el al. (1975) Nature 256: 495 or may be made by recombinant DNA methods (see, e.g., U.S. Pat. No. 4,816,567). The ’‘monoclonal antibodies” may also be isolated from phage antibody libraries using the techniques described in Clackson et al. (1991) Nature 352: 624-628 and Marks et al. (1991) J. Mol. Biol. 222: 581-597, for example. See also Presta (2005) J. Allergy Clin. Immunol. 116:731.

[0065] “N on-responder patient”, when referring to a specific anti-tumor response to treatment with a combination therapy described herein, means the patient did not exhibit the antitumor response.

[0066] “ORR” or “objective response rate” refers in some embodiments to CR + PR, and ORR refers to CR and PR measured using irRECIST in each patient in a cohort after anti-cancer treatment.

[0067] “Patient” or “subject” refers to any single subject for which therapy is desired or that is participating in a clinical trial, epidemiological study or used as a control, including humans and mammalian veterinary patients such as cattle, horses, dogs, and cats.

[0068] “PD-1 antagonist” means any chemical compound or biological molecule that blocks binding of PD-L1 expressed on a cancer cell to PD-1 expressed on an immune cell (T cell, B cell or NKT cell) and preferably also blocks binding of PD-L2 expressed on a cancer cell to the immune-cell expressing PD-1. Alternative names or synonyms for PD-1 and its ligands include: PDCD1, PD1, CD279 and SLEB2 for PD-1 ; PDCD1L1, PDL1, B7H1, B7-4, CD274 and B7-H for PD-L1; and PDCD1L2, PDL2, B7-DC, Btdc and CD273 for PD-L2. In any of the treatment methods, combinations, pharmaceutical compositions, medicaments and uses of the present invention in which a human individual is being treated, the PD-1 antagonist blocks binding of human PD-L1 to human PD-1. and preferably blocks binding of both human PD-L1 and PD-L2 to human PD-1. Human PD-1 amino acid sequences can be found in NCBI Locus No.:NP 005009. Human PD-L1 and PD-L2 amino acid sequences can be found in NCBI Locus No.: NP_054862 and NP_079515, respectively.

[0069] “Pembrolizumab” also know n as MK-3475. is a humanized IgG4 mAb with the structure described in WHO Drug Information, Vol. 27, No. 2, pages 161-162 (2013). Pembrolizumab corresponds with the monoclonal antibody that consists of two heavy chain and two light chain sequences of SEQ ID NO: 10 and SEQ ID NO: 5, respectively. When recombinantly expressed in host cells (e g., CHO cells), or during purification / storage, post- translational modifications can occur (e.g., C-terminal Lysine clipping in the heavy chain, conversion of the N-terminal glutamine to pyroglutamate or pyroglutamic acid in the heavy chain).

[0070] As used herein, a “pembrolizumab variant” means a monoclonal antibody that comprises heavy chain and light chain sequences that are substantially identical to those in pembrolizumab, except for having three, two or one conservative amino acid substitutions in the light chain at positions that are located outside of the light chain CDRs and / or six, five, four, three, two or one conservative amino acid substitutions in the heavy chain that are located outside of the heavy chain CDRs, e g., the variant positions are located in the FR regions or the constant region. In other words, pembrolizumab and a pembrolizumab variant comprise identical CDR sequences, but differ from each other due to having a conservative amino acid substitution at no more than three or six other positions in their full length light and heavy chain sequences, respectively. A pembrolizumab variant is substantially the same as pembrolizumab with respect to the following properties: binding affinity to PD-1 and ability to block the binding of each of PD-L1 and PD-L2 to PD-1.

[0071] “RECIST 1.1 Response Criteria” as used herein means the definitions set forth in Eisenhauer et al.. E. A. et al.. Eur. J Cancer 45:228-247 (2009) for target lesions or nontarget lesions, as appropriate based on the context in which response is being measured.

[0072] “Responder patient” when referring to a specific anti-tumor response to treatment with a combination therapy described herein, means the patient exhibited the anti-tumor response.

[0073] “Sustained response” means a sustained therapeutic effect after cessation of treatment with a therapeutic agent, or a combination therapy described herein. In some embodiments, the sustained response has a duration that is at least the same as the treatment duration, or in specific embodiments at least 1.5, 2.0, 2.5 or 3 times longer than the treatment duration.

[0074] “Tissue Section” refers to a single part or piece of a tissue sample, e.g.. a thin slice of tissue cut from a sample of a normal tissue or of a tumor.

[0075] “Treat” or “treating” cancer as used herein means to administer a combination therapy of the invention to a subj ect having cancer, or diagnosed with cancer, to achieve at least one positive therapeutic effect on the cancer, such as for example, reduced number of cancer cells, reduced tumor size, reduced rate of cancer cell infiltration into peripheral organs, or reduced rate of tumor metastasis or tumor growth. Positive therapeutic effects in cancer can be measured in a number of w ays (See, W. A. Weber, J. Nucl. Med. 50: 1S-10S (2009)). For example, with respect to tumor growth inhibition, according to NCI standards, a T / C ^42% is the minimum level of anti -tumor activity. A T / C < 10% is considered a high anti-tumor activity level, with T / C (%) = Median tumor volume of the treated / Median tumor volume of the control x 100. In some embodiments, response to a combination therapy described herein is assessed usingRECIST 1.1 criteria or irRC (bidimensional or unidimensional) and the treatment achieved by a combination of the invention is any of PR, CR, OR, PFS, DFS and OS. PFS, also referred to as “Time to Tumor Progression” indicates the length of time during and after treatment that the cancer does not grow and includes the amount of time patients have experienced a CR or PR, as well as the amount of time patients have experienced SD. DFS refers to the length of time during and after treatment that the patient remains free of disease. OS refers to a prolongation in life expectancy as compared to naive or untreated individuals or patients. In some embodiments, response to a combination therapy of the invention is any of PR, CR, PFS, DFS, OR and OS that is assessed using RECIST 1.1 response criteria. The treatment regimen for a combination therapy of the invention that is effective to treat a cancer patient may vary according to factors such as the disease state, age, and weight of the patient, and the ability of the therapy to elicit an anticancer response in the subject. While an embodiment of any of the aspects of the invention may not be effective in achieving a positive therapeutic effect on the cancer in every subject, it should do so in a statistically significant number of subjects as determined by any statistical test known in the art such as the Student’s t-test, the chi2-test, the U-test according to Mann and Whitney, the Kruskal-Wallis test (H-test), Jonckheere-Terpstra-test and the Wilcoxon-test.

[0076] “In need thereof’ when used in reference to an individual in need of treatment refers to the fact that the individual has cancer or has been diagnosed with cancer.

[0077] The terms “treatment regimen”, “dosing protocol” and “dosing regimen” are used interchangeably to refer to the dose and timing of administration of each therapeutic agent in a combination of the invention.

[0078] “Treatment cycle” refers to a repeating dosing interval between completion of the first doses of the PD-1 antagonist and LAG3 antagonist and completion of the second doses of the PD-1 antagonist and LAG3 antagonist.

[0079] “Tumor” as it applies to a subject diagnosed with, or suspected of having, cancer refers to a malignant or potentially malignant neoplasm or tissue mass of any size and includes primary tumors and secondary neoplasms. A solid tumor is an abnormal growth or mass of tissue that usually does not contain cysts or liquid areas. Different types of solid tumors are named for the type of cells that form them. Examples of solid tumors are sarcomas, carcinomas, and lymphomas. Leukemias (cancers of the blood) generally do not form solid tumors (National Cancer Institute, Dictionary of Cancer Terms).

[0080] “Tumor burden” also referred to as “tumor load”, refers to the total amount of tumor material distributed throughout the body. Tumor burden refers to the total number of cancer cells or the total size of tumor(s), throughout the body, including lymph nodes and bone marrow'.Tumor burden can be determined by a variety of methods know n in the art. such as, e.g.. by measuring the dimensions of tumor(s) upon removal from the subject, e g., using calipers, or while in the body using imaging techniques, e.g., ultrasound, bone scan, computed tomography (CT) or magnetic resonance imaging (MRI) scans.

[0081] The term “tumor size” refers to the total size of the tumor which can be measured as the length and width of a tumor. Tumor size may be determined by a variety of methods known in the art, such as, e.g., by measuring the dimensions of tumor(s) upon removal from the subject, e.g., using calipers, or while in the body using imaging techniques, e.g., bone scan, ultrasound, CT or MRI scans.

[0082] ‘Unidimensional irRC” refers to the set of criteria described in Nishino M, Giobbie- Hurder A, Gargano M, Suda M, Ramaiya NH, Hodi FS. Developing a Common Language for Tumor Response to Immunotherapy : Immune-related Response Criteria using Uni dimensional measurements. Clin Cancer Res. 2013; 19(14): 3936— 3943). These criteria utilize the longest diameter (cm) of each lesion.

[0083] “Neoadjuvant” as used herein refers to the treatment (e.g., combination therapy comprising the PD-1 antagonist, LAG3 antagonist, and all-trans retinoic acid) before surgical resection of a tumor in the patient.

[0084] “Adjuvant” as used herein refers to treatment after surgical resection of a tumor in the patient.

[0085] “Variable regions” or “V region” as used herein means the segment of IgG chains which is variable in sequence betw een different antibodies. Typically, it extends to Kabat residue 109 in the light chain and 113 in the heavy chain.PD-1 ANTAGONISTS AND LAG3 ANTAGONISTS

[0086] PD-1 antagonists useful in the treatment methods, combinations, pharmaceutical compositions, medicaments and uses of the present invention include a monoclonal antibody (mAb), or antigen binding fragment thereof, that specifically binds to PD-1 or PD-L1, and preferably specifically binds to human PD-1 or human PD-L1. The mAb may be a human antibody, a humanized antibody or a chimeric antibody, and may include a human constant region. In some embodiments the human constant region is selected from the group consisting of IgGl, IgG2, IgG3 and IgG4 constant regions, and in preferred embodiments, the human constant region is an IgGl or IgG4 constant region. In some embodiments, the antigen binding fragment is selected from the group consisting of Fab, Fab’-SH, F(ab’)2, scFv and Fv fragments.

[0087] Examples of mAbs that bind to human PD-1, and useful in the treatment methods, combinations, pharmaceutical compositions, medicaments and uses of the present invention, are described in U.S. patent nos. US7488802, US7521051, US8008449, US8354509, and US 8168757, and International application publn. Nos. W02004 / 004771, W02004 / 072286, W02004 / 056875, and US2011 / 0271358. Specific anti-human PD-1 mAbs useful as the PD-1 antagonist in the treatment methods, combinations, pharmaceutical compositions, medicaments and uses of the present invention include: pembrolizumab (also known as MK-3475), a humanized IgG4 mAb with the structure described in WHO Drug Information, Vol. 27, No. 2, pages 161-162 (2013) and that comprises the heavy and light chain amino acid sequences shown in Table 3; nivolumab (BMS-936558), a human IgG4 mAb with the structure described in WHO Drug Information, Vol. 27, No. 1, pages 68-69 (2013) and that comprises the heavy and light chain amino acid sequences shown in Table 3; the humanized antibodies h409Al l, h409A16 and h409A17, which are described in WO2008 / 156712, and AMP-514, which is being developed by Medlmmune; cemiplimab; camrelizumab; sintilimab; tislelizumab; and toripalimab. Additional anti-PD-1 antibodies contemplated for use herein include MEDI0680 (U.S. Patent no. 8609089), BGB-A317 (U.S. Patent publ. no. 2015 / 0079109), INCSHR1210 (SHR-1210) (PCT International application publ. no. WO2015 / 085847), REGN-2810 (PCT International application publ. no. WO2015 / 112800). PDR001 (PCT International application publ. no. WO2015 / 112900), TSR- 042 (ANB011) (PCT International application publ. no. WO2014 / 179664) and STI-11 10 (PCT International application publ. no. WO2014 / 194302).

[0088] Examples of mAbs that bind to human PD-L1, and useful in the treatment methods, combinations, pharmaceutical compositions, medicaments and uses of the present invention, are described in US8383796. Specific anti-human PD-L1 mAbs useful as the PD-1 antagonist in the treatment methods, combinations, pharmaceutical compositions, medicaments and uses of the present invention include BMS-936559, MEDI4736, and MSB0010718C.

[0089] Other PD-1 antagonists useful in the treatment methods, combinations, pharmaceutical compositions, medicaments and uses of the present invention include an immunoadhesin that specifically binds to PD-1 or PD-L1, and preferably specifically binds to human PD-1 or human PD-L1, e.g., a fusion protein containing the extracellular or PD-1 binding portion of PD-L1 or PD-L2 fused to a constant region such as an Fc region of an immunoglobulin molecule. Examples of immunoadhesion molecules that specifically bind to PD- 1 are described in PCT International application public. Nos. WO2010 / 027827 and WO2011 / 066342. Specific fusion proteins useful as the PD-1 antagonist in the treatment methods, combinations, pharmaceutical compositions, medicaments and uses of the presentinvention include AMP -224 (also known as B7-DClg), which is a PD-L2-FC fusion protein and binds to human PD-1.

[0090] In some preferred embodiments of the treatment methods, combinations, pharmaceutical compositions, medicaments and uses of the present invention, the PD-1 antagonist is a monoclonal antibody, or antigen binding fragment thereof, that comprises: (a) a light chain variable region comprising light chain CDR1, CDR2 and CDR3 of SEQ ID NOs: 1, 2 and 3, respectively and (b) a heavy chain variable region comprising heavy chain CDR1, CDR2 and CDR3 of SEQ ID NOs: 6, 7 and 8, respectively. In one embodiment, the PD-1 antagonist is a humanized antibody.

[0091] In other preferred embodiments of the treatment methods, combinations, pharmaceutical compositions, medicaments and uses of the present invention, the PD-1 antagonist is a monoclonal antibody, or antigen binding fragment thereof, that specifically binds to human PD-1 and comprises (a) a heavy chain variable region comprising SEQ ID NO:9 or a variant thereof, and (b) a light chain variable region comprising SEQ ID NO:4 or a variant thereof. A variant of a heavy chain variable region sequence is identical to the reference sequence except having up to six conservative amino acid substitutions in the framework region (i.e., outside of the CDRs). A variant of a light chain variable region sequence is identical to the reference sequence except having up to three conservative amino acid substitutions in the framework region (i.e., outside of the CDRs).

[0092] In another preferred embodiment of the treatment methods, combinations, pharmaceutical compositions, medicaments and uses of the present invention, the PD-1 antagonist is a monoclonal antibody that specifically binds to human PD-1 and comprises (a) a heavy chain comprising SEQ ID NO: 10 and (b) a light chain comprising SEQ ID NO:5. In one embodiment, the PD-1 antagonist is an anti-PD-1 antibody that comprises a heavy chain and a light chain, and wherein the heavy and light chains comprise the amino acid sequences in SEQ ID NO:10 and SEQ ID NO:5, respectively. In another embodiment, the PD-1 antagonist is an anti-PD-1 antibody that comprises two heavy chains and two light chains, and wherein the heavychain comprises SEQ ID NO: 10 and the light chain comprises SEQ ID NO:5. In a further embodiment, the PD-1 antagonist is pembrolizumab. In yet a further embodiment, the PD-1 antagonist is a pembrolizumab variant.

[0093] In yet another preferred embodiment of the treatment methods, combinations, pharmaceutical compositions, medicaments and uses of the present invention, the PD-1 antagonist is a monoclonal antibody^ that specifically binds to human PD-1 and comprises (a) aheavy chain comprising SEQ ID NO: 12 and (b) a light chain comprising SEQ ID NO: 11. In another embodiment, the PD-1 antagonist is nivolumab.

[0094] In all of the above treatment methods, combinations, pharmaceutical compositions, medicaments and uses, the PD-1 antagonist inhibits the binding of PD-L1 to PD-1, and preferably also inhibits the binding of PD-L2 to PD-1. In some embodiments of the above treatment methods, combinations, pharmaceutical compositions, medicaments and uses, the PD-1 antagonist is a monoclonal antibody, or an antigen binding fragment thereof, that specifically binds to PD-1 or to PD-L1 and blocks the binding of PD-L1 to PD-1.

[0095] Table 3 below provides a list of the amino acid sequences of exemplary- anti-PD-1 mAbs for use in the treatment methods, combinations, pharmaceutical compositions, medicaments and uses of the present invention.Table 3. Exemplary- PD-1 Antibody Sequences

[0096] LAG3 antagonists useful in the treatment methods, combinations, pharmaceutical compositions, medicaments and uses of the present invention include a monoclonal antibody (mAb), or antigen binding fragment thereof, that specifically binds to LAG3. The mAb may be a human antibody, a humanized antibody or a chimeric antibody, and may include a human constant region. In some embodiments the human constant region is selected from the group consisting of IgGl , IgG2, IgG3 and IgG4 constant regions, and in preferred embodiments, the human constant region is an IgGl or IgG4 constant region. In some embodiments, the antigen binding fragment is selected from the group consisting of Fab, Fab'-SH, F(ab')2, scFv and Fv fragments.

[0097] In one embodiment, the anti-LAG3 antibody is favezelimab.

[0098] Favezelimab light chain immunoglobulin amino acid sequence:DIVMTQTPLSLSVTPGQPASISCKASQSLDYEGDSDMNWYLQKPGQPPQLLIYGASNLES GVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCQQSTEDPRTFGGGTKVEIKRTVAAPSVF IFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSL SSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ ID NO: 22); and

[0099] Favezelimab heavy chain immunoglobulin amino acid sequence:QMQLVQSGPEVKKPGTSVKVSCKASGYTFTDYNVDWVRQARGQRLEWIGDINPNDGG TIYAQKFQERVTITVDKSTSTAYMELSSLRSEDTAVYYCARNYRWFGAMDHWGQGTTV TVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLG GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQ FNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPS QEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVD KSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO: 23).

[0100] Favezelimab light chain immunoglobulin variable domain amino acid sequence: DIVMTQTPLSLSVTPGQPASISCKASQSLDYEGDSDMNWYLQKPGQPPQLLIYGASNLES GVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCQQSTEDPRTFGGGTKVEIK (SEQ ID NO: 24); and

[0101] Favezelimab heavy chain immunoglobulin variable domain amino acid sequence: QMQLVQSGPEVKKPGTSVKVSCKASGYTFTDYNVDWVRQARGQRLEWIGDINPNDGG TIYAQKFQERVTITVDKSTSTAYMELSSLRSEDTAVYYCARNYRWFGAMDHWGQGTTV TVSS (SEQ ID NO: 25).

[0102] Favezelimab CDRs:CDR-L1: KASQSLDYEGDSDMN (SEQ ID NO: 26);CDR-L2: GASNLES (SEQ ID NO: 27);CDR-L3: QQSTEDPRT (SEQ ID NO: 28);CDR-H1: DYNVD (SEQ ID NO: 29);CDR-H2: DINPNDGGTIYAQKFQE (SEQ ID NO: 30); andCDR-H3: NYRWFGAMDH (SEQ ID NO: 31)

[0103] In some preferred embodiments of the treatment methods, combinations, pharmaceutical compositions, medicaments and uses of the present invention, the LAG3 antagonist is a monoclonal antibody, or antigen binding fragment thereof, that comprises: (a) light chain CDRs 1, 2, and 3 of SEQ ID NOs: 26, 27 and 28, respectively, and (b) heavy chain CDRs 1, 2. and 3 of SEQ ID NOs: 29. 30 and 31, respectively.

[0104] In other preferred embodiments of the treatment methods, combinations, pharmaceutical compositions, medicaments and uses of the present invention, the LAG3 antagonist is a monoclonal antibody, or antigen binding fragment thereof, that specifically bindsto human LAG3 and comprises (a) a heavy chain variable region comprising SEQ ID NO:25 or a variant thereof, and (b) a light chain variable region comprising SEQ ID NO:24 or a variant thereof. A variant of a heavy chain variable region sequence is identical to the reference sequence except having up to 5 conservative amino acid substitutions in the framework region (i.e., outside of the CDRs). A variant of a light chain variable region sequence is identical to the reference sequence except having up to three conservative amino acid substitutions in the framework region (i.e., outside of the CDRs).

[0105] In another preferred embodiment of the treatment methods, combinations, pharmaceutical compositions, medicaments and uses of the present invention, the LAG3 antagonist is a monoclonal antibody that specifically binds to human LAG3 and comprises (a) a heavy chain comprising SEQ ID NO: 23 and (b) a light chain comprising SEQ ID NO:22. In another embodiment, the LAG3 antagonist is an anti-LAG3 antibody that comprises two heavy chains and two light chains, and wherein the heavy chain comprises SEQ ID NO:23 and the light chain comprises SEQ ID NO:22. In another preferred embodiment of the treatment methods, combinations, pharmaceutical compositions, medicaments and uses of the present invention, the LAG3 antagonist is a monoclonal antibody that specifically binds to human LAG3 and comprises (a) a heavy chain variable region comprising SEQ ID NO: 25 and (b) a light chain variable region comprising SEQ ID NO:24. In one embodiment, the LAG3 antagonist is favezelimab. In another embodiment, the LAG3 antagonist is a favezelimab variant.

[0106] Other examples of mAbs that bind to human LAG3, and are useful in the treatment methods, combinations, pharmaceutical compositions, medicaments and uses of the present invention, are relatlimab disclosed in International patent application publication no. W02014 / 008218 as LAG3.5 (WHO Drug Information. Vol. 32, No. 2. 2018), IMP731, 1MP701, and the anti-LAG3 antibodies disclosed in U.S. patent application publication no. US2017101472. In one embodiment, the LAG3 antagonist is relatlimab. In another embodiment, the LAG3 antagonist is fianlimab. Other LAG3 antagonists useful in the treatment methods, combinations, pharmaceutical compositions, medicaments and uses of the present invention include an immunoadhesin that specifically binds to human LAG3, e.g., a fusion protein containing the extracellular LAG3 fused to a constant region such as an Fc region of an immunoglobulin molecule.

[0107] In one embodiment, each of the anti-PD-1 or anti-LAG3 antibodies or antigenbinding fragments thereof comprises a heavy chain constant region, e.g., a human constant region, such as yl, y2, y3, or y4 human heavy chain constant region or a variant thereof. In another embodiment, each of the anti-PD-1 or anti-LAG3 antibodies or antigen-bindingfragments thereof comprises a light chain constant region, e.g., a human light chain constant region, such as lambda or kappa human light chain region or a variant thereof. By way of example, and not limitation, the human heavy7chain constant region can be y4 and the human light chain constant region can be kappa. In an alternative embodiment, the Fc region of the antibody is y4 with a Ser228Pro mutation (Schuurman, J et al., Mol. Immunol. 38: 1-8. 2001).

[0108] In some embodiments, different constant domains may be appended to humanized VL and VH regions derived from the CDRs provided herein. For example, if a particular intended use of an antibody (or fragment) of the present invention were to call for altered effector functions, a heavy7chain constant domain other than human IgGl may be used, or hybrid IgGl / IgG4 may be utilized.

[0109] Although human IgGl antibodies provide for long half-life and for effector functions, such as complement activation and antibody-dependent cellular cytotoxicity7, such activities may not be desirable for all uses of the antibody. In such instances a human IgG4 constant domain, for example, may be used. The present invention includes the use of anti-PD-1 antibodies or anti- LAG3 antibodies and antigen-binding fragments thereof which comprise an IgG4 constant domain. In one embodiment, the IgG4 constant domain can differ from the native human IgG4 constant domain (Swiss-Prot Accession No. P01861.1) at a position corresponding to position 228 in the EU system and position 241 in the KABAT system, where the native Serl08 is replaced with Pro, in order to prevent a potential inter-chain disulfide bond between Cysl06 and Cysl09 (corresponding to positions Cys 226 and Cys 229 in the EU system and positions Cys 239 and Cys 242 in the KABAT system) that could interfere with proper intra-chain disulfide bond formation. See Angal et al. (1993) Mol. Imunol. 30: 105. In other instances, a modified IgGl constant domain which has been modified to increase half-life or reduce effector function can be used.METHODS, USES AND MEDICAMENTS

[0110] In one embodiment, the invention provides a method for treating cancer in an individual comprising co-administering to the individual a PD-1 antagonist, LAG3 antagonist and all-trans retinoic acid or a pharmaceutically acceptable salt thereof. In another embodiment, the invention provides a method for treating cancer in an individual comprising administering to the individual a composition comprising a PD-1 antagonist and a LAG3 antagonist, and a composition comprising all-trans retinoic acid or a pharmaceutically acceptable salt thereof. [OHl] In another embodiment, the invention provides a medicament comprising a PD-1 antagonist for use in combination with a LAG3 antagonist and all-trans retinoic acid or apharmaceutically acceptable salt thereof for treating cancer. In yet another embodiment, the invention provides a medicament comprising a LAG3 antagonist for use in combination with a PD-1 antagonist and all-trans retinoic acid or a pharmaceutically acceptable salt thereof for treating cancer. In yet another embodiment, the invention provides a medicament comprising all- trans retinoic acid or a pharmaceutically acceptable salt thereof for use in combination with a PD-1 antagonist and LAG3 antagonist for treating cancer.

[0112] In another embodiment, the invention provides for the use of a PD-1 antagonist in the manufacture of a medicament for treating cancer in an individual when administered in combination with a LAG3 antagonist and all-trans retinoic acid or a pharmaceutically acceptable salt thereof. In one embodiment, the invention provides for the use of a LAG3 antagonist in the manufacture of a medicament for treating cancer in an individual when administered in combination with a PD-1 antagonist and all-trans retinoic acid or a pharmaceutically acceptable salt thereof. In another embodiment, the invention provides for the use of all-trans retinoic acid or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating cancer in an individual when administered in combination with a LAG3 antagonist and PD-1 antagonist.

[0113] Other embodiments provide a LAG3 antagonist for use in the treatment of cancer, wherein the use is in combination with a PD-1 antagonist and all-trans retinoic acid or a pharmaceutically acceptable salt thereof; a PD-1 antagonist for use in the treatment of cancer, wherein the use is in combination with a LAG3 antagonist and all-trans retinoic acid or a pharmaceutically acceptable salt thereof; all-trans retinoic acid or a pharmaceutically acceptable salt thereof for use in the treatment of cancer, wherein the use is in combination with a PD-1 antagonist and a LAG3 antagonist.

[0114] In a still further embodiment, the invention provides use of a PD-1 antagonist and a LAG3 antagonist in the manufacture of a medicament for treating cancer in an individual when administered in combination with all-trans retinoic acid or a pharmaceutically acceptable salt thereof. In yet another embodiment, the invention provides a medicament comprising a PD-1 antagonist and a LAG3 antagonist for use in combination with all-trans retinoic acid or a pharmaceutically acceptable salt thereof for treating cancer.

[0115] In the foregoing methods, medicaments and uses, in one embodiment, the PD-1 antagonist and LAG3 antagonist are co-formulated and administered via intravenous infusion or subcutaneous injection and co-administered with all-trans retinoic acid. In another embodiment, the PD-1 antagonist and LAG3 antagonist are co-administered via intravenous infusion or subcutaneous injection and co-administered with all-trans retinoic acid.

[0116] In the foregoing methods, combinations, pharmaceutical compositions, medicaments and uses, in one embodiment, the PD-1 antagonist and LAG3 antagonist and all-trans retinoic acid or a pharmaceutically acceptable salt thereof are administered before surgical resection of a tumor, and optionally followed by treatment with a PD-1 antagonist after surgery. In the foregoing methods, combinations, pharmaceutical compositions, medicaments and uses, in one embodiment, the PD-1 antagonist, LAG3 antagonist and all-trans retinoic acid or a pharmaceutically acceptable salt thereof are administered after surgical resection of a tumor, and optionally a PD-1 antagonist is administered before surgery. In the foregoing methods, combinations, pharmaceutical compositions, medicaments and uses, in one embodiment, the PD- 1 antagonist, LAG3 antagonist and all-trans retinoic acid or a pharmaceutically acceptable salt thereof are administered before and after surgical resection of a tumor.

[0117] In one embodiment, the PD-1 antagonist is an anti-PD-1 antibody that blocks the binding of PD-1 to PD-L1 and PD-L2. In one embodiment, the PD-1 antagonist is an anti-PD-Ll antibody. In one embodiment, the LAG3 antagonist is an anti-LAG3 antibody that blocks the binding of LAG3 to MHC Class II. In one embodiment, the PD-1 antagonist is pembrolizumab and the LAG3 antagonist is favezelimab. In another embodiment, the PD-1 antagonist is nivolumab and the LAG3 antagonist is relatlimab. In one embodiment, the PD-1 antagonist is cemiplimab and the LAG3 antagonist is fianlimab.

[0118] Cancers that may be treated by the methods, combinations, pharmaceutical compositions, medicaments and uses of the invention include, but are not limited to: Cardiac cancers: sarcoma (angiosarcoma, fibrosarcoma, rhabdomyosarcoma, liposarcoma), myxoma, rhabdomyoma, fibroma, lipoma and teratoma; Lung cancers: bronchogenic carcinoma (squamous cell, undifferentiated small cell, undifferentiated large cell, adenocarcinoma), alveolar (bronchiolar) carcinoma, bronchial adenoma, sarcoma, lymphoma, chondromatous hamartoma, mesothelioma; Gastrointestinal cancers: esophagus (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), stomach (carcinoma, lymphoma, leiomyosarcoma), pancreas (ductal adenocarcinoma, insulinoma, glucagonoma, gastrinoma, carcinoid tumors, vipoma), small bowel (adenocarcinoma, lymphoma, carcinoid tumors, Karposi's sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), large bowel (adenocarcinoma, tubular adenoma, villous adenoma, hamartoma, leiomyoma) colorectal; Genitourinary' tract cancers: kidney (adenocarcinoma, Wilm's tumor (nephroblastoma), lymphoma, leukemia), bladder and urethra (squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma), prostate (adenocarcinoma, sarcoma), testis (seminoma, teratoma, embryonal carcinoma, teratocarcinoma, choriocarcinoma, sarcoma, interstitial cell carcinoma, fibroma, fibroadenoma, adenomatoidtumors, lipoma); Liver cancers: hepatoma (hepatocellular carcinoma), cholangiocarcinoma, hepatoblastoma, angiosarcoma, hepatocellular adenoma, hemangioma; Bone cancers: osteogenic sarcoma (osteosarcoma), fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma (reticulum cell sarcoma), malignant giant cell tumor chordoma, osteochronfroma (osteocartilaginous exostoses), benign chondroma, chondroblastoma, chondromyxofibroma, osteoid osteoma and giant cell tumors; Nervous system cancers: skull (osteoma, hemangioma, granuloma, xanthoma, osteitis deformans), meninges (meningioma, meningiosarcoma, gliomatosis), brain (astrocytoma, medulloblastoma, glioma, ependymoma, germinoma (pinealoma), glioblastoma multiform, oligodendroglioma, schwannoma, retinoblastoma, congenital tumors), spinal cord neurofibroma, meningioma, glioma, sarcoma); Gynecological cancers: uterus (endometrial carcinoma), cervix (cervical carcinoma, pre-tumor cervical dysplasia), ovaries (ovarian carcinoma (serous cystadenocarcinoma, mucinous cystadenocarcinoma, unclassified carcinoma), granulosa-thecal cell tumors, Sertoli-Leydig cell tumors, dysgerminoma. malignant teratoma), vulva (squamous cell carcinoma, intraepithelial carcinoma, adenocarcinoma, fibrosarcoma, melanoma), vagina (clear cell carcinoma, squamous cell carcinoma, botry oid sarcoma (embryonal rhabdomyosarcoma), fallopian tubes (carcinoma), breast; tumors of mesenchymal origin, including fibrosarcoma and rhabdomyosarcoma; tumors of the central and peripheral nervous system, including astrocytoma, neuroblastoma, glioma, and schwannomas; and other tumors, including melanoma, skin (non-melanomal) cancer, mesothelioma (cells), seminoma, teratocarcinoma, osteosarcoma, xenoderoma pigmentosum, keratoctanthoma, thyroid follicular cancer and Kaposi's sarcoma. In one embodiment, the forgoing cancers are resectable. In one embodiment, the forgoing cancers are unresectable. In one embodiment, the cancer is Stage IV. In another embodiment, the cancer is Stage III. In one embodiment, the cancer is Stage II. In one embodiment, the cancer is Stage I. In another embodiment, the cancer is without distant metastasis. In another embodiment, the cancer is stage I, II, III or IV without distant metastasis.

[0119] In another embodiment, cancers that may be treated by the methods, combinations, pharmaceutical compositions, medicaments and uses of the invention include, but are not limited to: cutaneous squamous cell carcinoma, melanoma, breast cancer, non-small cell lung cancer, bladder cancer, esophageal cancer, renal cell cancer, endometrial cancer and colorectal cancer. In one embodiment, the colorectal cancer or endometrial cancer is non-microsatellite instability- high (non-MSI-H) or proficient mismatch repair (pMMR). In one embodiment, the cancer is resectable. In one embodiment, the cancer is Stage IV. In another embodiment, the cancer isStage III. In one embodiment, the cancer is unresectable Stage III or IV. In one embodiment, thecancer is resectable Stage III or IV. In one embodiment, the cancer is Stage II. In one embodiment, the cancer is Stage I. In another embodiment, the cancer is without distant metastasis. In another embodiment, the cancer is stage I, II, III or IV without distant metastasis.

[0120] In specific embodiments, cancers that may be treated by the methods, combinations, pharmaceutical compositions, medicaments and uses of the invention is selected from the group consisting of: melanoma, lung cancer, head and neck cancer, bladder cancer, breast cancer, gastrointestinal cancer, multiple myeloma, hepatocellular cancer, Merkel cell carcinoma, cutaneous squamous cell carcinoma, lymphoma, renal cancer, mesothelioma, ovarian cancer, esophageal cancer, anal cancer, biliary tract cancer, colorectal cancer, endometrial cancer, cervical cancer, thyroid cancer, salivary cancer, prostate cancer (e.g., hormone refractory prostate adenocarcinoma), pancreatic cancer, colon cancer, liver cancer, thyroid cancer, glioblastoma, glioma, and other neoplastic malignancies. In another embodiment, the cancer is selected from the group consisting of: melanoma, non-small cell lung cancer, small cell lung cancer, head and neck squamous cell cancer, classical Hodgkin lymphoma, primary mediastinal large B-cell lymphoma, urothelial carcinoma, microsatellite instability -high or mismatch repair deficient cancer, gastric cancer, esophageal cancer, cervical cancer, hepatocellular carcinoma, Merkel cell carcinoma, renal cell carcinoma, endometrial carcinoma, a cancer characterized by a tumor having a high mutational burden, cutaneous squamous cell carcinoma, and triple negative breast cancer. In one embodiment, the cancer is resectable. In one embodiment, the cancer is Stage IV. In another embodiment, the cancer is Stage III. In one embodiment, the cancer is unresectable Stage III or IV. In one embodiment, the cancer is resectable Stage III or IV. In one embodiment, the cancer is Stage II. In one embodiment, the cancer is Stage I. In another embodiment, the cancer is without distant metastasis. In another embodiment, the cancer is stage 1, II. Ill or IV without distant metastasis.

[0121] In a further embodiment, cancers that may be treated by the methods, combinations, pharmaceutical compositions, medicaments or uses of the invention include cancers selected from the group consisting of: cutaneous squamous cell carcinoma, melanoma, esophageal cancer, breast cancer, non-small cell lung cancer and bladder cancer. In one embodiment, the cancer is cutaneous squamous cell carcinoma. In one embodiment, the cancer is melanoma. In one embodiment, the cancer is triple negative breast cancer. In one embodiment, the cancer is Her2 positive breast cancer. In one embodiment, the cancer is non-small cell lung cancer. In one embodiment, the cancer is non-muscle invasive bladder cancer. In one embodiment, the cancer is resectable. In one embodiment, the cancer is Stage IV. In another embodiment, the cancer isStage III. In one embodiment, the cancer is unresectable Stage III or IV. In one embodiment, thecancer is resectable Stage III or IV. In one embodiment, the cancer is Stage II. In one embodiment, the cancer is Stage I. In another embodiment, the cancer is without distant metastasis. In another embodiment, the cancer is stage I, II, III or IV without distant metastasis. In one embodiment, the cancer is unresectable Stage III or IV melanoma. In one embodiment, the cancer is resectable Stage III or IV melanoma. In another embodiment, the cancer is stage I, II, III or IV melanoma without distant metastasis.

[0122] In one aspect of the foregoing embodiments, the patient with cancer did not relapse after anti-PD-1 or anti-PD-Ll treatment. In one embodiment, the patient with cancer has not received prior anti-PD-1 or anti-PD-Ll treatment. In one embodiment, the patient with cancer has received prior immunotherapy treatment. In one embodiment, the combination therapy of the PD-1 antagonist, LAG3 antagonist and all-trans retinoic acid or a pharmaceutically acceptable salt thereof is for first line treatment.

[0123] In some embodiments, a combination therapy of the invention is administered to a patient who has not been previously treated with a biotherapeutic or chemotherapeutic agent, i.e., is treatment-naive.

[0124] The methods, combinations, pharmaceutical compositions, medicaments and uses of the invention may also comprise one or more additional therapeutic agents. The additional therapeutic agent may be, e.g.. a chemotherapeutic, a biotherapeutic agent, an immunogenic agent (for example, attenuated cancerous cells, tumor antigens, antigen presenting cells such as dendritic cells pulsed with tumor derived antigen or nucleic acids, immune stimulating cytokines (for example, IL-2, IFNa2, GM-CSF), and cells transfected with genes encoding immune stimulating cytokines such as but not limited to GM-CSF). The specific dosage and dosage schedule of the additional therapeutic agent can further vary, and the optimal dose, dosing schedule and route of administration will be determined based upon the specific therapeutic agent that is being used.

[0125] Each therapeutic agent in the methods, combinations, pharmaceutical compositions, medicaments and uses of the invention may be administered either alone or in a medicament (also referred to herein as a pharmaceutical composition) that comprises the therapeutic agent and one or more pharmaceutically acceptable carriers, excipients and diluents, according to standard pharmaceutical practice.

[0126] Each therapeutic agent in the methods, combinations, pharmaceutical compositions, medicaments and uses of the invention may be administered simultaneously (i.e., in the same medicament), concurrently (i.e., in separate medicaments administered one right after the other in any order) or sequentially in any order. Sequential administration is particularly useful when thetherapeutic agents in the combination therapy are in different dosage forms (one agent is a tablet or capsule and another agent is a sterile liquid) and / or are administered on different dosing schedules, e.g., a chemotherapeutic that is administered at least daily and a biotherapeutic that is administered less frequently, such as once weekly, once every two weeks, or once every three weeks.

[0127] In some embodiments, the LAG3 antagonist is administered before administration of the PD-1 antagonist, while in other embodiments, the LAG3 antagonist is administered after administration of the PD-1 antagonist. In another embodiment, the LAG3 antagonist is administered concurrently with the PD- 1 antagonist. In one embodiment, the all-trans retinoic acid or pharmaceutically acceptable salt thereof is administered before the LAG3 antagonist and the PD-1 antagonist.

[0128] In some embodiments, at least one of the therapeutic agents in the methods, combinations, pharmaceutical compositions, medicaments and uses of the invention is administered using the same dosage regimen (dose, frequency and duration of treatment) that is typically employed when the agent is used as monotherapy for treating the same cancer. In other embodiments, the patient receives a lower total amount of at least one of the therapeutic agents in the methods, combinations, pharmaceutical compositions, medicaments and uses than when the agent is used as monotherapy, e.g., smaller doses, less frequent doses, and / or shorter treatment duration.

[0129] Each small molecule therapeutic agent in the methods, combinations, pharmaceutical compositions, medicaments and uses of the invention can be administered orally or parenterally, including the intravenous, intramuscular, intraperitoneal, subcutaneous, rectal, topical, and transdermal routes of administration.

[0130] A combination therapy of the invention can be administered to a human patient who has a cancer that tests positive for one or both of PD-L1 and PD-L2, and preferably tests positive for PD-L1 expression. In some preferred embodiments, PD-L1 expression is detected using a diagnostic anti -human PD-L1 antibody, or antigen binding fragment thereof, in an IHC assay on an FFPE or frozen tissue section of a tumor sample removed from the patient. Typically, the patient's physician would order a diagnostic test to determine PD-L1 expression in a tumor tissue sample removed from the patient prior to initiation of treatment with the PD-1 antagonist, the LAG3 antagonist but it is envisioned that the physician could order the first or subsequent diagnostic tests at any time after initiation of treatment, such as for example after completion of a treatment cycle. In one embodiment, the PD-L1 expression is measured by the PD-L1 IHC 22C3 pharmDx assay. In another embodiment, the patient has a Mononuclear Inflammatory DensityScore for PD-L1 expression >2. In another embodiment, the patient has a Mononuclear Inflammatory Density Score for PD-L1 expression >3. In another embodiment, the patient has a Mononuclear Inflammatory Density Score for PD-L1 expression >4. In another embodiment, Tumor Proportion Score for PD-L1 expression is used for selection of non-small cell lung cancer patients. In another embodiment, the patient has a Tumor Proportion Score for PD-L1 expression >1%. In another embodiment, the patient has a Tumor Proportion Score for PD-L1 expression >10%. In another embodiment, the patient has a Tumor Proportion Score for PD-L1 expression>20%. In another embodiment, the patient has a Tumor Proportion Score for PD-L1 expression>30%. In another embodiment, the patient has a Tumor Proportion Score for PD-L1 expression>50%. In a further embodiment, the patient has a Combined Positive Score for PD-L1 expression >1%. In a further embodiment, the patient has a Combined Positive Score for PD-L1 expression between 1 and 20 %. In a further embodiment, the patient has a Combined Positive Score for PD- L1 expression > 2%. In a further embodiment, the patient has a Combined Positive Score for PD- L1 expression > 5%. In yet a further embodiment, the patient has a Combined Positive Score for PD-L1 expression > 10%. In a further embodiment, the patient has a Combined Positive Score for PD-L1 expression > 15%. In yet a further embodiment, the patient has a Combined Positive Score for PD-L1 expression > 20%.

[0131] Selecting a dosage regimen (also referred to herein as an administration regimen) for a combination therapy of the invention depends on several factors, including the serum or tissue turnover rate of the entity, the level of symptoms, the immunogenicity of the entity, and the accessibility of the target cells, tissue or organ in the individual being treated. Preferably, a dosage regimen maximizes the amount of each therapeutic agent delivered to the patient consistent with an acceptable level of side effects. Accordingly, the dose amount and dosing frequency of each biotherapeutic and chemotherapeutic agent in the combination depends in part on the particular therapeutic agent, the severity of the cancer being treated, and patient characteristics. Guidance in selecting appropriate doses of antibodies, cytokines, and small molecules are available. See. e.g., Wawrzynczak (1996) Antibody Therapy. Bios Scientific Pub. Ltd, Oxfordshire, UK; Kresina (ed.) (\99\) Monoclonal Antibodies. Cytokines and Arthritis , Marcel Dekker, New York, NY; Bach (ed.) (1993) Monoclonal Antibodies and Peptide Therapy in Autoimmune Diseases, Marcel Dekker, New York, NY; Baert et al. (2003) New Engl. J. Med. 348:601-608; Milgrom er ? / . (1999) New Engl. J. Med. 341 : 1966-1973; Slamon er a / . (2001) New Engl. J. Med. 344:783-792; Beniaminovitz et al. (2000) New Engl. J. Med. 342:613-619; Ghosh et al. (2003) New Engl. J. Med. 348:24-32; Lipsky et al. (2000) New Engl. J. Med. 343: 1594-1602; Physicians' Desk Reference 2003 (Physicians' Desk Reference, 57th Ed);Medical Economics Company; ISBN: 1563634457; 57th edition (November 2002).Determination of the appropriate dosage regimen may be made by the clinician, e.g., using parameters or factors known or suspected in the art to affect treatment or predicted to affect treatment, and will depend, for example, the patient's clinical history (e.g., previous therapy), the type and stage of the cancer to be treated and biomarkers of response to one or more of the therapeutic agents in the combination therapy.

[0132] Biotherapeutic agents in a combination therapy of the invention may be administered by continuous infusion, or by doses at intervals of, e.g., daily, every' other day, three times per week, or one time each week, two weeks, three weeks, monthly, bimonthly, etc. A total weekly dose is generally at least 0.05 pg / kg, 0.2 pg / kg, 0.5 pg / kg, 1 pg / kg, 10 pg / kg, 100 pg / kg, 0.2 mg / kg, 1.0 mg / kg, 2.0 mg / kg, 10 mg / kg, 25 mg / kg, 50 mg / kg body weight or more. See, e.g., Yang et al. (2003) New Engl. J. Med. 349:427-434; Herold et al. (2002) New Engl. J. Med. 346: 1692-1698; Liu et al. (1999) Neurol. Neurosurg. Psych. 67:451-456; Portielji et al. (20003) Cancer Immunol. Immunother. 52: 133-144.

[0133] In some embodiments that employ an anti -human PD-1 mAb as the PD-1 antagonist in the methods, combinations, pharmaceutical compositions, medicaments and uses of the invention, the dosing regimen will comprise administering the anti-human PD-1 mAb at a dose of 1, 2, 3, 5 or lOmg / kg at intervals of about 14 days (± 2 days) or about 21 days (± 2 days) or about 30 days (± 2 days) throughout the course of treatment.

[0134] In other embodiments that employ an anti-human PD-1 mAb as the PD-1 antagonist in the methods, combinations, pharmaceutical compositions, medicaments and uses of the invention, the dosing regimen will comprise administering the anti-human PD-1 mAb at a dose of from about 0.005 mg / kg to about 10 mg / kg, with intra-patient dose escalation. In other escalating dose embodiments, the interval between doses will be progressively shortened, e g., about 30 days (± 2 days) between the first and second dose, about 14 days (± 2 days) between the second and third doses. In certain embodiments, the dosing interval will be about 14 days (± 2 days), for doses subsequent to the second dose.

[0135] In certain embodiments, a subject will be administered an intravenous (IV) infusion or subcutaneous injection of a medicament comprising any of the PD-1 antagonists described herein.

[0136] In one preferred embodiment of the invention, the PD-1 antagonist in the combination therapy is nivolumab, which is administered intravenously at a dose selected from the group consisting of: 1 mg / kg Q2W, 2 mg / kg Q2W, 3 mg / kg Q2W, 5 mg / kg Q2W, 10 mg Q2W, 1 mg / kg Q3 W, 2 mg / kg Q3W, 3 mg / kg Q3 W, 5 mg / kg Q3 W, and 10 mg / kg Q3W.

[0137] In another preferred embodiment of the invention, the PD-1 antagonist in the combination therapy is pembrolizumab, or a pembrolizumab variant, that is administered in a liquid medicament at a dose selected from the group consisting of 1 mg / kg Q2W, 2 mg / kg Q2W, 3 mg / kg Q2W, 5 mg / kg Q2W, 10 mg / kg Q2W, 1 mg / kg Q3W, 2 mg / kg Q3W, 3 mg / kg Q3W, 5 mg / kg Q3W. 10 mg / kg Q3W and flat-dose equivalents of any of these doses, i.e., such as 200 mg Q3W or 400 mg Q6W. In some embodiments, pembrolizumab is provided as a liquid medicament that comprises 25 mg / ml pembrolizumab, 7% (w / v) sucrose, 0.02% (w / v) polysorbate 80 in 10 mM histidine buffer pH 5.5. In other embodiments, pembrolizumab is provided as a liquid medicament that comprises about 125 to about 200 mg / mL of pembrolizumab, or an antigen binding fragment thereof; about 10 mM histidine buffer; about 10 mM L-methionine, or a pharmaceutically acceptable salt thereof; about 7% (w / v) sucrose; and about 0.02 % (w / v) polysorbate 80.

[0138] In some embodiments, the selected dose of pembrolizumab is administered by IV infusion. In one embodiment, the selected dose of pembrolizumab is administered by IV infusion over a time period of between 25 and 40 minutes, or about 30 minutes. In other embodiments, the selected dose of pembrolizumab is administered by subcutaneous injection.

[0139] In one embodiment, the invention provides a method for treating cancer in a patient comprising administering via intravenous infusion to the individual a composition comprising 200 mg of pembrolizumab or pembrolizumab variant and 800 mg of anti-LAG3 antibody favezelimab or favezelimab variant every three weeks, and orally administering 100-200 mg / m2of all-trans retinoic acid or a pharmaceutically acceptable salt thereof daily for three days every' three weeks. In one embodiment, the invention provides a method for treating cancer in a patient comprising administering via intravenous infusion to the individual a composition comprising 200 mg of pembrolizumab or pembrolizumab variant and 800 mg of anti-LAG3 antibody favezelimab or favezelimab variant every three weeks, and orally administering 100-150 mg / m2of all-trans retinoic acid or a pharmaceutically acceptable salt thereof daily for three days every three weeks. In another embodiment, the invention provides a method for treating cancer in a patient comprising administering via intravenous infusion to the individual a composition comprising 200 mg of pembrolizumab or pembrolizumab variant and 800 mg of anti-LAG3 antibody favezelimab or favezelimab variant on every three weeks, and orally administering 100 mg / m2of all-trans retinoic acid or a pharmaceutically acceptable salt thereof daily for three days every three weeks. In another embodiment, the invention provides a method for treating cancer in a patient comprising administering via intravenous infusion to the individual a composition comprising 200 mg of pembrolizumab or pembrolizumab variant and 800 mg of anti-LAG3antibody favezelimab or favezelimab variant on every three weeks, and orally administering 150 mg / m2of all-trans retinoic acid or a pharmaceutically acceptable salt thereof daily for three days every three weeks. In one aspect of the foregoing embodiments, favezelimab, pembrolizumab and all-trans retinoic acid are administered. In another aspect of the foregoing embodiments, the favezelimab and pembrolizumab are administered on Day 1 of the 3-week cycle and the all-trans retinoic acid is administered on Days 1, 2 or 3, or Days -1, 1 or 2 of the 3-week cycle.

[0140] In one aspect of the invention, the patient is treated with the combination therapy of the PD-1 antagonist, LAG3 antagonist and all-trans retinoic acid or a pharmaceutically acceptable salt thereof before and after surgical resection of a tumor. In one embodiment, the patient is treated with the combination therapy of the PD-1 antagonist, LAG3 antagonist and all- trans retinoic acid or a pharmaceutically acceptable salt thereof for one to five treatment cycles before surgical resection of a tumor, e.g., one to five 3-week cycles. In some embodiments, the patient is treated with the combination therapy of the PD-1 antagonist, LAG3 antagonist and all- trans retinoic acid or a pharmaceutically acceptable salt thereof for one to four treatment cycles before surgical resection of a tumor, e g., one to four 3-week cycles. In some embodiments, the patient is treated with the combination therapy of the PD-1 antagonist, LAG3 antagonist and all- trans retinoic acid or a pharmaceutically acceptable salt thereof for one to three treatment cycles before surgical resection of a tumor, e.g., one to three 3-week cycles. In some embodiments, the patient is treated with the combination therapy of the PD-1 antagonist, LAG3 antagonist and all- trans retinoic acid or a pharmaceutically acceptable salt thereof for one to two treatment cycles before surgical resection of a tumor, e.g., one to two 3-week cycles. In some embodiments, the patient is treated with the combination therapy of the PD-1 antagonist, LAG3 antagonist and all- trans retinoic acid or a pharmaceutically acceptable salt thereof for 1 treatment cycle before surgical resection of a tumor, e g., one 3-week cycle. In one embodiment, the patient is treated with the combination therapy of the PD-1 antagonist, LAG3 antagonist and all-trans retinoic acid or a pharmaceutically acceptable salt thereof for five treatment cy cles before surgical resection of a tumor, e.g.. five 3-week cycles. In some embodiments, the patient is treated with the combination therapy of the PD-1 antagonist, LAG3 antagonist and all-trans retinoic acid or a pharmaceutically acceptable salt thereof for four treatment cycles before surgical resection of a tumor, e.g.. four 3-week cycles. In some embodiments, the patient is treated with the combination therapy of the PD-1 antagonist, LAG3 antagonist and all-trans retinoic acid or a pharmaceutically acceptable salt thereof for three treatment cycles before surgical resection of a tumor, e.g., three 3-week cycles. In some embodiments, the patient is treated with the combination therapy of the PD-1 antagonist, LAG3 antagonist and all-trans retinoic acid or a pharmaceutically acceptablesalt thereof for two treatment cycles before surgical resection of a tumor, e.g., two 3-week cycles. In some embodiments, the patient is treated with the combination therapy of the PD-1 antagonist, LAG3 antagonist and all-trans retinoic acid or a pharmaceutically acceptable salt thereof for 1 treatment cycle before surgical resection of a tumor, e.g., one 3-week cycle.

[0141] The surgical resection can take place any time after completion of a treatment cycle of the combination therapy of the PD-1 antagonist, LAG3 antagonist and all-trans retinoic acid or a pharmaceutically acceptable salt thereof. In one embodiment, the surgical resection is performed within 12 weeks of first administration of the PD-1 antagonist, LAG3 antagonist and all-trans retinoic acid or a pharmaceutically acceptable salt thereof. In one embodiment, the surgical resection is performed within 18 weeks of first administration of the combination therapy of the PD-1 antagonist, LAG3 antagonist and all-trans retinoic acid or a pharmaceutically acceptable salt thereof. In one embodiment, the surgical resection is performed within 14 weeks of first administration of the combination therapy of the PD-1 antagonist. LAG3 antagonist and all-trans retinoic acid or a pharmaceutically acceptable salt thereof.

[0142] Following surgical resection, administration of the PD-1 antagonist, LAG3 antagonist and all-trans retinoic acid or a pharmaceutically acceptable salt thereof can take place any time thereafter. In one embodiment, for patients with residual disease or microscopic positive margin involvement on pathology, re-resection is performed before adjuvant administration of the PD-1 antagonist, LAG3 antagonist and all-trans retinoic acid or a pharmaceutically acceptable salt thereof. In one embodiment, after surgical resection, a second surgical resection of the tumor is performed before administration of the anti-PD-1 antibody, anti-LAG3 antibody and all-trans retinoic acid or a pharmaceutically acceptable salt thereof.

[0143] After surgical resection (or re-resection) of the tumor, if margins are still positive on pathology, in one embodiment, patients receive adjuvant radiation therapy prior to receiving adjuvant administration of the PD-1 antagonist, LAG3 antagonist and all-trans retinoic acid or a pharmaceutically acceptable salt thereof. In one embodiment, after surgical resection or a second surgical resection, radiation therapy is administered before administration of the anti-PD-1 antibody, anti-LAG3 antibody and all-trans retinoic acid or a pharmaceutically acceptable salt thereof. In another embodiment, the anti-PD-1 antibody, anti-LAG3 antibody and all-trans retinoic acid or a pharmaceutically acceptable salt thereof administration starts at least 4 weeks and up to 12 weeks after surgical resection or last dose of radiation therapy.

[0144] In one embodiment, adjuvant radiation therapy is administered for one to six weeks. In one embodiment, adjuvant radiation therapy is administered for six weeks for a total of 60 to70 Gy. In one embodiment, adjuvant radiation therapy is administered for three to four weeks for a total of 45 to 50 Gy.

[0145] Typically, it is recommended to give 60 Gy in 30 fractions over 6 weeks to the site of resected disease; for patients with smaller lesions, or in cases where shorter fractionation is preferred and optimal long-term cosmesis is not a clinical priority, there is the option of hypofractionated schedules, in which radiotherapy is given as a dose of 50 Gy in 20 fractions over 4 weeks or as 45 Gy in 15 fractions over 3 weeks. In the presence of positive microscopic margins, a dose of 66 Gy in 2 Gy fractions may be recommended (20).

[0146] In further embodiments of the foregoing embodiments, the patient is treated with the combination therapy of the PD-1 antagonist, LAG3 antagonist and all-trans retinoic acid or a pharmaceutically acceptable salt thereof for one to eighteen treatment cycles after surgical resection of a tumor, e.g., one to eighteen 3-week cycles. In some embodiments, the patient is treated with the combination therapy of the PD-1 antagonist, LAG3 antagonist and all-trans retinoic acid or a pharmaceutically acceptable salt thereof for one to fourteen treatment cycles after surgical resection of a tumor, e g., one to fourteen 3-week cycles. In some embodiments, the patient is treated with the combination therapy of the PD-1 antagonist, LAG3 antagonist and all- trans retinoic acid or a pharmaceutically acceptable salt thereof for fourteen treatment cycles after surgical resection of a tumor, e.g., fourteen 3-week cycles. In a further embodiment, the combination therapy of the PD-1 antagonist, LAG3 antagonist and all-trans retinoic acid or a pharmaceutically acceptable salt thereof is administered at a total of seventeen treatment (3- week) cycles before and after surgical resection of the tumor. In a further embodiment, the combination therapy of the PD-1 antagonist, LAG3 antagonist and all-trans retinoic acid or a pharmaceutically acceptable salt thereof is administered at a total of eighteen treatment (3-week) cycles before and after surgical resection of the tumor.

[0147] In further embodiments of the foregoing embodiments, the patient is treated with the combination therapy of the PD-1 antagonist, LAG3 antagonist and all-trans retinoic acid or a pharmaceutically acceptable salt thereof for eighteen treatment cycles after surgical resection of a tumor, e.g., eighteen 3-week cycles. In some embodiments, the patient is treated with the combination therapy of the PD-1 antagonist, LAG3 antagonist and all-trans retinoic acid or a pharmaceutically acceptable salt thereof for seventeen treatment cycles after surgical resection of a tumor, e.g.. seventeen 3-week cycles. In some embodiments, the patient is treated with the combination therapy of the PD-1 antagonist, LAG3 antagonist and all-trans retinoic acid or a pharmaceutically acceptable salt thereof for sixteen treatment cycles after surgical resection of a tumor, e.g., sixteen 3-week cycles. In some embodiments, the patient is treated with thecombination therapy of the PD-1 antagonist, LAG3 antagonist and all-trans retinoic acid or a pharmaceutically acceptable salt thereof for fifteen treatment cycles after surgical resection of a tumor, e.g., fifteen 3-week cycles.

[0148] Pharmaceutically acceptable excipients of the present disclosure include for instance, solvents, bulking agents, buffering agents, tonicity adjusting agents, and preservatives (see, e.g.. Pramanick et al., Pharma Times, 45:65-77, 2013). In some embodiments the pharmaceutical compositions may comprise an excipient that functions as one or more of a solvent, a bulking agent, a buffering agent, and a tonicity adjusting agent (e.g., sodium chloride in saline may serve as both an aqueous vehicle and a tonicity adjusting agent).

[0149] In some embodiments, the pharmaceutical compositions comprise an aqueous vehicle as a solvent. Suitable vehicles include for instance sterile water, saline solution, phosphate buffered saline, and Ringer's solution. In some embodiments, the composition is isotonic.

[0150] The pharmaceutical compositions may comprise a bulking agent. Bulking agents are particularly useful when the pharmaceutical composition is to be lyophilized before administration. In some embodiments, the bulking agent is a protectant that aids in the stabilization and prevention of degradation of the active agents during freeze or spray drying and / or during storage. Suitable bulking agents are sugars (mono-, di- and polysaccharides) such as sucrose, lactose, trehalose, mannitol, sorbital, glucose and raffinose.

[0151] The pharmaceutical compositions may comprise a buffering agent. Buffering agents control pH to inhibit degradation of the active agent during processing, storage and optionally reconstitution. Suitable buffers include for instance salts comprising acetate, citrate, phosphate or sulfate. Other suitable buffers include for instance amino acids such as arginine, glycine, histidine, and lysine. The buffering agent may further comprise hydrochloric acid or sodium hydroxide. In some embodiments, the buffering agent maintains the pH of the composition within a range of 4 to 9. In some embodiments, the pH is greater than (lower limit) 4, 5, 6, 7 or 8. In some embodiments, the pH is less than (upper limit) 9, 8, 7, 6 or 5. That is, the pH is in the range of from about 4 to 9 in which the lower limit is less than the upper limit.

[0152] The pharmaceutical compositions may comprise a tonicity adjusting agent. Suitable tonicity adjusting agents include for instance dextrose, glycerol, sodium chloride, glycerin and mannitol.

[0153] The pharmaceutical compositions may comprise a preservative. Suitable preservatives include for instance antioxidants and antimicrobial agents. However, in preferred embodiments, the pharmaceutical composition is prepared under sterile conditions and is in a single use container, and thus does not necessitate inclusion of a preservative.

[0154] In some embodiments, a medicament comprising an anti-PD-1 antibody as the PD-1 antagonist may be provided as a liquid formulation or prepared by reconstituting a lyophilized powder with sterile water for injection prior to use. PCT International application publ. no. WO 2012 / 135408 describes the preparation of liquid and lyophilized medicaments comprising pembrolizumab that are suitable for use in the present invention. In some embodiments, a medicament comprising pembrolizumab is provided in a glass vial that contains about 100 mg of pembrolizumab in 4 ml of solution. Each 1 mL of solution contains 25 mg of pembrolizumab and is formulated in: L-histidine (1.55 mg), polysorbate 80 (0.2 mg), sucrose (70 mg), and Water for Injection, USP. The solution requires dilution for IV infusion.

[0155] In some embodiments, a medicament comprising an anti-LAG3 antibody as the LAG3 antagonist may be provided as a liquid formulation or prepared by reconstituting a lyophilized powder with sterile water for injection prior to use. In one embodiment, the liquid formulation comprises about 25 mg / mL anti-LAG3 antibody; about 50 mg / mL sucrose; about 0.2 mg / mL polysorbate 80; about 10 mM L-histidine buffer at about pH 5.8-6.0; about 70 mM L- Arginine-HCl thereof; and optionally about 10 mM L-methionine.

[0156] In other aspects, the medicament is a co-formulation of an anti-LAG3 antibody or antigen binding fragment and an anti-PD-1 antibody or antigen binding fragment with 20 mg / mL of favezelimab or favezelimab variant, 5 mg / mL or pembrolizumab or pembrolizumab variant, 56 mM L- Arginine HC1, 5.4% sucrose, 8.0 mM methionine, 0.02% PS-80, and 10 mM Histidine buffer.

[0157] The medicaments described herein may be provided as a kit that comprises a first container, a second container and a package insert or label. The medicaments described herein may also be provided as a kit which compnses a first container, a second container, and a package insert or label. The first container contains at least one dose of a medicament comprising a PD-1 antagonist and at least one dose of a medicament comprising a LAG3 antagonist and a second container that contains an all-trans retinoic acid or a pharmaceutically acceptable salt thereof, and the package insert or label, that comprises instructions for treating a patient for cancer using the medicaments. The first and second containers may be comprised of the same or different shapes (e.g., vials, syringes and bottles) and / or material (e.g., plastic or glass). The kit may further comprise other materials that may be useful in administering the medicaments, such as diluents, filters, IV bags and lines, needles and syringes. In some preferred embodiments of the kit, the PD-1 antagonist is an anti-PD-1 antibody and the instructions state that the medicaments are intended for use in treating a patient having cancer that tests positive for PD-L1 expression by an IHC assay.GENERAL METHODS

[0158] Standard methods in molecular biology are described Sambrook, Fritsch and Maniatis (1982 & 1989 2ndEdition, 2001 3rdEdition) Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; Sambrook and Russell (2001) Molecular Cloning, 3rded., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; Wu (1993) Recombinant DNA, Vol. 217, Academic Press, San Diego, CA). Standard methods also appear in Ausbel, et al. (2001) Current Protocols in Molecular Biology, Vols.1-4, John Wiley and Sons, Inc. New York, NY. which describes cloning in bacterial cells and DNA mutagenesis (Vol. 1), cloning in mammalian cells and yeast (Vol. 2), glycoconjugates and protein expression (Vol. 3), and bioinformatics (Vol. 4).

[0159] Methods for protein purification including immunoprecipitation, chromatography, electrophoresis, centrifugation, and cry stallization are described (Coligan, et al. (2000) Current Protocols in Protein Science, Vol. 1. John Wiley and Sons, Inc., New York). Chemical analysis, chemical modification, post-translational modification, production of fusion proteins, glycosylation of proteins are described (see, e.g., Coligan, et al. (2000) Current Protocols in Protein Science, Vol. 2, John Wiley and Sons, Inc., New York; Ausubel, et al. (2001) Current Protocols in Molecular Biology, Vol. 3, John Wiley and Sons, Inc., NY, NY, pp. 16.0.5-16.22.17; Sigma-Aldrich, Co. (2001) Products for Life Science Research. St. Louis, MO; pp. 45-89;Amersham Pharmacia Biotech (2001) BioDirectory, Piscataway, N.J., pp. 384-391). Production, purification, and fragmentation of polyclonal and monoclonal antibodies are described (Coligan, et al. (2001) Current Protcols in Immunology, Vol. 1, John Wiley and Sons, Inc., New York;Harlow and Lane (1999) Using Antibodies , Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; Harlow and Lane, supra). Standard techniques for characterizing hgand / receptor interactions are available (see, e.g., Coligan, et al. (2001) Current Protocols in Immunology, Vol. 4, John Wiley, Inc., New York).

[0160] Monoclonal, polyclonal, and humanized antibodies can be prepared (see, e.g., Sheperd and Dean (eds.) (2000) Monoclonal Antibodies, Oxford Univ. Press. New York. NY; Kontermann and Dubel (eds.) (2001) Antibody Engineering, Springer-Verlag, New York; Harlow and Lane (1988) Antibodies A Laboratory Manual, Cold Spring Harbor Laboratory' Press, Cold Spring Harbor, NY, pp. 139-243; Carpenter, et al. (2000) J. Immunol. 165:6205; He, et al. (1998) J. Immunol. 160: 1029; Tang et al. (1999) J. Biol. Chem. 274:27371-27378: Baca et al. (1997) J. Biol. Chem. 272: 10678-10684; Chothia et al. (1989) Nature 342:877-883; Foote and Winter (1992) J. Mol. Biol. 224:487-499; U.S. Pat. No. 6,329,511).

[0161] An alternative to humanization is to use human antibody libraries displayed on phage or human antibody libraries in transgenic mice (Vaughan et al. (1996) Nature Biotechnol.14:309-314; Barbas (1995) Nature Me dicine 1 :837-839; Mendez et al. (1997) Nature Genetics 15: 146-156; Hoogenboom and Chames (2000) Immunol. Today 21 :371-377; Barbas et al. (2001) Phage Display: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York; Kay et al. (1996) Phage Display of Peptides and Proteins: A Laboratory Manual, Academic Press, San Diego, CA; de Bruin et al. (1999) Nature Biotechnol. 17:397-399).

[0162] Purification of antigen is not necessary for the generation of antibodies. Animals can be immunized with cells bearing the antigen of interest. Splenocytes can then be isolated from the immunized animals, and the splenocytes can fuse with a myeloma cell line to produce a hybridoma (see, e.g., Meyaard et al. (1997) Immunity 7:283-290; Wright et al. (2000) Immunity 13:233-242; Preston et al., supra,' Kaithamana et al. (1999) J. Immunol. 163:5157-5164).

[0163] Antibodies can be conjugated, e.g., to small drug molecules, enzymes, liposomes, polyethylene glycol (PEG). Antibodies are useful for therapeutic, diagnostic, kit or other purposes, and include antibodies coupled, e.g., to dyes, radioisotopes, enzymes, or metals, e.g., colloidal gold (see, e.g., Le Doussal et al. (1991) J. Immunol. 146: 169-175; Gibellini et al. (1998) J. Immunol. 160:3891-3898; Hsing and Bishop (1999) J. Immunol. 162:2804-2811; Everts et al. (2002) J. Immunol. 168:883-889).

[0164] Methods for flow cytometry, including fluorescence activated cell sorting (FACS), are available (see, e.g., Owens, et al. (1994) Flow Cytometry Principles for Clinical Laboratory Practice, John Wiley and Sons, Hoboken, NJ; Givan (2001) Flow Cytometry, 2nded:, Wiley-Liss, Hoboken, NJ; Shapiro (2003) Practical Flow Cytometry, John Wiley and Sons, Hoboken, NJ). Fluorescent reagents suitable for modifying nucleic acids, including nucleic acid primers and probes, polypeptides, and antibodies, for use, e.g., as diagnostic reagents, are available (Molecular Probesy (2003) Catalogue, Molecular Probes, Inc., Eugene, OR; Sigma- Aldrich (2003) Catalogue, St. Louis, MO).

[0165] Standard methods of histology of the immune system are described (see, e.g., Muller- Harmelink (ed.) (1986) Human Thymus: Histopathology and Pathology, Springer Verlag, New York, NY; Hiatt, et al. (2000) Color Atlas of Histology, Lippincott, Williams, and Wilkins, Phila, PA; Louis, et al. (2002) Basic Histology: Text and Atlas, McGraw-Hill, New York, NY).

[0166] Software packages and databases for determining, e.g., antigenic fragments, leader sequences, protein folding, functional domains, glycosylation sites, and sequence alignments, are available (see, e.g., GenBank, Vector NTI® Suite (Informax, Inc, Bethesda, MD); GCG Wisconsin Package (Accelrys, Inc., San Diego, CA); DeCypher® (TimeLogic Corp., CrystalBay, Nevada); Menne. et al. (2000) Bioinformatics 16: 741-742; Menne, et al. (2000) Bioinformatics Applications Note 16:741-742; Wren, et al. (2002) Comput. Methods Programs Biomed. 68: 177-181; von Heijne (1983) Eur. J. Biochem. 133: 17-21; von Heijne (1986) Nucleic Acids Res. 14:4683-4690).EXAMPLES

[0167] Example 1: A Phase 1 / 2 Open-Label Rolling-Arm Clinical Study of co-formulated Favezelimab and Pembrolizumab in Participants With First Line (IL) Melanoma

[0168] Advanced melanoma patients (Stage III or IV with unresectable melanoma) are to receive MK-4280A (favezelimab 800 mg co-formulated with pembrolizumab 200 mg) every 3 weeks (Q3W) for up to 35 cycles, and ATRA 150 mg / m2 / day orally (for 3 days surrounding each infusion of MK-4280A, including Days 1, 2, and 3 of Cycle 1 and on Days -1, 1, and 2 of all subsequent cycles). ATRA is administered at 75 mg / m2twice daily in the morning and evening. MK-4280A is administered on Day I of each cycle Q3W. On those days where both ATRA and MK-4280A are administered, MK-4280A may be infused approximately 30 minutes after the ATRA morning dose. If ATRA 150 mg / m2 / day is not tolerated (Grade 3 or 4 toxicity), the dose will be de-escalated to 100 mg / m2 / day.

[0169] A participant will be eligible for inclusion in the study if the participant has histologically or cytologically confirmed melanoma; has unresectable Stage III or Stage IV melanoma, not amenable to local therapy; has the presence of at least 1 measurable lesion by CT or MRI per RECIST 1.1 as confirmed by BICR; has been untreated for advanced disease except as follows: BRAF V600 mutation-positive melanoma may have received SOC targeted therapy as IL therapy for advanced disease (e.g., BRAF / MEK inhibitor, alone or in combination).Prior IL therapy for advanced disease with targeted therapy is permitted if it was completed at least 4 weeks before randomization, disease progression has been documented radiologically and all related AEs have either returned to baseline or stabilized (resolution of toxic effect(s) to Grade 1 or less [except alopecia and Grade 2 neuropathy]). Prior adjuvant or neoadjuvant therapy, with targeted therapy or immunotherapy (such as anti-PD-1 therapy, anti-CTLA-4 or Interferon) is permitted. Prior anti-PD-1 therapy will only be permitted if relapse did not occur during treatment or within months of treatment discontinuation.

[0170] This study will use objective response as the primary endpoint. ORR is defined as the proportion of participants who have best response as CR or PR. Responses are based on BICR using RECIST 1.1 (modified to follow a maximum of 10 target lesions and a maximum of 5 target lesions per organ). DOR is used as a secondary endpoint. Duration of response is definedas the time from the earliest date of qualifying response until the earliest date of disease progression or death from any cause, whichever comes first. Duration of response per RECIST 1.1, modified to follow a maximum of 10 target lesions and a maximum of 5 target lesions per organ, assessed by BICR will serve as an additional measure of efficacy. PFS is used as an exploratory endpoint. PFS is defined as the time from date of randomization until the first date of disease progression or death from any cause, whichever comes first, based on RECIST 1.1 criteria as assessed by BICR. Images will be read by a BICR to minimize bias in the response assessments. PFS can reflect tumor grow th and be assessed before the determination of a survival benefit. Additionally, this study will use OS as an exploratory endpoint. OS is defined as the time from date of randomization to date of death due to any cause. Combining ATRA with MK- 4280A is expected to decrease the frequency of myeloid derived suppressor cells (MDSCs) and improve the efficacy of MK-4280A in the treatment of melanoma.REFERENCES1. Sharpe, A.H, Wherry, E.J., Ahmed R., and Freeman G.J. The function of programmed cell death 1 and its ligands in regulating autoimmunity and infection. Nature Immunology (2Q0iy, 8:239-245.2. Dong H et al. Tumor-associated B7-H1 promotes T-cell apoptosis: a potential mechanism of immune evasion. Nat Med. 2002 Aug;8(8):793-800.3. Yang et al. PD-1 interaction contributes to the functional suppression of T-cell responses to human uveal melanoma cells in vitro. Invest Ophthalmol Vis Sci. 2008 Jun;49(6 (2008): 49: 2518-2525.4. Ghebeh et ai. The B7-H1 (PD-L1) T lymphocyte-inhibitory molecule is expressed in breast cancer patients with infiltrating ductal carcinoma: correlation with important high-risk prognostic factors. Neoplasia (2006) 8: 190-198.5. Hamanishi J et al. Programmed cell death 1 ligand 1 and tumor-infiltrating CD8+ T lymphocytes are prognostic factors of human ovarian cancer. Proceeding of the National Academy of Sciences (IQIfiy 104: 3360-3365.6. Thompson RH et al. Significance of B7-H1 overexpression in kidney cancer. Clinical genitourin Cancer (2006): 5: 206-211.7. Nomi, T. Sho, M._ Akahori, T., et al. Clinical significance and therapeutic potential of the programmed death- 1 ligand / programmed death- 1 pathway in human pancreatic cancer. Clinical Cancer Research (2007);13:2151-2157.8. Ohigashi Y et al. Clinical significance of programmed death- 1 hgand-1 and programmed death-1 ligand 2 expression in human esophageal cancer. Clin. Cancer Research (2005): 1 1: 2947-2953.9. Inman et al. PD-L1 (B7-H1) expression by urothelial carcinoma of the bladder and BCG- induced granulomata: associations with localized stage progression. Cancer (2007): 109: MOO- ISOS.10. Shimauchi T et al. Augmented expression of programmed death- 1 in both neoplasmatic and nonneoplastic CD4+ T-cells in adult T-cell Leukemia / Lymphoma. Int. J. Cancer (2007): 121:2585-2590.11. Gao et a\. Overexpression of PD-L1 significantly associates with tumor aggressiveness and postoperative recurrence in human hepatocellular carcinoma. Clinical Cancer Research (2009) 15: 971-979.12. Nakanishi J. Overexpression of B7-H1 (PD-L1) significantly associates with tumor grade and postoperative prognosis in human urothelial cancers. Cancer Immunol Immunother. (2007) 56: 1 173- 1182.13. Hino et al. Tumor cell expression of programmed cell death-1 is a prognostic factor for malignant melanoma. Cancer (2010): 00: 1-9.14. Ghebeh H. Foxp3+ tregs and B7-H1+ / PD-1+ T lymphocytes co-infiltrate the tumor tissues of high-risk breast cancer patients: implication for immunotherapy. BMC Cancer. 2008 Feb 23;8:57.15. Ahmadzadeh M et al. Tumor antigen-specific CD8 T cells infiltrating the tumor express high levels of PD-1 and are functionally impaired. Blood (2009) 114: 1537-1544.16. Thompson RH et al. PD-1 is expressed by tumor infiltrating cells and is associated with poor outcome for patients with renal carcinoma. Clinical Cancer Research (2007) 15: 1757-1761.Nefedova Y, Fishman M, Sherman S, Wang X, Beg AA, Gabrilovich DI. Mechanism of all-trans retinoic acid effect on tumor-associated myeloid-derived suppressor cells. Cancer Res. 2007 Nov 15:67(22): 11021-8.

[0171] All references cited herein are incorporated by reference to the same extent as if each individual publication, database entry (e.g., Genbank sequences or GenelD entries), patent application, or patent, was specifically and individually indicated to be incorporated by reference each and every individual publication, database entry (e.g., Genbank sequences or GenelD entries), patent application, or patent, each of which is clearly identified in compliance with 37 C.F.R. §1.57(b)(2), even if such citation is not immediately adjacent to a dedicated statement of incorporation by reference. The inclusion of dedicated statements of incorporation by reference,if any, within the specification does not in any way weaken this general statement of incorporation by reference. Citation of the references herein is not intended as an admission that the reference is pertinent prior art, nor does it constitute any admission as to the contents or date of these publications or documents. To the extent that the references provide a definition for a claimed term that conflicts with the definitions provided in the instant specification, the definitions provided in the instant specification shall be used to interpret the claimed invention.

Claims

WHAT IS CLAIMED IS:

1. A method for treating cancer in an individual comprising administering to an individual a PD-1 antagonist, a LAG3 antagonist, and all-trans retinoic acid or a pharmaceutically acceptable salt thereof.

2. The method of claim 1, wherein the human PD-1 antagonist and human LAG3 antagonist and all-trans retinoic acid or a pharmaceutically acceptable salt thereof are administered before and / or after surgical resection of a tumor.

3. The method of claim 1 or 2, wherein the PD-1 antagonist is a monoclonal antibody, or an antigen binding fragment thereof, that specifically binds to human PD- 1 and blocks the binding of human PD-L1 to human PD-1.

4. The method of claim 3, wherein the PD-1 antagonist also blocks binding of human PD-L2 to human PD-1.

5. The method of claim 4, wherein the PD-1 antagonist is an antibody, or antigen binding fragment thereof, that comprises: (a) a light chain variable region comprising light chain CDR1, CDR2 and CDR3 amino acid sequences of SEQ ID NOs: 1, 2 and 3, respectively and (b) a heavy chain variable region comprising heavy chain CDR1, CDR2 and CDR3 amino acid sequences of SEQ ID NOs: 6, 7 and 8, respectively.

6. The method of claim 4, wherein the PD-1 antagonist is an anti-PD-1 antibody that comprises a heavy chain and a light chain, and wherein the heavy chain comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:9 and the light chain comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 4.

7. The method of claim 4, wherein the PD-1 antagonist is an anti-PD-1 antibody that comprises two heavy chains and two light chains, and wherein the heavy chain comprises the amino acid sequence of SEQ ID NOTO and the light chain comprises the amino acid sequence of SEQ ID NO:5.

8. The method of claim 4, wherein the PD-1 antagonist is pembrolizumab.

9. The method of claim 4, wherein the PD-1 antagonist is a pembrolizumab variant.

10. The method of any one of claims 1 to 4, wherein the PD-1 antagonist is nivolumab and the LAG3 antagonist is relatlimab.

11. The method of any one of claims 1 to 10, wherein the LAG3 antagonist is a monoclonal antibody, or an antigen binding fragment thereof, that blocks binding of LAG3 to MHC Class II molecules.

12. The method of any one of claims 1 to 9 and 11, wherein the LAG3 antagonist is an antibody, or antigen binding fragment thereof, that comprises: (a) a light chain variable region comprising light chain CDR1, CDR2 and CDR3 amino acid sequences of SEQ ID NOs: 26, 27 and 28. respectively and (b) a heavy chain variable region comprising heavy chain CDR1, CDR2 and CDR3 amino acid sequences of SEQ ID NOs: 29, 30 and 31, respectively.

13. The method of any one of claims 1 to 9 and 11, wherein the LAG3 antagonist is an anti- LAG3 monoclonal antibody that comprises a heavy chain and a light chain, and wherein the heavy chain comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:25 and the light chain comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 24.

14. The method of any one of claims 1 to 9 and 11, wherein the LAG3 antagonist is an anti- LAG3 antibody that comprises two heavy chains and two light chains, and wherein the heavy chain comprises the amino acid sequence of SEQ ID NO:23 and the light chain comprises the amino acid sequence of SEQ ID NO:22.

15. The method of any one of claims 1 to 9 and 11, wherein the LAG3 antagonist is favezelimab.

16. The method of any one of claims 1 to 9 and 11, wherein the LAG3 antagonist is favezelimab variant.

17. The method of any one of claims 1 to 2. wherein the PD-1 antagonist is cemiplimab and the LAG3 antagonist is fianlimab.

18. The method of any one of claims 1 to 3, wherein the PD-1 antagonist is a humanized anti- PD-1 antibody that comprises a heavy chain and a light chain, and wherein the heavy chain comprises a heavy chain variable region comprising heavy chain CDR1, CDR2 and CDR3 amino acid sequences of SEQ ID NOs: 6, 7 and 8, respectively and the light chain comprises a light chain variable region comprising light chain CDR1, CDR2 and CDR3 amino acid sequences of SEQ ID NOs: 1, 2 and 3, respectively; and the LAG3 antagonist is a humanized anti-LAG3 antibody that comprises a heavy chain and a light chain, and wherein the heavy chain comprises a heavy chain variable region comprising heavy chain CDR1, CDR2 and CDR3 amino acid sequences of SEQ ID NOs: 29, 30 and 31, respectively and the light chain comprises a light chain variable region comprising light chain CDR1. CDR2 and CDR3 amino acid sequences of SEQ ID NOs: 26, 27 and 28. respectively.

19. The method of any one of claims 1 to 18, wherein the PD-1 antagonist and LAG3 antagonist are co-formulated.

20. The method of any one of claims 1 to 19, comprising administering to the human individual an anti-PD-1 antibody, an anti-LAG3 antibody and all-trans retinoic acid for 1 treatment cycle, 1-2 treatment cycles, 1-3 treatment cycles, or 1-4 treatment cycles before surgical resection.

21. The method of any one of claims 1 to 20, comprising administering via intravenous infusion to the human individual a composition comprising 200 mg of the anti-PD-1 antibody and 800 mg of the anti-LAG3 antibody every 3 weeks for a treatment cycle, and orally administering 100-150 mg / m2all-trans retinoic acid for 3 days in the treatment cycle.

22. The method of any one of claims 1 to 20, comprising administering via intravenous infusion to the human individual a composition comprising 200 mg of the anti-PD-1 antibody and 800 mg of the anti-LAG3 antibody on Day 1 every 3 weeks of a treatment cycle, and orally administering 100 or 150 mg / m2all-trans retinoic acid on Days 1, 2, and 3 of the treatment cycle.

23. The method of claim 22. wherein all-trans retinoic acid is administered on Days 1, 2. and3 of the treatment cycle 1 and on Days -1, 1, and 2 of all subsequent treatment cycles.

24. The method of any one of claims 22 to 23. wherein the all-trans retinoic acid is orally administered twice daily at 50 or 75 mg / m225. The method of claim 23, on the days when the anti-PD-1 antibody and anti-LAG3 antibody and all-trans retinoic acid are administered, the first dose of all-trans retinoic acid is administered before the anti-PD-1 antibody and anti-LAG3 antibody.

26. The method of any one of claims 1 to 25, comprising administering to the individual the anti-PD-1 antibody , the anti-LAG3 antibody and all-trans retinoic acid 1 to 18, 1 to 17, 1 to 16, 1 to 15, or 1 to 14 treatment cycles after surgical resection.

27. The method of any one of claims 1 to 26, wherein the cancer is melanoma, non-small cell lung cancer, small cell lung cancer, head and neck squamous cell cancer, classical Hodgkin lymphoma, primary mediastinal large B-cell lymphoma, urothelial carcinoma, microsatellite instability -high or mismatch repair deficient cancer, gastric cancer, esophageal cancer, cervical cancer, hepatocellular carcinoma, Merkel cell carcinoma, renal cell carcinoma, endometrial carcinoma, a cancer characterized by a tumor having a high mutational burden, cutaneous squamous cell carcinoma, and triple negative breast cancer.

28. The method of any one of claims 1 to 26, wherein the cancer is melanoma.

29. The method of any one of claims 1 to 28, wherein the cancer is stage III or IV.

30. The method of any one of claims 1, 3 to 28, wherein the cancer is stage III or IV unresectable melanoma.

31. The method of any one of claims 1 to 28, wherein the cancer is stage III or IV resectable melanoma.

32. The method of any one of claims 1 to 28, wherein the cancer is stage I, II, III or IV melanoma without distant metastasis.

33. The combination of a PD-1 antagonist, a LAG-3 antagonist and all-trans retinoic acid or a pharmaceutically acceptable salt thereof, for use in any one of the methods of claims 1 to 32.

34. The combination of a PD-1 antagonist, a LAG-3 antagonist and an all-trans retinoic acid or a pharmaceutically acceptable salt thereof, for use in the manufacture of a medicament in any one of the methods of claims 1 to 32.

35. Use of the combination of a PD-1 antagonist, a LAG-3 antagonist and an all-trans retinoic acid or a pharmaceutically acceptable salt thereof in any one of the methods of claims 1 to 32.

36. A pharmaceutical composition comprising a PD-1 antagonist for use in combination with a LAG-3 antagonist and an all-trans retinoic acid or a pharmaceutically acceptable salt thereof in any one of the methods of claims 1 to 32.

37. A pharmaceutical composition comprising a LAG-3 antagonist for use in combination with a PD-1 antagonist and an all-trans retinoic acid or a pharmaceutically acceptable salt thereof in any one of the methods of claims 1 to 32.

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