Method of treating cancer using immune checkpoint inhibitor

Abstract

To provide a method for treating human papilloma virus (HPV) positive squamous cell carcinoma of the head and neck.SOLUTION: A method for treating a subject afflicted with a tumor derived from a human papilloma virus (HPV) positive squamous cell carcinoma head and neck cancer (SCCHN), the method comprising administering to the subject a therapeutically effective amount of an antibody or an antigen-binding portion thereof that binds specifically to a Programmed Death-1 receptor (PD-1) or Programmed Death-Ligand 1 (PD-L1) and inhibits PD-1 activity ("anti-PD-1 antibody") or PD-L1 activity ("anti-PD-L1 antibody"), respectively.SELECTED DRAWING: None

Description

[Technical Field]

[0001] Throughout this specification, various publications are cited by author and date in parentheses or by patent or patent publication number. Full citations for these publications can be found at the end of the specification immediately preceding the claims. The disclosures of these publications are incorporated herein by reference in their entireties in order to more fully describe the state of the art known to those skilled in the art at the time the invention is described and claimed herein. However, the citation of any reference herein should not be construed as an admission that such reference is antedate the present invention. [Background technology]

[0002] Background of the Invention Human cancers harbor numerous genetic and epigenetic alterations, producing neoantigens that can potentially be recognized by the immune system (Sjoblom et al. (2006) Science 314:268-74). The adaptive immune system, including T and B lymphocytes, possesses potent anticancer potential with broad capacity and exquisite specificity to respond to diverse tumor antigens. Furthermore, immunity exhibits considerable plasticity and a memory component. The successful utilization of all of these attributes of the adaptive immune system makes immunotherapy unique among all cancer treatment modalities.

[0003] Until recently, cancer immunotherapy has focused considerable effort on attempts to enhance antitumor immune responses by adoptive transfer of activated effector cells, immunization against relevant antigens, or provision of nonspecific immune stimulators such as cytokines. However, in the past decade, intensive efforts to develop specific immune checkpoint pathway inhibitors have led to the development of ipilimumab (Yervoy®), an antibody (Ab) that binds to and inhibits CTLA-4 for the treatment of patients with advanced melanoma (Hodi et al. (2010) N Engl J Med 363:711-23), and the development of antibodies such as nivolumab and pembrolizumab (formerly lambrolizumab; USAN Council Statement (2013) Pembrolizumab: Statement on a nonproprietary name adopted by the USAN Council (ZZ-165), November 27, 2013), which specifically bind to the programmed death-1 (PD-1) receptor and block the inhibitory PD-1 / PD-1 ligand pathway (Topalian et al. (2012a) N Engl J Med 366:2443-54; Topalian et al. (2012b) Curr Opin Immunol 24:207-12; Topalian et al. (2014) J Clin Oncol 32(10):1020-30; Hamid et al. (2013) N Engl J Med 369:134-144; Hamid and Carvajal (2013) Expert Opin Biol Ther 13(6):847-61; McDermott and Atkins (2013) Cancer Med 2(5):662-73).

[0004] PD-1 is an important immune checkpoint receptor expressed by activated T and B cells and mediates immunosuppression. Nivolumab (previously named 5C4, BMS-936558, MDX-1106, or ONO-4538) is a fully human IgG4(S228P) PD-1 immune checkpoint inhibitor antibody that selectively blocks interaction with PD-1 ligands (PD-L1 and PD-L2), thereby blocking downregulation of antitumor T cell function (U.S. Patent No. 8,008,449; Wang et al. (2014) In vitro characterization of the anti-PD-1 antibody nivolumab, BMS-936558, and in vivo toxicology in non-human primates, Cancer Imm Res, in press). Nivolumab is approved for the treatment of unresectable or metastatic melanoma and patients with disease progression on ipilimumab and, if BRAF V600 mutation-positive, BRAF inhibitors, and for the treatment of squamous non-small cell lung cancer. Summary of the Invention

[0005] Summary of the Invention The present invention provides methods for treating a subject having a tumor derived from human papillomavirus (HPV)-positive squamous cell carcinoma of the head and neck (SCCHN), comprising administering to the subject a therapeutically effective amount of an antibody or antigen-binding portion thereof that specifically binds to programmed death-1 receptor (PD-1) or programmed death-ligand 1 (PD-L1) and inhibits PD-1 activity (an "anti-PD-1 antibody") or PD-L1 activity (an "anti-PD-L1 antibody"), respectively.

[0006] The present invention also provides a method of treating a subject with an HPV-positive SCCHN-derived tumor, comprising: (i) measuring the level of HPV in a sample from the subject, wherein the subject is positive for HPV; and (ii) administering to the subject a therapeutically effective amount of an anti-PD-1 antibody, or antigen-binding portion thereof, or an anti-PD-L1 antibody, or antigen-binding portion thereof.

[0007] The present invention also provides a method for identifying a subject with an HPV-positive SCCHN-derived tumor suitable for anti-PD-1 antibody or anti-PD-L1 antibody treatment, comprising measuring the level of HPV in a sample from the subject, wherein the subject is positive for HPV, and administering a therapeutically effective amount of an anti-PD-1 antibody, or antigen-binding portion thereof, or an anti-PD-L1 antibody, or antigen-binding portion thereof.

[0008] The present invention also provides a method for identifying a subject with an HPV-positive SCCHN-derived tumor suitable for anti-PD-1 antibody or anti-PD-L1 antibody treatment, comprising: (i) measuring the level of HPV in a sample from the subject, wherein the subject is positive for HPV; and (ii) administering to the subject a therapeutically effective amount of an anti-PD-1 antibody, or antigen-binding portion thereof, or an anti-PD-L1 antibody, or antigen-binding portion thereof.

[0009] In certain embodiments, the method further comprises administering one or more additional anti-cancer agents. In certain embodiments, the anti-cancer agent is selected from the group consisting of an antibody or antigen-binding portion thereof that specifically binds to CTLA-4 and inhibits CTLA-4 activity, a chemotherapeutic agent, a platinum-based doublet chemotherapy, a tyrosine kinase inhibitor, an anti-VEGF inhibitor, or any combination thereof.

[0010] The present invention also provides methods for treating a subject having a tumor derived from human papillomavirus (HPV)-negative squamous cell carcinoma of the head and neck (SCCHN), comprising administering to the subject a therapeutically effective amount of an antibody or antigen-binding portion thereof that specifically binds to programmed death-1 receptor (PD-1) or programmed death-ligand 1 (PD-L1) and inhibits PD-1 activity (an "anti-PD-1 antibody") or PD-L1 activity (an "anti-PD-L1 antibody"), respectively.

[0011] The present invention also provides a method of treating a subject with an HPV-negative SCCHN-derived tumor, comprising: (i) measuring the level of HPV in a sample from the subject, wherein the subject is negative for HPV; and (ii) administering to the subject a therapeutically effective amount of an anti-PD-1 antibody, or antigen-binding portion thereof, or an anti-PD-L1 antibody, or antigen-binding portion thereof.

[0012] The present invention also provides a method for identifying a subject having an HPV-negative SCCHN-derived tumor suitable for anti-PD-1 antibody or anti-PD-L1 antibody treatment, comprising measuring the level of HPV in a sample from the subject, wherein the subject is negative for HPV, and administering a therapeutically effective amount of an anti-PD-1 antibody, or antigen-binding portion thereof, or an anti-PD-L1 antibody, or antigen-binding portion thereof.

[0013] The present invention also provides a method for identifying a subject having an HPV-negative SCCHN-derived tumor suitable for anti-PD-1 antibody or anti-PD-L1 antibody treatment, comprising: (i) measuring the level of HPV in a sample from the subject, wherein the subject is negative for HPV; and (ii) administering to the subject a therapeutically effective amount of an anti-PD-1 antibody, or antigen-binding portion thereof, or an anti-PD-L1 antibody, or antigen-binding portion thereof.

[0014] The present invention also provides kits for treating a subject having an HPV-positive SCCHN-derived tumor, the kit comprising: (a) an anti-PD-1 antibody, or antigen-binding portion thereof, or an anti-PD-L1 antibody, or antigen-binding portion thereof; and (b) instructions for determining HPV positivity of the tumor; and if the tumor is positive for HPV, administering to the subject the anti-PD-1 antibody, or antigen-binding portion thereof, or anti-PD-L1 antibody, or antigen-binding portion thereof, in a manner described herein.

[0015] The invention also provides kits for treating a subject having an HPV-negative SCCHN-derived tumor, the kit comprising: (a) an anti-PD-1 antibody, or antigen-binding portion thereof, or an anti-PD-L1 antibody, or antigen-binding portion thereof; and (b) instructions for determining HPV negativity of the tumor; and if the tumor is negative for HPV, administering to the subject an anti-PD-1 antibody, or antigen-binding portion thereof, or an anti-PD-L1 antibody, or antigen-binding portion thereof, in a manner described herein. [Brief explanation of the drawings]

[0016] [Figure 1]

[0013] Figure 1 provides a schematic diagram of a non-comparative, two-cohort, single-arm, open-label, Phase 1 / 2 clinical trial of nivolumab in subjects with selected virus-positive and virus-negative tumors, conducted to evaluate the safety, tolerability, and efficacy of nivolumab in subjects with virus-positive and virus-negative solid tumors. The trial is designed to have two cohorts: a neoadjuvant cohort ("Neoadjuvant") and a metastatic / recurrent cohort ("Metastatic"). DETAILED DESCRIPTION OF THE INVENTION

[0017] Detailed Description of the Invention term So that the present invention may be more readily understood, certain terms are first defined. As used herein, unless expressly indicated to the contrary herein, each of the following terms will have the meaning indicated below. Additional definitions are set forth throughout the specification.

[0018] "Administering" refers to the physical introduction of a composition containing a therapeutic agent using any of a variety of methods and delivery systems known to those skilled in the art. Routes of administration of immune checkpoint inhibitors, e.g., anti-PD-1 antibodies or anti-PD-L1 antibodies, include intravenous, intramuscular, subcutaneous, intraperitoneal, spinal, or other parenteral routes, e.g., by injection or infusion. As used herein, the term "parenteral administration" refers to modes of administration other than enteral and topical administration, typically by injection, and includes, but is not limited to, intravenous, intramuscular, intra-arterial, intrathecal, intralymphatic, intralesional, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intra-articular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal injection and infusion, and in vivo electroporation. In certain embodiments, the immune checkpoint inhibitor, e.g., anti-PD-1 antibodies or anti-PD-L1 antibodies, is administered non-parenterally, in certain embodiments, orally. Other non-injection routes include topical, epithelial or mucosal administration routes, e.g., intranasal, vaginal, rectal, sublingual or topical. Administration can also be carried out, e.g., once, multiple times and / or over one or more chronic periods.

[0019] As used herein, an "adverse event" (AE) is any untoward and generally unintended or undesirable sign (including abnormal laboratory findings), symptom, or disease associated with the use of a drug therapy. For example, an adverse event may be associated with activation of the immune system or expansion of immune system cells (e.g., T cells) in response to treatment. A drug therapy may be associated with one or more AEs, each of which may be of the same or different levels of severity. A method that can "modify an adverse event" refers to a treatment regimen that reduces the incidence and / or severity of one or more AEs with the use of a different treatment regimen.

[0020] "Antibody" (Ab) includes, but is not limited to, a glycoprotein immunoglobulin or antigen-binding portion thereof that specifically binds to an antigen and comprises at least two heavy (H) chains and two light (L) chains interconnected by disulfide bonds. Each H chain comprises a heavy chain variable region (herein referred to as V H The heavy chain constant region comprises three constant domains: C H1 , C H2 and C H3 Each light chain contains a light chain variable region (herein V L The light chain constant region contains one constant domain, C L Includes V H and V L The regions are further subdivided into regions of hypervariability called complementarity determining regions (CDRs) from which there are more conserved regions called framework regions (FRs). H and V L Each antibody contains three CDRs and four FRs, arranged from the amino terminus to the carboxy terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable regions of the heavy and light chains contain binding domains that interact with antigens. The constant regions of the antibody may mediate the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (C1q) of the classical complement system.

[0021] Immunoglobulins can be derived from any of the commonly known isotypes, including, but not limited to, IgA, secretory IgA, IgG, and IgM. IgG subclasses are also well known in the art and include, but are not limited to, human IgG1, IgG2, IgG3, and IgG4. "Isotype" refers to the antibody class or subclass (e.g., IgM or IgG1) encoded by the heavy chain constant region genes. In certain embodiments, one or more amino acids of an isotype can be mutated to alter effector function. The term "antibody" includes, by way of example, both naturally occurring and non-naturally occurring Abs, monoclonal and polyclonal Abs, chimeric and humanized Abs, human or non-human Abs, fully synthetic Abs, and single-chain antibodies. Non-human antibodies can be humanized by recombinant methods to reduce immunogenicity in humans. Unless expressly stated and unless otherwise indicated by context, the term "antibody" also includes antigen-binding fragments or portions of any of the above immunoglobulins, including monovalent and bivalent fragments or portions and single-chain antibodies.

[0022] An "isolated antibody" refers to an antibody that is substantially free of other antibodies having different antigenic specificities (e.g., an isolated antibody that specifically binds to PD-1 is substantially free of antibodies that specifically bind to antigens other than PD-1). An isolated antibody that specifically binds to PD-1 may, however, have cross-reactivity with other antigens, such as PD-1 molecules from different species. Moreover, an isolated antibody may be substantially free of other cellular material and / or chemicals.

[0023] The term "monoclonal antibody" ("mAb") refers to a non-naturally occurring preparation of antibody molecules of single molecular composition, i.e., antibody molecules which are essentially identical in primary sequence and which display a single binding specificity and affinity for a particular epitope. mAbs are examples of isolated antibodies. MAbs can be produced by hybridoma, recombinant, transgenic, or other techniques known to those skilled in the art.

[0024] A "human" antibody (HuMAb) refers to an antibody having variable regions in which both the framework and CDR regions are derived from human germline immunoglobulin sequences. Furthermore, if the antibody contains a constant region, the constant region is also derived from human germline immunoglobulin sequences. The human antibodies of the invention may include amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutation in vivo). However, as used herein, the term "human antibody" is not intended to include antibodies in which CDR sequences derived from the germline of other mammalian species, such as a mouse, have been grafted onto human framework region sequences. The terms "human" antibody and "fully human" antibody are used interchangeably.

[0025] A "humanized antibody" refers to an antibody in which some, most, or all of the amino acids outside the CDR domains of a non-human antibody are replaced with corresponding amino acids from a human immunoglobulin. In some embodiments of a humanized form of an antibody, some, most, or all of the amino acids outside the CDR domains are substituted with amino acids from a human immunoglobulin, while some, most, or all of the amino acids within one or more CDR regions remain unchanged. Small additions, deletions, insertions, substitutions, or modifications of amino acids are permissible as long as they do not abolish the ability of the antibody to bind to a particular antigen. A "humanized" antibody retains antigen specificity similar to that of the original antibody.

[0026] "Chimeric antibody" refers to an antibody in which the variable region is derived from one species and the constant region is derived from another species, such as an antibody in which the variable region is derived from a mouse antibody and the constant region is derived from a human antibody.

[0027] An "anti-antigen" antibody refers to an antibody that specifically binds to that antigen. For example, an anti-PD-1 antibody specifically binds to PD-1, and an anti-CTLA-4 antibody specifically binds to CTLA-4.

[0028] An "antigen-binding portion" (also called an "antigen-binding fragment") of an antibody refers to one or more fragments of an antibody that retain the ability to specifically bind to the antigen bound by the whole antibody.

[0029] "Cancer" refers to a broad group of diverse diseases characterized by the uncontrolled growth of abnormal cells in the body. Uncontrolled cell division and proliferation leads to the formation of malignant tumors that can invade nearby tissues and even metastasize to distant parts of the body via the lymphatic system or bloodstream. As used herein, "cancer" refers to primary, metastatic, and recurrent cancers. In one embodiment, the cancer is head and neck cancer. In a further embodiment, the cancer is head and neck squamous cell carcinoma. In one embodiment, the cancer is human papillomavirus (HPV)-positive cancer. In another embodiment, the cancer is HPV-negative cancer. In one embodiment, the cancer is HPV-positive head and neck squamous cell carcinoma. In another embodiment, the cancer is HPV-negative head and neck squamous cell carcinoma. In yet another embodiment, the cancer is nasopharyngeal carcinoma. In a different embodiment, the cancer is Merkel cell carcinoma. In one embodiment, the Merkel cell carcinoma is associated with polyomavirus. In another embodiment, the cancer is cervical, vaginal, or vulvar cancer. In one embodiment, the cancer is HPV-positive cervical, vaginal, or vulvar cancer. In other embodiments, the cancer is HPV-negative cervical, vaginal, or vulvar cancer. In certain embodiments, the cancer is Epstein-Barr virus (EBV)-positive cancer. In other embodiments, the cancer is EBV-negative cancer. In certain embodiments, the cancer is gastric cancer, including gastroesophageal junction cancer (including adenocarcinoma from the lower esophagus). In further embodiments, the cancer is EBV-positive gastric cancer. In yet another embodiment, the cancer is EBV-negative gastric cancer. In other embodiments, the cancer is EBV-positive nasopharyngeal carcinoma. In further embodiments, the cancer is EBV-negative nasopharyngeal carcinoma.

[0030] "Cytotoxic T-lymphocyte antigen-4" (CTLA-4) refers to an immunoinhibitory receptor belonging to the CD28 family. CTLA-4 is expressed exclusively on T cells in vivo and binds to two ligands, CD80 and CD86 (also known as B7-1 and B7-2, respectively). As used herein, the term "CTLA-4" includes human CTLA-4 (hCTLA-4), variants, isoforms, and species homologs of hCTLA-4, as well as analogs that share at least one epitope with hCTLA-4. The complete hCTLA-4 sequence can be found under GenBank Accession No. AAB59385.

[0031] As used herein, "dosing interval" refers to the time elapsed between administration of multiple doses of the formulations disclosed herein to a subject. Dosing intervals may therefore be expressed as ranges.

[0032] As used herein, the term "dosing frequency" refers to the frequency with which a formulation disclosed herein is administered over a given period of time. Dosing frequency may be expressed as the number of doses per period of time, e.g., once per week or once every two weeks.

[0033] As used herein, "EBV positive" refers to a subject with cancer that is positive for Epstein-Barr virus (EBV; also known as human herpesvirus 4). As used herein, "EBV negative" refers to a subject with cancer that is negative for EBV. In certain embodiments, a subject's EBV status is determined using EBER (Epstein-Barr virus-encoded small RNA) in situ hybridization to detect EBV-specific small RNA, polymerase chain reaction (PCR) to amplify and detect EBV-specific DNA, or any other method known in the art. In some embodiments, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 99% or at least about 100% or more tumor cells show EBER nuclear staining in in situ hybridization, and the tumor is considered EBV positive.In some embodiments, at least about 5% or more tumor cells show EBER nuclear staining in in situ hybridization, and the tumor is considered EBV positive.In other embodiments, at least about 50% or more tumor cells show EBER nuclear staining in in situ hybridization, and the tumor is considered EBV positive. In other embodiments, about 5%, about 4%, about 3%, about 2%, or about 1% or less of the tumor cells show nuclear staining for EBER in in situ hybridization, and the tumor is considered EBV-negative. In further embodiments, 5% or less of the tumor cells show nuclear staining for EBER in in situ hybridization, and the tumor is considered EBV-negative.

[0034] As used herein, "polyomavirus-positive" or "polyomavirus-associated" refers to a subject with cancer that is positive for polyomavirus. Polyomaviruses are small, unenveloped DNA viruses, some of which have been found to be associated with certain types of cancer. Merkel cell polyomavirus (MCPyV) has been found in tumor cells of cutaneous squamous cell carcinoma, basal cell carcinoma, Bowen's disease, non-small cell lung cancer, and cervical cancer. In some embodiments, the polyomavirus status of a subject is determined using PCR (including, but not limited to, real-time PCR and reverse transcription PCR) using primers for polyomavirus DNA, immunohistochemistry for polyomavirus-specific proteins, or any other method known in the art. In certain embodiments, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 99%, or at least about 100% of the tumor cells exhibit strong and diffuse nuclear and / or cytoplasmic staining by immunohistochemistry for one or more polyomavirus-specific antigens, and the tumor is considered polyomavirus-positive. In certain embodiments, at least about 5% of the tumor cells exhibit strong and diffuse nuclear and / or cytoplasmic staining by immunohistochemistry for one or more polyomavirus-specific antigens, and the tumor is considered polyomavirus-positive. In other embodiments, at least about 50% of the tumor cells exhibit strong and widespread nuclear and / or cytoplasmic staining by immunohistochemistry for one or more polyomavirus-specific antigens, and the tumor is considered polyomavirus-positive. In other embodiments, no more than about 5%, about 4%, about 3%, about 2%, or about 1% of the tumor cells exhibit strong and widespread nuclear and / or cytoplasmic staining by immunohistochemistry for one or more polyomavirus-specific antigens, and the tumor is considered polyomavirus-negative.In further embodiments, a tumor is considered polyomavirus negative when 5% or less of the tumor cells show strong and widespread nuclear and / or cytoplasmic staining by immunohistochemistry for one or more polyomavirus-specific antigens.

[0035] The term "flat dose" refers to a dose administered to a patient without consideration of the patient's weight or body surface area (BSA). Thus, a flat dose is not provided as a mg / kg dose, but rather as an absolute dose of an agent (e.g., an anti-PD-1 antibody). For example, a 60 kg person and a 100 kg person would receive the same dose of antibody (e.g., 240 mg of an anti-PD-1 antibody).

[0036] The term "fixed dose" with respect to compositions of the invention means that two or more different antibodies are present in a single composition in a specific (fixed) ratio to one another. In certain embodiments, the fixed dose is based on the weight (e.g., mg) of the antibody. In certain embodiments, the fixed dose is based on the concentration (e.g., mg / ml) of the antibody. In certain embodiments, the ratio is at least about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, about 1:30, about 1:40, about 1:50, about 1:60, about 1:70, about 1:80, about 1:90, about 1:100, about 1:120, about 1:140, about 1:160, about 1:180 ...30, about 1:40, about 1:50, about 1:30, about 1:40, about 1:50, about The ratio of mg of first antibody to mg of second antibody is about 0, about 200:1, about 180:1, about 160:1, about 140:1, about 120:1, about 100:1, about 90:1, about 80:1, about 70:1, about 60:1, about 50:1, about 40:1, about 30:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, or about 2:1. For example, a 3:1 ratio of first antibody and second antibody may indicate that a vial may contain about 240 mg of first antibody and 80 mg of second antibody, or about 3 mg / ml of first antibody and 1 mg / ml of second antibody.

[0037] The term "weight-based dose" as used herein means that the dose administered to a patient is calculated based on the patient's weight. For example, if a patient weighing 60 kg requires a combination of 3 mg / kg of anti-PD-1 antibody and 1 mg / kg of anti-CTLA-4 antibody, a 3:1 ratio fixed dose formulation of anti-PD-1 antibody and anti-CTLA-4 antibody can be used to derive appropriate amounts of anti-PD-1 antibody (i.e., 180 mg) and anti-CTLA-4 antibody (i.e., 60 mg) at one time.

[0038] As used herein, "HPV" refers to human papillomavirus. HPV is a group of over 200 viruses. Low-risk HPVs do not cause cancer. High-risk HPVs can cause cancer. In some embodiments, the HPV subtype can be any type of HPV. In some embodiments, the HPV is HPV subtype 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, 68, or any combination thereof.

[0039] As used herein, "HPV positive" refers to a subject with a cancer that is positive for HPV. As used herein, "HPV negative" refers to a subject with a cancer that is negative for HPV. In certain embodiments, determining the HPV status of a subject comprises determining whether the cancer tumor expresses one or more proteins from HPV or nucleotide sequences encoding said one or more proteins. In certain embodiments, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 99%, or at least about 100% of the tumor cells exhibit strong and widespread nuclear and cytoplasmic staining by immunohistochemistry for p16, and the tumor is considered HPV positive. In certain embodiments, at least about 70% of the tumor cells exhibit strong and widespread nuclear and cytoplasmic staining by immunohistochemistry for p16, and the tumor is considered HPV positive. In certain embodiments, no more than about 30%, 25%, 20%, 15%, 10%, 5%, or 1% of the tumor cells show strong and widespread nuclear and cytoplasmic staining by immunohistochemistry for p16, and the tumor is considered HPV negative. In further embodiments, no more than 30% of the tumor cells show strong and widespread nuclear and cytoplasmic staining by immunohistochemistry for p16, and the tumor is considered HPV negative.

[0040] The term "immunotherapy" refers to the treatment of a subject having a disease or at risk of developing or recurring a disease by methods that involve eliciting, enhancing, suppressing, or otherwise modifying the immune response. "Treatment" or "treatment" of a subject refers to any type of intervention or process or administration of an active agent administered to a subject with the intent to reverse, alleviate, ameliorate, arrest, delay, or prevent the onset, progression, development, severity, or recurrence of symptoms, complications, or conditions or biochemical manifestations associated with a disease.

[0041] As used herein, "PD-L1 positive" can be used interchangeably with "at least about 1% PD-L1 expression." In one embodiment, PD-L1 expression can be measured by any method known in the art. In other embodiments, PD-L1 expression is measured by automated IHC. Thus, a PD-L1-positive tumor has at least about 1%, at least about 2%, at least about 5%, at least about 10%, or at least about 20% of tumor cells expressing PD-L1, as measured by automated IHC. In one embodiment, "PD-L1 positive" means that there are at least 100 cells that express PD-L1 on the cell surface.

[0042] "Programmed death-1 (PD-1)" refers to an immunoinhibitory receptor belonging to the CD28 family. PD-1 is expressed in vivo primarily on preactivated T cells and binds to two ligands, PD-L1 and PD-L2. As used herein, the term "PD-1" includes human PD-1 (hPD-1), variants, isoforms, and species homologs of hPD-1, as well as analogs that share at least one epitope with hPD-1. The complete hPD-1 sequence can be found under GenBank Accession No. U64863.

[0043] "Programmed death-ligand-1 (PD-L1)" is one of two cell surface glycoprotein ligands for PD-1 (the other is PD-L2) that downregulates T cell activation and cytokine secretion upon binding to PD-1. As used herein, the term "PD-L1" includes human PD-L1 (hPD-L1), variants, isoforms, and species homologs of hPD-L1, and analogs that share at least one epitope with hPD-L1. The complete hPD-L1 sequence can be found in GenBank Accession No. Q9NZQ7.

[0044] A "subject" includes any human or non-human animal. The term "non-human animal" includes, but is not limited to, vertebrates such as non-human primates, sheep, dogs, and rodents such as mice, rats, and guinea pigs. In some embodiments, the subject is a human. The terms "subject" and "patient" are used interchangeably herein.

[0045] A "therapeutically effective amount" or "therapeutically effective dose" of a drug or therapeutic agent is any amount of drug that, when used alone or in combination with other therapeutic agents, protects a subject from the onset of disease or promotes disease regression, as evidenced by a decrease in the severity of disease symptoms, an increase in the frequency and duration of disease-free symptom intervals, or prevention of functional impairment or disability due to disease morbidity. The ability of a therapeutic agent to promote disease regression can be assessed using a variety of methods known to the skilled practitioner, such as assaying the activity of the agent in human subjects during clinical trials, in animal model systems that are predictive of efficacy in humans, or in in vitro assays.

[0046] As used herein, a "sub-therapeutic dose" refers to a dose of a therapeutic compound (e.g., an antibody) that is lower than the usual or typical dose of the therapeutic compound when administered alone to treat a hyperproliferative disease (e.g., cancer).

[0047] By way of example, an "anti-cancer agent" promotes cancer regression or prevents further tumor growth in a subject. In certain embodiments, a therapeutically effective amount of a drug promotes cancer regression to the point of eliminating the cancer. "Promoting cancer regression" means that administration of an effective amount of a drug, alone or in combination with an anti-neoplastic agent, results in a reduction in tumor growth or size, tumor necrosis, a reduction in the severity of at least one disease symptom, an increase in the frequency and duration of disease-free symptom intervals, or prevention of functional impairment or disability due to disease morbidity. Furthermore, the terms "effective" and "efficacy" with respect to treatment include both pharmacological efficacy and physiological safety. Pharmacological efficacy refers to the ability of a drug to promote cancer regression in a patient. Physiological safety refers to the level of toxicity or other adverse physiological effects (adverse effects) at the cellular, organ, and / or organismal level resulting from administration of a drug.

[0048] As an example of tumor treatment, a therapeutically effective amount of an anti-cancer agent can inhibit cell proliferation or tumor growth by at least about 10%, at least about 20%, at least about 40%, at least about 60%, or at least about 80% compared to untreated subjects, or, in certain embodiments, compared to patients treated with standard therapy. In other embodiments of the invention, tumor regression can be observed and sustained for a period of at least about 20 days, at least about 40 days, or at least about 60 days. Regardless of these ultimate indicators of therapeutic efficacy, evaluation of immunotherapeutic drugs must also take into account "immune-related" response patterns.

[0049] The term "immune-related" response pattern refers to a clinical response pattern frequently observed in cancer patients treated with immunotherapeutic agents that produce antitumor effects by eliciting a cancer-specific immune response or by modifying natural immune processes. This response pattern is characterized by a beneficial therapeutic effect followed by an initial increase in tumor burden or the appearance of new lesions, which in the field of traditional chemotherapy would be classified as disease progression and would be synonymous with treatment failure. Therefore, proper evaluation of immunotherapeutic agents may require long-term monitoring of the effects of these agents on the target disease.

[0050] A therapeutically effective amount of a drug includes a "prophylactically effective amount," which is any amount of drug that, when administered alone or in combination with an anti-neoplastic agent to a subject at risk of developing cancer (e.g., a subject with a pre-malignant condition) or having cancer recurrence, prevents the onset or recurrence of cancer. In some embodiments, a prophylactically effective amount completely prevents the onset or recurrence of cancer. "Preventing" the onset or recurrence of cancer means reducing the likelihood of cancer onset or recurrence or completely preventing the onset or recurrence of cancer.

[0051] The use of the alternative (e.g., "or") should be understood to mean either one, both, or any combination of the alternatives. As used herein, the terms "a," "an," and "the" should be understood to mean "one or more" of any of the cited or listed components.

[0052] The terms "about" or "essentially comprising" refer to a value or composition that is within an acceptable error range for a particular value or composition as determined by one of ordinary skill in the art, which depends in part on how the value or composition is measured or determined, i.e., the detection limit of the measurement system. For example, "about" or "essentially comprising" can mean within one or more standard deviations, as is customary in the art. Alternatively, "about" or "essentially comprising" can mean within a range of up to 10% or 20% (i.e., ±10% or ±20%). For example, about 3 mg can include any value between 2.7 mg and 3.3 mg (if 10%) or 2.4 mg and 3.6 mg (if 20%). Furthermore, particularly in biological systems or processes, the term can refer to up to an order of magnitude or up to a five-fold difference in value. When a particular value or composition is provided in the specification and claims, unless otherwise indicated, the meaning of "about" or "essentially comprising" should be assumed to be within an acceptable error range for that particular value or composition.

[0053] As used herein, the terms "approximately once a week," "approximately once every two weeks," or any other similar administration interval expression refer to approximate numbers. "Approximately once a week" can include every 7 days ± 1 day, i.e., every 6 to 8 days. "Approximately once every two weeks" includes every 14 days ± 3 days, i.e., every 11 to 17 days. Similar approximations apply to, for example, approximately once every 3 weeks, approximately once every 4 weeks, approximately once every 5 weeks, approximately once every 6 weeks, and approximately once every 12 weeks. In certain embodiments, an administration interval of approximately once every 6 weeks or approximately once every 12 weeks means that the first dose may be administered on any day of the first week, and then the next dose may be administered on any day of the 6th or 12th week, respectively. In other embodiments, a dosing interval of approximately once every 6 weeks or approximately once every 12 weeks means that the first dose is administered on a particular day of the week (e.g., Monday) in the first week, followed by the next dose on the same day of the week (i.e., Monday) in the 6th or 12th week, respectively. Similar principles apply to terms including, but not limited to, "approximately once every 2 weeks," "approximately once a month," etc.

[0054] As described herein, any concentration range, percentage range, ratio range, or integer range should be understood to include, unless otherwise specified, every integer value within the stated range, and fractions thereof, where appropriate (e.g., 1 / 10 and 1 / 100 of an integer).

[0055] Various aspects of the invention are described in further detail in the following sections.

[0056] Methods of the Invention The present invention provides methods for treating cancers associated with, resulting from, or caused by viruses using one or more immune checkpoint inhibitors (e.g., an anti-PD-1 antibody, or an antigen-binding portion thereof, or an anti-PD-L1 antibody, or an antigen-binding portion thereof) as monotherapy or in combination with other anti-cancer agents. In certain embodiments, the cancer is a solid tumor. In other embodiments, the cancer is a primary cancer. In other embodiments, the cancer is a metastatic or recurrent cancer. In certain embodiments, the subject is a human patient. In certain embodiments, the subject is a chemotherapy-naive patient (e.g., a patient who has not previously received any chemotherapy). In other embodiments, the subject has received other cancer treatments (e.g., chemotherapy), but is resistant or refractory to such other cancer treatments.

[0057] In some embodiments, the present invention relates to a method for treating a subject with a tumor associated with, derived from, or caused by HPV, the method comprising administering a therapeutically effective amount of an immune checkpoint inhibitor (e.g., an anti-PD-1 antibody or an antigen-binding portion thereof, or an anti-PD-L1 antibody or an antigen-binding portion thereof) as monotherapy or in combination with one or more anti-cancer agents. In some embodiments, the present invention relates to a method for treating HPV-positive tumors, comprising administering an immune checkpoint inhibitor (e.g., an anti-PD-1 antibody or an antigen-binding portion thereof, or an anti-PD-L1 antibody or an antigen-binding portion thereof). HPV contributes as a causative agent for several types of cancer, including cervical cancer, anal cancer, head and neck cancer (oropharyngeal cancer), vaginal cancer, vulvar cancer, and penile cancer. Five percent of all cancers worldwide are caused by HPV. In some embodiments, the method of the present invention relates to a method for identifying a subject with an HPV-positive cancer. There are over 200 related viruses in the HPV family, including subtypes 6, 11, 16, 18, 30, 31, 33, 34, 35, 39, 40, 42, 43, 44, 45, 51, 52, 53, 54, 55, 56, 57, 58, 59, 66, 68, and other unidentified subtypes. In some embodiments, the HPV subtype is HPV subtype 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, 68, or any combination thereof. In some embodiments, the HPV subtype is HPV subtype 16. In some embodiments, the HPV subtype is HPV subtype 18. In some embodiments, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 99% or at least about 100% of tumor cells show strong and widespread nuclear and cytoplasmic staining by immunohistochemistry for p16, and the tumor is considered HPV positive. In certain embodiments, at least about 70% of tumor cells show strong and widespread nuclear and cytoplasmic staining by immunohistochemistry for p16, and the tumor is considered HPV positive.

[0058] In certain embodiments, the present invention provides a method for treating a subject with a tumor derived from HPV-positive squamous cell carcinoma of the head and neck (SCCHN), comprising administering to the subject a therapeutically effective amount of an immune checkpoint inhibitor, e.g., an anti-PD-1 antibody or an anti-PD-L1 antibody. In certain embodiments, the present invention relates to a method for treating a subject with a tumor derived from HPV-positive SCCHN, comprising: (i) measuring the level of HPV in a sample from the subject, wherein the subject is positive for HPV; and (ii) administering to the subject a therapeutically effective amount of an immune checkpoint inhibitor, e.g., an anti-PD-1 antibody, or an antigen-binding portion thereof, or an anti-PD-L1 antibody, or an antigen-binding portion thereof. In certain embodiments, the anti-PD-1 antibody is nivolumab. In other embodiments, the anti-PD-1 antibody competes for binding with nivolumab. In a further embodiment, the anti-PD-1 antibody is pembrolizumab. In yet a further embodiment, the anti-PD-1 antibody competes for binding with pembrolizumab. In other embodiments, the anti-PD-1 antibody is BGB-A317 or competes for binding with BGB-A317. In some embodiments, the PD-L1 antibody is BMS-936559, MPDL3280A, MEDI4736, or MSB0010718C, or competes for binding with BMS-936559, MPDL3280A, MEDI4736, or MSB0010718C.

[0059] In certain embodiments, the present invention relates to a method for identifying a subject having an HPV-positive SCCHN-derived tumor suitable for immune checkpoint inhibitor therapy, e.g., anti-PD-1 antibody or anti-PD-L1 antibody therapy, comprising: (i) measuring the level of HPV in a sample from the subject, wherein the subject is positive for HPV, and (ii) administering to the subject a therapeutically effective amount of an immune checkpoint inhibitor, e.g., an anti-PD-1 antibody or antigen-binding portion thereof, or an anti-PD-L1 antibody or antigen-binding portion thereof.

[0060] In some embodiments, the cancer treated by the methods disclosed herein is HPV-negative. In some embodiments, the present invention relates to a method for treating a subject with an HPV-negative tumor, comprising administering a therapeutically effective amount of an immune checkpoint inhibitor (e.g., an anti-PD-1 antibody or an antigen-binding portion thereof, or an anti-PD-L1 antibody or an antigen-binding portion thereof) as monotherapy or in combination with one or more anti-cancer agents. In some embodiments, the methods of the present invention relate to a method for identifying a subject with an HPV-negative cancer. In some embodiments, the tumor is considered HPV-negative when no more than about 30%, about 25%, about 20%, about 15%, about 10%, about 5%, or about 1% of the tumor cells show strong and widespread nuclear and cytoplasmic staining by immunohistochemistry for p16. In further embodiments, the tumor is considered HPV-negative when no more than 30% of the tumor cells show strong and widespread nuclear and cytoplasmic staining by immunohistochemistry for p16.

[0061] In certain embodiments, the present invention provides a method for treating a subject with an HPV-negative SCCHN-derived tumor, comprising administering to the subject a therapeutically effective amount of an immune checkpoint inhibitor, such as an anti-PD-1 antibody or anti-PD-L1 antibody. In certain embodiments, the present invention relates to a method for treating a subject with an HPV-negative SCCHN-derived tumor, comprising (i) measuring the HPV level in the subject's sample, wherein the subject is negative for HPV, and (ii) administering to the subject a therapeutically effective amount of an immune checkpoint inhibitor, such as an anti-PD-1 antibody or antigen-binding portion thereof, or an anti-PD-L1 antibody or antigen-binding portion thereof. In certain embodiments, the anti-PD-1 antibody is nivolumab. In other embodiments, the anti-PD-1 antibody competes for binding with nivolumab. In a further embodiment, the anti-PD-1 antibody is pembrolizumab. In yet a further embodiment, the anti-PD-1 antibody competes for binding with pembrolizumab. In other embodiments, the anti-PD-1 antibody is BGB-A317 or competes for binding with BGB-A317. In other embodiments, the PD-L1 antibody is BMS-936559, MPDL3280A, MEDI4736, or MSB0010718C, or competes for binding with BMS-936559, MPDL3280A, MEDI4736, or MSB0010718C.

[0062] In certain embodiments, the present invention relates to a method for identifying a subject having an HPV-negative SCCHN-derived tumor suitable for immune checkpoint inhibitor therapy, e.g., anti-PD-1 antibody or anti-PD-L1 antibody therapy, comprising: (i) measuring the level of HPV in a sample from the subject, wherein the subject is negative for HPV, and administering to the subject a therapeutically effective amount of immune checkpoint inhibitor therapy, e.g., an anti-PD-1 antibody or antigen-binding portion thereof, or an anti-PD-L1 antibody or antigen-binding portion thereof. In certain embodiments, the present invention relates to a method for identifying a subject having an HPV-negative SCCHN-derived tumor suitable for immune checkpoint inhibitor therapy, e.g., anti-PD-1 antibody or anti-PD-L1 antibody therapy, comprising: (i) measuring the level of HPV in a sample from the subject, wherein the subject is negative for HPV, and (ii) administering to the subject a therapeutically effective amount of immune checkpoint inhibitor therapy, e.g., an anti-PD-1 antibody or antigen-binding portion thereof, or an anti-PD-L1 antibody or antigen-binding portion thereof.

[0063] In certain embodiments, measuring HPV as disclosed herein comprises identifying the expression of one or more proteins from HPV or nucleotide sequences encoding said one or more proteins. In further embodiments, the one or more proteins from HPV comprise p16, Ki-67, cyclin D1, p53, ProEx C, E6, E7, or any combination thereof. In yet other embodiments, the nucleotide sequences encode p16, Ki-67, cyclin D1, p53, ProEx C, E6, E7, or any combination thereof.

[0064] In other embodiments, one or more proteins derived from HPV are identified by immunohistochemistry, ELISA, Western blot, or protein array, or any other assay known in the art. In other embodiments, the nucleotide sequence is identified by in situ hybridization, DNA or RNA array, or nucleotide hybridization techniques, tumor sequencing techniques, or quantitative polymerase chain reaction (PCR), or any other assay known in the art. See, e.g., Burd, E., Clin. Microbiol. Rev. 2003 Jan; 16(1): 1-17. In some embodiments, HPV DNA is detected by type-specific PCR, universal primer PCR, or liquid hybridization. In some embodiments, mRNA is tested using the In-Cell (Invirion, Frankfurt, Mich.) viral load test. In further embodiments, HPV measurement includes monolayer cytology or histopathology. In yet further embodiments, HPV measurement is performed using any assay known in the art. In some embodiments, the sample comprises a primary tumor or a metastatic lymph node.

[0065] In some embodiments, the cancer treated by the present invention is an EBV-positive cancer. In some embodiments, the present invention relates to the use of an immune checkpoint inhibitor (e.g., a PD-1 antibody or an antigen-binding portion thereof, or a PD-L1 antibody or an antigen-binding portion thereof) for the treatment of EBV-positive cancer, either as a monotherapy or in combination with other anticancer agents. EBV is an enveloped virus and a member of the herpesvirus family. Cancers thought to be associated with EBV include, but are not limited to, nasopharyngeal carcinoma (NPC), gastric adenocarcinoma, high-grade lymphomas (e.g., Burkitt's lymphoma and certain large B-cell lymphomas), certain Hodgkin's lymphoma tumors, lymphomatoid granulomatosis, angioimmunoblastic T-cell lymphoma, peripheral T-cell lymphoma, NK cell neoplasms and leukemias, and inflammatory pseudotumor-like follicular dendritic cell neoplasms. Treatment of EBV-associated tumors may include antiviral agents (including ganciclovir, famciclovir, acyclovir, valacyclovir, foscarnet, and cidofovir) and immunotherapy (including the use of monoclonal antibodies, such as those directed against CD-20). In certain embodiments, EBV-positive tumors treatable by the present invention include EBV-associated gastric cancer, including adenocarcinoma originating from the lower esophagus, and EBV-positive gastroesophageal junction cancer. In certain embodiments of the present invention, subjects with EBV-associated gastric cancer and EBV-positive gastroesophageal junction cancer are administered an anti-PD-1 antibody or antigen-binding portion thereof, or an anti-PD-L1 antibody or antigen-binding portion thereof, as monotherapy or in any combination disclosed herein. Surgical resection of cancerous tissue is the standard of care treatment for EBV-associated gastric cancer. In certain embodiments, patients with EBV-associated gastric cancer are administered nivolumab in addition to surgical resection. In other embodiments, patients with EBV-positive gastroesophageal junction cancer are administered nivolumab in addition to surgical resection.

[0066] In some embodiments, the cancer treated by the present invention is a polyomavirus-positive cancer. In some embodiments, the present invention relates to the use of an immune checkpoint inhibitor (e.g., a PD-1 antibody or an antigen-binding portion thereof or a PD-L1 antibody or an antigen-binding portion thereof) for the treatment of a polyomavirus-positive cancer, either as a monotherapy or in combination with other anticancer agents. Polyomaviruses are DNA-based, unenveloped viruses associated with various tumors in humans. For example, Merkel cell polyomavirus (MCPyV) has been found in cutaneous squamous cell carcinoma, basal cell carcinoma, Bowen's disease, non-small cell lung cancer, and cervical cancer. Merkel cell carcinoma (MCC), or cutaneous neuroendocrine carcinoma, is a type of skin cancer that typically appears on the face, head, or neck of patients. The standard treatment for patients with MCC is surgical resection for early-stage disease, followed by radiation therapy and chemotherapy for advanced disease. Chemotherapy drugs that can be used to treat MCC include, but are not limited to, platinum-based drugs, including etoposide and carboplatin. In one embodiment, a subject with an MCC tumor is administered an anti-PD-1 antibody, or antigen-binding portion thereof, or an anti-PD-L1 antibody, or antigen-binding portion thereof, as monotherapy or in any combination disclosed herein. In one embodiment, a patient with an MCC tumor is administered nivolumab in addition to surgical resection, radiation therapy, chemotherapy, or a combination thereof.

[0067] Cervical, vaginal, and vulvar cancers may also be polyomavirus and / or HPV positive. In certain embodiments, the present invention relates to the use of immune checkpoint inhibitors (e.g., PD-1 antibodies or antigen-binding portions thereof, or PD-L1 antibodies or antigen-binding portions thereof) for the treatment of cervical, vaginal, and vulvar cancers, either as monotherapy or in combination with other anti-cancer agents. These tumors generally develop in the squamous cell population of the respective tissues. Cervical cancer can include squamous cell carcinoma of the cervix and adenocarcinoma of the cervix, which affect the glandular and columnar cells that support the cervix and cervical opening. In certain embodiments, patients with cervical, vaginal, or vulvar tumors are administered an anti-PD-1 antibody or antigen-binding portion thereof, or an anti-PD-L1 antibody or antigen-binding portion thereof, as monotherapy or in any combination disclosed herein. The standard therapeutic treatment for patients with cervical cancer depends on the stage of the cancer but often includes a radical hysterectomy (surgical removal of the uterus, cervix, and part of the vagina). In some cases, patients may receive radiation therapy in combination with chemotherapy. The standard treatment for vaginal and vulvar cancer depends on the stage of treatment, but may include surgical resection for early stage disease, and chemotherapy in addition to radiation for late stage tumors.Chemotherapy includes but is not limited to platinum-based drugs such as 5-FU or cisplatin.In some embodiments, nivolumab is administered to patients with polyomavirus or HPV positive cervical, vaginal or vulvar tumors in addition to surgical resection and chemotherapy.

[0068] Nasopharyngeal carcinoma (NPC) is a cancer that develops in the nasopharynx. Prior exposure to Epstein-Barr virus (EBV) has been shown to increase a patient's chance of developing NPC. In one embodiment, the present invention relates to the use of immune checkpoint inhibitors (e.g., PD-1 antibodies or antigen-binding portions thereof, or PD-L1 antibodies or antigen-binding portions thereof) as monotherapy or in combination with other anti-cancer agents for the treatment of NPC. In one embodiment, a subject with NPC is administered an anti-PD-1 antibody or antigen-binding portion thereof, or an anti-PD-L1 antibody or antigen-binding portion thereof, either as monotherapy or in any combination disclosed herein. Early-stage NPC is generally treated with radiation therapy, and later-stage tumors are supplemented with chemotherapy, most often including platinum-based drugs such as 5-FU and cisplatin. In one embodiment, a patient with EPV-positive NPC is administered nivolumab in addition to radiation therapy and / or chemotherapy.

[0069] In other embodiments, the tumor further expresses PD-L1. The PD-L1 status of the tumor in a subject can be measured prior to administration of any composition or use of any method disclosed herein. In certain embodiments, the PD-L1 expression level of the tumor is at least about 1%, at least about 2%, at least about 3%, at least about 4%, at least about 5%, at least about 6%, at least about 7%, at least about 8%, at least about 9%, at least about 10%, at least about 11%, at least about 12%, at least about 13%, at least about 14%, at least about 15%, at least about 20%, or greater than at least about 20%. In other embodiments, the PD-L1 status of the tumor is at least about 1%. In other embodiments, the PD-L1 status of the subject is at least about 5%. In certain embodiments, the PD-L1 status of the tumor is at least about 10%. Determining PD-L1 status can be performed using antibodies, in situ mRNA hybridization, automated IHC methods, or as described in Taube et al., "Colocalization of inflammatory response with B7-h1 expression in human melanocytic lesions supports an adaptive resistance mechanism of immune escape," Sci. Transl. Med. 4(127):127ra37 (2012), or U.S. Provisional Application Nos. 62 / 152,669, 62 / 153,954, and 62 / 167,674.

[0070] In certain embodiments, a treatment of the invention (e.g., administration of an anti-PD-1 antibody or anti-PD-L1 antibody and, optionally, other anti-cancer agents) effectively extends the survival of a subject. In certain embodiments, an anti-PD-1 antibody treatment or anti-PD-L1 antibody treatment of the invention extends the survival of a subject compared to standard of care treatment. In certain embodiments, a treatment of the invention extends the overall survival of a subject. In certain embodiments, a subject exhibits an overall survival of at least about 10 months, at least about 11 months, at least about 12 months, at least about 13 months, at least about 14 months, at least about 15 months, at least about 16 months, at least about 17 months, at least about 18 months, at least about 19 months, at least about 20 months, at least about 21 months, at least about 22 months, at least about 23 months, at least about 2 years, at least about 3 years, at least about 4 years, or at least about 5 years after administration. In certain embodiments, the survival or overall survival of a subject is increased by at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, or at least about 75% when compared to other subjects treated only with standard therapy. In other embodiments, the survival or overall survival of a subject is increased by at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 6 months, at least about 1 year, at least about 18 months, at least about 2 years, at least about 3 years, at least about 4 years, or at least about 5 years when compared to other subjects treated only with standard therapy.

[0071] In certain embodiments, the treatments of the invention effectively extend the progression-free survival of a subject. For example, the progression-free survival of a subject is extended by at least about 2 weeks, at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 6 months, or at least about 1 year, compared to other subjects treated only with standard therapy. In certain embodiments, after administration of the anti-PD-1 antibody or anti-PD-L1 antibody treatment, the subject exhibits an overall response rate that is improved by at least about 30%, 35%, 36%, 37%, 39%, 40%, 45%, or 50%, compared to the response rate after administration of the standard therapy.

[0072] Immune checkpoint inhibitors (e.g., anti-PD-1 and anti-PD-L1 antibodies) Immune checkpoint inhibitors suitable for use in the methods of the present disclosure include anti-PD-1 antibodies that bind to PD-1 with high specificity and affinity, block binding to PD-L1, and inhibit the immunosuppressive effects of the PD-1 signaling pathway. In any of the therapeutic methods disclosed herein, an anti-PD-1 or anti-PD-L1 "antibody" comprises an antigen-binding portion that binds to the PD-1 or PD-L1 receptor, respectively, and exhibits functional properties similar to those of whole antibodies in ligand binding inhibition and immune system upregulation. In certain embodiments, the anti-PD-1 antibody, or antigen-binding portion thereof, cross-competes with nivolumab for binding to human PD-1. In other embodiments, the anti-PD-L1 antibody, or antigen-binding portion thereof, competes with the binding of BMS-936559, MPDL3280A, MEDI4736, or MSB0010718C for binding to human PD-L1.

[0073] In other embodiments, the anti-PD-1 antibody, anti-PD-L1 antibody, or antigen-binding portion thereof, is a chimeric, humanized, or human monoclonal antibody, or portion thereof. In certain embodiments for treating a human subject, the antibody is a humanized antibody. In other embodiments for treating a human subject, the antibody is a human antibody. Antibodies of the IgG1, IgG2, IgG3, or IgG4 isotype can be used.

[0074] In some embodiments, the anti-PD-1 antibody, anti-PD-L1 antibody, or antigen-binding portion thereof, comprises a heavy chain constant region of the human IgG1 or IgG4 isotype. In some embodiments, the sequence of the IgG4 heavy chain constant region of the anti-PD-1 antibody, anti-PD-L1 antibody, or antigen-binding portion thereof, contains a S228P mutation, which replaces a serine residue in the hinge region with a proline residue normally found at the corresponding position in IgG1 isotype antibodies. This mutation, present in nivolumab, prevents Fab arm exchange with endogenous IgG4 antibodies while maintaining the low affinity for activating Fc receptors associated with wild-type IgG4 antibodies (Wang et al., 2014 Cancer Immunol Res. 2(9):846-56). In yet other embodiments, the antibody comprises a light chain constant region that is a human kappa or lambda constant region. In other embodiments, the anti-PD-1 antibody, anti-PD-L1 antibody, or antigen-binding portion thereof, is a mAb or antigen-binding portion thereof.

[0075] HuMAbs that specifically bind to PD-1 with high affinity are disclosed in U.S. Patent No. 8,008,449. Other anti-PD-1 mAbs are described, for example, in U.S. Patent Nos. 6,808,710, 7,488,802, 8,168,757, and 8,354,509 and PCT Publication WO 2012 / 145493. Each of the anti-PD-1 HuMAbs disclosed in U.S. Patent No. 8,008,449 has been shown to exhibit one or more of the following characteristics: (a) 1×10 binding to human PD-1 as determined by surface plasmon resonance using a Biacore biosensor system; -7 K below M D(b) does not substantially bind to human CD28, CTLA-4, or ICOS; (c) increases T cell proliferation in a mixed lymphocyte reaction (MLR) assay; (d) increases interferon-γ production in an MLR assay; (e) increases IL-2 secretion in an MLR assay; (f) binds to human PD-1 and cynomolgus PD-1; (g) inhibits the binding of PD-L1 and / or PD-L2 to PD-1; (h) stimulates an antigen-specific memory response; (i) stimulates an antibody response; and (j) inhibits tumor cell growth in vivo. Anti-PD-1 antibodies useful in the present invention include mAbs that specifically bind to human PD-1 and exhibit at least one, and in some embodiments, at least five, of the above characteristics. In one embodiment, the anti-PD-1 antibody is nivolumab. In one embodiment, the anti-PD-1 antibody is pembrolizumab.

[0076] In one embodiment, the anti-PD-1 antibody is nivolumab. Nivolumab (also known as "Opdivo®" and formerly named 5C4, BMS-936558, MDX-1106, or ONO-4538) is a fully human IgG4 (S228P) PD-1 immune checkpoint inhibitor antibody that selectively inhibits interaction with PD-1 ligands (PD-L1 and PD-L2), thereby blocking downregulation of anti-tumor T cell function (U.S. Patent No. 8,008,449; Wang et al. In vitro characterization of the anti-PD-1 antibody nivolumab, BMS-936558, and in vivo toxicology in non-human primates, Cancer Imm Res, 2(9):846-56 (2014)).

[0077] In other embodiments, the anti-PD-1 antibody or fragment thereof cross-competes with nivolumab. In other embodiments, the anti-PD-1 antibody or fragment thereof binds to the same epitope as nivolumab. In some embodiments, the anti-PD-1 antibody has the same CDRs as nivolumab.

[0078] In other embodiments, the anti-PD-1 antibody or fragment thereof cross-competes with pembrolizumab. In certain embodiments, the anti-PD-1 antibody or fragment thereof binds to the same epitope as pembrolizumab. In certain embodiments, the anti-PD-1 antibody has the same CDRs as pembrolizumab. In other embodiments, the anti-PD-1 antibody is pembrolizumab. Pembrolizumab ("Keytruda") (登録商標) ", also known as lambrolizumab and MK-3475), is a humanized monoclonal IgG4 antibody directed against the human cell surface receptor PD-1 (programmed death-1 or programmed cell death-1). Pembrolizumab is described, e.g., in U.S. Patent Nos. 8,354,509 and 8,900,587; see also http: / / www.cancer.gov / drugdictionary?cdrid=695789 (last accessed: December 14, 2014). Pembrolizumab is approved by the FDA for the treatment of recurrent or refractory melanoma.

[0079] In other embodiments, the anti-PD-1 antibody or fragment thereof cross-competes with MEDI0608. In yet other embodiments, the anti-PD-1 antibody or fragment thereof binds to the same epitope as MEDI0608. In certain embodiments, the anti-PD-1 antibody has the same CDRs as MEDI0608. In other embodiments, the anti-PD-1 antibody is the monoclonal antibody MEDI0608 (formerly AMP-514). MEDI0608 is described, for example, in U.S. Patent No. 8,609,089 B2 or http: / / www.cancer.gov / drugdictionary?cdrid=756047 (last accessed December 14, 2014).

[0080] In some embodiments, the first antibody is an anti-PD-1 antagonist. One example of an anti-PD-1 antagonist is AMP-224, a B7-DC Fc fusion protein. AMP-224 is described in U.S. Publication No. 2013 / 0017199 or http: / / www.cancer.gov / publications / dictionaries / cancer-drug?cdrid=700595 (last accessed July 8, 2015).

[0081] In other embodiments, the anti-PD-1 antibody or fragment thereof cross-competes with BGB-A317. In certain embodiments, the anti-PD-1 antibody or fragment thereof binds to the same epitope as BGB-A317. In certain embodiments, the anti-PD-1 antibody has the same CDRs as BGB-A317. In certain embodiments, the anti-PD-1 antibody is the humanized monoclonal antibody BGB-A317. BGB-A317 is described in U.S. Publication No. 2015 / 0079109.

[0082] Anti-PD-1 antibodies useful in the methods of the present disclosure also include isolated antibodies that specifically bind to human PD-1 and cross-compete with nivolumab for binding to human PD-1 (see, e.g., U.S. Patent Nos. 8,008,449 and 8,779,105; WO 2013 / 173223). The ability of antibodies to cross-compete for binding to an antigen indicates that these antibodies bind to the same epitope region of the antigen and sterically interfere with the binding of other cross-competing antibodies to that particular epitope region. Because these cross-competing antibodies bind to the same epitope region of PD-1, they are expected to have functional properties very similar to nivolumab. Cross-competing antibodies can be readily identified based on their ability to cross-compete with nivolumab in standard PD-1 binding assays, such as Biacore analysis, ELISA assays, or flow cytometry (see, e.g., WO 2013 / 173223).

[0083] In some embodiments, the antibody that cross-competes with nivolumab for binding to human PD-1 or binds to the same epitope region of human PD-1 is a mAb. For administration to human subjects, these cross-competing antibodies are chimeric antibodies or humanized or human antibodies. Such chimeric, humanized, or human mAbs can be produced and isolated by methods well known in the art.

[0084] Anti-PD-1 antibodies useful in the methods of the invention also include antigen-binding portions of such antibodies. It is well established that the antigen-binding function of an antibody can be performed by fragments of a full-length antibody. Examples of binding fragments encompassed by the term "antigen-binding portion" of an antibody include: (i) V L , V H , C L and C H1 (ii) a Fab fragment, which is a monovalent fragment consisting of two Fab fragments linked by disulfide bridges at the hinge region; (iii) a V H and C H1 (iv) a single-arm V fragment of an antibody, L and V H It contains an Fv fragment consisting of domains.

[0085] Anti-PD-1 antibodies suitable for use in the disclosed compositions are those that bind to PD-1 with high specificity and affinity, block binding of PD-L1 and / or PD-L2, and inhibit the immunosuppressive effects of the PD-1 signaling pathway. In any of the compositions or methods disclosed herein, an anti-PD-1 "antibody" includes an antigen-binding portion or fragment that binds to the PD-1 receptor and exhibits functional properties similar to those of the whole antibody in ligand binding inhibition and immune system upregulation. In certain embodiments, the anti-PD-1 antibody or antigen-binding portion thereof cross-competes with nivolumab for binding to human PD-1. In other embodiments, the anti-PD-1 antibody or antigen-binding portion thereof is a chimeric, humanized, or human monoclonal antibody or portion thereof. In certain embodiments, the antibody is a humanized antibody. In other embodiments, the antibody is a human antibody. Antibodies of the IgG1, IgG2, IgG3, or IgG4 isotype can be used.

[0086] In some embodiments, the anti-PD-1 antibody, or antigen-binding portion thereof, comprises a heavy chain constant region of the human IgG1 or IgG4 isotype. In some embodiments, the sequence of the IgG4 heavy chain constant region of the anti-PD-1 antibody, or antigen-binding portion thereof, comprises a S228P mutation, replacing a serine residue in the hinge region with a proline residue normally found at the corresponding position in IgG1 isotype antibodies. This mutation, present in nivolumab, prevents Fab arm exchange with endogenous IgG4 antibodies while maintaining the low affinity for activating Fc receptors associated with wild-type IgG4 antibodies (Wang et al. (2014)). In yet other embodiments, the antibody comprises a light chain constant region that is a human kappa or lambda constant region. In other embodiments, the anti-PD-1 antibody, or antigen-binding portion thereof, is a mAb, or antigen-binding portion thereof.

[0087] In certain embodiments of any of the methods of treatment described herein comprising administration of an anti-PD-1 antibody, the anti-PD-1 antibody is nivolumab. In other embodiments, the anti-PD-1 antibody is pembrolizumab. In other embodiments, the anti-PD-1 antibody is selected from human antibodies 17D8, 2D3, 4H1, 4A11, 7D3, and 5F4, described in U.S. Patent No. 8,008,449. In yet other embodiments, the anti-PD-1 antibody is MEDI0608 (formerly AMP-514), AMP-224, or BGB-A317.

[0088] Anti-PD-1 antibodies useful in the methods of the invention also include antigen-binding portions of such antibodies. It is well established that the antigen-binding function of an antibody can be performed by fragments of a full-length antibody. Examples of binding fragments encompassed by the term "antigen-binding portion" of an antibody include: (i) V L , V H , C L and C H1 (ii) a Fab fragment, which is a monovalent fragment consisting of two Fab fragments linked by disulfide bridges at the hinge region; (iii) a V H and C H1 (iv) a single-arm V fragment of an antibody, L and V H It contains an Fv fragment consisting of domains.

[0089] In certain embodiments, the anti-PD-1 antibody used in the methods can be replaced with another PD-1 or anti-PD-L1 antagonist. For example, an anti-PD-L1 antibody can be substituted for the use of an anti-PD-1 antibody in the methods disclosed herein, because anti-PD-L1 antibodies block the interaction between PD-1 and PD-L1, thereby exerting a similar effect on the PD-1 signaling pathway. Thus, in certain embodiments, the present invention relates to a method for treating a subject with an HPV-positive SCCHN-derived tumor, comprising administering to the subject a therapeutically effective amount of an anti-PD-L1 antibody.

[0090] In one embodiment, the anti-PD-L1 antibody is BMS-936559 (formerly 12A4 or MDX-1105) (see, e.g., U.S. Patent No. 7,943,743; WO2013 / 173223).

[0091] In other embodiments, the anti-PD-L1 antibody is MPDL3280A (also known as RG7446) (see, e.g., Herbst et al. (2013) J Clin Oncol 31(suppl):3000. Abstract.; U.S. Patent No. 8,217,149).

[0092] In other embodiments, the anti-PD-L1 antibody is MEDI4736 (also known as durvalumab; see Khleif (2013) In: Proceedings from the European Cancer Congress 2013; September 27-October 1, 2013; Amsterdam, The Netherlands. Abstract 802, U.S. Patent No. 8,779,108, or US2014 / 0356353 filed May 6, 2014).

[0093] In a further embodiment, the anti-PD-L1 antibody is MSB0010718C (also known as avelumab; see US 2014 / 0341917).

[0094] Because anti-PD-1 and anti-PD-L1 target the same signaling pathway and have been shown to exhibit similar levels of efficacy in clinical trials across a wide variety of cancers, including RCC (see Brahmer et al. (2012) N Engl J Med 366:2455-65; Topalian et al. (2012a) N Engl J Med 366:2443-54; WO2013 / 173223), an anti-PD-L1 antibody may be substituted for an anti-PD-1 antibody in any of the treatment methods disclosed herein. In one embodiment, the anti-PD-L1 antibody is BMS-936559 (formerly 12A4 or MDX-1105) (see, e.g., U.S. Patent No. 7,943,743; WO2013 / 173223). In other embodiments, the anti-PD-L1 antibody is MPDL3280A (also known as RG7446) (see, e.g., Herbst et al. (2013) J Clin Oncol 31(suppl):3000. Abstract; U.S. Patent No. 8,217,149) or MEDI4736 (Khleif (2013) In: Proceedings from the European Cancer Congress 2013; September 27-October 1, 2013; Amsterdam, The Netherlands. Abstract 802). In one embodiment, the antibody that cross-competes with the above-mentioned control PD-L1 antibody for binding to human PD-L1 or binds to an epitope region of human PD-L1 is a mAb. For administration to human subjects, these cross-competing antibodies may be chimeric, humanized, or human. Such chimeric, humanized, or human mAbs can be produced and isolated by methods well known in the art.

[0095] In certain embodiments, the immune checkpoint inhibitor used in the present invention, e.g., an anti-PD-1 antagonist, is a PD-1 Fc fusion protein.

[0096] Combination therapy with anti-PD-1 or anti-PD-L1 antibodies In some embodiments, the immune checkpoint inhibitor (e.g., an anti-PD-1 antibody or anti-PD-L1 antibody) is administered in combination with one or more other anti-cancer agents. In some embodiments, the one or more anti-cancer agents have been administered to the subject prior to administration of the anti-PD-1 or anti-PD-L1 antibody or prior to combination with the anti-PD-1 or anti-PD-L1 antibody. In some embodiments, the one or more anti-cancer agents have not been effective in treating cancer. In some embodiments, the other anti-cancer agent is any anti-cancer agent described herein or known in the art. In some embodiments, the other anti-cancer agent is an anti-CTLA-4 antibody. In some embodiments, the other anti-cancer agent is a chemotherapeutic agent or a platinum-based doublet chemotherapeutic agent (PT-DC). In some embodiments, the other anti-cancer agent is an EGFR-targeted tyrosine kinase inhibitor (TKI). In some embodiments, the other anti-cancer agent is an anti-VEGF antibody. In other embodiments, the anti-cancer agent is a platinum agent (e.g., cisplatin, carboplatin), a mitotic inhibitor (e.g., paclitaxel, albumin-bound paclitaxel, docetaxel, taxotere, docecad), a fluorinated vinca alkaloid (e.g., vinflunine, javlor), vinorelbine, vinblastine, etoposide, or pemetrexed gemcitabine. In certain embodiments, the other anti-cancer agent is 5-fluorouracil (5-FU). In certain embodiments, the other anti-cancer agent is any other anti-cancer agent known in the art. In certain embodiments, two or more additional anti-cancer agents are administered in combination with an anti-PD-1 or anti-PD-L1 antibody. In certain embodiments, a PD-1 or PD-L1 antibody is combined with surgical resection and / or radiation therapy.

[0097] In some embodiments, anti-PD-1 antibodies or anti-PD-L1 antibodies can be combined with other immunotherapies. In some embodiments, immunotherapies involving immune checkpoint blockade are administered as monotherapy. In other embodiments, immunotherapies involving immune checkpoint blockade are administered in combination with other treatments. In some embodiments, HPV-positive SCCHN patients can benefit from a combination of various immunotherapeutic agents.

[0098] Anti-CTLA-4 antibody In one embodiment, an anti-PD-1 antibody or an anti-PD-L1 antibody is combined with an anti-CTLA-4 antibody. Anti-CTLA-4 antibodies useful in the combination are capable of binding to human CTLA-4 in a manner that disrupts the interaction between CTLA-4 and human B7 receptor. Because the interaction between CTLA-4 and B7 transmits a signal that leads to the inactivation of T cells that bear the CTLA-4 receptor, disrupting this interaction effectively induces, enhances, or prolongs the activation of such T cells, thereby inducing, enhancing, or prolonging an immune response.

[0099] HuMAbs that specifically bind to CTLA-4 with high affinity are disclosed in U.S. Patent Nos. 6,984,720 and 7,605,238. Other CTLA-4 mAbs are disclosed, for example, in U.S. Patent Nos. 5,977,318, 6,051,227, 6,682,736, and 7,034,121. The CTLA-4 HuMAbs disclosed in U.S. Patent Nos. 6,984,720 and 7,605,238 have been shown to exhibit one or more of the following characteristics: (a) a specific affinity of at least about 10 as determined by Biacore analysis; 7 M -1 or about 10 9 M -1 or about 10 10 M -1 ~10 11 M -1 or higher equilibrium binding constant (K a ), (b) specifically binds human CTLA-4 with a binding affinity reflected by at least about 10 3 , about 10 4 or about 10 5 m -1 s -1 The kinetic binding constant (k a ), (c) at least about 10 3 , about 10 4 or about 10 5 m -1 s -1 The kinetic dissociation constant (k d) and (d) inhibit the binding of CTLA-4 to B7-1 (CD80) and B7-2 (CD86). Anti-CTLA-4 antibodies useful in the present invention are mAbs that specifically bind human CTLA-4 and exhibit at least one, at least two, or, in some embodiments, at least three of the above characteristics. An example of a clinical anti-CTLA-4 antibody is human mAb 10D1 (now known as ipilimumab and commercially available as Yervoy®), disclosed in U.S. Patent No. 6,984,720. Ipilimumab is an anti-CTLA-4 antibody for use in the methods disclosed herein. Another anti-CTLA-4 antibody useful in the methods is tremelimumab.

[0100] An example of a clinical anti-CTLA-4 antibody useful in combination is the human mAb 10D1 (now known as ipilimumab and commercially available as Yervoy®), disclosed in U.S. Patent No. 6,984,720. Ipilimumab is an anti-CTLA-4 antibody for use in the methods disclosed herein. Ipilimumab is a fully human, IgG1 monoclonal antibody that blocks the binding of CTLA-4 to its B7 ligand, thereby stimulating T cell activation and improving overall survival (OS) in patients with advanced melanoma.

[0101] Another anti-CTLA-4 antibody useful in the present methods is tremelimumab (also known as CP-675,206). Tremelimumab is a human IgG2 monoclonal anti-CTLA-4 antibody. Tremelimumab is described in WO / 2012 / 122444, U.S. Publication No. 2012 / 263677, or WO Publication No. 2007 / 113648A2.

[0102] The anti-CTLA-4 antibody useful in the method of the present disclosure is also an isolated antibody that specifically binds to human CTLA-4 and cross-competes with ipilimumab or tremelimumab for binding to human CTLA-4 or binds to the same epitope region of human CTLA-4 as ipilimumab or tremelimumab. In some embodiments, the antibody that cross-competes with ipilimumab or tremelimumab for binding to human CTLA-4 or binds to the same epitope region of human CTLA-4 is an antibody that comprises a heavy chain of human IgG1 isotype. For administration to human subjects, these cross-competing antibodies are chimeric antibodies or humanized or human antibodies. Useful anti-CTLA-4 antibodies also include antigen-binding portions of the above antibodies, such as Fab, F(ab')2, Fd, or Fv fragments.

[0103] Ipilimumab (Yervoy®) is a fully human, IgG1 monoclonal antibody that blocks the binding of CTLA-4 to its B7 ligand, thereby stimulating T cell activation and improving overall survival (OS) in patients with advanced melanoma (Hodi et al. (2010) N Engl J Med 363:711-23). ​​Simultaneous therapy with nivolumab and ipilimumab in a phase 1 clinical trial resulted in rapid and profound tumor regression in a significant proportion of patients with advanced melanoma and was significantly more effective than either antibody alone (Wolchok et al. (2013) N Engl J Med 369(2):122-33; WO2013 / 173223). However, it was previously unknown whether this combination of immunoregulatory antibodies would be similarly effective in other tumor types.

[0104] Anti-VEGF antibody In other embodiments, an anti-PD-1 antibody or anti-PD-L1 antibody is combined with an anti-VEGF antagonist, e.g., an anti-VEGF antibody. Vascular endothelial growth factor ("VEGF") is an endothelial cell-specific mitogen and inducer of angiogenesis. VEGF plays a prominent role in angiogenesis and tumor growth and development. In some embodiments of the invention, an anti-PD-1 antibody is administered in combination with an anti-VEGF antagonist. In some embodiments, the anti-VEGF antagonist is an anti-VEGF antibody, antigen-binding molecule, or fragment thereof. In certain embodiments, the anti-VEGF antibody is bevacizumab (described in U.S. Patent No. 7,169,901) or any other VEGF antibody, including ranibizumab (U.S. Patent No. 7,297,334), VGX-100 (U.S. Patent No. 7,423,125), r84 (U.S. Patent No. 8,034,905), aflibercept (U.S. Patent No. 5,952,199), IMC-18F1 (U.S. Patent No. 7,972,596), IMC-1C11 (PCT / US2000 / 02180), and ramucirumab (U.S. Patent No. 7,498,414).

[0105] Chemotherapeutic agents and platinum-based chemotherapy agents In some embodiments, the anti-PD-1 antibody is administered in combination with any chemotherapeutic agent known in the art. In some embodiments, the chemotherapeutic agent is a platinum-based chemotherapeutic agent. The platinum-based chemotherapeutic agent is a platinum coordination complex. In some embodiments, the platinum-based chemotherapeutic agent is a platinum doublet chemotherapeutic agent. In some embodiments, the chemotherapeutic agent is administered at a dose approved for the particular indication. In other embodiments, the chemotherapeutic agent is administered at any dose disclosed herein. In some embodiments, the platinum-based chemotherapeutic agent is cisplatin, carboplatin, oxaliplatin, satraplatin, picoplatin, nedaplatin, triplatin, lipoplatin, or a combination thereof. In some embodiments, the platinum-based chemotherapeutic agent is any other platinum-based chemotherapeutic agent known in the art. In some embodiments, the chemotherapeutic agent is the nucleotide analog gemcitabine. In some embodiments, the chemotherapeutic agent is an antifolate. In some embodiments, the antifolate is pemetrexed. In some embodiments, the chemotherapeutic agent is a taxane. In other embodiments, the taxane is paclitaxel. In other embodiments, the chemotherapeutic agent is a nucleoside analog. In some embodiments, the nucleoside analog is gemcitabine. In some embodiments, the chemotherapeutic agent is any other chemotherapeutic agent known in the art. In some embodiments, at least one, at least two, or more chemotherapeutic agents are administered in combination with an anti-PD-1 antibody, an anti-PD-L1 antibody, or an antigen-binding portion thereof. In some embodiments, an anti-CTLA-4 antibody is further administered.

[0106] tyrosine kinase inhibitors In some embodiments, the anti-PD-1 antibody or anti-PD-L1 antibody is administered in combination with a tyrosine kinase inhibitor. In some embodiments, the tyrosine kinase inhibitor is gefitinib, erlotinib, a combination thereof, or any other tyrosine kinase inhibitor known in the art. In some embodiments, the tyrosine kinase inhibitor acts on epidermal growth factor receptor (EGFR). In some embodiments, an anti-CTLA-4 antibody is further administered.

[0107] Pharmaceutical Compositions and Dosages The therapeutic agents of the present invention can be formulated into compositions, e.g., pharmaceutical compositions, comprising an antibody and a pharmaceutically acceptable carrier. As used herein, "pharmaceutically acceptable carrier" includes any and all physiologically compatible solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like. In some embodiments, the carrier for an antibody-containing composition is suitable for intravenous, intramuscular, subcutaneous, parenteral, spinal, or epidermal administration (e.g., by injection or infusion), while the carrier for a TKI-containing composition is suitable for non-injectable, e.g., oral, administration. The pharmaceutical compositions of the present invention can include one or more pharmaceutically acceptable salts, antioxidants, aqueous and non-aqueous carriers, and / or adjuvants such as preservatives, wetting agents, emulsifying agents, and dispersing agents.

[0108] The dosing regimen is adjusted to provide the optimal desired response, e.g., the maximum therapeutic response and / or the minimum adverse effects. In certain embodiments, the methods of the invention can be used using a flat dose or a weight-based dose. In a further embodiment, the anti-PD-1 antibody, anti-PD-L1 antibody, or antigen-binding portion thereof, is administered as a flat dose. In a further embodiment, the anti-PD-1 antibody, anti-PD-L1 antibody, or antigen-binding portion thereof, is administered as a weight-based dose. For administration of anti-PD-1 antibodies as monotherapy or in combination with other anti-cancer agents, the dose can be in the range of about 0.01 to about 20 mg / kg, about 0.1 to about 10 mg / kg, about 0.1 to about 5 mg / kg, about 1 to about 5 mg / kg, about 2 to about 5 mg / kg, about 7.5 to about 12.5 mg / kg, or about 0.1 to about 30 mg / kg of the subject's body weight, or in the range of about 80 mg to at least 800 mg, about 80 mg to at least 700 mg, about 80 mg to at least 600 mg, about 80 mg to at least 500 mg, about 80 mg to at least 400 mg, about 80 mg to at least 300 mg, about 100 mg to at least 300 mg, or about 200 mg to about 300 mg. For example, the dose can be about 0.1 mg / kg body weight, about 0.3 mg / kg body weight, about 1 mg / kg body weight, about 2 mg / kg body weight, about 3 mg / kg body weight, about 5 mg / kg body weight, or about 10 mg / kg body weight, or about 0.3 mg / kg body weight, about 1 mg / kg body weight, about 2 mg / kg body weight, about 3 mg / kg body weight, or about 5 mg / kg body weight, or about 80 mg, about 100 mg, about 160 mg, about 200 mg, about 240 mg, about 300 mg, about 320 mg, about 400 mg, about 500 mg, about 600 mg, about 700 mg, or about 800 mg. Dosing schedules are generally based on the typical pharmacokinetic properties of antibodies and are designed to achieve an exposure that results in sustained receptor occupancy (RO). Exemplary treatment regimens involve administration approximately once a week, approximately once every two weeks, approximately once every three weeks, approximately once every four weeks, approximately once a month, approximately once every three to six months, or longer. In one embodiment, an anti-PD-1 antibody, such as nivolumab, is administered to a subject approximately once every two weeks. In another embodiment, the antibody is administered approximately once every three weeks. Dosage and schedule may vary during the course of treatment.For example, dosing schedules for anti-PD-1 monotherapy include administration of Ab (i) approximately every 2 weeks in an approximately 6-week cycle, (ii) approximately every 4 weeks for about 6 doses, followed by approximately every 3 months, (iii) approximately every 3 weeks, or (iv) about 3 to about 10 mg / kg once, followed by about 1 mg / kg about every 2 to 3 weeks. Considering that the half-life of IgG4 antibodies is generally 2 to 3 weeks, dosing regimens for anti-PD-1 antibodies of the present invention include intravenous administration of at least about 0.3 to at least about 10 mg / kg body weight, at least about 1 to at least about 5 mg / kg body weight, or at least about 1 to at least about 3 mg / kg body weight, or at least about 80 to at least about 800 mg, of the antibody, administered approximately every 14 to 21 days for up to about 6 weeks or in an approximately 12-week cycle until complete remission or confirmed disease progression. In one embodiment, anti-PD-1 monotherapy is administered at 3 mg / kg every 2 weeks until disease progression or unacceptable toxicity occurs. In other embodiments, anti-PD-1 monotherapy is administered at 240 mg every two weeks until disease progression or unacceptable toxicity. In certain embodiments, the antibody treatment disclosed herein or any combination treatment is continued for at least about 1 month, at least about 3 months, at least about 6 months, at least about 9 months, at least about 1 year, at least about 18 months, at least about 24 months, at least about 3 years, at least about 5 years, or at least about 10 years.

[0109] When used in combination with other cancer drugs, the dose of the anti-PD-1 antibody may be reduced compared to the monotherapy dose. A typical subtherapeutic dose of nivolumab is less than 3 mg / kg, but not less than 0.001 mg / kg. A subtherapeutic dose of an anti-PD-1 antibody used in the methods herein is greater than 0.001 mg / kg and less than 3 mg / kg. In some embodiments, the subtherapeutic dose is about 0.001 mg / kg to about 1 mg / kg, about 0.01 mg / kg to about 1 mg / kg, about 0.1 mg / kg to about 1 mg / kg, or about 0.001 mg / kg to about 0.1 mg / kg body weight. In some embodiments, a subtherapeutic dose is at least about 0.001 mg / kg, at least about 0.005 mg / kg, at least about 0.01 mg / kg, at least about 0.05 mg / kg, at least about 0.1 mg / kg, at least about 0.5 mg / kg, or at least about 1.0 mg / kg body weight. In some embodiments, the subtherapeutic flat dose is less than about 240 mg every two weeks, for example, about 160 mg or about 80 mg every two weeks. Receptor occupancy data from 15 subjects receiving nivolumab at doses ranging from 0.3 mg / kg to 10 mg / kg indicate that PD-1 occupancy is dose-independent within this dose range. Across all doses, the mean occupancy was 85% (range, 70%-97%), with a mean plateau occupancy of 72% (range, 59%-81%) (Brahmer et al. (2010) J Clin Oncol 28:3167-75). In some embodiments, a 0.3 mg / kg dose may allow sufficient exposure to achieve maximal biological activity.

[0110] In one embodiment, the dose of the anti-PD-1 antibody (or anti-PD-L1 antibody) is a fixed dose in the pharmaceutical composition. In other embodiments, the methods of the present invention can be used with flat doses (doses given to patients regardless of the patient's weight). For example, a flat dose of nivolumab can be about 240 mg. For example, a flat dose of pembrolizumab can be about 200 mg. In one embodiment, the anti-PD-1 antibody or antigen-binding portion thereof is administered at a dose of about 240 mg. In one embodiment, the anti-PD-1 antibody or antigen-binding portion thereof is administered at a dose of about 360 mg. In one embodiment, the anti-PD-1 antibody or antigen-binding fragment thereof is administered at a dose of about 480 mg. In one embodiment, 360 mg of the anti-PD-1 antibody or antigen-binding fragment is administered once every three weeks. In another embodiment, 480 mg of the anti-PD-1 antibody or antigen-binding fragment is administered once every four weeks.

[0111] For administration of anti-PD-L1 antibodies, either as monotherapy or in combination with other anti-cancer agents, the dose can be in the range of about 0.01 to about 20 mg / kg, about 0.1 to about 10 mg / kg, about 0.1 to about 5 mg / kg, about 1 to about 5 mg / kg, about 2 to about 5 mg / kg, about 7.5 to about 12.5 mg / kg, or about 0.1 to about 30 mg / kg of the subject's body weight, or about 80 mg to at least 800 mg, about 80 mg to at least 700 mg, about 80 mg to at least 600 mg, about 80 mg to at least 500 mg, about 80 mg to at least 400 mg, about 80 mg to at least 300 mg, about 100 mg to at least 300 mg, or about 200 mg to about 300 mg. For example, the dose can be about 0.1 mg / kg body weight, about 0.3 mg / kg body weight, about 1 mg / kg body weight, about 2 mg / kg body weight, about 3 mg / kg body weight, about 5 mg / kg body weight, or about 10 mg / kg body weight, or about 0.3 mg / kg body weight, about 1 mg / kg body weight, about 2 mg / kg body weight, about 3 mg / kg body weight, or about 5 mg / kg body weight, or about 80 mg, about 100 mg, about 160 mg, about 200 mg, about 240 mg, about 300 mg, about 320 mg, about 400 mg, about 500 mg, about 600 mg, about 700 mg, or about 800 mg. Dosing schedules are generally based on typical pharmacokinetics of antibodies and are designed to achieve an exposure that results in sustained receptor occupancy (RO). Examples of treatment regimens entail administration about once a week, about once every two weeks, about once every three weeks, about once every four weeks, about once a month, about once every three to six months or longer.

[0112] Although high nivolumab monotherapy dosing of approximately 10 mg / kg every 2 weeks has been achieved without reaching the maximum tolerated dose (MTD), significant toxicity has been reported in other trials of checkpoint inhibitors plus antiangiogenic therapeutics (e.g., Johnson et al. (2013) Cancer Immunol Res 1:373-77; Rini et al. (2011) Cancer 117:758-67), supporting the selection of nivolumab doses lower than 10 mg / kg.

[0113] In certain embodiments, the dose of the anti-PD-1 antibody (or anti-PD-L1 antibody) is a fixed dose in the pharmaceutical composition.

[0114] Ipilimumab (Yervoy®) is approved for the treatment of melanoma at 3 mg / kg intravenously every three weeks for four doses. In certain embodiments, the dose of the anti-CTLA-4 antibody is a flat dose given to the patient regardless of body weight. In a specific embodiment, the flat dose of the anti-CTLA-4 antibody is about 80 mg.

[0115] Thus, in some embodiments, about 3 mg / kg is the maximum dose of ipilimumab used in combination with an anti-PD-1 antibody, but in some embodiments, when combined with nivolumab, an anti-CTLA-4 antibody such as ipilimumab can be administered at a dose ranging from about 0.3 to about 10 mg / kg, about 0.5 to about 10 mg / kg, about 0.5 to about 5 mg / kg, or about 1 to about 5 mg / kg body weight approximately every 2 or 3 weeks. In other embodiments, ipilimumab is administered on a different dosing schedule than nivolumab. In some embodiments, ipilimumab is administered approximately every week, approximately every 2 weeks, approximately every 3 weeks, approximately every 4 weeks, approximately every 5 weeks, approximately every 6 weeks, approximately every 7 weeks, approximately every 8 weeks, approximately every 9 weeks, approximately every 10 weeks, approximately every 11 weeks, approximately every 12 weeks, or approximately every 15 weeks.

[0116] A typical ipilimumab dose is less than 3 mg / kg every three weeks, but not less than 0.001 mg / kg. A subtherapeutic dose of an anti-CTLA-4 antibody used in the methods herein is greater than 0.001 mg / kg and less than 3 mg / kg. In some embodiments, the subtherapeutic dose is about 0.001 mg / kg to about 1 mg / kg, about 0.01 mg / kg to about 1 mg / kg, about 0.1 mg / kg to about 1 mg / kg, or about 0.001 mg / kg to about 0.1 mg / kg body weight. In some embodiments, the subtherapeutic dose is at least about 0.001 mg / kg, at least about 0.005 mg / kg, at least about 0.01 mg / kg, at least about 0.05 mg / kg, at least about 0.1 mg / kg, at least about 0.5 mg / kg, or at least about 1.0 mg / kg body weight. It has been shown that administration of a combination of 3 mg / kg nivolumab and 3 mg / kg ipilimumab exceeds the MTD in melanoma populations, while combinations of 1 mg / kg nivolumab and 3 mg / kg ipilimumab or 3 mg / kg nivolumab and 1 mg / kg ipilimumab have been found to be tolerable in melanoma patients (Wolchok et al. (2013) N Engl J Med 369(2):122-33). Thus, nivolumab is tolerated up to 10 mg / kg intravenously every 2 weeks, but in some embodiments, the dose of the anti-PD-1 antibody, when combined with ipilimumab, does not exceed about 3 mg / kg. In certain embodiments, based on risk-benefit and PK-PD assessments, the doses used include a combination of about 1 mg / kg nivolumab and about 3 mg / kg ipilimumab, about 3 mg / kg nivolumab and about 1 mg / kg ipilimumab, or about 3 mg / kg nivolumab and about 3 mg / kg ipilimumab, each administered approximately once every 2 to 4 weeks, and in certain embodiments, about once every 2 weeks or about once every 3 weeks.In certain embodiments, nivolumab is administered in combination with ipilimumab administered at a dose of about 0.1 mg / kg, about 0.3 mg / kg, about 1 mg / kg, about 2 mg / kg, about 3 mg / kg, or about 5 mg / kg approximately every two weeks, about once every three weeks, or about once every four weeks. In further embodiments, the doses used include a combination of about 240 mg nivolumab and about 80 mg ipilimumab, about 240 mg nivolumab and about 240 mg ipilimumab, or about 80 mg nivolumab and about 240 mg ipilimumab, each administered approximately once every two to four weeks, in certain embodiments, at a dosing frequency of about once every two weeks or about once every three weeks. In certain embodiments, nivolumab is administered approximately once every two weeks, approximately once every three weeks, or approximately once every four weeks at a dose of about 40 mg, about 80 mg, about 100 mg, about 160 mg, about 200 mg, about 240 mg, about 320 mg, or about 400 mg, in combination with ipilimumab at a dose of about 40 mg, about 80 mg, about 160 mg, about 240 mg, about 320 mg, or about 400 mg.

[0117] In one embodiment, the combination of an anti-PD-1 antibody or anti-PD-L1 antibody and an anti-CTLA-4 antibody is administered intravenously to a subject once, twice, three times, or four times approximately every two or three weeks during an induction phase. In one embodiment, the combination of an anti-PD-1 antibody and an anti-PD-L1 antibody is administered intravenously approximately once every two or three weeks for about four times during the induction phase. The induction phase is followed by a maintenance phase, during which the anti-PD-1 antibody or anti-PD-L1 antibody alone is administered to a subject at a dose of about 0.1, about 0.3, about 1, about 2, about 3, about 5, or about 10 mg / kg, or about 40 mg, about 80 mg, about 100 mg, about 160 mg, about 200 mg, about 240 mg, about 320 mg, or about 400 mg approximately every two or three weeks, as long as treatment proves effective or until unmanageable toxicity or disease progression occurs. In certain embodiments, nivolumab is administered at a dose of about 3 mg / kg body weight or about 240 mg approximately every two weeks during the maintenance phase.

[0118] In certain embodiments, the dose of the anti-PD-1 antibody or anti-PD-L1 antibody is a fixed dose in a pharmaceutical composition with the second anti-cancer agent. In certain embodiments, the anti-PD-1 antibody or anti-PD-L1 antibody and the anti-CTLA-4 antibody are formulated as a single composition, wherein the dose of the anti-PD-1 antibody or anti-PD-L1 antibody and the dose of the anti-CTLA-4 antibody are combined in a ratio of 1:50, 1:40, 1:30, 1:20, 1:10, 1:5, 1:3, 1:1, 3:1, 5:1, 10:1, 20:1, 30:1, 40:1, or 50:1.

[0119] For the combination of nivolumab and other anti-cancer drugs, these drugs are administered at their approved doses. Treatment is continued as long as clinical benefit is observed or until unacceptable toxicity or disease progression occurs. Nevertheless, in some embodiments, the doses of these anti-cancer drugs administered are significantly lower than the approved doses, i.e., subtherapeutic doses of the drugs are administered in combination with anti-PD-1 antibodies or anti-PD-L1 antibodies. Anti-PD-1 antibodies or anti-PD-L1 antibodies can be administered at doses that have been shown to produce the highest efficacy as monotherapy in clinical trials, such as about 3 mg / kg nivolumab, administered approximately once every three weeks (Topalian et al. (2012a) N Engl J Med 366:2443-54; Topalian et al. (2012b) Curr Opin Immunol 24:207-12), at significantly lower doses, i.e., subtherapeutic doses or flat doses, i.e., 240 mg. In certain embodiments, the anti-PD-1 antibody is administered at about 3 mg / kg or 240 mg approximately once every two weeks.

[0120] In some embodiments, the anti-PD-1 antibody or anti-PD-L1 antibody is administered in combination with standard treatment for certain types of cancer. In further embodiments, the anti-PD-1 antibody or anti-PD-L1 antibody is administered in combination with chemotherapy including 5-FU, etoposide, and platinum-based drugs, such as carboplatin or cisplatin. In some embodiments, the anti-PD-1 antibody or anti-PD-L1 antibody is administered before, simultaneously with, or after radiation therapy. In some embodiments, the anti-PD-1 antibody or anti-PD-L1 antibody is administered before, simultaneously with, or after surgical resection.

[0121] Dosage and frequency vary depending on the half-life of the antibody in the subject. Generally, human antibodies have the longest half-life, followed by humanized antibodies, chimeric antibodies, and non-human antibodies. Dosage and frequency of administration can vary depending on whether the treatment is preventive or therapeutic. In preventive applications, relatively low doses are generally administered less frequently over a long period of time. Some patients continue to receive treatment for the rest of their lives. In therapeutic applications, relatively high doses and relatively short intervals may be required until disease progression is reduced or stopped, or until the patient shows partial or complete improvement in disease symptoms. Thereafter, the patient can be administered a preventive regimen.

[0122] The actual dosage level of one or more active ingredients in the pharmaceutical compositions of the present invention may be varied to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration without causing undue toxicity to the patient. The selected dosage level will depend on a variety of pharmacokinetic factors, including the activity of the particular composition of the present invention used, the route of administration, the timing of administration, the excretion rate of the particular compound used, the duration of treatment, other drugs, compounds, and / or substances used in combination with the particular composition used, the age, sex, weight, condition, general health, and past medical history of the patient being treated, and similar factors well known in the medical field. The compositions of the present invention are administered in a Administration can be by one or more routes of administration using one or more of a wide variety of methods well known in the art. As will be appreciated by those skilled in the art, the route and / or mode of administration will depend on the desired results.

[0123] kit Also within the scope of the present invention are kits containing an immune checkpoint inhibitor (e.g., an anti-PD-1 antibody or / and an anti-PD-L1 antibody) for therapeutic use and, optionally, other anti-cancer agents. The kit generally includes a label indicating the intended use and instructions for use of the contents of the kit. Here, a label includes any writing or recording medium provided on or with the kit, or that otherwise accompanies the kit. In certain embodiments, the present invention relates to a kit for treating a subject with an HPV-positive SCCHN-derived tumor, the kit including (a) an immune checkpoint inhibitor (e.g., an anti-PD-1 antibody, or an antigen-binding portion thereof, or an anti-PD-L1 antibody, or an antigen-binding portion thereof), and (b) instructions for determining HPV positivity of the tumor, and if the tumor is positive for HPV, the immune checkpoint inhibitor (e.g., an anti-PD-1 antibody, or an antigen-binding portion thereof, or an anti-PD-L1 antibody, or an antigen-binding portion thereof) is administered to the subject using the methods disclosed herein. In certain embodiments, the kit further includes a reagent for HPV positivity determination. In certain embodiments, HPV positivity is measured by HPV p16 expression. In one embodiment, the present invention relates to a kit for treating a subject with an HPV-negative SCCHN-derived tumor, the kit comprising: (a) an immune checkpoint inhibitor (e.g., an anti-PD-1 antibody, or an antigen-binding portion thereof, or an anti-PD-L1 antibody, or an antigen-binding portion thereof); and (b) instructions for determining HPV negativity of the tumor. If the tumor is negative for HPV, the immune checkpoint inhibitor (e.g., an anti-PD-1 antibody, or an antigen-binding portion thereof, or an anti-PD-L1 antibody, or an antigen-binding portion thereof) is administered to the subject in a manner disclosed herein. In one embodiment, the kit further comprises reagents for determining HPV negativity. In one embodiment, HPV negativity is measured by HPV p16 expression. In a further embodiment, the kit comprises one or more additional anti-cancer agents, such as an anti-CTLA-4 antibody and / or a TKI. In one embodiment, the anti-PD-1 antibody, anti-PD-L1 antibody, anti-CTLA-4 antibody, and / or a TKI can be co-packaged in a unit-dose form.In certain embodiments for the treatment of human patients, the kit comprises an anti-human PD-1 or anti-human PD-L1 antibody disclosed herein, e.g., nivolumab or pembrolizumab. In other embodiments, the kit comprises an anti-human CTLA-4 antibody disclosed herein, e.g., ipilimumab or tremelimumab.

[0124] This invention is further illustrated by the following examples, which should not be construed as further limiting. All references cited throughout this specification are expressly incorporated herein by reference. [Example]

[0125] A non-comparative, two-cohort, single-arm, open-label, phase 1 / 2 clinical trial of nivolumab will be conducted in subjects with selected virally positive and virally negative solid tumors to evaluate their safety, tolerability, and efficacy. Subjects will receive a flat dose of 240 mg nivolumab administered intravenously over 30 minutes every 2 weeks until progression or unacceptable toxicity. Efficacy will be measured by objective response rate and duration of response.

[0126] The clinical trial includes two cohorts: a neoadjuvant cohort and a metastatic / recurrent cohort. The primary objective of the neoadjuvant cohort is to investigate the safety and tolerability of neoadjuvant nivolumab in the following tumor types: HPV-positive squamous cell carcinoma of the head and neck (SCCHN), HPV-negative SCCHN, Merkel cell carcinoma (MCC), and cervical, vaginal, or vulvar cancer (GYN). The primary objective of the metastatic / recurrent cohort is to evaluate the investigator-assessed objective response rate (ORR) of nivolumab monotherapy (a flat dose of 240 mg administered intravenously every 2 weeks) in subjects with the following disease: metastatic or recurrent nasopharyngeal carcinoma (NPC), metastatic or recurrent EBV-associated gastric cancer, metastatic or recurrent Merkel cell carcinoma, metastatic or recurrent cervical, vaginal, or vulvar cancer, and metastatic or recurrent HPV-positive SCCHN.

[0127] Secondary objectives of the neoadjuvant cohort are to determine the percent change from baseline in immune cells and virus-specific T cell select immune activating / inhibitory molecules in tumor-specific subsets of nivolumab-treated subjects.Secondary objectives of the metastatic / recurrent cohort are to assess progression-free survival and overall survival in subjects treated with nivolumab monotherapy.

[0128] Patients in the neoadjuvant cohort will be evaluated for progression-free survival up to one year after neoadjuvant administration of nivolumab. Patients will be further monitored to determine percent change from baseline in tumor volume after two flat doses of 240 mg neoadjuvant nivolumab. Pathologic complete tumor response will also be determined in subjects who have undergone surgical resection after two doses of neoadjuvant nivolumab in SCCHN, resectable Merkel cell carcinoma, and cervical, vaginal, or vulvar cancer.

[0129] In the metastatic / recurrent cohort, the safety and tolerability, as defined by adverse events and specific laboratory adverse event rates (worst CTC grade per subject), of nivolumab monotherapy in subjects with metastatic or recurrent virally mediated tumors will also be monitored.

[0130] In both cohorts, changes in antiviral and antitumor immune responses at the tumor site will be evaluated using proliferative and / or functional assays. Investigators will also investigate potential correlations between selected biomarker measurements, including PD-L1, in peripheral blood and tumor tissues, measured by gene expression, flow cytometry, immunohistochemistry, and soluble factor assays, and safety and clinical efficacy measures and pharmacodynamic activity of nivolumab in peripheral blood and tumor tissues. Subjects will be further monitored to examine the effect of nivolumab on viral antigen-specific T cell responsiveness in peripheral blood, as well as potential correlations between the number of tumor mutations and neoantigens and clinical efficacy measures, and whether tumor antigen-specific T cells are present in the periphery. Additionally, pre- and post-treatment Epstein-Barr virus (EBV) DNA levels will be assessed in subjects with EBV-positive gastric and nasopharyngeal cancer.

[0131] Additionally, all subjects will be assessed for global health status as assessed by EQ-5D and cancer-specific health-related quality of life as assessed by EORTC QLQ-C30.In all cases, the immunogenicity and pharmacokinetics of nivolumab monotherapy will be characterized and, where possible, exposure-response relationships will be explored.

[0132] subject All eligible patients have histopathologically confirmed Merkel cell carcinoma, EBV-positive gastric or esophagogastric junction cancer (including adenocarcinoma arising from the lower esophagus), nasopharyngeal carcinoma, squamous cell carcinoma of the cervix, vagina, or vulva, or SCCHN. For subjects with Merkel cell carcinoma, Merkel cell polyomavirus (MCPyV) status will be determined after enrollment. For subjects in the metastatic cohort with gastric tumor types, EBV positivity will be defined by EBV-encoded RNA (EBER) in situ hybridization. For subjects in the metastatic cohort with nasopharyngeal carcinoma tumor types, EBV positivity will be defined by EBER in situ hybridization, and viral testing will be performed retrospectively only if prior test results are not available. For subjects in the metastatic cohort with gynecological tumors, HPV positivity will be defined by in situ hybridization, real-time PCR, or immunohistochemistry (IHC). High-risk HPV positivity includes the following subtypes: 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, and 68. Viral testing is performed retrospectively only if prior test results are unavailable. For subjects in the virus-positive neoadjuvant and metastatic cohorts with head and neck squamous cell carcinoma, HPV positivity is defined by HPV p16 in situ hybridization, IHC, or tumor sequencing. HPV p16 IHC should be interpreted as positive if >70% intense and widespread nuclear and cytoplasmic staining is specific to tumor cells. HPV-positive status can be obtained from the primary tumor or metastatic lymph nodes. For subjects in the virus-negative neoadjuvant cohort, HPV negativity should be documented by HPV p16 in situ hybridization, IHC, or tumor sequencing.

[0133] Subjects in the neoadjuvant cohort may include those with SCCHN for whom surgical resection is planned. These subjects must have newly diagnosed, histologically or cytologically confirmed squamous cell carcinoma or oropharyngeal undifferentiated carcinoma, and must be determined to have resectable disease. Biopsies can be obtained from the primary tumor or metastatic lymph nodes. In addition, these subjects must have a T2 or larger primary tumor, N2 or larger nodal disease, and a smoking history of more than 10 pack years.

[0134] Neoadjuvant cohort squamous cell cervical, vulvar, or vaginal cancer subjects include those with stage II-IVA cervical cancer for which surgical staging or chemotherapy / radiation treatment is planned or stage II-IVA vulvar or vaginal cancer for which radical resection or chemotherapy / radiation treatment is planned.

[0135] Subjects with Merkel cell carcinoma in the neoadjuvant cohort must have tumors available for pretreatment biopsy (core needle), and posttreatment biopsy includes surgical specimens. Additionally, subjects have resectable disease characterized by primary tumors ≥ 2 cm or stage IIA-IIIB disease of any size with palpable regional lymph node metastases or resectable migrating metastases, stage IV disease with resectable limited metastases, or locally / regionally recurrent disease defined as a total burden ≥ 1 cm in diameter with resectable disease as defined by local or institutional surgical practice.

[0136] Subjects in the metastatic / recurrent cohort include those with progressive metastatic or recurrent disease who have been treated with no more than two prior systemic therapies. These subjects have measurable disease by CT or MRI according to RECIST 1.1 criteria (radiological tumor assessment must be performed within 35 days prior to the first dose). Subjects who actively refuse chemotherapy or other standard treatment for unresectable or metastatic disease (advanced stage III or stage IV) may also be included in this cohort. Subjects in the metastasis / recurrence cohort will be selected from those with histologically confirmed gastric or gastroesophageal junction cancer (including adenocarcinoma arising from the lower esophagus) that is EBV-positive, excluding HPV-associated nasopharyngeal carcinoma and keratinizing squamous cell carcinoma (WHO type I); histologically confirmed HPV-positive SCCHN (oral cavity, pharynx, larynx) that is not suitable for curative-intent local treatment (surgery or radiation therapy with or without chemotherapy); histologically confirmed HPV-positive cervical, vulvar, or vaginal cancer including those that have not received prior systemic treatment; histologically confirmed Merkel cell carcinoma; and histologically confirmed EBV-positive nasopharyngeal carcinoma.

[0137] Subjects in both cohorts were required to be 18 years of age or older and have an Eastern Cooperative Oncology Group (ECOG) performance status of 0 or 1. Subjects must be willing to provide tumor tissue (archived or fresh biopsy specimens) for PD-L1 expression analysis and other biomarker correlation studies. Biopsies can be excision, incision, or core needle. Fine needle aspiration is insufficient.

[0138] Subjects with active brain or leptomeningeal metastases will be excluded from the study unless the brain metastases are treated and progression-free by MRI documented at least 4 weeks after completion of treatment and within 28 days prior to first study drug administration. Subjects must not require immunosuppressive doses of systemic corticosteroids (>10 mg / day prednisone equivalent) for at least 2 weeks prior to study drug administration.

[0139] Subjects are further excluded if they meet the following criteria:Any serious or uncontrolled medical disorder that, in the opinion of the investigator, may increase the risks associated with study participation or study drug administration, impair the subject's ability to receive protocol treatment, or interfere with the interpretation of study results; active, known, or suspected autoimmune disease (excluding vitiligo, type 1 diabetes, residual hypothyroidism due to an autoimmune condition requiring hormone replacement only, or a condition not expected to recur in the absence of an external trigger); corticosteroids (>10 mg daily prednisolone equivalent) or other immunosuppressive medications within 14 days of study drug administration. conditions requiring systemic treatment with steroids (inhaled or topical steroids and adrenal replacement doses are acceptable in the absence of active autoimmune disease), primary tumor or nodal metastases fixed to the carotid artery, skull base, or cervical spinal cord, experimental antitumor vaccines such as anti-PD-1, anti-PD-L1, anti-PD-L2, anti-CD137, or anti-CTLA-4 antibodies, including ipilimumab or other medications that specifically target T cells, prior treatment with any T cell costimulatory or checkpoint pathway, toxicity and fatigue due to prior anticancer therapy other than alopecia were grade 1 (NCI 2016) or higher before administration of the study drug. CTCAE version 4) or resolved to baseline (subjects with toxicities related to prior anticancer therapy that are not expected to resolve and result in persistent sequelae, such as neuropathy after platinum-based therapy, are permitted to enroll); prior treatment with any chemotherapy, radiation therapy, biologic, or investigational therapy for cancer within 28 days of the first dose of investigational treatment (subjects with prior cytotoxic or investigational products <4 weeks before treatment must be eligible, after discussion between the investigator and sponsor, if the toxicity from the prior treatment is Grade 1 (NCI CTCAE version 4) and resolved); and a positive hepatitis B virus surface antigen (HBV sAg) or hepatitis C virus (ribonucleic acid or HCV antibody) test indicating acute or chronic infection; a known history of a positive human immunodeficiency virus (HIV) test or known acquired immunodeficiency syndrome (AIDS); a history of allergy to any investigational drug component; a severe hypersensitivity reaction to any monoclonal antibody; women of childbearing potential, pregnant, or lactating; or women with a positive pregnancy test prior to enrollment.

[0140] Study design and treatment This is an open-label, multicenter, Phase 1 / 2 study to investigate the safety and efficacy of nivolumab as a single agent in selected virus-positive and virus-negative solid tumors. The neoadjuvant cohort includes 84 subjects with three tumor types (Figure 1). This cohort will help investigate the safety and tolerability of neoadjuvant nivolumab administration.

[0141] Enrollment for each tumor type in the neoadjuvant cohort will be paused after the enrollment of the first 10 subjects for safety evaluations to determine the number of subjects whose chemotherapy / radiation (GYN patients, if appropriate) or surgery (SCCHN, MCC, and GYN patients) will be delayed by more than 4 weeks from the planned date (Figure 1). If ≥ 3 of the first 10 subjects for a single tumor type experience a delay of more than 4 weeks from the planned surgery date or the planned start date of concurrent chemoradiation therapy due to a labeled nivolumab immune-related adverse event, that particular tumor cohort will be closed. The remaining tumor types in the neoadjuvant cohort will not be closed, and enrollment must be tumor type closed due to nivolumab-induced surgery delays. If the first 8 patients for a single tumor type experience no delays, no enrollment pause is required.

[0142] The neoadjuvant cohort will enroll three tumor types: HPV-positive and HPV-negative SCCHN, HPV-positive cervical / vaginal / vulvar cancer, and polyomavirus-associated Merkel cell carcinoma. SCCHN tumor types require prospective (pre-trial drug assignment) testing for viral positivity. 21 viral-positive and 21 viral-negative SCCHN subjects will be enrolled. Other tumor types, given their high (>85% positivity) susceptibility, do not require prospective viral testing for entry. The viral-negative group will serve as the control group for biological analysis. Subjects will be initially biopsied and will receive two flat doses of 240 mg nivolumab administered by intravenous (IV) infusion over 30 minutes on days 1 and 15, followed by surgical resection or chemotherapy / radiation (Figure 1, Table 1). No other preoperative treatments will be permitted. [Table 1]

[0143] Selected subjects initially treated with nivolumab will then receive standard of care treatment. After standard of care treatment, subjects may receive relapse treatment with a flat dose of 240 mg nivolumab IV every 2 weeks until disease progression, toxicity, or study discontinuation (Table 1).

[0144] The recurrent / metastatic cohort includes 115 subjects with 5 tumor types (Figure 1). This cohort will help evaluate investigator-assessed ORR for nivolumab monotherapy.

[0145] The metastatic cohort will enroll subjects with metastatic or recurrent disease. This cohort includes five tumor types: EBV-associated gastric, EBV-associated nasopharyngeal, HPV-positive SCCHN, HPV-positive cervical / vulvar / vaginal, and polyomavirus-associated Merkel cell carcinoma. SCCHN and gastric tumor types require prospective testing for viral positivity. A flat dose of 240 mg nivolumab will be administered IV over 30 minutes every 2 weeks until unacceptable toxicity or disease progression as defined by RECIST 1.1 (Table 1).

[0146] evaluation Baseline safety assessments include a medical history obtained to capture relevant underlying conditions. Baseline examinations, including signs and symptoms, weight, height, ECOG performance status, blood pressure (BP), heart rate (HR), temperature, respiratory rate, and oxygen saturation by pulse oximetry at rest and after exercise, must be performed within 14 days prior to the first dose. Concomitant medications must also be collected within 14 days prior to the first dose and throughout the duration of study treatment. Baseline safety laboratory assessments must be performed within 14 days prior to the first dose.

[0147] If subjects are receiving any investigational drug, safety must be assessed. Toxicity assessments will be performed continuously throughout the treatment period. Intra-investigational assessments will include weight, height, ECOG performance status, BP, HR, temperature, respiratory rate, and oxygen saturation by pulse oximetry at rest and after exercise. Intra-investigational safety laboratory assessments will also be performed.

[0148] Efficacy will be assessed by tumor imaging for ongoing study treatment decisions by the investigator using RECIST (Response Evaluation Criteria in Solid Tumors) 1.1 criteria.

[0149] statistical analysis Sample size determination was not based on statistical power calculations. For the neoadjuvant cohort, the SCCHN tumor type included 21 HPV-positive and 21 HPV-negative subjects. The MCC and HPV-positive cervical, vaginal, or vulvar cancer tumor types each included 21 subjects. A sample size of 21 would provide greater than 66% and 89% probability of detecting safety events occurring at 5% and 10%, respectively. Assuming pathologic complete response rates of 10%, 15%, and 20%, a sample size of 21 would provide greater than 89%, 97%, and 99% probability of detecting at least one pathologic complete response, respectively.

[0150] For the recurrence / metastasis cohort, each specific disease or tumor type in the recurrence / metastasis cohort included 23 subjects. Table 2 shows the probability of observing 0, 1, or 2 responders and ≥3 responders, assuming true response rates of 5%, 20%, and 30% for ORR. Table 3 shows the two-sided 95% exact CI using the Clopper-Pearson method based on the observation of 3, 4, and 5 responders among 23 subjects. [Table 2] [Table 3]

[0151] Endpoints and Analysis The primary goal of the neoadjuvant cohort is safety assessment. All reported adverse events will be listed and tabulated by system organ class, preferred period, and cohort / arm and coded according to the latest version of MedDRA. The incidence of adverse events will be reviewed for potential significance and clinical importance. Vital signs and laboratory results will be listed and summarized by cohort / arm. Any significant physical examination findings and laboratory results will be listed. The incidence of infusion reactions will be reviewed to assess the safety and tolerability of reduced nivolumab infusions. The proportion of subjects in the biopsy / neoadjuvant cohort whose surgery was delayed by >4 weeks due to drug-related AEs will be reported for each tumor type.

[0152] The primary objective of the metastatic / recurrent cohort is the determination of investigator-assessed ORR, defined as the number of subjects with a confirmed complete response (CR) or partial response (PR) best overall response (BOR) divided by the number of treated subjects. BOR is defined as the best response title recorded between the first dose and the date of objectively reported tumor progression by investigator assessment using RECIST 1.1 criteria or the date of the last tumor assessment before subsequent treatment.

[0153] Investigator-assessed ORR in the metastatic cohort is summarized by the binomial response rate using the Clopper-Pearson method and its corresponding two-sided 95% exact CI. DOR summarizes subjects who achieved a confirmed PR or CR. DOR is defined as the time between the first confirmed response (CR or PR) and the first objectively reported date of tumor progression or death from any cause, as determined by investigator assessment using RECIST 1.1 criteria, whichever occurs first. For subjects who do not progress or die, DOR is censored at the date of the last evaluable tumor assessment. DOR is assessed only in subjects with an objective response of CR or PR.

[0154] The distribution of time to events will be estimated using the Kaplan-Meier method. This will be performed for PFS (based on investigator assessment) and OS. The median PFS or OS and 95% CI will be constructed based on the log-log transformed CI for the survival function. Rates at several fixed time points will be derived using Kaplan-Meier estimation, and the corresponding confidence intervals will be derived based on the Greenwood formula for variance and the applied log-log transformation for the survival function.

[0155] The pharmacodynamic effects of nivolumab on select biomarkers will be assessed using simple statistics aggregated across time and cohorts and corresponding changes (or percent changes) from baseline. Additionally, the time course of biomarker outcomes will be explored graphically using summary plots or individual subject plots. If meaningful pharmacodynamic trends are demonstrated, methods such as linear mixed models can be used to characterize patterns of change over time. Potential correlations between PD-L1 expression levels (IHC) and clinical efficacy measures will be assessed using Fisher's exact test or other methods, as appropriate.

[0156] The potential influence of various biomarker measurements on pharmacokinetics, safety, and clinical efficacy measurements will be investigated based on available data, and these correlations can be assessed using methods such as, but not limited to, logistic regression analysis and graphing.

[0157] This application claims priority to U.S. Provisional Application No. 62 / 192,396, filed July 14, 2015, which is incorporated herein by reference in its entirety.

Claims

1. A method of treating a subject having a tumor derived from human papillomavirus (HPV)-positive squamous cell carcinoma of the head and neck (SCCHN), comprising administering to the subject a therapeutically effective amount of an antibody or antigen-binding portion thereof that specifically binds to programmed death-1 receptor (PD-1) or programmed death-ligand 1 (PD-L1) and inhibits PD-1 activity (an "anti-PD-1 antibody") or PD-L1 activity (an "anti-PD-L1 antibody"), respectively.

2. (i) measuring the level of HPV in a sample from a subject, wherein the subject is positive for HPV; and (ii) administering to the subject a therapeutically effective amount of an anti-PD-1 antibody, or an antigen-binding portion thereof, or an anti-PD-L1 antibody, or an antigen-binding portion thereof.

10. A method of treating a subject having an HPV-positive SCCHN-derived tumor, comprising:

3. A method for identifying a subject having an HPV-positive SCCHN-derived tumor suitable for anti-PD-1 antibody or anti-PD-L1 antibody treatment, comprising measuring the level of HPV in a sample from the subject, wherein the subject is positive for HPV, and administering to the subject a therapeutically effective amount of an anti-PD-1 antibody, or an antigen-binding portion thereof, or an anti-PD-L1 antibody, or an antigen-binding portion thereof.

4. (i) measuring the level of HPV in a sample from a subject, wherein the subject is positive for HPV; and (ii) administering to the subject a therapeutically effective amount of an anti-PD-1 antibody, or an antigen-binding portion thereof, or an anti-PD-L1 antibody, or an antigen-binding portion thereof. A method for identifying a subject having an HPV-positive SCCHN-derived tumor suitable for anti-PD-1 antibody or anti-PD-L1 antibody treatment, comprising:

5. The method of any one of claims 1 to 4, wherein the HPV-positive SCCHN comprises a tumor that expresses one or more proteins or nucleotide sequences encoding one or more proteins from HPV.

6. The method according to any one of claims 2 to 5, wherein the measuring comprises identifying the expression of one or more proteins from HPV or nucleotide sequences encoding said one or more proteins.

7. 7. The method of claim 5 or 6, wherein the one or more proteins derived from HPV include p16, Ki-67, cyclin D1, p53, ProEx C, E6, E7, or any combination thereof.

8. 8. The method of claim 7, wherein one or more proteins from HPV are identified by immunohistochemistry, ELISA, Western blot, protein array, or any combination thereof.

9. 7. The method of claim 5 or 6, wherein the nucleotide sequence encodes p16, Ki-67, cyclin D1, p53, ProEx C, E6, E7, or any combination thereof.

10. 10. The method of claim 8 or 9, wherein the nucleotide sequence is identified by in situ hybridization, DNA or RNA array or nucleotide hybridization techniques, tumor sequencing techniques, quantitative polymerase chain reaction (PCR), or any combination thereof.

11. 11. The method of any of claims 1 to 10, wherein the HPV comprises HPV subtypes 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, 68, or any combination thereof.

12. The method of any one of claims 1 to 11, wherein more than about 70% of the tumor cells show strong and widespread nuclear and cytoplasmic staining by immunohistochemistry for p16.

13. The method of any one of claims 2 to 12, wherein the sample comprises a primary tumor or a metastatic lymph node.

14. The method of any one of claims 1 to 13, wherein the tumor further expresses PD-L1.

15. The method of any of claims 1-14, wherein the anti-PD-1 antibody, or antigen-binding portion thereof, cross-competes with nivolumab for binding to human PD-1.

16. The method of any of claims 1 to 15, wherein the anti-PD-1 antibody or antigen-binding portion thereof is a chimeric, humanized, or human monoclonal antibody or portion thereof.

17. The method of any of claims 1-16, wherein the anti-PD-1 antibody, or antigen-binding portion thereof, comprises a heavy chain constant region that is of the human IgG1 or IgG4 isotype.

18. The method of any one of claims 1 to 17, wherein the anti-PD-1 antibody is nivolumab.

19. The method of any one of claims 1 to 17, wherein the anti-PD-1 antibody is pembrolizumab.

20. 15. The method of any of claims 1-14, wherein the anti-PD-L1 antibody, or antigen-binding portion thereof, competes with the binding of BMS-936559, MPDL3280A, MEDI4736, or MSB0010718C for binding to human PD-L1.

21. The method of any of claims 1 to 14 and 20, wherein the anti-PD-L1 antibody or antigen-binding portion thereof is a chimeric, humanized, or human monoclonal antibody or portion thereof.

22. 22. The method of any of claims 1-14 and 20-21, wherein the anti-PD-L1 antibody, or antigen-binding portion thereof, comprises a heavy chain constant region that is of the human IgG1 or IgG4 isotype.

23. The method of any one of claims 1 to 14 and 20 to 22, wherein the anti-PD-L1 antibody is BMS-936559.

24. The method of any one of claims 1 to 14 and 20 to 22, wherein the anti-PD-L1 antibody is MPDL3280A.

25. The method of any one of claims 1 to 14 and 20 to 22, wherein the anti-PD-L1 antibody is MEDI4736.

26. The method of any one of claims 1 to 14 and 20 to 22, wherein the anti-PD-L1 antibody is MSB0010718C.

27. 27. The method of any of claims 1 to 26, wherein the anti-PD-1 antibody, or antigen-binding portion thereof, or anti-PD-L1 antibody, or antigen-binding portion thereof, is administered at a dose ranging from at least about 80 mg to at least about 800 mg, or from at least about 0.1 mg / kg to at least about 10.0 mg / kg of body weight.

28. 28. The method of claim 27, wherein the anti-PD-1 antibody, or antigen-binding portion thereof, or anti-PD-L1 antibody, or antigen-binding portion thereof, is administered in a dose of at least about 3 mg / kg body weight or 240 mg approximately once every two weeks.

29. 29. The method of any of claims 1-28, wherein the anti-PD-1 antibody or antigen-binding portion thereof, or anti-PD-L1 antibody or antigen-binding portion thereof, is administered for as long as treatment proves effective or until unmanageable toxicity or disease progression occurs.

30. 30. The method of any of claims 14 to 29, wherein the tumor has PD-L1 expression that is at least about 1%.

31. The method of any of claims 14 to 30, wherein the tumor has PD-L1 expression that is at least about 5% or about 10%.

32. 32. The method of any of claims 1-31, wherein the subject exhibits an overall survival of at least about 10 months, at least about 11 months, at least about 12 months, at least about 13 months, at least about 14 months, at least about 15 months, at least about 16 months, at least about 17 months, at least about 18 months, at least about 19 months, at least about 20 months, at least about 21 months, at least about 22 months, at least about 23 months, at least about 2 years, at least about 3 years, at least about 4 years, or at least about 5 years after administration.

33. The method of any one of claims 1 to 32, wherein the anti-PD-1 antibody or portion thereof or the anti-PD-L1 antibody or portion thereof is formulated for intravenous administration.

34. 34. The method of any of claims 1 to 33, wherein the anti-PD-1 antibody, or antigen-binding portion thereof, or the anti-PD-L1 antibody, or antigen-binding portion thereof, is administered in a subtherapeutic dose.

35. The method of any one of claims 1 to 34, further comprising the administration of one or more additional anti-cancer agents.

36. 36. The method of claim 35, wherein the anti-cancer agent is selected from the group consisting of an antibody or antigen-binding portion thereof that specifically binds to CTLA-4 and inhibits CTLA-4 activity, a chemotherapeutic agent, a platinum-based doublet chemotherapy, a tyrosine kinase inhibitor, an anti-VEGF inhibitor, or any combination thereof.

37. 36. The method of claim 35, wherein the anti-cancer agent is an antibody or antigen-binding portion thereof that specifically binds to CTLA-4 and inhibits CTLA-4 activity.

38. 1. A kit for treating a subject having an HPV-positive SCCHN-derived tumor, comprising: (a) an anti-PD-1 antibody, or an antigen-binding portion thereof, or an anti-PD-L1 antibody, or an antigen-binding portion thereof; (b) instructions for determining HPV positivity of the tumor, and if the tumor is positive for HPV, administering to the subject an anti-PD-1 antibody, or antigen-binding portion thereof, or an anti-PD-L1 antibody, or antigen-binding portion thereof, in accordance with any one of claims 1 to 37. kit.

39. 39. The kit of claim 38, further comprising a reagent for determining the HPV positivity of a tumor.

40. 40. The kit of claim 39, wherein HPV positivity is determined by expression of HPV p16.

41. A method of treating a subject having a tumor derived from human papillomavirus (HPV)-negative squamous cell carcinoma of the head and neck (SCCHN), comprising administering to the subject a therapeutically effective amount of an antibody or antigen-binding portion thereof that specifically binds to programmed death-1 receptor (PD-1) or programmed death-ligand 1 (PD-L1) and inhibits PD-1 activity (an "anti-PD-1 antibody") or PD-L1 activity (an "anti-PD-L1 antibody"), respectively.

42. (i) measuring the level of HPV in a sample from a subject, wherein the subject is negative for HPV; and (ii) administering to the subject a therapeutically effective amount of an anti-PD-1 antibody, or an antigen-binding portion thereof, or an anti-PD-L1 antibody, or an antigen-binding portion thereof.

10. A method of treating a subject having an HPV-negative SCCHN-derived tumor, comprising:

43. A method for identifying a subject having an HPV-negative SCCHN-derived tumor suitable for anti-PD-1 antibody or anti-PD-L1 antibody treatment, comprising measuring the level of HPV in a sample from the subject, wherein the subject is negative for HPV, and administering a therapeutically effective amount of an anti-PD-1 antibody, or an antigen-binding portion thereof, or an anti-PD-L1 antibody, or an antigen-binding portion thereof.

44. (i) measuring the level of HPV in a sample from a subject, wherein the subject is negative for HPV; and (ii) administering to the subject a therapeutically effective amount of an anti-PD-1 antibody, or an antigen-binding portion thereof, or an anti-PD-L1 antibody, or an antigen-binding portion thereof. A method for identifying a subject having an HPV-negative SCCHN-derived tumor suitable for anti-PD-1 antibody or anti-PD-L1 antibody treatment, comprising:

45. The method of any of claims 41 to 44, wherein the HPV-negative SCCHN comprises a tumor that expresses one or more proteins or nucleotide sequences encoding one or more proteins from HPV.

46. The method according to any one of claims 42 to 45, wherein the measuring comprises identifying the expression of one or more proteins from HPV or a nucleotide sequence encoding said one or more proteins.

47. 47. The method of claim 45 or 46, wherein the one or more proteins derived from HPV comprise p16, Ki-67, cyclin D1, p53, ProEx C, E6, E7, or any combination thereof.

48. 48. The method of claim 47, wherein one or more proteins from HPV are identified by immunohistochemistry, ELISA, Western blot, protein array, or any combination thereof.

49. 47. The method of claim 45 or 46, wherein the nucleotide sequence encodes p16, Ki-67, cyclin D1, p53, ProEx C, E6, E7, or any combination thereof.

50. 50. The method of claim 48 or 49, wherein the nucleotide sequence is identified by in situ hybridization, DNA or RNA array or nucleotide hybridization techniques, tumor sequencing techniques, quantitative polymerase chain reaction (PCR), or any combination thereof.

51. 51. The method of any of claims 41 to 50, wherein the HPV comprises HPV subtypes 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, 68, or any combination thereof.

52. 52. The method of any one of claims 41 to 51, wherein no more than about 30% of the tumor cells show strong and widespread nuclear and cytoplasmic staining by immunohistochemistry for p16.

53. The method of any of claims 42 to 52, wherein the sample comprises a primary tumor or a metastatic lymph node.

54. The method of any one of claims 41 to 53, wherein the tumor further expresses PD-L1.

55. The method of any of claims 41-54, wherein the anti-PD-1 antibody, or antigen-binding portion thereof, cross-competes with nivolumab for binding to human PD-1.

56. 56. The method of any of claims 41-55, wherein the anti-PD-1 antibody, or antigen-binding portion thereof, is a chimeric, humanized, or human monoclonal antibody, or portion thereof.

57. 57. The method of any of claims 41-56, wherein the anti-PD-1 antibody, or antigen-binding portion thereof, comprises a heavy chain constant region that is of the human IgG1 or IgG4 isotype.

58. The method of any one of claims 41 to 57, wherein the anti-PD-1 antibody is nivolumab.

59. The method of any one of claims 41 to 57, wherein the anti-PD-1 antibody is pembrolizumab.

60. 55. The method of any of claims 41-54, wherein the anti-PD-L1 antibody, or antigen-binding portion thereof, competes with the binding of BMS-936559, MPDL3280A, MEDI4736, or MSB0010718C for binding to human PD-L1.

61. 61. The method of any of claims 41 to 54 and 60, wherein the anti-PD-L1 antibody or antigen-binding portion thereof is a chimeric, humanized, or human monoclonal antibody or portion thereof.

62. 62. The method of any of claims 41-54 and 60-61, wherein the anti-PD-L1 antibody, or antigen-binding portion thereof, comprises a heavy chain constant region that is of the human IgG1 or IgG4 isotype.

63. The method of any one of claims 41 to 54 and 60 to 62, wherein the anti-PD-L1 antibody is BMS-936559.

64. The method of any one of claims 41 to 54 and 60 to 62, wherein the anti-PD-L1 antibody is MPDL3280A.

65. The method of any one of claims 41 to 44 and 60 to 62, wherein the anti-PD-L1 antibody is MEDI4736.

66. The method of any one of claims 41 to 54 and 60 to 62, wherein the anti-PD-L1 antibody is MSB0010718C.

67. 67. The method of any of claims 41-66, wherein the anti-PD-1 antibody, or antigen-binding portion thereof, or the anti-PD-L1 antibody, or antigen-binding portion thereof, is administered at a dose ranging from at least about 80 mg to at least about 800 mg, or from at least about 0.1 mg / kg to at least about 10.0 mg / kg of body weight.

68. 68. The method of claim 67, wherein the anti-PD-1 antibody, or antigen-binding portion thereof, or anti-PD-L1 antibody, or antigen-binding portion thereof, is administered in a dose of at least about 3 mg / kg body weight or 240 mg approximately once every two weeks.

69. 69. The method of any of claims 41-68, wherein the anti-PD-1 antibody or antigen-binding portion thereof, or anti-PD-L1 antibody or antigen-binding portion thereof, is administered for as long as treatment proves effective or until unmanageable toxicity or disease progression occurs.

70. 70. The method of any of claims 41-69, wherein the tumor has PD-L1 expression that is at least about 1%.

71. 71. The method of any of claims 41 to 70, wherein the tumor has PD-L1 expression that is at least about 5% or about 10%.

72. 72. The method of any of claims 41-71, wherein the subject exhibits an overall survival of at least about 10 months, at least about 11 months, at least about 12 months, at least about 13 months, at least about 14 months, at least about 15 months, at least about 16 months, at least about 17 months, at least about 18 months, at least about 19 months, at least about 20 months, at least about 21 months, at least about 22 months, at least about 23 months, at least about 2 years, at least about 3 years, at least about 4 years, or at least about 5 years after administration.

73. The method of any one of claims 41 to 72, wherein the anti-PD-1 antibody or portion thereof or the anti-PD-L1 antibody or portion thereof is formulated for intravenous administration.

74. 74. The method of any of claims 41-73, wherein the anti-PD-1 antibody, or antigen-binding portion thereof, or the anti-PD-L1 antibody, or antigen-binding portion thereof, is administered in a subtherapeutic dose.

75. 75. The method of any of claims 41 to 74, further comprising the administration of one or more additional anti-cancer agents.

76. 76. The method of claim 75, wherein the anti-cancer agent is selected from the group consisting of an antibody or antigen-binding portion thereof that specifically binds to CTLA-4 and inhibits CTLA-4 activity, a chemotherapeutic agent, a platinum-based doublet chemotherapy, a tyrosine kinase inhibitor, an anti-VEGF inhibitor, or any combination thereof.

77. 76. The method of claim 75, wherein the anti-cancer agent is an antibody or antigen-binding portion thereof that specifically binds to CTLA-4 and inhibits CTLA-4 activity.

78. 1. A kit for treating a subject having an HPV-negative SCCHN-derived tumor, comprising: (a) an anti-PD-1 antibody, or an antigen-binding portion thereof, or an anti-PD-L1 antibody, or an antigen-binding portion thereof; (b) instructions for HPV negativity determination of the tumor, and if the tumor is negative for HPV, administering to the subject an anti-PD-1 antibody, or antigen-binding portion thereof, or an anti-PD-L1 antibody, or antigen-binding portion thereof, in accordance with any of claims 41-77. kit.

79. 79. The kit of claim 78, further comprising a reagent for determining the HPV negativity of a tumor.

80. 80. The kit of claim 79, wherein HPV negativity is determined by expression of HPV p16.

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