Methods of Treating Skin Cancer by Administering PD-1 Inhibitors

Abstract

To provide pharmaceutical compositions for treating cancer in a subject, ameliorating at least one symptom or indication of the cancer, or inhibiting the growth of the cancer.SOLUTION: A pharmaceutical composition for use in a method of treating a tumor or inhibiting the growth of a tumor comprises an isolated antibody or antigen-binding fragment thereof that specifically binds programmed death 1 (PD-1). The method comprises: (a) selecting a patient with a brain cancer; and (b) administering to the patient a therapeutically effective amount of the antibody or antigen-binding fragment thereof that specifically binds PD-1.SELECTED DRAWING: None

Description

[Technical field]

[0001] This application was filed as a PCT international patent application on May 12, 2017, and claims the benefit of priority to U.S. Provisional Patent Applications Nos. 62 / 335,743, filed May 13, 2016; 62 / 340,142, filed May 23, 2016; 62 / 348,546, filed June 10, 2016; 62 / 350,305, filed June 15, 2016; 62 / 364,920, filed July 21, 2016; 62 / 374,020, filed August 12, 2016; and 62 / 451,274, filed January 27, 2017, the disclosures of each of which are incorporated herein by reference in their entireties.

[0002] Sequence Listing This application contains a sequence listing in electronic format as a text file entitled "Sequence Listing", which was created on May 12, 2017, and has a size of 8.63 kilobytes (KB). The contents of the text file "Sequence Listing" are incorporated herein by reference.

[0003] The present invention relates to a method of treating skin cancer comprising administering to a subject in need thereof a therapeutically effective amount of an antibody that specifically binds to the Programmed Cell Death 1 (PD-1) receptor. [Background technology]

[0004] Skin cancer is the most common cancer in the United States (Non-Patent Document 1). In 2012, an estimated 5.4 million cases of non-melanoma skin cancer, including basal cell carcinoma and squamous cell carcinoma, were diagnosed in the United States (Non-Patent Document 2). Cutaneous squamous cell carcinoma (CSCC) is the second most common malignant tumor in the United States after basal cell carcinoma (BCC) (Non-Patent Document 3). Risk factors for CSCC include UV exposure, advanced age, and immunosuppression (Non-Patent Document 4; Non-Patent Document 5). Although the majority of individuals with a diagnosis of CSCC or BCC have a very favorable prognosis, CSCC is more invasive and prone to recurrence than BCC. Individuals diagnosed with CSCC, unlike those diagnosed with BCC, have an increased mortality rate compared to age-matched controls (Non-Patent Document 6).

[0005] Surgical resection is the mainstay of clinical management of CSCC. The primary goal is complete resection of the cancer, with acceptable cosmetic outcomes being a secondary goal. Factors associated with poor prognosis in CSCC include tumor size >2 cm, tumor depth >2 mm, perineural invasion, host immunosuppression, and recurrent disease. For the minority of patients who develop unresectable locally recurrent or metastatic disease, treatment options are limited. Patients may be administered postoperative radiation therapy. Chemotherapy is not an attractive option for many patients due to safety and tolerability concerns.

[0006] The most common clinical subtype is nodular BCC. The next most common clinical subtypes are superficial, sclerosing (fibrous), and fibroepithelial. Most patients are cured by surgery, but a minority develop unresectable locally advanced or metastatic disease. Virtually all BCCs are characterized by aberrant signaling of the Hedgehog signaling pathway, most commonly due to sporadic loss-of-function mutations in the gene encoding the tumor suppressor protein patched homologue (PTCH). PTCH mutations result in loss of patched-mediated inhibition of the G protein-coupled receptor smoothened (SMO), which enhances downstream signaling and leads to uncontrolled cell proliferation (Non-Patent Document 7). Oncogenic Role of SMO in BCC This recognition led to the development of orally available SMO inhibitors, vismodegib and sonidegib, commonly referred to as hedgehog inhibitors (HHIs). In addition to the adverse side effects of HHIs, patients who progressed on one HHI (vismodegib) were found to have no tumor inhibition upon subsequent treatment with another HHI (sonidegib) (Non-Patent Document 8). There are no agents approved for BCC in patients who have experienced disease progression during HHI treatment or who are intolerant to previous HHI treatments. [Prior art documents] [Non-patent literature]

[0007] [Non-Patent Document 1] Guy et al., 2015, Am.J.Prev.Med.48:183~7 [Non-Patent Document 2] Rogers et al., 2015, JAMA Dermatol., electronic publication, April 30, 2015. [Non-Patent Document 3] Karia et al., 2013, J. Am. Acad. Dermatol. 68:957-966 [Non-Patent Document 4] Alam et al., 2001, New Engl. J. Med. 344 (975-983) [Non-Patent Document 5] Madan 2010, Lancet 375:673~685 [Non-Patent Document 6] Rees et al., 2015, Int. J. Cancer 137:878-84 [Non-Patent Document 7] Sekulic et al., 2016, Cell 164:831 [Non-Patent Document 8] Danial et al., 2016, Clin. Cancer Res. 22:1325-29 Summary of the Invention [Problem to be solved by the invention]

[0008] Therefore, there is a need for safe and effective systemic treatments for skin cancers, including CSCC and BCC. [Means for solving the problem]

[0009] In certain embodiments, the present invention provides a method of treating, ameliorating at least one symptom or indication of, or inhibiting the growth of cancer in a subject. The method according to this aspect of the invention comprises administering to a subject in need thereof a therapeutically effective amount of an antibody or antigen-binding fragment thereof that specifically binds to programmed cell death 1 (PD-1), optionally in combination with radiation therapy.

[0010] In certain embodiments, the invention includes a method of treating cancer, including solid tumors, comprising selecting a subject with cancer and administering one or more doses of an anti-PD-1 antibody in combination with one or more doses of radiation therapy. In certain embodiments, administration of the combination provides enhanced therapeutic or anti-tumor efficacy compared to administration of either the antibody or radiation alone.

[0011] In certain embodiments of the invention, methods are provided for treating cancer, alleviating at least one symptom or indication of cancer, or inhibiting the growth of cancer in a subject. In certain embodiments of the invention, methods are provided for slowing tumor growth or preventing tumor recurrence. In certain embodiments of the invention, methods are provided for extending overall survival or progression-free survival of a cancer patient. The method according to this aspect of the invention comprises sequentially administering one or more doses of a therapeutically effective amount of an antibody or antigen-binding fragment thereof that specifically binds to PD-1. In one embodiment, the anti-PD-1 antibody is administered in combination with radiation therapy. The drugs are administered in combination.

[0012] In certain embodiments, the cancer or tumor is a solid tumor or malignant tumor. In certain embodiments, the solid tumor is selected from the group consisting of colorectal cancer, ovarian cancer, prostate cancer, breast cancer, brain cancer, cervical cancer, bladder cancer, anal cancer, uterine cancer, colon cancer, liver cancer, pancreatic cancer, lung cancer, endometrial cancer, bone cancer, testicular cancer, skin cancer, kidney cancer, stomach cancer, esophageal cancer, head and neck cancer, salivary gland cancer, and myeloma.

[0013] In certain embodiments, the anti-PD-1 antibody is administered as a "first line" treatment to a cancer patient who has not previously received systemic treatment for the cancer. In certain embodiments, the anti-PD-1 antibody is administered as a "second line" treatment to a cancer (e.g., metastatic cancer) patient who has previously been treated with "standard of care," including but not limited to chemotherapy, surgery, and radiation.

[0014] One embodiment of the present invention relates to an anti-PD-1 antibody for use in treating skin cancer. In certain embodiments, the skin cancer is non-melanoma skin cancer, including but not limited to cutaneous squamous cell carcinoma and basal cell carcinoma. The anti-PD-1 antibody may be administered to patients with metastatic or locally advanced cutaneous squamous cell carcinoma, as described herein. In certain embodiments, the anti-PD-1 antibody is administered to patients with advanced basal cell carcinoma who are intolerant to hedgehog pathway inhibitors (e.g., vismodegib, sonidegib) or who have been treated with hedgehog pathway inhibitors but show progression, as described herein.

[0015] In certain embodiments, each dose of the anti-PD-1 antibody comprises 0.1-20 mg per kg of the subject's body weight. In certain embodiments, each dose of the anti-PD-1 antibody comprises 0.3, 1, 3, 5, or 10 mg per kg of the subject's body weight. In certain embodiments, each dose of the anti-PD-1 antibody comprises 20-600 mg. In one embodiment, each dose of the anti-PD-1 antibody comprises about 200 mg. In one embodiment, each dose of the anti-PD-1 antibody comprises about 250 mg. In one embodiment, each dose of the anti-PD-1 antibody comprises about 350 mg.

[0016] In certain embodiments, radiation therapy is administered in one or more doses. In certain embodiments, each dose of radiation therapy comprises 2 to 100 Gray (Gy). In certain embodiments, radiation therapy is hypofractionated radiation therapy. In certain embodiments, radiation therapy comprises 2 to 12 fractions.

[0017] In certain embodiments, the methods of the invention involve administering a therapeutically effective amount of an anti-PD-1 antibody prior to, concurrently with, or following radiation therapy, hi one embodiment, the methods of the invention involve administering an anti-PD-1 antibody prior to a dose of radiation therapy.

[0018] In certain embodiments, the methods of the invention comprise administering 0-50 therapeutic doses each of an anti-PD-1 antibody, with each dose being administered 0.5-12 weeks after the immediately preceding dose. In one embodiment, each dose is administered 1 week after the immediately preceding dose. In one embodiment, each dose is administered 2 weeks after the immediately preceding dose. In one embodiment, each dose is administered 3 weeks after the immediately preceding dose.

[0019] In certain embodiments, one or more doses of an anti-PD-1 antibody, and optionally radiation therapy, are included in a treatment cycle. The method according to this aspect of the invention comprises administering to a subject in need thereof at least one treatment cycle comprising one or more doses of an anti-PD-1 antibody. In certain embodiments, up to 12 treatment cycles are administered to a subject in need thereof. In certain embodiments, at least A treatment cycle further comprises one or more doses of radiation therapy. In certain embodiments, radiation therapy is administered in only one treatment cycle. In certain embodiments, radiation therapy is hypofractionated radiation therapy. In certain embodiments, the anti-PD-1 antibody is administered prior to radiation therapy.

[0020] In certain embodiments, the anti-PD-1 antibody and radiation therapy are administered in combination with an additional therapeutic agent or treatment (e.g., cyclophosphamide, or any agent or treatment disclosed herein).

[0021] In certain embodiments, the treatment results in one or more therapeutic effects selected from the group consisting of tumor regression, abscopal effect, inhibition of tumor metastasis, reduction in metastatic lesions over time, reduction in the use of chemotherapy or cytotoxic agents, reduction in tumor burden, prolonged progression-free survival, prolonged overall survival, complete response, partial response, and stable disease.

[0022] According to certain embodiments, an anti-PD-1 antibody or antigen binding protein comprises a heavy chain complementarity determining region (HCDR) of a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 1, and a light chain CDR of a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 2. One such type of antigen binding protein that can be used in the context of the methods of the invention is an anti-PD-1 antibody, such as REGN2810.

[0023] In certain embodiments, the present invention provides the use of an anti-PD-1 antibody or an antigen-binding fragment thereof in the manufacture of a medicament for treating or inhibiting the growth of cancer in a subject, including a human. In certain embodiments, the cancer is a solid tumor. In certain embodiments, the cancer is colorectal cancer, ovarian cancer, prostate cancer, breast cancer, brain cancer, cervical cancer, bladder cancer, anal cancer, uterine cancer, colon cancer, liver cancer, pancreatic cancer, lung cancer, endometrial cancer, bone cancer, testicular cancer, skin cancer, kidney cancer, gastric cancer, esophageal cancer, head and neck cancer, salivary gland cancer, or myeloma.

[0024] In certain embodiments, the present invention provides the use of an anti-PD-1 antibody or an antigen-binding fragment thereof in the manufacture of a medicament for treating or inhibiting the growth of cancer in a subject, including a human, in combination with radiation therapy. In certain embodiments, the cancer is a solid tumor. In certain embodiments, the cancer is colorectal cancer, ovarian cancer, prostate cancer, breast cancer, brain cancer, cervical cancer, bladder cancer, anal cancer, uterine cancer, colon cancer, liver cancer, pancreatic cancer, lung cancer, endometrial cancer, bone cancer, testicular cancer, skin cancer, kidney cancer, gastric cancer, esophageal cancer, head and neck cancer, salivary gland cancer, or myeloma.

[0025] In one aspect, the invention provides a kit for treating a subject suffering from cancer, the kit comprising: (a) a dose of an antibody or antigen-binding fragment thereof that specifically binds and inhibits PD-1; and (b) instructions for using the anti-PD-1 antibody to treat the subject according to the methods disclosed herein. In certain embodiments, the cancer is selected from the group consisting of colorectal cancer, ovarian cancer, prostate cancer, breast cancer, brain cancer, cervical cancer, bladder cancer, anal cancer, uterine cancer, colon cancer, liver cancer, pancreatic cancer, lung cancer, endometrial cancer, bone cancer, testicular cancer, skin cancer, kidney cancer, gastric cancer, esophageal cancer, head and neck cancer, salivary gland cancer, and myeloma.

[0026] Other embodiments of the invention will become apparent upon review of the following detailed description. [Brief description of the drawings]

[0027] [Figure 1] FIG. 1 shows the study design involving administration of anti-PD-1 antibody and radiation (XRT) in mice implanted with MC38 tumors (study described in Example 1 herein). [Diagram 2] FIG. 1 shows the mean tumor growth in mice treated with isotype control antibody (●), anti-PD-1 antibody (■), isotype control + radiation (XRT) (▲), or anti-PD-1 antibody + XRT (▼) in the study described in Example 1 herein. [Diagram 3] FIG. 1 shows overall survival of mice treated with isotype control antibody (●), anti-PD-1 antibody (■), isotype control + radiation (XRT) (▲), or anti-PD-1 antibody + XRT (▼) in the study described in Example 1 herein. [Figure 4] FIG. 1 shows the study design involving administration of anti-PD-1 antibody and radiation (XRT) in mice implanted with B16F10.9 tumors (study described in Example 2 herein). [Diagram 5]FIG. 1 shows the mean tumor growth in mice treated with isotype control antibody (●), anti-PD-1 antibody (■), isotype control + radiation (XRT) (◆), or anti-PD-1 antibody + XRT (○) in the study described in Example 2 herein. [Figure 6] FIG. 1 shows overall survival of mice treated with isotype control antibody (●), anti-PD-1 antibody (■), isotype control + radiation (XRT) (◆), or anti-PD-1 antibody + XRT (○) in the study described in Example 2 herein. [Figure 7] FIG. 1 shows the study design involving administration of anti-PD-1 antibody and radiation (XRT) in mice implanted with MC38 tumors (study described in Example 4 herein). [Figure 8] FIG. 1 shows the mean growth of primary tumors in mice treated with isotype control antibody (●), anti-PD-1 antibody (■), isotype control + radiation (XRT) (▲), or anti-PD-1 antibody + XRT (▼) in the study described in Example 4 herein. [Figure 9] FIG. 1 shows overall survival of mice treated with isotype control antibody (●), anti-PD-1 antibody (■), isotype control + radiation (XRT) (▲), or anti-PD-1 antibody + XRT (▼) in the study described in Example 4 herein. [Figure 10] FIG. 1 shows the growth of secondary tumors in mice treated with isotype control antibody (●), anti-PD-1 antibody (■), isotype control + radiation (XRT) (▲), or anti-PD-1 antibody + XRT (▼) in the study described in Example 4 herein. [Figure 11] FIG. 1 shows the study design involving administration of anti-PD-1 antibody, anti-GITR antibody and radiation (XRT) in mice implanted with MC38 tumors (study described in Example 5 herein). [Figure 12]FIG. 1 shows the mean tumor growth in mice treated with an isotype control antibody (●), anti-PD-1 antibody (■), anti-GITR antibody (▲), a combination of anti-PD-1 and anti-GITR antibodies (▼), isotype control + radiation (XRT) (◆), anti-PD-1 antibody + XRT (○), anti-GITR antibody + XRT (□), or a combination of anti-PD-1 antibody, anti-GITR antibody + XRT (△) in the study described in Example 5 herein. [Figure 13] FIG. 1 shows overall survival of mice treated with an isotype control antibody (●), anti-PD-1 antibody (■), anti-GITR antibody (▲), a combination of anti-PD-1 and anti-GITR antibodies (▼), isotype control + radiation (XRT) (◆), anti-PD-1 antibody + XRT (○), anti-GITR antibody + XRT (□), or a combination of anti-PD-1 antibody, anti-GITR antibody + XRT (△) in the study described in Example 5 herein. [Figure 14A] FIG. 1 shows x-ray images of lung metastases in a patient with basal cell carcinoma (BCC) indicated by arrows at baseline, left, and at week 24, right. [Figure 14B] FIG. 1 shows x-ray images of a neck tumor mass in a patient with cutaneous squamous cell carcinoma (CSCC) at baseline, left, and at week 16, right. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0028] Before describing the present invention, it is to be understood that the present invention is not limited to the particular methods and experimental conditions described, as such methods and conditions may vary. Similarly, it is to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, as the scope of the present invention will be limited only by the appended claims.

[0029] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this invention belongs.As used herein, the term "about" when used with reference to a specific recited numerical value means that the value may vary from the recited value by 1% or less.For example, as used herein, the expression "about 100" includes 99 and 101, and all values ​​therebetween (for example, 99.1, 99.2, 99.3, 99.4, etc.).

[0030] Although any methods and materials similar or equivalent to those described herein can be used in the practice of the present invention, the preferred methods and materials are described below. All publications mentioned herein are incorporated by reference herein as if set forth in their entirety.

[0031] Methods for Treating or Inhibiting Cancer Growth The present invention includes a method of treating cancer, ameliorating or reducing the severity of at least one symptom or indication of cancer, or inhibiting the growth of cancer in a subject. The method according to this aspect of the invention includes administering to a subject in need thereof a therapeutically effective amount of an antibody or antigen-binding fragment thereof that specifically binds to PD-1. In certain embodiments, the anti-PD-1 antibody is administered in combination with an anti-tumor therapy (described elsewhere herein). In one embodiment, the anti-tumor therapy is radiation therapy. As used herein, the terms "treat," "treating," and the like refer to alleviating symptoms, temporarily or permanently eliminating the cause of symptoms, slowing or inhibiting tumor growth, reducing tumor cell mass or burden, promoting tumor regression, causing tumor shrinkage, necrosis, and / or disappearance, preventing tumor recurrence, preventing or inhibiting metastasis, inhibiting metastatic tumor growth, and / or prolonging the survival of a subject.

[0032] As used herein, the term "subject in need thereof" refers to a human or non-human mammal that exhibits one or more symptoms or indications of cancer and / or has been diagnosed with cancer, including solid tumors, and is in need of treatment for cancer. In many embodiments, the term "subject" can be used interchangeably with the term "patient." For example, a human subject may be diagnosed with a primary or metastatic tumor and / or with one or more symptoms or indications, including, but not limited to, unexplained weight loss, general weakness, persistent fatigue, loss of appetite, fever, night sweats, bone pain, shortness of breath, abdominal distension, chest pain / tenderness, enlarged spleen, and elevated levels of cancer-related biomarkers (e.g., CA125). This term includes subjects with primary or established tumors. In certain embodiments, the term includes human subjects with and / or in need of treatment for solid tumors, such as colon cancer, breast cancer, lung cancer, prostate cancer, skin cancer, liver cancer, bone cancer, ovarian cancer, cervical cancer, pancreatic cancer, head and neck cancer, and brain cancer. The term includes subjects with primary or metastatic tumors (advanced malignancies). In certain embodiments, the phrase "subject in need thereof" includes patients with solid tumors that are resistant or refractory to previous therapies (e.g., treatment with anti-cancer drugs) or that are inadequately controlled. For example, the phrase includes subjects who have been treated with one or more previous therapies, such as treatment with chemotherapy (e.g., carboplatin or docetaxel). In certain embodiments, the phrase "subject in need thereof" includes patients with solid tumors that have been treated with one or more previous therapies but have subsequently relapsed or metastasized. For example, patients with solid tumors who have been treated with one or more anti-cancer drugs that result in tumor regression, but who subsequently experience a relapse of the cancer that is resistant to the one or more anti-cancer drugs (e.g., chemotherapy-resistant cancer) are treated by the methods of the present invention. The term also includes subjects with solid tumors for whom conventional anti-cancer treatment is not advisable, for example, due to toxic side effects. For example, the term includes patients who have been treated with one or more cycles of chemotherapy that have toxic side effects.

[0033] In certain embodiments, the methods of the present invention may be used to treat patients who exhibit elevated levels of one or more cancer-associated biomarkers (e.g., programmed cell death ligand 1 (PD-L1), CA125, CA19-9, prostate-specific antigen (PSA), lactate dehydrogenase, KIT, carcinoembryonic antigen, epidermal growth factor receptor (EGFR), ALK gene rearrangement). For example, the methods of the present invention include administering a therapeutically effective amount of an anti-PD-1 antibody in combination with radiation therapy to a patient with elevated levels of PD-L1 and / or EGFR. In a preferred embodiment, the methods of the present invention are used in cancer patients selected based on PD-L1 expression in cancer tissue. In certain embodiments, the methods of the present invention are used to treat cancer patients selected based on at least 1%, at least 2%, at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, or at least 50% PD-L1 expression in cancer tissue and / or immune cells. Methods for determining PD-L1 expression in cancer tissue and / or immune cells are well known in the art. In certain embodiments, the expression of PD-L1 in tumor tissue is determined by any assay known in the art, such as an ELISA assay or an immunohistochemistry (IHC) assay, as described in PCT Publication No. WO 2016124558 or WO 2016191751 or US Patent Publication No. 20160305947. In certain embodiments, the expression of PD-L1 is determined by quantifying RNA expression, such as by in situ hybridization, or RT-PCR. In certain embodiments, the expression of PD-L1 is determined by imaging with a labeled anti-PD-L1 antibody, such as by immunopositron emission tomography or iPET (see, e.g., The Oncologist, 12:1379 (2007); Journal of Nuclear Medicine, 52(8):1171 (2011); US Provisional Patent Application No. 62 / 428,672, filed December 1, 2016).

[0034] In certain embodiments, the methods of the invention are used in subjects with solid tumors. The terms "tumor," "cancer," and "malignant tumor" are used interchangeably herein.

[0035] The term "solid tumor" as used herein refers to an abnormal mass of tissue that does not usually contain cysts or liquid areas. Solid tumors can be benign (not cancerous) or malignant (cancer). For the purposes of the present invention, the term "solid tumor" refers to a malignant solid tumor. The term includes various types of solid tumors named for the cell type that forms them, namely sarcomas, carcinomas, and lymphomas. However, the term does not include leukemias. In various embodiments, the term "solid tumor" includes cancers arising from connective or supportive tissues (e.g., bone or muscle) (called sarcomas), cancers arising from the glandular cells of the body and epithelial cells lining the inside of body tissues (called carcinomas), and cancers of lymphatic organs such as lymph nodes, spleen, and thymus (called lymphomas). Cells of the lymphatic system are present in almost every tissue of the body, and thus lymphomas can develop in a wide variety of organs. In certain embodiments, the term "solid tumor" includes cancers including, but not limited to, colorectal cancer, ovarian cancer, prostate cancer, breast cancer, brain cancer, cervical cancer, bladder cancer, anal cancer, uterine cancer, colon cancer, liver cancer, pancreatic cancer, lung cancer, endometrial cancer, bone cancer, testicular cancer, skin cancer, kidney cancer, stomach cancer, esophageal cancer, head and neck cancer, salivary gland cancer, and myeloma. The term "solid tumor" includes cancers including, but not limited to, hepatocellular carcinoma, non-small cell lung cancer, squamous cell carcinoma of the head and neck, basal cell carcinoma, breast cancer, squamous cell carcinoma of the skin, chondrosarcoma, angiosarcoma, cholangiocarcinoma, soft tissue sarcoma, colorectal cancer, melanoma, Merkel cell carcinoma, and glioblastoma multiforme. In certain embodiments, the term "solid tumor" includes more than one solid tumor lesion that exists apart from each other in a subject in need of treatment, such as 2, more than 2, more than 5, more than 10, more than 15, more than 20, or more than 25 lesions. In certain embodiments, more than one lesion exists distal to each other in the same organ. In certain other embodiments, the tumor lesions may exist in different organs.

[0036] In certain embodiments, the present invention includes methods of treating or inhibiting the growth of cancer, including, but not limited to, colorectal cancer, ovarian cancer, prostate cancer, breast cancer, brain cancer, cervical cancer, bladder cancer, anal cancer, uterine cancer, colon cancer, liver cancer, pancreatic cancer, lung cancer, endometrial cancer, bone cancer, testicular cancer, skin cancer, kidney cancer, gastric cancer, esophageal cancer, head and neck cancer, salivary gland cancer, and myeloma. In certain embodiments, the present invention includes methods of treating or inhibiting the growth of cancer, including, but not limited to, hepatocellular carcinoma, non-small cell lung cancer, squamous cell carcinoma of the head and neck, basal cell carcinoma, squamous cell carcinoma of the skin, chondrosarcoma, angiosarcoma, cholangiocarcinoma, soft tissue sarcoma, colorectal cancer, melanoma, Merkel cell carcinoma, and glioblastoma multiforme. In certain embodiments, the present invention includes a method of treating advanced solid tumors, including but not limited to metastatic cutaneous squamous cell carcinoma (CSCC), unresectable locally advanced CSCC, metastatic colorectal cancer, advanced or metastatic hepatocellular carcinoma, advanced non-small cell lung cancer, basal cell carcinoma, recurrent glioblastoma multiforme, castration-recurrent prostate cancer, and any advanced solid tumor that is resistant to first-line therapy. The method according to this aspect includes administering a therapeutically effective amount of an anti-PD-1 antibody. In certain embodiments, the method includes administering a therapeutically effective amount of an anti-PD-1 antibody in combination with an anti-tumor treatment. The anti-tumor treatment includes but is not limited to conventional anti-tumor treatments such as chemotherapy, radiation, and surgery. Other anti-tumor treatments are described elsewhere herein. In one embodiment, the anti-tumor treatment includes radiation therapy. In certain embodiments, one or more doses of an anti-PD-1 antibody are administered to a subject in need thereof, with each dose being administered 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 weeks after the immediately preceding dose. In certain embodiments, each dose contains 0.1-10 mg / kg of subject body weight (e.g., 0.3 mg / kg, 1 mg / kg, 3 mg / kg, or 10 mg / kg). In certain other embodiments, each dose contains 20-600 mg of anti-PD-1 antibody, e.g., 50 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 400 mg, or 500 mg of anti-PD-1 antibody.

[0037] In certain embodiments, the present invention includes methods of treating or inhibiting the growth of cancers with microsatellite instability (MSI). As used herein, the term "microsatellite instability," also known as "MSI," refers to genetic hypermutability caused by alterations in microsatellite repeat sequences or defects in DNA mismatch repair in tumor cells. Microsatellites, also known as simple repeat sequences, are repetitive sequences of DNA that contain repeat units of 1-6 base pairs in length. The length of a microsatellite is highly variable from person to person, contributing to a DNA fingerprint, but each individual has a certain length of microsatellite. MSI results from the inability of mismatch repair (MMR) proteins to repair DNA replication errors. MSI involves DNA polymorphisms in which the replication errors differ in length, not sequence. MSI involves frameshift mutations, either through insertions or deletions, or hypermethylation, which results in gene silencing. It is known in the art that microsatellite instability can lead to colon cancer, gastric cancer, endometrial cancer, ovarian cancer, hepatic bile duct cancer, urinary tract cancer, brain cancer, and skin cancer. The present invention provides a method for treating cancers with MSI, comprising administering to a patient in need thereof a therapeutically effective amount of an anti-PD-1 antibody, optionally in combination with radiation therapy. The present invention includes methods of treatment.

[0038] One embodiment of the present invention relates to an anti-PD-1 antibody (e.g., REGN2810) for use in treating advanced solid tumors with MSI, including but not limited to metastatic colorectal cancer with MSI, metastatic endometrial cancer with MSI, and castration-recurrent prostate cancer with MSI. In certain embodiments, one or more doses of the anti-PD-1 antibody are administered to a subject with an advanced solid tumor with MSI, each dose comprising 0.1-20 mg / kg of the subject's body weight, and each dose administered 0.5-4 weeks after the immediately preceding dose. In certain embodiments, one or more doses of the anti-PD-1 antibody are administered to a subject with an advanced solid tumor with MSI, each dose comprising 20-600 mg of the anti-PD-1 antibody, and each dose administered 0.5-4 weeks after the immediately preceding dose.

[0039] The term "radiotherapy" as used herein, also referred to as "XRT", refers to the use of ionizing radiation to kill cancer cells, generally as part of anti-cancer treatment. X-rays, gamma rays, or charged particles (e.g., protons or electrons) are used to generate ionizing radiation. Radiation therapy may be delivered by a device placed outside the patient's body (external beam radiation therapy) or a source placed inside the patient's body (internal beam radiation therapy or brachytherapy), or by whole-body radioisotopes delivered intravenously or orally (total body radioisotope therapy). Radiation therapy may be planned and administered in conjunction with imaging-based techniques such as computed tomography (CT), magnetic resonance imaging (MRI) to precisely determine the dose and location of radiation to be administered. In various embodiments, radiation therapy is selected from the group consisting of total body radiation therapy, conventional external beam radiation therapy, stereotactic radiosurgery, stereotactic radiation therapy, three-dimensional conformal radiation therapy, intensity-modulated radiation therapy, image-guided radiation therapy, tomotherapy, brachytherapy, and total body radiation therapy. In certain embodiments, depending on its intent, radiation therapy is curative, supportive, or palliative. In certain embodiments, the term "radiation therapy" refers to hypofractionated radiation therapy. Hypofractionated radiation therapy refers to radiation therapy in which the radiation dose is contained in two or more fractions. In various embodiments, each fraction contains 2-20 Gy. For example, a radiation dose of 50 Gy may be divided into 10 fractions, each containing 5 Gy. In certain embodiments, two or more fractions are administered consecutively or on consecutive days. In certain other embodiments, two or more fractions are administered once every 2 days, once every 3 days, once every 4 days, once every 5 days, once every 6 days, once every 7 days, or combinations thereof.

[0040] According to certain embodiments, the invention includes a method of treating a tumor or slowing or inhibiting tumor growth. In certain embodiments, the invention includes a method of promoting tumor regression. In certain embodiments, the invention includes a method of reducing tumor cell mass or reducing tumor burden. In certain embodiments, the invention includes a method of preventing tumor recurrence. The method according to this aspect of the invention includes administering a therapeutically effective amount of an anti-PD-1 antibody in combination with radiation therapy continuously to a subject in need thereof, where the antibody is administered to the subject in multiple doses, for example, as part of a specific therapeutic dosing regimen. For example, the therapeutic dosing regimen may include administering one or more doses of an anti-PD-1 antibody to a subject about once a day, once every two days, once every three days, once every four days, once every five days, once every six days, once a week, once every two weeks, once every three weeks, once every four weeks, once a month, once every two months, once every three months, once every four months, or less frequently. In certain embodiments, one or more doses of an anti-PD-1 antibody are administered in combination with one or more doses of radiation therapy, where the one or more doses of radiation are administered to a subject about once every day, once every two days, once every three days, once every four days, once every five days, once every six days, once every week, once every two weeks, once every three weeks, once every four weeks, once every month, once every two months, once every three months, once every four months, or less frequently.

[0041] In certain embodiments, one or more doses are included in a treatment cycle. The method according to this embodiment includes administering at least one treatment cycle to a subject in need thereof, the at least one treatment cycle including 1-10 doses of an anti-PD-1 antibody and optionally including one or more doses of radiation therapy. In certain embodiments, 2-12 treatment cycles are administered to a subject in need thereof.

[0042] In certain embodiments, the invention provides methods of increasing anti-tumor efficacy or increasing tumor inhibition. The method according to this aspect of the invention comprises administering a therapeutically effective amount of an anti-PD-1 antibody to a subject having a solid tumor prior to administering a dose of radiation, where the anti-PD-1 antibody may be administered about 1 day, more than 1 day, more than 2 days, more than 3 days, more than 4 days, more than 5 days, more than 6 days, more than 7 days, or more than 8 days prior to the radiation therapy. In certain embodiments, the method provides an increase in tumor inhibition of, for example, about 20%, more than 20%, more than 30%, more than 40%, more than 50%, more than 60%, more than 70%, or more than 80%, compared to a subject administered a dose of radiation prior to the anti-PD-1 antibody. In certain embodiments, the radiation therapy comprises hypofractionated radiation therapy.

[0043] In certain embodiments, the present invention provides a method of treating cancer, comprising: selecting a subject having a first tumor lesion and at least a second tumor lesion; and administering one or more doses of an anti-PD-1 antibody in combination with radiation therapy, such that both lesions are treated. In certain embodiments, the method comprises administering radiation therapy to the first tumor lesion, but not to the second tumor lesion, whereby both tumor lesions experience tumor regression (abscopal effect). In certain embodiments, the method comprises selecting a subject having a first tumor lesion and at least a second tumor lesion; and administering one or more doses of an anti-PD-1 antibody in combination with hypofractionated radiation therapy, whereby hypofractionated radiation therapy is administered to the first lesion, but not to the second lesion, whereby both lesions are treated by such administration. In certain embodiments, the anti-PD-1 antibody is administered prior to radiation therapy.

[0044] In certain embodiments, the present invention includes a method of treating cancer, comprising administering one or more sub-therapeutic doses of an anti-PD-1 antibody in combination with one or more anti-tumor therapies, such as radiation therapy, to a subject in need thereof. As defined elsewhere herein, the term "sub-therapeutic dose" refers to a dose that is lower than a therapeutic dose and can be used to reduce the toxicity of the administered treatment. In certain embodiments, administering a sub-therapeutic dose of an anti-PD-1 antibody in combination with radiation therapy provides therapeutic anti-tumor efficacy compared to administering a sub-therapeutic dose of an anti-PD-1 antibody alone. In certain other embodiments, the method of the present invention includes administering a therapeutically effective amount of an anti-PD-1 antibody in combination with a sub-therapeutic dose of an anti-tumor treatment, such as chemotherapy or radiation. For example, a therapeutically effective amount of an anti-PD-1 antibody may be administered in combination with a sub-therapeutic dose of cyclophosphamide to increase efficacy compared to either monotherapy.

[0045] In certain embodiments, the invention includes a method of inhibiting, slowing, or stopping tumor metastasis or tumor invasion to peripheral organs. The method according to this aspect includes administering to a subject in need thereof a therapeutically effective amount of an anti-PD-1 antibody. In certain embodiments, the anti-PD-1 antibody is administered in combination with radiation. In one embodiment, the radiation is hypofractionated radiation. In one embodiment, the radiation is administered after administration of one or more doses of the anti-PD-1 antibody.

[0046] In certain embodiments, the methods of the invention comprise administering a therapeutically effective amount of an anti-PD-1 antibody to a subject having an advanced solid tumor. In certain embodiments, the advanced solid tumor is metastatic lung cancer, head and neck cancer, hepatocellular carcinoma, or breast cancer. In certain other embodiments, the advanced solid tumor is cutaneous squamous cell carcinoma. In certain embodiments, the advanced solid tumor is a cutaneous squamous cell carcinoma. In certain embodiments, the advanced solid tumor is indolent or aggressive. In certain embodiments, the subject has not responded to previous treatment or has relapsed after previous treatment (e.g., with carboplatin). In certain further embodiments, the subject has an advanced solid tumor that is resistant to first-line chemotherapy. In certain further embodiments, the method of the invention further comprises administering radiation and / or cyclophosphamide to the subject with an advanced solid tumor.

[0047] In certain embodiments, the present invention includes methods of treating or inhibiting the growth of cancer, including but not limited to colorectal cancer, ovarian cancer, prostate cancer, breast cancer, brain cancer, cervical cancer, bladder cancer, anal cancer, uterine cancer, colon cancer, liver cancer, pancreatic cancer, lung cancer, endometrial cancer, bone cancer, testicular cancer, skin cancer, kidney cancer, gastric cancer, esophageal cancer, head and neck cancer, salivary gland cancer, and myeloma. In certain embodiments, the present invention includes methods of treating or inhibiting the growth of cancer, including but not limited to hepatocellular carcinoma, non-small cell lung cancer, head and neck squamous cell carcinoma, basal cell carcinoma, cutaneous squamous cell carcinoma, chondrosarcoma, angiosarcoma, cholangiocarcinoma, soft tissue sarcoma, colorectal cancer, melanoma, Merkel cell carcinoma, and glioblastoma multiforme. In certain embodiments, the present invention comprises a method of treating advanced solid tumors, including but not limited to metastatic cutaneous squamous cell carcinoma (CSCC), unresectable locally advanced CSCC, metastatic colorectal cancer, advanced or metastatic hepatocellular carcinoma, advanced non-small cell lung cancer, recurrent glioblastoma multiforme, newly diagnosed glioblastoma multiforme, post-castration recurrent prostate cancer, and any advanced solid tumor that is refractory to first line therapy.

[0048] According to one embodiment, the present invention comprises a method of treating or inhibiting tumor growth comprising the steps of: (a) selecting a patient with cutaneous squamous cell carcinoma (CSCC), selected based on an attribute selected from the group consisting of: (i) the patient has locally advanced CSCC; (ii) the patient has metastatic CSCC; (iii) the tumor is unresectable; (iv) the patient has been previously treated with at least one anti-tumor therapy; (v) the patient has disease considered to be inoperable; (vi) surgery and / or radiation is contraindicated; (vii) the patient has previously been treated with radiation and the tumor is resistant or non-responsive to radiation; (viii) the patient has locally advanced CSCC and is not a candidate for curative surgery; (ix) the tumor contains uv-induced DNA damage; and (x) the patient exhibits PD-L1 expression of 1% or more, 5% or more, or 10% or more in tumor cells; and (b) administering a therapeutically effective amount of an anti-PD-1 antibody to a patient in need thereof. In certain embodiments, one or more doses of anti-PD-1 antibody are administered 1-12 weeks after the immediately preceding dose, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 weeks after the immediately preceding dose. In certain embodiments, each dose of anti-PD-1 antibody comprises 0.1, 1, 0.3, 3, 4, 5, 6, 7, 8, 9, or 10 mg per kg of patient body weight. In certain embodiments, each dose comprises 50-500 mg of anti-PD-1 antibody, for example, 200 mg, 250 mg, or 350 mg of anti-PD-1 antibody, and each dose is administered 0.5, 1, 2, 3, or 4 weeks after the immediately preceding dose. In one embodiment, the anti-PD-1 antibody is REGN2810.

[0049] According to one embodiment, the present invention provides a method of treating or inhibiting tumor growth, wherein: (a) (i) the patient has locally advanced BCC; (ii) the patient has metastatic BCC; (iii) the tumor is unresectable; (iv) the patient has been previously treated with at least one anti-tumor therapy; (v) the patient has been previously treated with a Hedgehog pathway inhibitor (e.g., vismodegib, sonidegib) and has progressed during treatment; (vi) the patient is intolerant to a Hedgehog pathway inhibitor; (vii) the patient has disease that is considered inoperable or is not a candidate for curative surgery; (viii) surgery and / or radiation are contraindicated; (ix) the patient has been previously treated with radiation and the tumor is resistant or non-responsive to radiation; (x) the patient has a tumor in the tumor cells. and (xi) the tumor contains uv-induced DNA damage; and (b) administering to a patient in need thereof a therapeutically effective amount of an anti-PD-1 antibody. In certain embodiments, one or more doses of the anti-PD-1 antibody are administered 1-12 weeks after the immediately preceding dose, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 weeks after the immediately preceding dose. In certain embodiments, each dose of the anti-PD-1 antibody comprises 0.1, 1, 0.3, 3, 4, 5, 6, 7, 8, 9, or 10 mg per kg of patient body weight. In certain embodiments, each dose contains 50-500 mg of the anti-PD-1 antibody, for example 200 mg, 250 mg, or 350 mg of the anti-PD-1 antibody, and each dose is administered 0.5, 1, 2, 3, or 4 weeks after the immediately preceding dose. In one embodiment, the anti-PD-1 antibody is REGN2810.

[0050] In certain embodiments, each dose of anti-PD-1 antibody is administered 1 week, 2 weeks, 3 weeks, or 4 weeks after the immediately preceding dose, and each dose contains 50-600 mg of anti-PD-1 antibody, hi one embodiment, each dose contains 200, 250, 300, or 350 mg of anti-PD-1 antibody.

[0051] One embodiment of the present invention relates to an anti-PD-1 antibody (e.g., REGN2810) for use in treating cholangiocarcinoma. In certain embodiments, one or more doses of anti-PD-1 antibody are administered to a subject with cholangiocarcinoma, each dose comprising 0.1-20 mg per kg of the subject's body weight, and each dose being administered 0.5-4 weeks after the immediately preceding dose. In certain embodiments, each dose comprises 50-500 mg of anti-PD-1 antibody, e.g., 200 mg, 250 mg, or 350 mg of anti-PD-1 antibody, and each dose being administered 0.5, 1, 2, 3, or 4 weeks after the immediately preceding dose.

[0052] One embodiment of the present invention relates to an anti-PD-1 antibody (e.g., REGN2810) for use in treating advanced hepatocellular carcinoma (HCC). In certain embodiments, one or more doses of the anti-PD-1 antibody are administered to a subject with HCC, each dose comprising 0.1-20 mg / kg of the subject's body weight, and each dose being administered 0.5-4 weeks after the immediately preceding dose. In certain embodiments, each dose comprises 50-500 mg of the anti-PD-1 antibody, e.g., 200 mg, 250 mg, or 350 mg of the anti-PD-1 antibody, and each dose being administered 0.5, 1, 2, 3, or 4 weeks after the immediately preceding dose.

[0053] One embodiment of the present invention relates to an anti-PD-1 antibody (e.g., REGN2810) for use in treating soft tissue sarcoma. In certain embodiments, one or more doses of anti-PD-1 antibody are administered to a subject with soft tissue sarcoma, each dose comprising 0.1-20 mg per kg of the subject's body weight, and each dose being administered 0.5-4 weeks after the immediately preceding dose. In certain embodiments, each dose comprises 50-500 mg of anti-PD-1 antibody, e.g., 200 mg, 250 mg, or 350 mg of anti-PD-1 antibody, and each dose being administered 0.5, 1, 2, 3, or 4 weeks after the immediately preceding dose.

[0054] One embodiment of the present invention relates to an anti-PD-1 antibody (e.g., REGN2810) for use in treating non-small cell lung cancer (NSCLC). In certain embodiments, one or more doses of the anti-PD-1 antibody are administered to a subject with NSCLC, each dose comprising 0.1-20 mg / kg of the subject's body weight, and each dose being administered 0.5-4 weeks after the immediately preceding dose. In certain embodiments, one or more doses of the anti-PD-1 antibody are administered to a subject with NSCLC, each dose comprising 50-600 mg of the anti-PD-1 antibody, and each dose being administered 0.5-4 weeks after the immediately preceding dose.

[0055] According to one aspect, the present invention includes a method of treating a tumor or inhibiting tumor growth, comprising the steps of selecting a subject having brain cancer and administering a therapeutically effective amount of an anti-PD-1 antibody or antigen-binding fragment thereof to a subject in need thereof. In certain embodiments, the brain cancer is glioblastoma multiforme. In one embodiment, the subject has newly diagnosed glioblastoma multiforme. In one embodiment, the subject is 65 years of age or older. In one embodiment, the anti-PD-1 antibody is administered in one or more doses, each dose being administered 0.5-4 weeks after the immediately preceding dose. In one embodiment, each dose of the anti-PD-1 antibody comprises 1, 3, or 10 mg per kg of the subject's body weight. In certain embodiments, the anti-PD-1 antibody is administered in combination with radiation therapy. In one embodiment, the radiation therapy is hypofractionated radiation therapy. In one embodiment, the subject is administered 20-60 Gy in 2-20 fractions. In certain embodiments, one or more doses of the anti-PD-1 antibody are included in one or more treatment cycles, each treatment cycle including 1-6 doses of the anti-PD-1 antibody. In one embodiment, at least one treatment cycle further includes radiation therapy. In a further embodiment, the radiation therapy is hypofractionated radiation therapy. In certain embodiments, the subject is administered hypofractionated radiation therapy in a first treatment cycle, the hypofractionated radiation therapy including 2-20 fractions of 20-60 Gy. In one embodiment, the subject is administered hypofractionated radiation therapy one week after administration of the anti-PD-1 antibody in the first treatment cycle. In certain embodiments, if the subject develops intracranial edema following administration of the anti-PD-1 antibody, the method of the present invention further includes administering an anti-angiogenic agent to the subject. In one embodiment, the anti-angiogenic agent is a vascular endothelial growth factor (VEGF) inhibitor. In one embodiment, the anti-angiogenic agent is an angiopoietin-2 (Ang-2) inhibitor (e.g., an anti-Ang-2 antibody such as nesbacumab).In certain embodiments, the VEGF inhibitor is selected from the group consisting of a VEGF inhibitory fusion protein (e.g., a "VEGF-Trap" or other VEGF inhibitory fusion protein such as aflibercept described in U.S. Pat. No. 7,087,411), an anti-VEGF antibody (e.g., bevacizumab), and a small molecule kinase inhibitor of a VEGF receptor (e.g., sunitinib, sorafenib, or pazopanib).

[0056] In certain embodiments, the methods of the invention include administering an anti-PD-1 antibody in combination with radiation therapy to a subject in need thereof as a "first line" treatment (e.g., initial treatment). In other embodiments, the anti-PD-1 antibody in combination with radiation therapy is administered as a "second line" treatment (e.g., after previous treatment). For example, the anti-PD-1 antibody in combination with radiation therapy is administered as a "second line" treatment to a subject who has relapsed after previous treatment, for example, with chemotherapy.

[0057] According to certain embodiments, the methods of the invention include administering to a subject a therapeutically effective amount of an anti-PD-1 antibody and radiation in combination with an additional therapeutic agent or treatment regimen or procedure that may be administered to increase anti-tumor efficacy, reduce toxic effects of one or more therapies, and / or reduce the dosage of one or more therapies. In various embodiments, the additional therapeutic agent or treatment regimen or procedure is, for example, chemotherapy, cyclophosphamide, surgery, a cancer vaccine, a programmed death-ligand 1 (PD-L1) inhibitor (e.g., an anti-PD-L1 antibody), a lymphocyte-activation gene 3 (LAG3) inhibitor (e.g., an anti-LAG3 antibody), a cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) inhibitor (e.g., ipilimumab), a glucocorticoid-induced tumor necrosis factor receptor (GITR) inhibitor (e.g., an anti-GITR antibody), a T-cell immunoglobulin and mucin domain-containing protein 3 (TIM3) inhibitor, a B- and T-lymphocyte attenuator (BTLA) inhibitor, a T-cell immunoreceptor with Ig and ITIM domains (TIGIT) inhibitor, a CD47 inhibitor, an indoleamine-2,3-dioxygenase (IDO) inhibitor, a vascular endothelial growth factor (VEGF) antagonist, an angiopoietin-2 (Ang2) inhibitor, a transforming growth factor (GF) inhibitor, a vasopressin-1 (vasopressin-1 ...) inhibitor, a vasopressin-1) inhibitor, a vasopressin-1) inhibitor, a vasopressin-1) inhibitor, a vasopressin-1) inhibitor The therapeutic agent is selected from the group consisting of transforming growth factor beta (TGFβ) inhibitors, epidermal growth factor receptor (EGFR) inhibitors, antibodies against tumor-specific antigens (e.g., CA9, CA125, melanoma-associated antigen 3 (MAGE3), carcinoembryonic antigen (CEA), vimentin, tumor-M2-PK, prostate-specific antigen (PSA), mucin-1, MART-1, and CA19-9), anti-CD3 / anti-CD20 bispecific antibodies, vaccines (e.g., bacillus Calmette-Guerin), granulocyte-macrophage colony-stimulating factor, cytotoxins, chemotherapeutic agents, cytokines such as IL-6R inhibitors, IL-4R inhibitors, IL-10 inhibitors, IL-2, IL-7, IL-21, and IL-15, anti-inflammatory agents such as corticosteroids and nonsteroidal anti-inflammatory agents, and nutritional supplements such as antioxidants. In certain embodiments, the anti-PD-1 antibody may be administered in combination with treatments including chemotherapeutic agents and surgery. As used herein, the term "in combination with" means that the anti-PD-1 antibody is administered to the subject simultaneously with, immediately before, or immediately after the administration of radiation therapy and an additional therapeutic agent. In certain embodiments, the additional therapeutic agent is administered in the same formulation as the anti-PD-1 antibody.

[0058] One embodiment of the present invention relates to a combination of an anti-PD-1 antibody (e.g., REGN2810), radiation therapy, cyclophosphamide, and GM-CSF for use in treating head and neck squamous cell carcinoma (HNSCC). In certain embodiments, one or more doses of anti-PD-1 antibody are administered to a subject with HNSCC, each dose comprising 0.1-20 mg per kg of the subject's body weight, and each dose is administered 0.5-4 weeks after the immediately preceding dose. In certain embodiments, each dose comprises 50-500 mg of anti-PD-1 antibody, e.g., 200 mg, 250 mg, or 350 mg of anti-PD-1 antibody, and each dose is administered 0.5, 1, 2, 3, or 4 weeks after the immediately preceding dose.

[0059] One embodiment of the present invention relates to a combination of an anti-PD-1 antibody (e.g., REGN2810), radiation therapy, and cyclophosphamide for use in treating breast cancer. In certain embodiments, one or more doses of the anti-PD-1 antibody are administered to a subject with breast cancer, each dose comprising 0.1-20 mg / kg of the subject's body weight, and each dose is administered 0.5-4 weeks after the immediately preceding dose.

[0060] One embodiment of the present invention relates to a combination of an anti-PD-1 antibody (e.g., REGN2810), radiation therapy, cyclophosphamide, and GM-CSF for use in treating advanced solid tumors in patients who have been previously treated with an anti-PD-1 antibody or an anti-PD-L1 antibody. In certain embodiments, one or more doses of the anti-PD-1 antibody are administered to a patient in need thereof, each dose comprising 0.1-20 mg / kg of the subject's body weight, and each dose is administered 0.5-4 weeks after the immediately preceding dose.

[0061] One embodiment of the present invention relates to a combination of an anti-PD-1 antibody (e.g., REGN2810), docetaxel, and optionally carboplatin for use in treating advanced solid tumors that are resistant to first-line chemotherapy. In certain embodiments, docetaxel is administered at a low dose. In certain embodiments, one or more doses of the anti-PD-1 antibody are administered to a subject in need thereof, each dose comprising 0.1-20 mg / kg of the subject's body weight, and each dose is administered 0.5-4 weeks after the previous dose.

[0062] One embodiment of the present invention relates to a combination of an anti-PD-1 antibody (e.g., REGN2810) and radiation therapy for use in treating newly diagnosed or recurrent glioblastoma multiforme (GBM). In certain embodiments, one or more doses of the anti-PD-1 antibody are administered to a subject in need thereof, each dose comprising 0.1-20 mg / kg of the subject's body weight, and each dose is administered 0.5-4 weeks after the immediately preceding dose. In certain embodiments, the radiation is hypofractionated radiation therapy as described herein.

[0063] Certain embodiments of the invention relate to a combination of an anti-PD-1 antibody (e.g., REGN2810) and radiation therapy for use in treating cervical squamous cell carcinoma, anal squamous cell carcinoma, Merkel cell carcinoma, small intestine adenocarcinoma, or ovarian serous carcinoma. In certain embodiments, one or more doses of the anti-PD-1 antibody are administered to a subject in need thereof, each dose comprising 0.1-20 mg per kg of the subject's body weight, and each dose is administered 0.5-4 weeks after the immediately preceding dose. In certain embodiments, the radiation is hypofractionated radiation therapy as described herein.

[0064] In certain embodiments, the present invention includes a method of treating large or advanced malignant tumors, comprising administering to a subject in need thereof an anti-PD-1 antibody in combination with radiation therapy and an additional therapeutic agent, wherein the additional therapeutic agent is administered to overcome regulatory T cell (Treg)-mediated immune suppression. In certain embodiments, the additional therapeutic agent is selected from the group consisting of an anti-GITR antibody, an anti-LAG3 antibody, cyclophosphamide, and GM-CSF.

[0065] The term "large tumor" as used herein refers to the size of a tumor. This typically correlates with high tumor burden or tumor mass. In certain embodiments, this correlates with the stage of a disease, such as an advanced malignant tumor. In certain embodiments, this correlates with an increased likelihood of metastasis.

[0066] In certain embodiments, the invention includes methods comprising administering one or more doses of an anti-PD-1 antibody in combination with radiation therapy and a sub-therapeutic dose of cyclophosphamide. As used herein, a sub-therapeutic dose of cyclophosphamide (also referred to herein as "low-dose cyclophosphamide") refers to an amount of cyclophosphamide that does not itself impart a therapeutic effect, but preferably does not cause toxicity. Exemplary doses of cyclophosphamide that are considered "sub-therapeutic" in the context of the present invention include 100 mg / m 2, 90 mg / m 2 , 80 mg / m 2 , or less.

[0067] In one aspect, the invention includes a method comprising administering a therapeutically effective amount of an anti-PD-1 antibody in combination with radiation to a subject undergoing a background anti-cancer treatment regimen. The background anti-cancer treatment regimen may include, for example, a series of administrations of a chemotherapeutic agent. The anti-PD-1 antibody in combination with radiation therapy may be added outside of the background anti-cancer treatment regimen. In some embodiments, the anti-PD-1 antibody is added as part of a "background step-down" scheme in which the background anti-cancer treatment is gradually tapered (stepwise) from the subject over time, but the anti-PD-1 antibody is administered to the subject at a constant dose or at increasing or decreasing doses over time. For example, the background anti-cancer treatment may include a chemotherapeutic agent administered at a low or sub-therapeutic dose. In certain embodiments, the invention includes a method of treating cancer comprising administering one or more doses of an anti-PD-1 antibody in combination with radiation therapy, and one or more doses of a chemotherapeutic agent, where the chemotherapeutic agent is administered at a sub-therapeutic dose.

[0068] In certain embodiments, radiation therapy is administered to a first tumor lesion but not to a second tumor lesion, and combined administration with an anti-PD-1 antibody results in tumor regression in both the first and second tumor lesions (abscopal effect). In certain embodiments, the methods of the invention include administration of an anti-PD-1 antibody in combination with radiation therapy to produce a sustained abscopal effect.

[0069] In certain embodiments, the methods of the invention comprise administering to a subject in need thereof a therapeutically effective amount of an anti-PD-1 antibody, optionally in combination with radiation therapy, where administration of the combination results in increased inhibition of tumor growth. Growth is inhibited by at least about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, or about 80% compared to untreated subjects or subjects administered either the antibody or radiation as monotherapy. In certain embodiments, administration of an anti-PD-1 antibody and / or radiation therapy results in tumor regression, increased tumor shrinkage and / or disappearance. In certain embodiments, administration of an anti-PD-1 antibody and / or radiation therapy results in a delay in tumor growth and development, e.g., tumor growth may be delayed by about 3 days, more than 3 days, about 7 days, more than 7 days, more than 15 days, more than 1 month, more than 3 months, more than 6 months, more than 1 year, more than 2 years, or more than 3 years compared to untreated subjects or subjects treated with either the antibody or radiation as monotherapy. In certain embodiments, administration of an anti-PD-1 antibody in combination with radiation therapy prevents tumor recurrence and / or extends the survival of a subject, e.g., by more than 15 days, more than 1 month, more than 3 months, more than 6 months, more than 12 months, more than 18 months, more than 24 months, more than 36 months, or more than 48 months, compared to an untreated subject or a subject receiving either the antibody or radiation as monotherapy. In certain embodiments, administration of an anti-PD-1 antibody in combination with radiation therapy extends progression-free survival or overall survival. In certain embodiments, administration of an anti-PD-1 antibody in combination with radiation therapy increases the response and duration of response in a subject, e.g., by more than 2%, more than 3%, more than 4%, more than 5%, more than 6%, more than 7%, more than 8%, more than 9%, more than 10%, more than 20%, more than 30%, more than 40%, or more than 50%, compared to an untreated subject or a subject receiving either the antibody or radiation as monotherapy. In certain embodiments, administration of an anti-PD-1 antibody and / or radiation therapy to a subject with cancer results in the complete disappearance of all evidence of tumor cells (a "complete response"). In certain embodiments, administration of an anti-PD-1 antibody and / or radiation therapy to a subject with cancer results in at least a 30% or greater reduction in tumor cells or tumor size (a "partial response").In certain embodiments, administration of anti-PD-1 antibodies and / or radiation therapy to a subject with cancer results in complete or partial disappearance of tumor cells / lesions, including new measurable lesions. Tumor reduction can be measured by any method known in the art, such as x-ray, positron emission tomography (PET), computed tomography (CT), magnetic resonance imaging (MRI), cytology, histology, or molecular genetic analysis.

[0070] In certain embodiments, the methods of the invention comprise administering a therapeutically effective amount of an anti-PD-1 antibody to a subject in need thereof, wherein administration of the anti-PD-1 antibody results in an increase in the patient's overall survival (OS) or progression-free survival (PFS) compared to patients administered a standard of care (SOC) therapy (e.g., chemotherapy, surgery, or radiation). In certain embodiments, PFS is increased by at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 1 year, at least 2 years, or at least 3 years compared to patients administered any one or more SOC therapies. In certain embodiments, OS is increased by at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 1 year, at least 2 years, or at least 3 years compared to patients administered any one or more SOC therapies.

[0071] The present invention also provides kits comprising anti-PD-1 antibodies for therapeutic use. The kit typically includes a label indicating the intended use of the contents of the kit and instructions for use. The term label includes any written or recorded material supplied with or accompanying the kit, or that otherwise accompanies the kit. Thus, the present disclosure provides kits for treating a subject suffering from cancer, comprising (a) a dose of an antibody or antigen-binding portion thereof that specifically binds to PD-1 and inhibits PD-1 activity, and (b) instructions for using the anti-PD-1 antibody in any of the therapeutic methods disclosed herein. In certain embodiments for treating a human patient, the kit comprises an anti-human PD-1 antibody disclosed herein, such as REGN2810. In other embodiments, the anti-PD-1 antibody can be any one of nivolumab, pembrolizumab, or any of the anti-PD-1 antibodies disclosed herein. In certain embodiments, the dose of the anti-PD-1 antibody ranges from 0.1 to 10 mg / kg body weight. In certain embodiments, the dose of the anti-PD-1 antibody comprises 50 to 600 mg.

[0072] Methods for suppressing regulatory T cells According to certain aspects, the present invention provides methods of suppressing or inhibiting activation and / or proliferation of regulatory T (Treg) cells. In certain embodiments, the present invention provides methods of suppressing the activity of Treg cells. The methods according to these aspects include selecting a subject having a solid tumor, and administering to the subject an anti-PD-1 antibody or antigen-binding fragment thereof in combination with at least one of: (i) radiation therapy, and (ii) a glucocorticoid-induced tumor necrosis factor receptor (GITR) antagonist. In certain embodiments, the method includes administering to a subject in need thereof an anti-PD-1 antibody or antigen-binding fragment thereof in combination with radiation therapy and a GITR antagonist.

[0073] In certain embodiments, the GITR antagonist is an anti-GITR antibody or an antigen-binding fragment thereof. According to certain exemplary embodiments of the present invention, the anti-GITR antibody or an antigen-binding fragment thereof comprises a heavy chain variable region (HCVR), a light chain variable region (LCVR), and / or a complementarity determining region (CDR) comprising any of the amino acid sequences of the anti-GITR antibodies described in U.S. Patent Application No. 62 / 256,922 (filed November 18, 2015), the contents of which are incorporated herein in their entirety. Other anti-GITR antibodies that can be used in the context of the method of the present invention include any of the anti-GITR antibodies described in, for example, U.S. Patent Nos. 9228016, 8709424, 8591886, 7812135, or U.S. Patent Application Publication No. 20150368349.

[0074] In certain embodiments, the present invention provides a method of suppressing or eliminating Treg activity comprising administering to a subject in need thereof an anti-PD-1 antibody or antigen-binding fragment thereof in combination with one or more doses of radiation and a cytotoxic T-lymphocyte antigen-4 (CTLA) antagonist. In certain embodiments, the CTLA antagonist is an anti-CTLA antibody (e.g., ipilimumab).

[0075] In certain embodiments, the present invention provides a method of suppressing or eliminating Treg activity, comprising administering to a subject in need thereof an anti-PD-1 antibody or an antigen-binding fragment thereof in combination with one or more doses of radiation and a lymphocyte activation gene 3 (LAG-3) antagonist. In certain embodiments, the LAG-3 antagonist is an anti-LAG-3 antibody. Anti-LAG-3 antibodies that can be used in the context of the methods of the present invention are disclosed in U.S. Patent Application No. 15 / 289,032 (filed October 7, 2016), the contents of which are incorporated herein in their entirety.

[0076] In certain embodiments, the invention provides a method of suppressing or eliminating Treg activity, comprising administering to a subject in need thereof an anti-PD-1 antibody, or antigen-binding fragment thereof, in combination with one or more doses of radiation and cyclophosphamide.

[0077] In one embodiment, the method of the invention comprises administering an anti-PD-1 antibody to a subject having a solid tumor in combination with radiation therapy and an additional therapeutic agent selected from the group consisting of a GITR antagonist, an anti-LAG-3 antibody, and cyclophosphamide, The administration results in an effect selected from the group consisting of inhibition of tumor growth, reduction in tumor size, slowing of tumor growth, inhibition of tumor metastasis, reduction in metastatic lesions over time, reduction in the use of chemotherapeutic or cytotoxic agents, prolonged survival, complete response, partial response, and stable disease. In certain embodiments, the administration results in a reduction in tumor burden in the subject. In certain embodiments, the subject has a large tumor. As defined elsewhere herein, the term "large tumor" refers to the size of the tumor and correlates with increased tumor burden and increased likelihood of developing metastases. In certain embodiments, the term refers to an advanced malignancy.

[0078] Anti-PD-1 antibodies and antigen-binding fragments thereof According to certain exemplary embodiments of the present invention, the method comprises administering a therapeutically effective amount of an anti-PD-1 antibody or an antigen-binding fragment thereof. As used herein, the term "antibody" includes immunoglobulin molecules comprising four polypeptide chains, i.e., two heavy (H) chains and two light (L) chains interconnected by disulfide bonds, as well as multimers thereof (e.g., IgM). In a typical antibody, each heavy chain comprises a heavy chain variable region (referred to herein as HCVR or V H The heavy chain constant region is made up of three domains: H 1. C H 2, and C H Each light chain comprises a light chain variable region (referred to herein as LCVR or V LThe light chain constant region comprises one domain (C L 1) is included. H and V L The regions can be further subdivided into regions of hypervariability, called complementarity determining regions (CDRs), interspersed between more conserved regions, called framework regions (FRs). H and V L is composed of three CDRs and four FRs arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. In different embodiments of the invention, the FRs of an anti-IL-4R antibody (or antigen-binding portion thereof) may be identical to human germline sequences or may be naturally or artificially modified. An amino acid consensus sequence may be defined based on a parallel analysis of two or more CDRs.

[0079] The term "antibody" as used herein also includes antigen-binding fragments of complete antibody molecules. As used herein, the terms "antigen-binding portion" of an antibody, "antigen-binding fragment" of an antibody, etc., include any naturally occurring, enzymatically obtainable, synthetic, or genetically engineered polypeptide or glycoprotein that specifically binds to an antigen to form a complex. Antigen-binding fragments of antibodies may be derived, for example, from complete antibody molecules using any suitable standard technique, such as proteolytic digestion, or recombinant genetic engineering techniques involving the manipulation and expression of DNA encoding the variable and optionally constant domains of the antibody. Such DNA is known and / or readily available, for example, from commercial sources, DNA libraries (including, for example, phage-antibody libraries), or can be synthesized. For example, the DNA may be sequenced and manipulated chemically or using molecular biology techniques to align one or more variable and / or constant domains into a suitable shape, or to introduce codons, create cysteine ​​residues, modify, add, or delete amino acids.

[0080] Non-limiting examples of antigen-binding fragments include: (i) Fab fragments; (ii) F(ab')2 fragments; (iii) Fd fragments; (iv) Fv fragments; (v) single chain Fv (scFv) molecules; (vi) dAb fragments; and (vii) minimal recognition units consisting of amino acid residues that mimic the hypervariable regions of an antibody (e.g., isolated complementarity determining regions (CDRs), such as CDR3 peptides), or limited FR3-CDR4-FR4 peptides. Domain-specific antibodies, single domain antibodies, domain deleted antibodies, chimeric antibodies, CDR grafted antibodies, diabodies, triabodies, tetrabodies, minibodies, nanobodies (e.g., monovalent nanobodies, bivalent nanobodies, etc.), small molecule modular immunopharmaceuticals (SMIPs), and other engineered molecules such as shark variable IgNAR domains are also encompassed within the term "antigen-binding fragments" as used herein.

[0081] Antigen-binding fragments of antibodies typically contain at least one variable domain. A variable domain may be of any size or amino acid composition and generally contains at least one CDR adjacent to or in frame with one or more framework sequences. L Domain-associated V H In an antigen-binding fragment having a domain, H and V L The domains can be in any suitable arrangement relative to each other. For example, the variable region can be a dimer, H -V H , V H -V L , or V L -V L Alternatively, the antigen-binding fragment of the antibody may comprise a monomeric V H or V L It may also include a domain.

[0082] In certain embodiments, an antigen-binding fragment of an antibody may comprise at least one variable domain covalently linked to at least one constant domain. Non-limiting exemplary configurations of variable and constant domains that may be found in an antigen-binding fragment of an antibody of the invention include: (i) a V H -C H 1;(ii)V H -C H 2;(iii)V H -C H 3;(iv)V H -C H 1-C H 2;(v)V H -C H 1-C H 2-C H 3;(vi)V H -C H 2-C H 3;(vii)V H -C L ;(viii)V L -C H 1;(ix)V L -C H 2;(x)V L -C H 3;(xi)V L -C H 1-C H 2;(xii)V L -C H 1-C H 2-C H 3;(xiii)V L -C H 2-C H 3; and (xiv) V L -C LIn any configuration of variable and constant domains, including any of the exemplary configurations above, the variable and constant domains may be directly linked to each other or may be linked by a complete or partial hinge or linker region. The hinge region may consist of at least two (e.g., 5, 10, 15, 20, 40, 60 or more than 60) amino acids such that a flexible or semi-flexible linkage occurs between adjacent variable and / or constant domains in a single polypeptide molecule. Moreover, antigen-binding fragments of antibodies of the present invention may be linked to each other and / or to one or more monovalent V H or V L It may comprise homodimers or heterodimers (or other multimers) of any of the above variable and constant domain configurations, with the domains non-covalently linked (eg, by disulfide bonds).

[0083] The term "antibody" as used herein also includes multispecific (e.g., bispecific) antibodies. Multispecific antibodies or antigen-binding fragments of antibodies typically contain at least two different variable domains, each of which can specifically bind to an individual antigen or a different epitope on the same antigen. Any multispecific antibody format may be adapted for use in the context of the antibodies or antigen-binding fragments of antibodies of the present invention using routine techniques available in the art. For example, the present invention includes methods that include the use of bispecific antibodies, in which one arm of the immunoglobulin is specific for PD-1 or a fragment thereof, and the other arm of the immunoglobulin is specific for a second therapeutic target or is conjugated to a therapeutic moiety. Exemplary bispecific formats that can be used in the context of the present invention include, for example, scFv-based or diabody-type bispecific formats, IgG-scFv fusions, dual variable domain (DVD)-Ig, quadroma, knobs-into-holes, common light chain (e.g., common light chain with knobs-into-holes), CrossMab, CrossFab, (SEED) antibodies, leucine zipper, Duobody, IgG1 / IgG2, biacting Fab (DAF)-IgG, and Mab2 Bispecific formats include, but are not limited to, bispecific formats (see, e.g., Klein et al., 2012, mAbs 4:6, pp. 1-11, and references cited therein, for a review of the aforementioned formats). Bispecific antibodies can also be constructed using peptide / nucleic acid conjugation, e.g., using unnatural amino acids with orthogonal chemical reactivity to generate site-specific antibody-oligonucleotide conjugates that then self-assemble into multimeric complexes with defined composition, valency, and shape (see, e.g., Kazane et al., J. Am. Chem. Soc. [Epub: December 4, 2012]).

[0084] The antibody used in the method of the present invention can be a human antibody. The term "human antibody" as used herein is intended to include antibodies with variable and constant regions derived from human germline immunoglobulin sequences. The human antibody of the present invention may nevertheless include amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by in vitro random or site-specific mutagenesis or in vivo somatic mutations), for example in CDRs and especially CDR3. However, the term "human antibody" as used herein is not intended to include antibodies in which CDR sequences derived from the germline of another mammalian species, such as mouse, are grafted onto human framework sequences.

[0085] The antibody used in the method of the present invention may be a recombinant human antibody. As used herein, the term "recombinant human antibody" is intended to include all human antibodies prepared, expressed, produced, or isolated by recombinant means, such as antibodies expressed using a recombinant expression vector transfected into a host cell (described in detail below), antibodies isolated from a recombinant combinatorial human antibody library (described in detail below), antibodies isolated from an animal (e.g., a mouse) that is transgenic for human immunoglobulin genes [see, e.g., Taylor et al. (1992) Nucl. Acids Res. 20:6287-6295], or antibodies prepared, expressed, produced, or isolated by any other means involving splicing of human immunoglobulin gene sequences to other DNA sequences. Such recombinant human antibodies have variable and constant regions derived from human germline immunoglobulin sequences. However, in certain embodiments, such recombinant human antibodies are subjected to in vitro mutagenesis (or in vivo somatic mutagenesis, when animals transgenic for human Ig sequences are used), thus improving the V H and V L The amino acid sequence of the region is H and V L These are sequences that are related by origin but do not naturally occur within the human antibody germline repertoire in vivo.

[0086] According to certain embodiments, the antibody used in the methods of the invention specifically binds to PD-1. The term "specifically binds" and the like means that the antibody or antigen-binding fragment thereof forms a complex with the antigen that is relatively stable under physiological conditions. Methods for determining whether an antibody specifically binds to an antigen are well known in the art and include, for example, equilibrium dialysis, surface plasmon resonance, and the like. For example, an antibody that "specifically binds" to PD-1 as used in the context of the present invention has a K of less than about 500 nM, less than about 300 nM, less than about 200 nM, less than about 100 nM, less than about 90 nM, less than about 80 nM, less than about 70 nM, less than about 60 nM, less than about 50 nM, less than about 40 nM, less than about 30 nM, less than about 20 nM, less than about 10 nM, less than about 5 nM, less than about 4 nM, less than about 3 nM, less than about 2 nM, less than about 1 nM, or less than about 0.5 nM as measured in a surface plasmon resonance assay. D and includes antibodies that bind to PD-1 or a portion thereof. However, an isolated antibody that specifically binds human PD-1 may have cross-reactivity to other antigens, such as PD-1 molecules from other (non-human) species.

[0087] According to certain exemplary embodiments of the invention, an anti-PD-1 antibody or antigen-binding fragment thereof comprises a heavy chain variable region (HCVR), a light chain variable region (LCVR), and / or a complementarity determining region (CDR) comprising the amino acid sequence of any of the anti-PD-1 antibodies set forth in U.S. Patent Application Publication No. 20150203579, which is incorporated herein in its entirety. In certain exemplary embodiments, an anti-PD-1 antibody or antigen-binding fragment thereof that can be used in the context of the methods of the invention comprises a heavy chain complementarity determining region (HCDR) of the heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO:1, and a light chain complementarity determining region (LCDR) of the light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO:2. In certain embodiments, an anti-PD-1 antibody or antigen-binding fragment thereof comprises three HCDRs (HCDR1, HCDR2, and HCDR3) and three LCDRs (LCDR1, LCDR2, and and LCDR3), wherein HCDR1 comprises the amino acid sequence of SEQ ID NO:3; HCDR2 comprises the amino acid sequence of SEQ ID NO:4; HCDR3 comprises the amino acid sequence of SEQ ID NO:5; LCDR1 comprises the amino acid sequence of SEQ ID NO:6; LCDR2 comprises the amino acid sequence of SEQ ID NO:7; and LCDR3 comprises the amino acid sequence of SEQ ID NO:8. In still other embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof comprises an HCVR comprising SEQ ID NO:1 and an LCVR comprising SEQ ID NO:2. In certain embodiments, the methods of the invention comprise the use of an anti-PD-1 antibody, wherein the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:9. In some embodiments, the anti-PD-1 antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO:10. An exemplary antibody comprising a heavy chain comprising the amino acid sequence of SEQ ID NO:9 and a light chain comprising the amino acid sequence of SEQ ID NO:10 is the fully human anti-PD-1 antibody known as REGN2810, also known as empleumab. According to certain exemplary embodiments, the methods of the invention comprise the use of REGN2810, or a biological equivalent thereof. The term "bioequivalent" as used herein refers to an anti-PD-1 antibody or PD-1 binding protein, or fragment thereof, that is a pharmaceutical equivalent or substitute whose rate and / or extent of absorption does not differ significantly from that of REGN2810 when administered at the same molar dose under similar experimental conditions, either in single or multiple doses. In the context of the present invention, the term refers to an antigen binding protein that binds to PD-1 that has no clinically meaningful differences from REGN2810 in its safety, purity, and / or potency.

[0088] According to certain embodiments of the invention, the anti-human PD-1 or antigen-binding fragment thereof comprises a HCVR having 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO:1.

[0089] According to certain embodiments of the invention, the anti-human PD-1 or antigen-binding fragment thereof comprises an LCVR having 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO:2.

[0090] According to certain embodiments of the invention, the anti-human PD-1 or antigen-binding fragment thereof comprises a HCVR comprising the amino acid sequence of SEQ ID NO: 1 with no more than five amino acid substitutions. According to certain embodiments of the invention, the anti-human PD-1 or antigen-binding fragment thereof comprises a LCVR comprising the amino acid sequence of SEQ ID NO: 2 with no more than two amino acid substitutions.

[0091] Sequence identity can be measured by any method known in the art (eg, GAP, BESTFIT, and BLAST).

[0092] The invention also includes the use of anti-PD-1 antibodies in methods for treating cancer, which include variants of any of the HCVR, LCVR, and / or CDR amino acid sequences disclosed herein having one or more conservative amino acid substitutions. For example, the invention includes the use of anti-PD-1 antibodies having HCVR, LCVR, and / or CDR amino acid sequences that have, e.g., 10 or fewer, 8 or fewer, 6 or fewer, 4 or fewer, etc., conservative amino acid substitutions compared to any of the HCVR, LCVR, and / or CDR amino acid sequences disclosed herein.

[0093] Other anti-PD-1 antibodies that can be used in the context of the methods of the invention include, for example, antibodies known in the art as nivolumab (U.S. Patent No. 8,008,449) and pembrolizumab (U.S. Patent No. 8,354,509), MEDI0608 (U.S. Patent No. 8,609,089), pidilizumab (U.S. Patent No. 8,686,119), or any of the anti-PD-1 antibodies described in U.S. Patent Nos. 6,808,710, 7,488,802, 8,168,757, 8,354,509, 8,779,105, or 8,900,587.

[0094] The anti-PD-1 antibody used in the context of the method of the present invention may have pH-dependent binding characteristics. For example, the anti-PD-1 antibody for use in the method of the present invention may exhibit reduced binding to PD-1 at acidic pH compared to neutral pH. Alternatively, the anti-PD-1 antibody of the present invention may exhibit enhanced binding to its antigen at acidic pH compared to neutral pH. The expression "acidic pH" includes pH values ​​less than about 6.2, such as about 6.0, 5.95, 5.9, 5.85, 5.8, 5.75, 5.7, 5.65, 5.6, 5.55, 5.5, 5.45, 5.4, 5.35, 5.3, 5.25, 5.2, 5.15, 5.1, 5.05, 5.0, or less. The expression "neutral pH" as used herein means a pH of about 7.0 to about 7.4. The expression "neutral pH" includes pH values ​​of about 7.0, 7.05, 7.1, 7.15, 7.2, 7.25, 7.3, 7.35, and 7.4.

[0095] In certain instances, "reduced binding to PD-1 at acidic pH compared to neutral pH" refers to the K D K value of antibody binding to PD-1 at neutral pH D For example, an antibody or antigen-binding fragment thereof may be expressed in terms of a ratio to a value (or vice versa). D When a ratio is presented, it can be considered for purposes of the present invention to indicate "reduced binding to PD-1 at acidic pH compared to neutral pH." In certain exemplary embodiments, the acidic / neutral K D The ratio can be about 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10.0, 10.5, 11.0, 11.5, 12.0, 12.5, 13.0, 13.5, 14.0, 14.5, 15.0, 20.0, 25.0, 30.0, 40.0, 50.0, 60.0, 70.0, 100.0, or greater than 100.0.

[0096] Antibodies with pH-dependent binding characteristics can be obtained, for example, by screening antibody populations for reduced (or enhanced) binding to a particular antigen at acidic pH compared to neutral pH. In addition, modification of the antigen-binding domain at the amino acid level can result in antibodies with pH-dependent characteristics. For example, one or more amino acids in the antigen-binding domain (e.g., in the CDR) can be replaced with histidine residues to obtain an antibody with reduced antigen binding at acidic pH compared to neutral pH. As used herein, the expression "acidic pH" refers to pH 6.0 or less than 6.0.

[0097] Combination Therapy The method of the invention according to certain embodiments includes administering a therapeutically effective amount of an anti-PD-1 antibody to a subject. In certain embodiments, the method of the invention includes administering radiation therapy in combination with an anti-PD-1 antibody for additive or synergistic activity to treat cancer. As used herein, the phrase "in combination with" means that radiation therapy is administered before, after, or simultaneously with the anti-PD-1 antibody. The term "in combination with" also includes sequential or simultaneous administration of an anti-PD-1 antibody and radiation therapy. For example, when administered "before" radiation therapy, the anti-PD-1 antibody may be administered more than 150 hours, about 150 hours, about 100 hours, about 72 hours, about 60 hours, about 48 hours, about 36 hours, about 24 hours, about 12 hours, about 10 hours, about 8 hours, about 6 hours, about 4 hours, about 2 hours, about 1 hour, or about 30 minutes, about 15 minutes, or about 10 minutes before the administration of radiation therapy. When administered "after" radiation therapy, the anti-PD-1 antibody may be administered about 10 minutes, about 15 minutes, about 30 minutes, about 1 hour, about 2 hours, about 4 hours, about 6 hours, about 8 hours, about 10 hours, about 12 hours, about 24 hours, about 36 hours, about 48 hours, about 60 hours, about 72 hours, or more than 72 hours after the administration of radiation therapy. Administration "concurrent with" radiation therapy means that the anti-PD-1 antibody is administered to the subject within less than 10 minutes of (before, after, or simultaneously with) the administration of radiation therapy.

[0098] In certain embodiments, the method of the present invention includes the administration of an additional therapeutic agent that is an anti-cancer agent. As used herein, "anti-cancer agent" refers to any agent that is useful for treating cancer, including but not limited to agents such as cytotoxins, antimetabolites, alkylating agents, anthracyclines, antibiotics, antimitotics, procarbazine, hydroxyurea, asparaginase, corticosteroids, mitotane (O,P'-(DDD)), biopharmaceuticals (e.g., antibodies and interferons), and radioactive agents. As used herein, "cytotoxins or cytotoxic agents" also refers to chemotherapeutic agents and refers to any agent that is harmful to cells. Examples include, but are not limited to, Taxol® (paclitaxel), temozolomide, cytochalasin B, gramicidin D, ethidium bromide, emetine, cisplatin, mitomycin, etoposide, teniposide, vincristine, vinblastine, colchicine, doxorubicin, daunorubicin, dihydroxyanthracin dione, mitoxantrone, mithramycin, actinomycin D, 1-dehydrotestosterone, glucocorticoids, procaine, tetracaine, lidocaine, propranolol, and puromycin, and analogs or homologs thereof.

[0099] In certain embodiments, the methods of the invention include treatment with any of the following agents: surgery, radiation, a programmed death-ligand 1 (PD-L1) inhibitor (e.g., an anti-PD-L1 antibody disclosed in U.S. Patent Application Publication No. 2015 / 0203580 or atezolizumab), a lymphocyte-activation gene 3 (LAG-3) inhibitor (e.g., an anti-LAG-3 antibody), a cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) inhibitor (e.g., ipilimumab), a glucocorticoid-induced tumor necrosis factor receptor (GITR) inhibitor (e.g., an anti-GITR antibody), a T-cell immunoglobulin and mucin domain-containing protein 3 (TIM3) inhibitor (e.g., an anti-GITR antibody), a cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) inhibitor (e.g., an anti-GITR antibody), a T-cell immunoglobulin and mucin domain-containing protein 3 (TIM3 ... inhibitors, B and T lymphocyte attenuator (BTLA) inhibitors, T cell immunoreceptor with Ig and ITIM domains (TIGIT) inhibitors, CD47 inhibitors, antagonists of another T cell co-inhibitor or ligand (e.g., antibodies against CD28, 2B4, LY108, LAIR1, ICOS, CD160, or VISTA), CD20 inhibitors (e.g., anti-CD20 antibodies, or bispecific CD3 / CD20 antibodies), indoleamine-2,3-dioxygenase (IDO) inhibitors, vascular endothelial growth factor (VEGF) antagonists [e.g., U.S. Pat. No. 7,087,VEGF-Trap" or other VEGF inhibitory fusion proteins such as aflibercept described in US Pat. No. 411, or anti-VEGF antibodies or antigen-binding fragments thereof (e.g., bevacizumab or ranibizumab), or small molecule kinase inhibitors of VEGF receptors (e.g., sunitinib, sorafenib, or pazopanib)], angiopoietin-2 (Ang2) inhibitors (e.g., nesvacumab), transforming growth factor beta (TGFβ) inhibitors, CD38 inhibitors, epidermal growth factor receptor (EGFR) inhibitors (e.g., erlotinib, cetuximab), agonists for costimulatory receptors (e.g., agonists for glucocorticoid-induced TNFR-related protein), tumor-specific antigens [e.g., CA9, CA125, melanoma-associated antigen 3 (MAGE3), carcinoembryonic antigen (CEA), vimentin, tumor-M2-PK, prostate-specific Antibodies against antigens (PSA, mucin-1, MART-1, and CA19-9), vaccines (e.g., bacillus Calmette-Guerin, cancer vaccines), cyclophosphamide, adjuvants to increase antigen presentation (e.g., granulocyte-macrophage colony-stimulating factor), cytotoxins, chemotherapeutic agents (e.g., dacarbazine, temozolomide, docetaxel, doxorubicin, daunorubicin, cisplatin, carboplatin, gemcitabine, methotrexate, mitoxantrone, oxaliplatin, paclitaxel, and vincristine), interleukin-6 receptor (IL-6R) inhibitors (e.g., sarilumab), IL-4R inhibitors (e.g., dupilumab), IL-10 inhibitors, cytokines such as IL-2, IL-7, IL-21, and IL-15, antibody-drug conjugates (ADCs) (e.g., anti-CD19-DAM4 ADCs, and anti-DS6-DM4 ADCs), chimeric antigen receptor T cells (e.g., CD19-targeted T cells), anti-inflammatory drugs (e.g., corticosteroids and nonsteroidal anti-inflammatory drugs), and nutritional supplements such as antioxidants. The present invention includes the administration of an additional therapeutic agent or treatment regimen or procedure selected from the group consisting of dietary supplements.

[0100] In certain embodiments, the methods of the invention include administering an anti-PD-1 antibody in combination with radiation therapy and optionally an anti-GITR antibody to generate long-term sustainable anti-tumor responses and / or extend the survival of cancer patients. In some embodiments, the methods of the invention include administering radiation therapy before, simultaneously with, or after administering an anti-PD-1 antibody and an anti-GITR antibody to a cancer patient. For example, radiation therapy may be administered in one or more doses to a tumor lesion after administration of one or more doses of the antibody. In some embodiments, radiation therapy may be administered locally to a tumor lesion after systemic administration of an anti-PD-1 antibody and / or an anti-GITR antibody to enhance the local immunogenicity of the patient's tumor (adjuvant radiation) and / or to kill tumor cells (ablative radiation). In certain embodiments, radiation therapy is administered to a first tumor lesion but not to a second tumor lesion, and administration in combination with an anti-PD-1 antibody results in tumor regression in both the first and second tumor lesions (abscopal effect). In certain embodiments, the methods of the invention comprise administering an anti-PD-1 antibody in combination with radiation therapy and optionally an anti-GITR antibody to produce a sustained abscopal effect.

[0101] In certain embodiments, the anti-PD-1 antibody may be administered in combination with radiation therapy and a chemotherapeutic agent (e.g., temozolomide or cyclophosphamide), a VEGF antagonist (e.g., aflibercept), or granulocyte-macrophage colony-stimulating factor.

[0102] Pharmaceutical Compositions and Administration The present invention includes a method comprising administering to a subject an anti-PD-1 antibody in combination with radiation, wherein the anti-PD-1 antibody is contained within a pharmaceutical composition. The pharmaceutical compositions of the present invention may be formulated with suitable carriers, excipients, and other agents that provide suitable transport, delivery, tolerability, and the like. A large number of suitable formulations can be found in a formulary known to all medicinal chemists: Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, PA. These formulations include, for example, powders, pastes, ointments, jellies, waxes, oils, lipids, lipid (cationic or anionic)-containing vesicles (such as LIPOFECTIN™), DNA conjugates, anhydrous absorption pastes, oil-in-water and water-in-oil emulsions, emulsions-carbowax (polyethylene glycols of various molecular weights), semi-solid gels, and semi-solid mixtures containing carbowax. See also Powell et al., "Compendium of excipients for parenteral formulations," PDA (1998) J Pharm Sci Technol 52:238-311.

[0103] Various delivery systems, such as encapsulation in liposomes, microparticles, microcapsules, recombinant cells capable of expressing mutant viruses, receptor-mediated endocytosis, are known and can be used to administer the pharmaceutical compositions of the present invention (see, e.g., Wu et al., 1987, J. Biol. Chem. 262:4429-4432). Methods of administration include, but are not limited to, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, and oral routes. The compositions may be administered by any convenient route, such as infusion or bolus injection, absorption through epithelial or mucocutaneous linings (e.g., oral mucosa, rectal and intestinal mucosa, etc.), and may be administered together with other bioactive agents.

[0104] The pharmaceutical compositions of the present invention can be delivered subcutaneously or intravenously via a standard needle and syringe. In addition, for subcutaneous delivery, pen delivery devices are easily adapted to deliver the pharmaceutical composition of the present invention. Such pen delivery devices may be reusable or disposable. Reusable pen delivery devices generally utilize a replaceable cartridge containing the pharmaceutical composition. Once all of the pharmaceutical composition in the cartridge has been administered and the cartridge is emptied, the empty cartridge can be easily discarded and replaced with a new cartridge containing the pharmaceutical composition. In this manner, the pen delivery device can be reused. In disposable pen delivery devices, there is no replaceable cartridge. Rather, disposable pen delivery devices are pre-filled with the pharmaceutical composition contained in a reservoir within the device. Once the reservoir is emptied of the pharmaceutical composition, the entire device is discarded.

[0105] In certain circumstances, the pharmaceutical composition can be delivered in a controlled release system. In one embodiment, a pump can be used. In another embodiment, a polymeric material can be used; see Medical Applications of Controlled Release, Langer and Wise (eds.), 1974, CRC Pres., Boca Raton, Florida. In yet another embodiment, a controlled release system can be placed proximal to the target of the composition, thus requiring only a fraction of the systemic dose (see, e.g., Goodson, 1984, "Medical Applications of Controlled Release," supra, vol. 2, pp. 115-138). Other controlled release systems are discussed in the review by Langer, 1990, Science 249:1527-1533.

[0106] The injectable preparations may include dosage forms for intravenous, subcutaneous, intradermal, and intramuscular injections, drip infusions, and the like. These injectable preparations may be prepared by known methods. For example, the injectable preparations may be prepared by dissolving, suspending, or emulsifying, for example, the above-mentioned antibody or its salt in a sterile aqueous or oily medium that is usually used for injections. Examples of aqueous media for injection include isotonic solutions containing physiological saline, glucose, and other auxiliary agents, and the like, which may be used in combination with suitable solubilizers such as alcohols (e.g., ethanol), polyhydric alcohols (e.g., propylene glycol, polyethylene glycol), nonionic surfactants [e.g., polysorbate 80, HCO-50 (polyoxyethylene (50 moles) adduct of hydrogenated castor oil)], and the like. Examples of oily media include sesame oil, soybean oil, and the like, which may be used in combination with solubilizers such as benzyl benzoate, benzyl alcohol, and the like. The injectable preparations thus prepared are preferably filled into suitable ampoules.

[0107] The above-mentioned pharmaceutical compositions for oral or parenteral use are advantageously prepared in a dosage form with a unit dose suitable for the dose of the active ingredient.Such dosage forms with a unit dose include, for example, tablets, pills, capsules, injections (ampoules), suppositories, etc.

[0108] In certain embodiments, the invention provides a pharmaceutical formulation comprising a therapeutic amount of an anti-PD-1 antibody and a pharmaceutical carrier, hi certain embodiments, the invention provides an anti-PD-1 antibody formulated into a pharmaceutical composition for use in intravenous administration.

[0109] Dosing regimen The invention includes methods comprising administering an anti-PD-1 antibody to a subject about 4 times per week, 2 times per week, 1 time per week, 1 time per 2 weeks, 1 time per 3 weeks, 1 time per 4 weeks, 1 time per 5 weeks, 1 time per 6 weeks, 1 time per 8 weeks, 1 time per 12 weeks, or less frequently, so long as a therapeutic response is achieved. In certain embodiments, the invention includes methods comprising administering an anti-PD-1 antibody to a subject about 4 times per week, 2 times per week, 1 time per 2 weeks, 1 time per 3 weeks, 1 time per 4 weeks, 1 time per 5 weeks, 1 time per 6 weeks, 1 time per 8 weeks, 1 time per 12 weeks, or less frequently, so long as a therapeutic response is achieved. The methods include administering radiation therapy to a subject about 7 times per week, about 4 times per week, 2 times per week, once per week, once per week, once per 2 weeks, once per 3 weeks, once per 4 weeks, once per 5 weeks, once per 6 weeks, once per 8 weeks, once per 12 weeks, or less, as long as a therapeutic response is achieved. In certain embodiments, the methods include administering an anti-PD-1 antibody in combination with radiation therapy about 7 times per week, about 4 times per week, 2 times per week, once per week, once per 2 weeks, once per 3 weeks, once per 4 weeks, once per 5 weeks, once per 6 weeks, once per 8 weeks, once per 9 weeks, once per 12 weeks, or less, as long as a therapeutic response is achieved.

[0110] In certain embodiments, the methods of the invention include administering radiation therapy that is hypofractionated radiation therapy. In certain embodiments, the hypofractionated radiation therapy includes 2-12 fractions. In certain embodiments, the 2-12 fractions are administered on consecutive days. In certain embodiments, the radiation therapy is administered after administration of one or more doses of an anti-PD-1 antibody. In certain embodiments, the anti-PD-1 antibody is administered 0.5-2 weeks prior to administration of one or more fractions of radiation therapy.

[0111] According to certain embodiments of the invention, multiple doses of anti-PD-1 antibody in combination with radiation therapy may be administered to the subject over a defined time course. The method according to this aspect of the invention comprises sequentially administering to the subject one or more doses of anti-PD-1 antibody in combination with one or more doses of radiation. As used herein, "sequentially administering" means that each dose of the antibody is administered to the subject at different times, e.g., on different days separated by a predefined interval (e.g., hours, days, weeks, or months). In certain embodiments, the method of the invention comprises sequentially administering one or more doses of anti-PD-1 antibody, each dose being administered 0.5-12 weeks after the immediately preceding dose. In certain further embodiments, the method further comprises administering radiation therapy. The radiation therapy may be hypofractionated radiation therapy. In certain embodiments, the radiation therapy comprises 2-12 fractions. In some embodiments, the fractions of radiation are administered on consecutive or alternating days. In certain embodiments, radiation fractions are administered once every 3 days, once every 4 days, once every 5 days, once every 6 days, once every 7 days, or a combination thereof.

[0112] In certain embodiments, the invention includes methods comprising sequentially administering to a patient a single initial dose of an anti-PD-1 antibody, followed by one or more second doses of an anti-PD-1 antibody, and optionally followed by one or more third doses of an anti-PD-1 antibody. In certain embodiments, the methods further comprise sequentially administering to a patient a single initial dose of radiation therapy, followed by one or more second doses of radiation therapy, and optionally one or more third doses of radiation therapy. In alternative embodiments, the methods further comprise sequentially administering one or more fractions of hypofractionated radiation therapy.

[0113] According to certain embodiments of the invention, multiple doses of anti-PD-1 antibody and radiation therapy may be administered to a subject over a defined time course. The method according to this aspect of the invention includes sequentially administering to a subject multiple doses of anti-PD-1 antibody and radiation. As used herein, "sequentially administering" means that each dose of anti-PD-1 antibody in combination with radiation therapy is administered to a subject at different time points, e.g., on different days separated by a defined interval (e.g., hours, days, weeks, or months).

[0114] The terms "first dose," "second dose," and "third dose" refer to the temporal order of administration. Thus, a "first dose" is the dose administered at the beginning of a treatment regimen (also called a "baseline dose"); a "second dose" is the dose administered after the first dose. and the "third dose" is the dose administered after the second dose. The first, second, and third doses may all contain the same amount of antibody (anti-PD-1 antibody). However, in certain embodiments, the amounts contained in the first, second, and / or third doses differ from each other during the course of treatment (e.g., adjusted up or down as needed). In certain embodiments, one or more (e.g., 1, 2, 3, 4, or 5) doses are administered as a "loading dose" at the beginning of the treatment regimen, followed by subsequent doses administered less frequently (e.g., "maintenance doses"). For example, an anti-PD-1 antibody can be administered to a cancer patient at a loading dose of about 1 to 3 mg / kg of patient body weight, followed by one or more maintenance doses of about 0.1 to about 20 mg / kg.

[0115] In one exemplary embodiment of the invention, each second and / or third dose is administered ½ to 14 weeks (e.g., ½, 1, ½, 2, ½, 3, ½, 4, ½, 5, ½, 6, ½, 7, ½, 8, ½, 9, ½, 10, ½, 11, ½, 12, ½, 13, ½, 14, ½, or more than 14 ½ weeks) after the immediately preceding dose. As used herein, the phrase "immediately preceding dose" refers to a dose of anti-PD-1 antibody (and / or radiation) that is administered to a patient prior to administration of the immediately next dose in the sequence, without any intervening doses.

[0116] The method according to this aspect of the invention may include administering any number of second and / or third doses of anti-PD-1 antibody (and / or radiation therapy) to the patient. For example, in certain embodiments, only a single second dose is administered to the patient. In other embodiments, two or more than two (e.g., 2, 3, 4, 5, 6, 7, 8, or more than eight) second doses are administered to the patient. Similarly, in certain embodiments, only a single third dose is administered to the patient. In other embodiments, two or more than two (e.g., 2, 3, 4, 5, 6, 7, 8, or more than eight) third doses are administered to the patient.

[0117] In embodiments involving multiple second doses, each second dose may be administered at the same time as the other second doses. For example, each second dose may be administered to the patient 1-2 weeks after the immediately preceding dose. Similarly, in embodiments involving multiple third doses, each third dose may be administered at the same time as the other third doses. For example, each third dose may be administered to the patient 2-4 weeks after the immediately preceding dose. Alternatively, the number of times the second and / or third doses are administered to the patient may vary over the course of the treatment regimen. The number of administrations may be adjusted by the physician during the course of treatment according to the needs of the individual patient after consultation.

[0118] In certain embodiments, one or more doses of anti-PD-1l antibody and / or radiation are administered at the start of a treatment regimen as a more frequent "induction dose" (twice per week, once per week, or once every two weeks), followed by subsequent doses ("consolidation doses" or "maintenance doses") administered less frequently (e.g., once every 2-12 weeks). In certain embodiments, one or more doses of anti-PD-1 antibody and / or radiation are administered more frequently (twice per week, once per week, or once every two weeks) as an "induction dose" at the start of a treatment regimen, followed by the next dose of anti-PD-1 antibody.

[0119] The invention includes methods comprising sequentially administering one or more doses of an anti-PD-1 antibody in combination with one or more doses of radiation therapy, where one or more doses are included in one or more treatment cycles.

[0120] According to certain embodiments of the invention, the method comprises administering to a subject one or more of the anti-PD-1 antibodies and optionally one or more doses of radiation therapy. In certain embodiments, the treatment cycle comprises 1-10 doses of anti-PD-1 antibody, with each dose of anti-PD-1 antibody being administered 0.5-8 weeks after the immediately preceding dose. In certain embodiments, the method of the invention comprises the administration of up to 6 or 8 treatment cycles. In certain other embodiments, the method of the invention comprises the administration of up to 12 or more treatment cycles, depending on therapeutic efficacy. In certain embodiments, at least one treatment cycle further comprises radiation therapy. In some embodiments, the radiation therapy is hypofractionated radiation therapy, and the hypofractionated radiation therapy comprises 2-12 fractions. In certain embodiments, the 2-12 fractions are administered on consecutive days.

[0121] The present invention includes methods that include sequential administration of an anti-PD-1 antibody to a patient in combination with radiation therapy to treat cancer (e.g., solid tumors). In some embodiments, the methods of the invention include administering one or more doses of an anti-PD-1 antibody followed by radiation therapy. In certain further embodiments, radiation therapy is administered in fractions (hypofractionated radiation therapy). In certain embodiments, the methods of the invention include a single dose of an anti-PD-1 antibody followed by 2-10 fractions of radiation therapy followed by administration of one or more doses of an anti-PD-1 antibody. In some embodiments, one or more doses of about 0.1 mg / kg to about 20 mg / kg of an anti-PD-1 antibody may be administered followed by radiation therapy to inhibit tumor growth and / or prevent tumor recurrence in a subject with cancer (e.g., solid tumor). In some embodiments, administration of one or more doses of an anti-PD-1 antibody is followed by radiation therapy, which results in increased anti-tumor efficacy (e.g., greater inhibition of tumor growth, increased prevention of tumor recurrence compared to untreated subjects or subjects administered either the antibody or radiation as monotherapy). Alternative embodiments of the invention relate to the co-administration of an anti-PD-1 antibody and radiation, administered at similar or different times compared to the anti-PD-1 antibody, hi some embodiments, radiation therapy is administered before, after, or simultaneously with the anti-PD-1 antibody.

[0122] dosage The amount of anti-PD-1 antibody administered to a subject according to the method of the present invention is generally a therapeutically effective amount. As used herein, the term "therapeutically effective amount" refers to an amount of antibody (anti-PD-1 antibody) that results in one or more of the following: (a) reducing the severity or duration of symptoms or indications of cancer, such as solid tumors; (b) inhibiting tumor growth or increasing tumor necrosis, tumor shrinkage and / or tumor disappearance; (c) slowing tumor growth and progression; (d) inhibiting tumor metastasis; (e) preventing recurrence of tumor growth; (f) prolonging the survival of subjects with cancer; and / or (g) reducing the use of conventional anti-cancer treatments (e.g., reducing or eliminating the use of chemotherapeutic or cytotoxic agents) compared to untreated subjects or subjects administered the antibody as a monotherapy.

[0123] In the case of an anti-PD-1 antibody, a therapeutically effective amount can be about 0.05 mg to about 600 mg, about 1 mg to about 500 mg, about 10 mg to about 450 mg, about 50 mg to about 400 mg, about 75 mg to about 350 mg, or about 100 mg to about 300 mg of the antibody. For example, in various embodiments, the amount of anti-PD-1 antibody can be about 0.05 mg, about 0.1 mg, about 1.0 mg, about 1.5 mg, about 2.0 mg, about 10 mg, about 20 mg, about 30 mg, about 40 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, about 100 mg, about 110 mg, about 120 mg, about 130 mg, about 140 mg, about 150 mg, about 160 mg, about 170 mg, about 180 mg, or about 200 mg of the anti-PD-1 antibody. 0mg, about 190mg, about 200mg, about 210mg, about 220mg, about 230mg, about 240mg, about 250mg, about 260mg, about 270mg, about 280mg, about 290mg, about 300mg, about 310 mg, about 320mg, about 330mg, about 340mg, about 350mg, about 360mg, about 370mg, about 380mg, about 390mg, about 400mg, about 410mg, about 420mg, about 430mg, about 440mg , about 450 mg, about 460 mg, about 470 mg, about 480 mg, about 490 mg, about 500 mg, about 510 mg, about 520 mg, about 530 mg, about 540 mg, about 550 mg, about 560 mg, about 570 mg, about 580 mg, about 590 mg, or about 600 mg. In one embodiment, 250 mg of an anti-PD-1 antibody is administered in accordance with the methods of the invention. In one embodiment, 200 mg of an anti-PD-1 antibody is administered in accordance with the methods of the invention. In one embodiment, 350 mg of an anti-PD-1 antibody is administered in accordance with the methods of the invention.

[0124] The amount of any anti-PD-1 antibody included in an individual dose can be expressed in terms of milligrams of antibody per kilogram of the subject's body weight (i.e., mg / kg). In certain embodiments, the anti-PD-1 antibody used in the methods of the invention can be administered to the subject at a dose of about 0.0001 to about 100 mg per kilogram of the subject's body weight. In certain embodiments, the anti-PD-1 antibody can be administered at a dose of about 0.1 mg to about 20 mg per kilogram of the subject's body weight. In certain embodiments, the methods of the invention include administration of an anti-PD-1 antibody at a dose of about 1 mg, 3 mg, 5 mg, or 10 mg per kilogram of the patient's body weight.

[0125] In certain embodiments, the amount of anti-PD-1 antibody administered to a patient may be less than a therapeutically effective amount, i.e., a sub-therapeutic dose. For example, if the therapeutically effective amount of anti-PD-1 antibody comprises 3 mg / kg, a sub-therapeutic dose comprises an amount less than 3 mg / kg, such as 2 mg / kg, 1.5 mg / kg, 1 mg / kg, 0.5 mg / kg, or 0.3 mg / kg. As defined herein, a "sub-therapeutic dose" refers to an amount of anti-PD-1 antibody that does not achieve a therapeutic effect alone. However, in certain embodiments, a sub-therapeutic dose of anti-PD-1 antibody is administered together with a second therapeutic agent and optionally a third therapeutic agent to promote a therapeutic effect.

[0126] In certain embodiments, the radiation therapy administered to a subject in need thereof comprises 2-100 Gray (Gy). In certain embodiments, the radiation therapy comprises 5, 7, 8, 9, 10, 11, 12, 15, 20, 23, 25, 27, 30, 35, 40, or 45 Gy. In certain other embodiments, the radiation therapy comprises 50-100, 60-90, or 70-80 Gy. In certain embodiments, the radiation therapy is administered in 2-12 fractions (hypofractic radiation therapy), each fraction comprising 2-10 Gy. For example, 30 Gy of radiation is administered in 5 fractions, each fraction comprising 6 Gy.

[0127] Selected Embodiments Selected embodiments of the present invention include:

[0128] In some embodiments, the disclosure provides a method of treating or inhibiting tumor growth in a subject, comprising: (a) selecting a subject with cancer; and (b) administering to a subject in need thereof one or more doses of radiation therapy in combination with one or more doses of a therapeutically effective amount of an antibody or antigen-binding fragment thereof that specifically binds to Programmed Cell Death 1 (PD-1), wherein administration of the combination results in enhanced therapeutic efficacy compared to administration of the antibody or radiation alone; The present invention provides a method comprising:

[0129] In one embodiment, each dose of the anti-PD-1 antibody contains 0.1-20 mg per kg of the subject's body weight.

[0130] In another embodiment, each dose of the anti-PD-1 antibody contains 0.3, 1, 3, 5, or 10 mg / kg of the subject's body weight.

[0131] In other embodiments, each dose of the anti-PD-1 antibody comprises 20 to 400 mg.

[0132] In some embodiments, each dose of the anti-PD-1 antibody comprises 200 mg.

[0133] In one embodiment, each dose of radiation comprises between 2 and 80 Gray (Gy).

[0134] In another embodiment, each dose of anti-PD-1 antibody comprises 1, 3, or 10 mg / kg of subject's body weight and each dose of radiation therapy comprises 20-50 Gy.

[0135] In other embodiments, the radiation therapy is fractionated radiation therapy.

[0136] In some embodiments, the fractionated radiation therapy comprises 2 to 10 fractions.

[0137] In one embodiment, the fractionated radiation therapy comprises 5 fractions of 30 Gy.

[0138] In another embodiment, the fractionated radiation therapy comprises 27 Gy in 3 fractions.

[0139] In other embodiments, between 4 and 50 doses of the anti-PD-1 antibody are administered, with each dose being administered 0.5 to 4 weeks after the immediately preceding dose.

[0140] In some embodiments, each dose of the anti-PD-1 antibody is administered two weeks after the immediately preceding dose.

[0141] In one embodiment, the anti-PD-1 antibody is administered before, concurrently with, or after radiation therapy.

[0142] In another embodiment, the anti-PD-1 antibody is administered prior to radiation therapy.

[0143] In other embodiments, the anti-PD-1 antibody is administered one week prior to radiation therapy.

[0144] In some embodiments, the enhanced therapeutic efficacy comprises an effect selected from the group consisting of tumor regression, abscopal effect, inhibition of tumor metastasis, reduction in metastatic lesions over time, reduction in the use of chemotherapeutic or cytotoxic agents, reduction in tumor burden, prolonged progression-free survival, prolonged overall survival, complete response, partial response, and stable disease.

[0145] In one embodiment, the enhanced efficacy of treatment comprises tumor regression in tumors distal to the irradiated tumor.

[0146] In another embodiment, tumor growth is inhibited by at least 50% compared to subjects receiving either the antibody or radiation alone.

[0147] In other embodiments, tumor growth is inhibited by at least 50% compared to subjects who received a dose of radiation prior to the anti-PD-1 antibody.

[0148] In some embodiments, the disclosure provides methods of treating a tumor comprising the steps of: (a) selecting a subject having cancer; and (b) administering to the subject at least one treatment cycle, wherein the at least one treatment cycle comprises 1 to 6 doses of an anti-PD-1 antibody, with each dose administered 2 weeks after the immediately preceding dose.

[0149] In one embodiment, each dose of the anti-PD-1 antibody contains 1, 3, 5, or 10 mg / kg of the subject's body weight.

[0150] In another embodiment, at least one treatment cycle further comprises radiation therapy.

[0151] In other embodiments, radiation therapy comprises about 20-50 Gy.

[0152] In some embodiments, the radiation therapy comprises about 27 Gy.

[0153] In one embodiment, the radiation therapy comprises about 30 Gy.

[0154] In another embodiment, the radiation therapy is fractionated radiation therapy.

[0155] In another embodiment, the fractionated radiation therapy comprises 2 to 6 fractions.

[0156] In some embodiments, the fractionated radiation therapy comprises three fractions.

[0157] In one embodiment, the fractionated radiation therapy comprises 5 fractions.

[0158] In another embodiment, the radiation therapy comprises about 27 Gy in three fractions.

[0159] In other embodiments, radiation therapy comprises about 30 Gy in 5 fractions.

[0160] In some embodiments, the divisions are administered on consecutive days.

[0161] In one embodiment, the anti-PD-1 antibody is administered one week prior to radiation therapy.

[0162] In another embodiment, up to 10 treatment cycles are administered to a subject in need thereof.

[0163] In other embodiments, six treatment cycles are administered to a subject in need thereof.

[0164] In some embodiments, radiation therapy is administered in the first treatment cycle.

[0165] In one embodiment, radiation therapy comprises about 20-50 Gy.

[0166] In another embodiment, the radiation therapy comprises hypofractionated radiation therapy.

[0167] In another embodiment, the fractionated radiation therapy comprises 2 to 6 fractions.

[0168] In some embodiments, radiation therapy comprises about 27 Gy in three fractions.

[0169] In one embodiment, radiation therapy comprises about 30 Gy in five fractions.

[0170] In another embodiment, the divisions are administered on consecutive days.

[0171] In other embodiments, the anti-PD-1 antibody is administered one week prior to radiation therapy.

[0172] In some embodiments, each treatment cycle comprises 4 doses of the anti-PD-1 antibody.

[0173] In one embodiment, the treatment results in a therapeutic effect selected from the group consisting of inhibition of tumor growth, tumor regression, reduction in tumor size, reduction in tumor cell count, slowing of tumor growth, abscopal effect, inhibition of tumor metastasis, reduction in metastatic lesions over time, reduction in the use of chemotherapeutic or cytotoxic agents, reduction in tumor burden, prolonged progression-free survival, prolonged overall survival, complete response, partial response, and stable disease.

[0174] In another embodiment, the treatment effect comprises tumor regression in a tumor distal to the irradiated tumor in the subject.

[0175] In another embodiment, tumor growth is delayed by at least 10 days compared to untreated controls.

[0176] In some embodiments, tumor growth is inhibited by at least 50% compared to untreated controls.

[0177] In one embodiment, tumor growth is inhibited by at least 50% compared to subjects receiving either the antibody or radiation alone.

[0178] In some embodiments, the disclosure provides a method of treating a tumor comprising: (a) selecting a subject having a first solid tumor lesion and a second solid tumor lesion, where the second solid tumor lesion is located distal to the first solid tumor lesion; and (b) administering an anti-PD-1 antibody, or antigen-binding fragment thereof, in combination with radiation therapy.

[0179] In other embodiments, radiation therapy is administered to a first tumor lesion but not to a second tumor lesion, and the administration results in tumor regression in both the first and second tumor lesions.

[0180] In one embodiment, the anti-PD-1 antibody is administered prior to radiation therapy.

[0181] In another embodiment, the subject is refractory or has responded inadequately to treatment, or has relapsed after previous treatment.

[0182] In other embodiments, the cancer is a recurrent or metastatic cancer.

[0183] In some embodiments, the method further comprises administering to the subject an additional therapeutic agent or treatment, the additional therapeutic agent or treatment being surgery, a chemotherapeutic agent, a cancer vaccine, a programmed death-ligand 1 (PD-L1) inhibitor, a lymphocyte-activation gene 3 (LAG3) inhibitor, a cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) inhibitor, a glucocorticoid-induced tumor necrosis factor receptor (GITR) inhibitor, a T-cell immunoglobulin and mucin domain-containing protein 3 (TIM3) inhibitor, a B- and T-lymphocyte attenuator (BTLA) inhibitor, a T-cell immunoreceptor with Ig and ITIM domains (TIGIT) inhibitor, a CD47 inhibitor, an indoleamine-2,3-dioxygenase inhibitor, or an indoleamine-2,3-dioxygenase inhibitor. and / or anti-CD3 / anti-CD20 antibodies, vascular endothelial growth factor (VEGF) antagonists, angiopoietin-2 (Ang2) inhibitors, transforming growth factor beta (TGFβ) inhibitors, epidermal growth factor receptor (EGFR) inhibitors, granulocyte-macrophage colony-stimulating factor (GM-CSF), cyclophosphamide, antibodies against tumor-specific antigens, Bacillus Calmette-Guerin vaccines, cytotoxins, interleukin-6 receptor (IL-6R) inhibitors, interleukin-4 receptor (IL-4R) inhibitors, IL-10 inhibitors, IL-2, IL-7, IL-21, IL-15, antibody-drug conjugates, anti-inflammatory drugs, and dietary supplements.

[0184] In one embodiment, the additional therapeutic agent is an anti-GITR antibody.

[0185] In another embodiment, the additional therapeutic agent is cyclophosphamide.

[0186] In other embodiments, the additional therapeutic agent is GM-CSF.

[0187] In some embodiments, the additional therapeutic agent is selected from the group consisting of docetaxel, carboplatin, paclitaxel, cisplatin, gemcitabine, and pemetrexed.

[0188] In one embodiment, the anti-PD-1 antibody is administered intravenously, subcutaneously, or intraperitoneally.

[0189] In another embodiment, the cancer comprises a solid tumor.

[0190] In other embodiments, the solid tumor is selected from the group consisting of colorectal cancer, ovarian cancer, prostate cancer, breast cancer, brain cancer, cervical cancer, bladder cancer, anal cancer, uterine cancer, colon cancer, liver cancer, pancreatic cancer, lung cancer, endometrial cancer, bone cancer, testicular cancer, skin cancer, kidney cancer, gastric cancer, esophageal cancer, head and neck cancer, salivary gland cancer, and myeloma.

[0191] In some embodiments, the solid tumor is selected from the group consisting of hepatocellular carcinoma, non-small cell lung cancer, squamous cell carcinoma of the head and neck, basal cell carcinoma, breast cancer, squamous cell carcinoma of the skin, chondrosarcoma, angiosarcoma, cholangiocarcinoma, soft tissue sarcoma, colorectal cancer, melanoma, Merkel cell carcinoma, and glioblastoma multiforme.

[0192] In one embodiment, the anti-PD-1 antibody, or antigen-binding fragment thereof, comprises a heavy chain complementarity determining region (HCDR1, HCDR2, and HCDR3) of a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO:1, and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) of a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO:2.

[0193] In another embodiment, the anti-PD-1 antibody or antigen-binding fragment thereof comprises three HCDRs (HCDR1, HCDR2, and HCDR3) and three LCDRs (LCDR1, LCDR2, and LCDR3), where HCDR1 comprises the amino acid sequence of SEQ ID NO:3; HCDR2 comprises the amino acid sequence of SEQ ID NO:4; HCDR3 comprises the amino acid sequence of SEQ ID NO:5; LCDR1 comprises the amino acid sequence of SEQ ID NO:6; LCDR2 comprises the amino acid sequence of SEQ ID NO:7; and LCDR3 comprises the amino acid sequence of SEQ ID NO:8.

[0194] In other embodiments, the HCVR comprises the amino acid sequence of SEQ ID NO:1 and the LCVR comprises the amino acid sequence of SEQ ID NO:2.

[0195] In another embodiment, the anti-PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:9 and a light chain comprising the amino acid sequence of SEQ ID NO:10.

[0196] In some embodiments, the disclosure provides a method of inhibiting activation and / or proliferation of regulatory T (Treg) cells, the method comprising: (a) selecting a subject having a solid tumor; and (b) administering to the subject at least one of: (i) an anti-PD-1 antibody or antigen-binding fragment thereof, (ii) radiation therapy, and (iii) an antibody that specifically binds to glucocorticoid-induced tumor necrosis factor receptor (GITR) or an antigen-binding fragment thereof, cyclophosphamide, GM-CSF, an anti-LAG3 antibody, docetaxel, or carboplatin.

[0197] In one embodiment, the subject has a large tumor.

[0198] In another embodiment, the dose of radiation is from 2 to 50 Gy.

[0199] In other embodiments, the administration results in at least one effect selected from the group consisting of inhibition of tumor growth, tumor regression, reduction in tumor size, reduction in tumor cell count, slowing of tumor growth, abscopal effect, inhibition of tumor metastasis, reduction in metastatic lesions over time, reduction in the use of chemotherapeutic or cytotoxic agents, reduction in tumor burden, prolonged progression-free survival, prolonged overall survival, complete response, partial response, and stable disease.

[0200] In some embodiments, the solid tumor is selected from the group consisting of colorectal cancer, ovarian cancer, prostate cancer, breast cancer, brain cancer, cervical cancer, bladder cancer, anal cancer, uterine cancer, colon cancer, liver cancer, pancreatic cancer, lung cancer, endometrial cancer, bone cancer, testicular cancer, skin cancer, kidney cancer, gastric cancer, esophageal cancer, head and neck cancer, salivary gland cancer, and myeloma.

[0201] In one embodiment, the anti-PD-1 antibody, or antigen-binding fragment thereof, comprises a heavy chain complementarity determining region (HCDR1, HCDR2, and HCDR3) of a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO:1, and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) of a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO:2.

[0202] In another embodiment, the anti-PD-1 antibody or antigen-binding fragment thereof comprises three HCDRs (HCDR1, HCDR2, and HCDR3) and three LCDRs (LCDR1, LCDR2, and LCDR3), where HCDR1 comprises the amino acid sequence of SEQ ID NO:3; HCDR2 comprises the amino acid sequence of SEQ ID NO:4; HCDR3 comprises the amino acid sequence of SEQ ID NO:5; LCDR1 comprises the amino acid sequence of SEQ ID NO:6; LCDR2 comprises the amino acid sequence of SEQ ID NO:7; and LCDR3 comprises the amino acid sequence of SEQ ID NO:8.

[0203] In other embodiments, the HCVR comprises the amino acid sequence of SEQ ID NO:1 and the LCVR comprises the amino acid sequence of SEQ ID NO:2.

[0204] In one embodiment, the anti-PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:9 and a light chain comprising the amino acid sequence of SEQ ID NO:10.

[0205] In some embodiments, the disclosure provides a method of treating or inhibiting tumor growth, comprising: (a) selecting a subject with skin cancer; and (b) administering to a subject in need thereof a therapeutically effective amount of an antibody or antigen-binding fragment thereof that specifically binds to PD-1; The present invention provides a method comprising:

[0206] In one embodiment, the antibody or antigen-binding fragment thereof that specifically binds to PD-1 is administered as a monotherapy.

[0207] In another embodiment, the skin cancer is a UV-related skin cancer.

[0208] In other embodiments, the skin cancer is selected from the group consisting of cutaneous squamous cell carcinoma (CSCC), basal cell carcinoma (BCC), Merkel cell carcinoma, and melanoma.

[0209] In some embodiments, however, the skin cancer is not head and neck squamous cell carcinoma.

[0210] In one embodiment, the skin cancer is metastatic, unresectable, and / or locally advanced.

[0211] In another embodiment, the skin cancer is BCC and the patient is intolerant to or has progressed following treatment with a hedgehog pathway inhibitor.

[0212] In other embodiments, the antibody or antigen-binding fragment thereof is administered in one or more doses, with each dose being administered 0.5 to 4 weeks after the immediately preceding dose.

[0213] In some embodiments, each dose is administered two weeks after the immediately preceding dose.

[0214] In one embodiment, each dose contains 1, 3, or 10 mg / kg of subject body weight.

[0215] In another embodiment, the antibody or antigen-binding fragment thereof that specifically binds to PD-1 is an antibody as defined in any one of the preceding embodiments.

[0216] In some embodiments, the disclosure provides a method of treating or inhibiting tumor growth in a subject, the method comprising: selecting a subject having brain cancer; and administering to a subject in need thereof a therapeutically effective amount of an antibody or antigen-binding fragment thereof that specifically binds PD-1.

[0217] In one embodiment, the subject has glioblastoma multiforme (GBM).

[0218] In another embodiment, the subject has newly diagnosed GBM.

[0219] In other embodiments, the subject is 65 years of age or older.

[0220] In one embodiment, the anti-PD-1 antibody or antigen-binding fragment thereof is administered in one or more doses, with each dose being administered 0.5 to 4 weeks after the immediately preceding dose.

[0221] In one embodiment, each dose is administered two weeks after the immediately preceding dose.

[0222] In another embodiment, each dose contains 1, 3, or 10 mg / kg of subject body weight.

[0223] In other embodiments, the method further comprises administering radiation therapy to a subject in need thereof.

[0224] In some embodiments, the radiation therapy is hypofractionated radiation therapy.

[0225] In one embodiment, the subject receives 20-50 Gy of radiation in 2-20 fractions.

[0226] In another embodiment, the subject is administered radiation therapy one week after the first dose of the anti-PD-1 antibody.

[0227] In other embodiments, one or more doses of the anti-PD-1 antibody are included in one or more treatment cycles, with each cycle containing 1 to 6 doses of the anti-PD-1 antibody.

[0228] In some embodiments, each treatment cycle comprises 4 doses of the anti-PD-1 antibody, and each dose is administered 2 weeks after the previous dose.

[0229] In one embodiment, each dose contains 1, 3, or 10 mg / kg of subject body weight.

[0230] In another embodiment, the first treatment cycle further comprises radiation therapy.

[0231] In other embodiments, the radiation therapy is hypofractionated radiation therapy.

[0232] In some embodiments, the subject receives 20-50 Gy of radiation in 2-20 fractions.

[0233] In one embodiment, subjects receive 30 Gy in 5 daily fractions.

[0234] In another embodiment, the radiation therapy is administered one week after administration of the anti-PD-1 antibody.

[0235] In other embodiments, if the subject develops intracranial edema following administration of the anti-PD-1 antibody, the method further comprises administering an anti-angiogenic agent to the subject.

[0236] In some embodiments, the anti-angiogenic agent is selected from the group consisting of a vascular endothelial growth factor (VEGF) inhibitor and an angiopoietin-2 (Ang-2) inhibitor.

[0237] In one embodiment, the antiangiogenic agent is bevacizumab or aflibercept.

[0238] In another embodiment, the antibody or antigen-binding fragment thereof that specifically binds to PD-1 is an antibody as defined in any one of the preceding embodiments. EXAMPLES

[0239] The following examples are set forth to provide those skilled in the art with a complete disclosure and description of how to make and use the methods and compositions of the present invention, and are not intended to limit the scope of what the inventors regard as the invention. Efforts have been made to ensure accuracy with respect to the numbers used (e.g., amounts, temperatures, etc.), but some experimental error and deviation should be allowed for. Unless otherwise indicated, amounts are parts by weight, molecular weights are average molecular weights, temperatures are in degrees Celsius, and pressures are at or near atmospheric pressure. EXAMPLES

[0240] In vivo efficacy of anti-PD-1 antibody in combination with radiotherapy against MC38 tumors In this example, the effect of PD-1 blockade in combination with radiation therapy was examined on established MC38 tumors in mice.

[0241] MC38 colon cancer cells 5 x 10 5 The tumors were subcutaneously implanted into the right flank of female C57BL / 6 mice (Jackson Laboratory). The mean tumor volume was approximately 100 mm. 3 Treatment began 9 days after implantation, when the tumor size reached 100 mg / kg. Mice were randomly assigned to receive either an isotype control (2A3, BioXcell) or a PD-1 blocking antibody (RMP1-14, BioXcell) at 5 mg / kg intraperitoneal injection twice weekly for a total of 5 injections. One day after the initiation of antibody treatment, mice assigned to the radiation therapy group received 12 Gy of radiation to the tumor in the right flank. Radiation therapy was performed using an RS2000 Biological A Research Irradiator (Rad Source) was used to irradiate a partial body with a fixture (Precision X-ray) and a sheet of lead (Imag The tumors were delivered to shielded and anesthetized mice (ketamine / xylazine) by a fluoroscopy (Esco Scientific Instruments). Tumor growth was assessed three times a week until days 70-80, after which all mice were euthanized. Figure 1 shows the study design of the experiment including administration of anti-PD-1 antibody and radiation.

[0242] FIG. 2 and Table 1 show the mean tumor volumes in mice treated with anti-PD-1 antibodies alone or in combination with radiation.

[0243] [Table 1]

[0244] Blockade of PD-1 (RMP1-14) synergized with focal radiation (XRT) to induce significant tumor regression in MC38 tumor-bearing mice (4 of 6 mice) compared to XRT + isotype control treated mice (2 of 6 mice). Tumor growth was inhibited or delayed in mice treated with anti-PD-1 antibody in combination with radiation. Mice treated with anti-PD-1 antibody and radiation grew 500 mm 3 compared with mice treated with monotherapy, which took less than 20 days to reach a tumor volume of 500 mm 3 It took more than 40 days to reach a regression of the tumor. Tumor regression lasted up to 4 weeks in the combo (XRT + anti-PD-1 antibody) treated group (1 of 4 mice that rejected the tumor relapsed at this time point) compared to 1.5 weeks in the XRT + isotype treated group (1 of 2 mice that rejected the tumor relapsed). Blockade of PD-1 as a monotherapy had no effect on primary tumor growth in this tumor model.

[0245] [Table 2]

[0246] The therapeutic efficacy of the combination treatment (XRT + anti-PD-1 antibody) was demonstrated by a statistical increase in overall survival in this group (50% alive at day 70 after tumor implantation) compared to all other treatment groups: isotype control (0% alive at day 70), anti-PD-1 antibody treated (0% alive at day 70), and XRT + isotype treated mice (17% alive at day 70) (Figure 3; Table 2). EXAMPLES

[0247] In vivo efficacy of anti-PD-1 antibody and radiotherapy against B16 tumors In this example, the anti-tumor effect of an anti-mouse PD-1 antibody in combination with radiation therapy was tested against established B16 tumors in mice.

[0248] B16F10.9 melanoma cells 2 × 10 5 The tumors were subcutaneously implanted into the right flank of female C57BL / 6 mice (Jackson Laboratory). The mean tumor volume was approximately 150 mm. 3 Treatment was initiated when tumor size reached 100%. Mice were randomly assigned to receive intraperitoneal injections of either isotype control (2A3, BioXcell) or PD-1 blocking antibody (RMP1-14, BioXcell) at 5 mg / kg twice weekly for a total of five injections. One day after the initiation of antibody treatment, mice assigned to the radiation therapy group received 8 Gy of radiation to the tumors in the right flank. Radiation therapy was delivered to anesthetized mice (ketamine / xylazine) using an RS2000 Biological Research Irradiator (Rad Source), shielded by a partial body irradiation fixture (Precision X-ray) and a sheet of lead (Images Scientific Instruments). Tumor growth was assessed three times a week until 70–80 days, after which all mice were euthanized. Figure 4 shows the study design of the experiment including administration of anti-PD-1 antibody and radiation.

[0249] PD-1 (RMP1-14) blocking antibody treatment in combination with localized radiation therapy (XRT) delayed the growth of primary B16 tumors compared with XRT or anti-PD-1 antibody monotherapy (Figure 5; Table 3).

[0250] [Table 3]

[0251] [Table 4]

[0252] The combination of XRT and anti-PD-1 antibody treatment increased overall survival (50% survival at day 50 post-transplant) compared to XRT alone (0% survival at day 50), anti-PD-1 antibody alone (0% survival at day 40), and isotype alone (0% survival at day 30) (Figure 6; Table 4). EXAMPLES

[0253] In vivo efficacy of anti-PD-1 antibody in combination with radiotherapy for metastatic lung tumors In this example, the effect of PD-1 blockade in combination with radiation therapy was investigated on established and metastatic tumors in mice.

[0254] 4T1 breast cancer cells 1.5×10 5 The tumors were subcutaneously implanted into the right flank of female Balb / c mice (Jackson Laboratory). The mean tumor volume was approximately 100 mm. 3 Treatment began 12 days after implantation, when tumor volume reached 100 μg / mL. Mice were randomly assigned to receive intraperitoneal injections of either isotype control (2A3, BioXcell) or PD-1 blocking antibody (RMP1-14, BioXcell) at 5 mg / kg twice weekly for a total of five injections. One day after the initiation of antibody treatment, tumors in the right flank of mice assigned to radiation therapy were 4T1 mice were irradiated with 8 Gy of radiation using a RS2000 Biological Research Irradiator (Rad Source) with a partial body irradiation fixture (Precision X-ray) and shielded with a sheet of lead (Images Scientific Instruments) and delivered to anesthetized mice (ketamine / xylazine). Tumor growth was assessed three times a week until 28 days, after which all mice were euthanized to assess lung metastatic burden using a clonogenic assay. Briefly, lung tissues were dissociated by DNase / Liberase TL (Roche) and cultured in medium supplemented with 60 μM 6-thioguanine. After 2 weeks of culture, plates were counterstained with methylene blue and colonies were counted (one colony represents one metastatic 4T1 cell).

[0255] Treatment with anti-PD-1 antibodies in combination with radiation is expected to promote tumor regression as well as mediate inhibition of metastatic growth. EXAMPLES

[0256] In vivo efficacy of anti-human PD-1 antibody combined with radiotherapy promotes abscopal effect against distant tumors In this example, the effect of PD-1 blockade in combination with radiation therapy was examined against primary and distant MC38 tumors in mice humanized for PD-1 using an anti-human PD-1 antibody.

[0257] The exemplary anti-PD-1 antibody used in this example was REGN2810 (also known as H4H7798N, as disclosed in U.S. Patent Application Publication No. 20150203579), which was a fully human monoclonal anti-PD-1 antibody comprising a heavy chain comprising the amino acid sequence of SEQ ID NO:9 and a light chain comprising the amino acid sequence of SEQ ID NO:10; a HCVR / LCVR amino acid sequence pair comprising SEQ ID NOs:1 / 2; and heavy and light chain CDR sequences comprising SEQ ID NOs:3-8.

[0258] Mice humanized for PD-1 were engineered using VelociGene® technology (Valenzuela et al 2003, Nat. Biotechnol. 21:652-659; US Patent Application Publication No. 2015 / 0366174).

[0259] MC38 colon cancer cells 5 x 10 5 were implanted subcutaneously into female humanized PD-1 / C57BL / 6 mice on day 0 (right flank primary tumor) and day 3 (left flank tumor; distal tumor). The mean primary tumor volume was approximately 150 mm 3 Treatment was initiated when the tumor size reached 100 mg / kg. Mice were randomly assigned to receive either isotype control or PD-1 blocking antibody (REGN2810) at 5 mg / kg intraperitoneally twice weekly for a total of eight injections. One day after the initiation of antibody treatment, mice assigned to the radiation therapy group received 8 Gy of radiation to the tumor in the right flank. Radiotherapy was delivered to anesthetized mice (ketamine / xylazine) using an RS2000 Biological Research Irradiator (Rad Source), shielded by a partial body irradiation fixture (Precision X-ray) and a sheet of lead (Images Scientific Instruments). Primary and secondary tumor growth was assessed three times a week until 70–80 days, after which all mice were euthanized. Figure 7 shows the study design of the experiment including administration of anti-PD-1 antibody and radiation.

[0260] result Primary tumors: PD-1 blockade (REGN2810) treatment synergized with localized radiation (XRT) in rejecting primary MC38 tumors (4 of 6 tumor-free mice) compared to XRT+isotype control treated mice (1 of 6 tumor-free mice). Tumor regression was consistent with the PD-1 blockade (REGN2810) treatment. The RT+isotype treatment group lasted for 3 weeks (rejected tumors relapsed at this time) whereas the combo treatment group lasted for 8 weeks until the end of the experiment (Figure 8; Table 5).

[0261] [Table 5]

[0262] PD-1 blockade as monotherapy mediated rejection in 2 out of 5 mice, but the 1 mouse that rejected its first tumor succumbed to second tumor growth, with only 1 mouse surviving at the end of the experiment. The strong therapeutic efficacy of the combination treatment (XRT+REGN2810) was evidenced by a statistically increased overall survival (approximately 67% survival at day 70 after tumor implantation) compared to all other groups: isotype control or XRT alone (0% survival at day 70), and REGN2810 as monotherapy (20% survival at day 70) (Figure 9; Table 6).

[0263] [Table 6]

[0264] Distant Tumors: REGN2810 in combination with XRT significantly promoted the abscopal effect (rejection of tumors implanted at distant sites) with 5 of 6 mice being tumor free compared to XRT alone (2 of 6 mice with no distant tumors), REGN2810 alone (1 of 6 mice with no distant tumors), and isotype control-treated mice (1 of 6 mice with no distant tumors) (Figure 10; Table 7).

[0265] [Table 7] EXAMPLES

[0266] In vivo efficacy of anti-PD-1 antibodies in combination with radiotherapy and GITR antagonists against MC38 tumors In this example, the effects of PD-1 blockade in combination with radiation therapy and a glucocorticoid-induced tumor necrosis factor receptor (GITR) antagonist (anti-GITR antibody) were examined on large established MC38 tumors in mice.

[0267] MC38 colon cancer cells 5 x 105 The tumors were subcutaneously transplanted into the right flank of female C57BL / 6 mice (Jackson Laboratory). The mean tumor volume was approximately 150-200 mm. 3 Once tumor size reached 100 mg / kg (classified as "large tumors"), treatment was initiated. Mice were randomly assigned to receive intraperitoneal injections of either isotype control antibody (2A3 or LTF-2; BioXcell), anti-PD-1 antibody (RMP1-14; BioXcell), anti-GITR antibody (DTA-1; BioXcell), or a combination of both anti-PD-1 and anti-GITR antibodies at 5 mg / kg twice weekly for a total of five injections. One day after the initiation of antibody treatment, mice assigned to the radiation therapy group received 8 Gy of radiation to the tumors in the right flank. Radiation therapy was delivered to anesthetized mice (ketamine / xylazine) using an RS2000 Biological Research Irradiator (Rad Source), shielded by a partial body irradiation fixture (Precision X-ray) and a sheet of lead (Images Scientific Instruments). Tumor growth was assessed three times weekly until 70–80 days, after which all mice were euthanized. FIG. 11 shows the study design of the experiment involving administration of anti-PD-1 antibody, anti-GITR antibody, and radiation.

[0268] Anti-PD-1 antibody (RMP1-14) treatment was associated with a significant improvement in rejection of large MC38 tumors compared with XRT + anti-GITR antibody (2 of 6 mice were tumor-free) and XRT + anti-PD-1 antibody (2 of 6 mice were tumor-free). The combination of PD-1 and anti-GITR antibodies mediated rejection in 1 out of 5 mice, and the combination of the two antibodies mediated rejection in 2 out of 5 mice. Tumor regression lasted up to 6.5 weeks after treatment initiation for the triple combo-treated mice, compared to 2 weeks for XRT+anti-GITR treated mice (Figure 12). Monotherapy (anti-PD-1 or anti-GITR antibodies) or combination treatment (anti-PD-1 + anti-GITR antibodies) had little effect on tumor growth, with anti-PD-1 or anti-GITR antibody treatment mediating rejection in 1 out of 5 mice, and the combination of the two antibodies mediated rejection in 2 out of 5 mice. Tumor regression lasted up to 6.5 weeks after treatment initiation for the triple combo-treated mice, compared to 2 weeks for XRT+anti-GITR treated mice (Figure 12).

[0269] [Table 8]

[0270] Table 8 and Figure 13 show the survival of mice treated with anti-PD-1 antibody in combination with radiation therapy and anti-GITR antibody. Furthermore, treatment with anti-PD-1 antibody + XRT reduced the survival of very large tumors (approximately 300 mm 3 ) caused tumor regression. EXAMPLES

[0271] In vivo efficacy of anti-PD-1 antibodies in combination with radiotherapy and GITR antagonists against B16 tumors In this example, the effects of PD-1 blockade in combination with radiation therapy and a GITR antagonist (anti-GITR antibody) were investigated on established B16 tumors in mice.

[0272] 2.5 × 10 B16F10.9 melanoma cells 5 The tumors were subcutaneously implanted into the right flank of female C57BL / 6 mice (Jackson Laboratory). The mean tumor volume was approximately 100 mm. 3Treatment was initiated when tumor size reached 100%. Mice were randomly assigned to receive intraperitoneal injections of either isotype control (2A3, LTF-2; BioXcell), anti-PD-1 antibody (RMP1-14; BioXcell), anti-GITR antibody (DTA-1; BioXcell), or a combination of both anti-PD-1 and anti-GITR antibodies at 5 mg / kg twice weekly for a total of five injections. One day after the initiation of antibody treatment, mice assigned to the radiation therapy group received 8 Gy of radiation to the tumors in the right flank. Radiation therapy was delivered to anesthetized mice (ketamine / xylazine) using an RS2000 Biological Research Irradiator (Rad Source), shielded by a partial body irradiation fixture (Precision X-ray) and a sheet of lead (Images Scientific Instruments). Tumor growth was assessed three times weekly until 70–80 days, after which all mice were euthanized.

[0273] Anti-GITR antibodies and anti-PD-1 antibodies in combination with radiation therapy have been used as monotherapy or Anti-PD-1 antibodies combined with radiation therapy are expected to promote tumor regression and slow tumor growth. EXAMPLES

[0274] Clinical Trial of Anti-PD-1 Antibody and Radiation Therapy in Patients with Advanced Solid Tumors This study was an open-label, multicenter, dose-escalation study with multiple dose-escalation and dose-expansion arms to investigate the efficacy, safety, and tolerability of an anti-PD-1 antibody, alone and in combination with other anticancer treatments (including radiation therapy), in adult patients with advanced solid tumors.

[0275] The exemplary anti-PD-1 antibody used in this study was REGN2810 (also known as H4H7798N, as disclosed in U.S. Patent Application Publication No. 20150203579), which was a fully human monoclonal anti-PD-1 antibody comprising a heavy chain comprising the amino acid sequence of SEQ ID NO:9 and a light chain comprising the amino acid sequence of SEQ ID NO:10; an HCVR / LCVR amino acid sequence pair comprising SEQ ID NOs:1 / 2; and heavy and light chain CDR sequences comprising SEQ ID NOs:3-8.

[0276] Purpose of the test The primary objective of the study is to characterize the safety, tolerability, and dose-limiting toxicities (DLTs) of REGN2810 administered intravenously (IV) as monotherapy or in combination with targeted radiation (intended to serve primarily as an immune stimulant rather than tumor ablation therapy), low-dose cyclophosphamide (a treatment shown to inhibit regulatory T-cell responses), granulocyte-macrophage colony-stimulating factor, carboplatin, docetaxel, or combinations thereof in patients with advanced malignancies.

[0277] Secondary objectives of this study were: (1) to determine the recommended phase 2 dose (RP2D) of REGN2810 as monotherapy and in combination with other anti-cancer therapies (targeted radiation, low-dose cyclophosphamide, or both); (2) to describe the preliminary anti-tumor activity of REGN2810 alone and with each combination partner; (3) to characterize the PK of REGN2810 as monotherapy and in combination with other anti-cancer therapies (targeted radiation, low-dose cyclophosphamide, or both); and (4) to evaluate the immunogenicity of REGN2810.

[0278] Justification for study design The 3+3 model of the dose-escalation phase of the study was designed to allow evaluation of the safety of REGN2810 as monotherapy at different dose levels and in combination with the immune-boosting treatment cyclophosphamide; limited, targeted radiation delivered in one of two dosing regimens; or a combination of radiation and cyclophosphamide.

[0279] Because the tolerability of REGN2810 has been established alone and in combination with radiation and / or cyclophosphamide, multiple expansion cohorts using various combinations or monotherapies in selected indications (NSCLC, BC, HNSCC, CSCC, tumors with MSI (colorectal, endometrial, prostate, or other tumor types), HCC, and other advanced solid tumors) were added to further confirm safety and evaluate enhanced antitumor activity. Granulocyte-macrophage colony-stimulating factor (GM-CSF), carboplatin, and / or docetaxel were added to some of these combinations.

[0280] Table 9 lists some of the cohorts using REGN2810 as a single agent and in combination with other disease modalities.

[0281] [Table 9]

[0282] Initial planned treatment with REGN2810 was every 14 days for up to 48 weeks with 24 weeks of observation. Radiation was administered 1 week after the first dose of REGN2810. Low-dose cyclophosphamide was administered 1 day prior to each of the first 4 doses of REGN2810. It was administered to patients assigned to phosphamide.

[0283] Test period Patients were treated for up to 48 weeks, followed by a follow-up period of 24 weeks. Patients were treated until the 48-week treatment period was completed or until progression, unacceptable toxicity, withdrawal of consent, or another study discontinuation criterion was met. Patients with a confirmed complete response (CR) after at least 24 weeks of treatment may decide to discontinue treatment and continue all relevant study evaluations (e.g., efficacy evaluations). Patients with a stable tumor burden assessment (SD) or partial response (PR), with three consecutive tumor assessments remaining unchanged after at least 24 weeks of treatment, may also decide to discontinue treatment and continue all relevant study evaluations (e.g., efficacy evaluations).

[0284] Study population The target population for this study included patients with advanced malignancies who were not candidates for standard therapy, were unwilling to undergo standard therapy, or for whom available treatments were not expected to convey clinical benefit, as well as patients with malignancies that were incurable and had not responded to standard therapy or had demonstrated tumor progression despite standard therapy.

[0285] Inclusion Criteria: Patients must meet the following criteria to be eligible for study participation: (1) proven progression of solid tumors with no alternative standard of care treatment options available; (2) at least one lesion to assess response. Patients assigned to radiation therapy must have at least one additional lesion that can be safely irradiated while sparing the index lesion and where a limited, palliative dose of radiation is deemed medically appropriate; (3) patients must have relapsed or be refractory after first-line treatment (and up to two prior treatments) in the setting of recurrent or metastatic disease and have disease for which palliative radiation therapy is indicated; (4) patients with metastatic cancer with microsatellite instability (MSI) that is refractory to up to two prior treatments; (5) patients with metastatic cancer with microsatellite instability that is refractory to up to two prior treatments; (6) patients with metastatic cancer with microsatellite instability that is refractory to up to two prior treatments; (7) patients with metastatic cancer with microsatellite instability that is refractory to up to two prior treatments; (8) patients with metastatic cancer with microsatellite instability that is refractory to up to two prior treatments; (9) patients with metastatic cancer with microsatellite instability that is refractory to up to two prior treatments; (10) patients with metastatic cancer with microsatellite instability that is refractory to up to two prior treatments; (11) patients with metastatic cancer with microsatellite instability that is refractory to up to two prior treatments; (12) patients with metastatic cancer with microsatellite instability that is refractory to up to two prior treatments; (13) patients with metastatic cancer with microsatellite instability that is refractory to up to two prior treatments; (14) patients with metastatic cancer with microsatellite instability that is refractory to up to two prior treatments; (15) patients with metastatic cancer with micro Eastern Cooperative Association of Group (ECOG) performance status ≤ 1; (6) age > 18 years; (7) liver function: a. total bilirubin ≤ 1.5 × upper limit of normal (ULN, if liver metastases ≤ 3 × ULN), b. transaminases ≤ 3 × ULN (or ≤ 5.0 × ULN if liver metastases), c. alkaline phosphatase (ALP) ≤ 2.5 × ULN (or ≤ 5.0 × ULN if liver metastases); (8) renal function: serum creatinine ≤ 1.5 × ULN; (9) neutrophil count (ANC) ≤ 1.5 × 10 9 c. Platelet count is 75 x 10 9 (10) able to provide signed informed consent; and (11) able and willing to consent to scheduled visits, treatment plans, laboratory tests, and other study-related procedures.

[0286] Test procedure REGN2810 is supplied as a liquid in sterile, single-use vials. Each vial contains sufficient volume to withdraw 10 mL of REGN2810 at a concentration of 25 mg / mL. REGN2810 was administered in an outpatient setting as a 30-minute IV infusion. Each patient's dose was dependent on their individual body weight. The dose of REGN2810 was adjusted each cycle if body weight changed by 10% or more. REGN2810 was administered alone or in combination with radiation and / or cyclophosphamide. Cyclophosphamide was administered at a dose of 200 mg / m 2 or low dose (100 mg / m 2 ) was administered.

[0287] Monotherapy REGN2810 will be administered to patients on an outpatient basis by IV infusion over 30 minutes every 14 days (i.e., days 1, 15±3, 29±3, and 43±3 of a 56-day cycle) for 48 weeks. The planned monotherapy regimens to be assigned may include: (i) 1 mg / kg IV infusion over 30 minutes every 14 days for 48 weeks; (ii) 3 mg / kg IV infusion over 30 minutes every 14 days for 48 weeks; (iii) 10 mg / kg IV infusion over 30 minutes every 14 days for 48 weeks; (iv) 0.3 mg / kg IV infusion over 30 minutes every 14 days for 48 weeks (if the MTD was determined to be less than 1 mg / kg); and (v) a fixed dose of 200 mg IV infusion over 30 minutes every 14 days for 48 weeks.

[0288] Combination Therapy Concomitant radiation therapy, cyclophosphamide, GM-CSF, carboplatin, and docetaxel were provided via the regimen and their use, doses, dose modifications, reductions, or delays, and any possible AEs due to their use were tracked along with REGN2810.

[0289] Concurrent administration of REGN2810 and radiation: REGN2810 was administered by IV infusion over 30 minutes every 14 days for 48 weeks in combination with radiation treatment from days 8 to 12. Planned combination REGN2810 and radiation therapy regimens may include: REGN2810 1 mg / kg infusion over 30 minutes every 14 days for 48 weeks plus 30 Gy of radiation therapy (6 Gy x 5 fractions / week; administered preferably on consecutive days, 1 week after the first dose of REGN2810) REGN2810 1 mg / kg infusion over 30 minutes every 14 days for 48 weeks plus 27 Gy of radiation therapy (9 Gy x 3 fractions / week; administered 1 week after the first dose of REGN2810, preferably on non-consecutive days) REGN2810 3 mg / kg infusion over 30 minutes every 14 days for 48 weeks plus 30 Gy of radiation therapy (6 Gy x 5 fractions / week; administered preferably on consecutive days, 1 week after the first dose of REGN2810) REGN2810 3 mg / kg infusion over 30 minutes every 14 days for 48 weeks plus 27 Gy of radiation therapy (9 Gy x 3 fractions / week; administered 1 week after the first dose of REGN2810, preferably on non-consecutive days)

[0290] Patients received either 30 Gy in 5 fractions of 6 Gy administered daily starting 1 week after the first dose of REGN2810, or 27 Gy in 3 fractions of 9 Gy administered every other day starting 1 week after the first dose of REGN2810. Lesions selected for radiation should be those that can be safely irradiated by local irradiation while sparing the index lesion and those for which irradiation at the contemplated limited palliative doses is deemed medically appropriate.

[0291] Coadministration of REGN2810 and cyclophosphamide: REGN2810 was administered in combination with low-dose cyclophosphamide at 100 mg / m 2 REGN2810 was administered by IV infusion over 30 minutes every 14 days (2 weeks) for 48 weeks in combination with IV infusion of 4 doses of cyclophosphamide every 14 days. Each of the four cyclophosphamide doses was administered one day before each of the first four REGN2810 doses (days -1, 14, 28, and 42 of the first 56-day cycle).

[0292] The planned REGN2810 and cyclophosphamide regimen combinations are: Cyclophosphamide 100mg / m 2 or 200 mg / m 2 IV every 14 days (days -1, 14, 28, and 42 of the first 56-day cycle) for a total of 4 doses; plus REGN2810 3 mg / kg infusion over 30 minutes every 14 days for 48 weeks (however, the monotherapy dose of 3 mg / kg is below the MTD; if 3 mg / kg is above the MTD, the dose is 1 mg / kg) It was.

[0293] Concurrent Administration of REGN2810, Radiation, and Cyclophosphamide: The planned combination REGN2810, radiation, and cyclophosphamide regimen is: Cyclophosphamide 100mg / m 2 (low dose) IV every 14 days (days -1, 14, 28, and 42 of the first 56-day cycle) for a total of 4 doses; plus 27 Gy of radiation therapy (9 Gy x 3 fractions / week; administered 7 or 8 days after the first dose of REGN2810, preferably on non-consecutive days), or 30 Gy of radiation therapy (6 Gy x 5 fractions / week; administered 7 or 8 days after the first dose of REGN2810, preferably on consecutive days); plus REGN2810 3 mg / kg infusion over 30 minutes every 14 days for 48 weeks (however, the monotherapy dose of 3 mg / kg is below the MTD; if 3 mg / kg is above the MTD, the dose is 1 mg / kg) Includes:

[0294] Concurrent Administration of REGN2810, Radiation, and GM-CSF: The planned combination REGN2810, radiation, and GM-CSF regimen is: GM-CSF 250 mcg SC daily for 7 days, given 7 days apart for 4 doses (days 1 to 7, 15 to 21, 29 to 35, and 43 to 49 of the first 56-day cycle); plus Radiation therapy of 27 Gy (9 Gy x 3 fractions per week; preferably administered on non-consecutive days 1 week after the first dose of REGN2810); plus REGN2810 3 mg / kg infusion over 30 minutes every 14 days for 48 weeks (however, the monotherapy dose of 3 mg / kg is below the MTD; if 3 mg / kg is above the MTD, the dose is 1 mg / kg) Includes:

[0295] Concomitant Administration of REGN2810, Radiation, GM-CSF, and Cyclophosphamide: The planned combination REGN2810, radiation, GM-CSF, and cyclophosphamide regimen is: GM-CSF 250 mcg SC daily for 7 days, given 7 days apart for 4 doses (days 1 to 7, 15 to 21, 29 to 35, and 43 to 49 of the first 56-day cycle); plus Radiation therapy of 27 Gy (9 Gy x 3 fractions per week; preferably administered on non-consecutive days 1 week after the first dose of REGN2810); plus Cyclophosphamide 100mg / m 2 or 200 mg / m 2 IV every 14 days (days -1, 14, 28, and 42 of the first 56-day cycle) for a total of 4 doses; plus REGN2810 3 mg / kg infusion over 30 minutes every 14 days for 48 weeks (however, the monotherapy dose of 3 mg / kg is below the MTD; if 3 mg / kg is above the MTD, the dose is 1 mg / kg) Includes:

[0296] Concurrent administration of REGN2810 and docetaxel with or without carboplatin: The proposed order of drug administration was docetaxel followed by carboplatin (if enrolled in a carboplatin-containing cohort), followed by REGN2810: Docetaxel 30mg / m 2 IV over approximately 1 hour on days 1, 8, 29, and 36 of the first 56-day cycle. Dexamethasone 8 mg IV is administered For subsequent docetaxel treatments, the dexamethasone premedication dose may be 8 mg or 4 mg, at the discretion of the investigator. Carboplatin AUC2 IV over approximately 30 minutes is given on days 1, 8, 29, and 36 of an initial 56-day cycle. Carboplatin should be dosed using the Calvert formula on the carboplatin label. Creatinine clearance should be calculated using the Cockcroft-Gault formula. REGN2810 3 mg / kg administered as an infusion over approximately 30 minutes every 14 days for 48 weeks.

[0297] Procedures and Evaluation Screening procedures performed included serum beta-HCG, brain MRI, and chest x-ray.

[0298] Safety procedures will include anamnesis, physical examination, vital signs, electrocardiogram (ECG), coagulation, immune safety assays (for patients treated with REGN2810), B symptom assessments and performance status assessments, clinical laboratories, AEs, and concomitant medications.

[0299] Efficacy procedures to be performed for tumor evaluation included CT or MRI scans, 18F-fluorodeoxyglucose-positron emission tomography (FDG-PET) scans, and / or tumor biopsies. CT or MRI for tumor evaluation was performed at the screening visit (within 28 days prior to infusion) and between cycles on day 56 ± 3 (approximately every 8 weeks), and if progression was suspected. In addition, for patients who did not progress on study, tumor evaluation was performed at follow-up visits 3, 5, and 7. After the choice to use CT scan or MRI was made, subsequent evaluations were performed using the same modality. Tumor response evaluation was performed according to the New Response Evaluation Guidelines for Solid Tumors (2019). The study was conducted according to the Clinical Trials Evaluation Criteria in Solid Tumors (RECIST) version 1.1 (Eisenhauer et al 2009, Eur. J. Cancer 45:228-247). Measurable lesions selected as target lesions for RECIST measurements were also included as index lesions for immune-related response criteria (irRC; Nishino et al 2013, Clin. Cancer Res. 19:3936-3943). RECIST response is the preferred statistical evaluation of response rate. For individual patients, the irRC may inform decisions regarding whether to continue treatment at the investigator's discretion due to the possibility of an unusual response.

[0300] Blood samples were collected for PK and anti-drug antibody (ADA) evaluation.

[0301] Test variables The primary variables of the study were the occurrence of DLTs throughout 48 weeks of treatment, and the occurrence and severity of TEAEs and abnormal clinical findings.

[0302] The subvariables are: Antitumor activity assessed using criteria appropriate for the indication (described elsewhere herein): Response Evaluation in Solid Tumors as Measured by CT or MRI (RECIST; Eisenhauer et al 2009, Eur. J. Cancer 45:228-247) Criteria Other evaluation criteria will be used for specific tumors where RECIST measurements are not standard The Immune-Related Response Criteria (irRC; Nishino et al. 2013, Clin. Cancer Res. 19:3936-3943). All In cases, RECIST (or other tumor-specific criteria) is the preferred tool for determining PD, SD, CR, or PR. irRCs were collected for clinical decision and information purposes. Incidence of anti-REGN2810 antibodies Antitumor activity measured by PFS and overall survival It was.

[0303] For the purposes of this study, patients were reassessed for response every 8 weeks. Confirmatory scans were also obtained 4 weeks after the first reported objective response or progression. Response and progression were evaluated in this study using the international criteria proposed by the revised Response Evaluation Criteria in Solid Tumors (RECIST) (version 1.1; Eisenhauer et al 2009, Eur. J. Cancer 45:228-247). In the case of malignant lymph nodes, the change in the longest diameter (unidimensional measurement) and shortest diameter of the tumor lesion was used in the RECIST criteria.

[0304] Lesion Selection Measurable Disease: Measurable lesions were defined as lesions that could be accurately measured in at least one dimension (longest diameter recorded) as ≥20 mm (≥2 cm) by chest x-ray or ≥10 mm (≥1 cm) by caliper by CT scan, MRI, or clinical examination. All tumor measurements must be recorded in millimeters (or decimals of a centimeter). Note: See below for evaluation of irradiated target lesions.

[0305] Malignant lymph nodes: To be considered pathologically enlarged and measurable, lymph nodes had to measure 15 mm or more (1.5 cm or more) in the short axis as assessed by CT scan (CT scan slice thickness of 5 mm [0.5 cm] or less is recommended). Only the short axis was measured and tracked at baseline and follow-up.

[0306] Nonmeasurable disease: All other lesions (or disease sites), including small lesions (<10 mm [<1 cm] in longest diameter or pathologic lymph nodes, ≥10 mm but <15 mm [≥1 cm but <1.5 cm] in short axis), were considered nonmeasurable disease. Bone lesions, leptomeningeal disease, ascites, pleural / pericardial effusions, cutaneous / pulmonary lymphangiopathy, inflammatory breast cancer, and abdominal tumor masses (not traceable by CT or MRI) were considered nonmeasurable. Note: Cystic lesions meeting the criteria for radiologically defined simple cysts should not be considered as malignant lesions (either measurable or nonmeasurable) since they are, by definition, simple cysts. "Cystic lesions" thought to represent cystic metastases can be considered as measurable lesions if they meet the definition of measurability above. However, if noncystic lesions are present in the same patient, these are preferred for selection as target lesions.

[0307] Target Lesions: All measurable lesions, up to two per organ and five overall, representative of all affected organs, should be identified as target lesions and recorded and measured at baseline. Target lesions were selected based on their size (longest diameter of the lesion) to represent all affected organs, but also to include lesions that could be reproducibly and repeatedly measured. Occasionally, the largest lesion could not be reproducibly measured, and in such situations, the next largest lesion that could be reproducibly measured was selected. The sum of the diameters of all target lesions (longest diameter for non-nodal lesions, short axis for nodal lesions) was calculated and reported as the baseline sum of longest diameters. If lymph nodes were included in the sum, only the short axis was added to the sum. The baseline sum of longest diameters was used as a reference to further characterize any objective tumor regression in the measurable dimensions of disease.

[0308] Non-target lesions: All other lesions (or sites of disease), including the five target lesions plus any measurable lesions, were identified as non-target lesions and recorded at baseline. Measurement of these lesions is not required, but their presence, absence, or, in rare cases, overt progression will be noted during follow-up.

[0309] How to assess measurable disease All measurements were made and recorded in metric units using a ruler or calipers. All baseline assessments should be performed at the start of treatment whenever possible, and not more than 4 weeks prior to the start of treatment. The same assessment method and the same technique should be used to characterize each identified and reported lesion during the baseline and follow-up periods. Imaging-based assessments are preferred over clinical examination assessments, except when the lesion to be followed cannot be imaged but can be assessed by clinical examination.

[0310] Clinical Lesions: Clinical lesions were considered measurable only if they were superficial (e.g., skin nodules or palpable lymph nodes) and were ≥10 mm (≥1 cm) in diameter when assessed using calipers (e.g., skin nodules). In the case of skin lesions, reporting by color photographs including a ruler to assess the size of the lesion was encouraged.

[0311] Chest X-ray: Lesions on chest X-ray were acceptable as measurable lesions if they were well-defined and surrounded by air-containing lung. However, CT is preferred.

[0312] Conventional CT and MRI: This guideline defines measurability of lesions on CT scans based on the assumption that the CT slice thickness is 5 mm (0.5 cm) or less. If the CT scan slice thickness is greater than 5 mm (0.5 cm), the minimum size of a measurable lesion should be twice the slice thickness. MRI is also acceptable in certain circumstances.

[0313] PET-CT: If the CT performed as part of the PET-CT is of the same diagnostic quality as the diagnostic CT (including IV and oral contrast), the CT portion of the PET-CT can be used for RECIST measurements and can be used interchangeably with regular CT to accurately measure cancer lesions over time.

[0314] Ultrasound: Ultrasound is not useful for assessing lesion size and should not be used as a method of measurement. If new lesions are identified by ultrasound during the course of the study, confirmation by CT or MRI is recommended. MRI may be used instead of CT in selected cases if there are concerns regarding radiation exposure during CT.

[0315] Endoscopy, laparoscopy: Utilizing these techniques for objective tumor evaluation is not recommended, however such techniques may be useful to obtain biopsies to confirm pathologic complete response or to determine relapse in clinical trials where complete response (CR) or recurrence after surgical resection is an endpoint.

[0316] Tumor markers: Tumor markers alone should not be used to assess response. If markers are initially above the upper limit of the normal range, they must normalize in order for the patient to be considered in clinical complete response.

[0317] Cytology, histology: These techniques can be used to distinguish between partial response (PR) and complete response (CR) in rare cases (e.g., residual disease in tumor types such as germ cell tumors where known residual benign tumor may remain). Any tumor that appears or worsens during treatment when measurable tumor meets criteria for response or stable disease. Cytological confirmation of the neoplastic origin of the effusion is essential to distinguish between response or stable disease (effusion may be a side effect of treatment) and progression.

[0318] FDG-PET: When FDG-PET response assessment requires additional testing, it is sometimes reasonable to incorporate the use of FDG-PET scans to supplement CT scans in the evaluation of progression (especially possible "new" disease). New lesions based on FDG-PET imaging can be identified according to the following algorithm: a. FDG-PET negative at baseline and FDG-PET positive at follow-up is indicative of PD based on new lesions. b. No FDG-PET at baseline and FDG-PET positive at follow-up: If the FDG-PET positivity at follow-up corresponds to a new site of disease confirmed by CT, this is PD. If the FDG-PET positivity at follow-up is not confirmed as a new site of disease on CT, a further follow-up CT scan is necessary to determine whether true progression has occurred at that site (in such cases, the date of PD is the date of the first abnormal FDG-PET scan). If the FDG-PET positivity at follow-up corresponds to an existing site of disease on CT that has not progressed based on anatomic imaging, this is not PD. c In cases where radiographic residual abnormalities are thought to represent fibrosis or scarring, FDG-PET may be used to upgrade the response to CR, similar to biopsy. The use of FDG-PET in these situations should be pre-specified in the protocol and supported by disease-specific medical literature on the indications. However, both approaches must recognize that false-positive CRs may occur due to the limitations of FDG-PET and the resolution / sensitivity of biopsy. Note: FDG-PET scan "positive" lesions refer to FDG-demanding lesions with uptake 2-fold greater than the surrounding tissue in attenuation-corrected images.

[0319] Efficacy criteria for evaluation of target lesions Complete Response (CR): Disappearance of all target lesions. Any pathological lymph nodes (target or non-target) must be reduced to less than 10 mm (less than 1 cm) in the short axis. Partial Response (PR): At least a 30% reduction in the sum of the longest diameters of target lesions compared to the baseline sum of the longest diameters. Progression (PD): At least a 20% increase in the sum of the longest diameters of the target lesions referenced to the study's smallest longest diameter sum (this includes the baseline longest diameter sum if it is the study's smallest value). In addition to the 20% relative increase, the sum must also demonstrate an absolute increase of at least 5 mm (0.5 cm). (Note: the appearance of one or more new lesions is also considered to be progression). Stable Disease (SD): Not enough reduction to qualify for PR or not enough increase to qualify for PD, based on the smallest sum of longest diameters in the study.

[0320] Response criteria for evaluation of non-target lesions Complete Response (CR): Disappearance of all non-target lesions and normalization of tumor marker levels. All lymph node size must be non-pathological (short axis <10 mm (<1 cm)). Note: If tumor markers are initially above the upper limit of the normal range, they must normalize for the patient to be considered in clinical complete response. Non-CR / Non-PD: persistence of one or more non-target lesions and / or maintenance of tumor markers at levels above the upper limit of normal. Progression (PD): Appearance of one or more new lesions and / or overt progression of existing non-target lesions. Overt progression should not usually be dependent on the status of target lesions. It should represent a change in the overall disease state, not an increase in a single lesion.

[0321] Immune-related response criteria Immune-related response criteria were the longest diameter of all target lesions and new lesions, if present. It differs from RECIST (version 1.1) in that it uses the sum of all disease states to determine response. The presence of new lesions does not in itself determine progression: the total tumor burden is taken into account.

[0322] Target Lesion Evaluation Complete Response (CR): Disappearance of all target lesions. Any pathological lymph nodes (target or non-target) must be reduced to less than 10 mm (less than 1 cm) in the short axis. Partial Response (PR): At least a 30% reduction in the sum of the longest diameters of target lesions, including new lesions, compared to the baseline sum of the longest diameters. Progression (PD): At least a 20% increase in the sum of the longest diameters of target lesions, including new lesions, referenced to the study's smallest longest diameter sum (which includes the baseline longest diameter sum, if that is the study's smallest value). In addition to the 20% relative increase, the sum must also demonstrate an absolute increase of at least 5 mm (0.5 cm). Stable Disease (SD): No sufficient shrinkage to qualify for PR or no sufficient increase to qualify for PD, including using the smallest sum of longest diameters during the study as a reference and including measurements of new lesions.

[0323] Evaluation of non-target lesions Complete Response (CR): Disappearance of all non-target lesions and normalization of tumor marker levels. All lymph node size must be non-pathological (short axis <10 mm (<1 cm)). Note: If tumor markers are initially above the upper limit of normal, they must normalize for the patient to be considered in clinical complete response. Non-CR / Non-PD: persistence of one or more non-target lesions and / or maintenance of tumor markers at levels above the upper limit of normal. Progression (PD): Overt progression of existing non-target lesions. Overt progression should not usually be dependent on the status of target lesions. It should represent a change in the overall disease state and not an increase in a single lesion. Overt progression of only "non-target" lesions is exceptional, but in such circumstances the opinion of the attending physician should prevail and the progression status should be confirmed at a later date.

[0324] Evaluation of the overall effectiveness criteria Best overall response is the best response recorded from the start of treatment to progression / recurrence (the reference for progression is the smallest measurement recorded after the start of treatment). Best response allocation for patients depends on reaching both measurement and confirmation criteria. The revised Response Evaluation Criteria in Solid Tumors (RECIST) version 1.1; (Eisenhauer et al 2009, Eur. J. Cancer 45:228-247) and Immune-Related Response Criteria (irRC; Nishino et al 2013, Clin. Cancer Res. 19:3936-3943) are summarized in Tables 10 and 11 below.

[0325] [Table 10]

[0326] [Table 11]

[0327] Evaluation of radiation target lesions Radiation target lesions were evaluated by a modified version of the international criteria proposed by the Response Evaluation Criteria in Solid Tumors (RECIST) Committee, version 1.1. To define local control, we incorporated the RECIST 1.1 guidelines, as well as additional definitions specific to this protocol.

[0328] The response criteria for irradiated lesions were as follows:

[0329] Local Enhancement (LE): At least a 20% increase in the LD of a target lesion, referenced to the smallest LD recorded since the start of treatment. Ideally, this determination is made based on CT imaging evaluation.

[0330] Local recurrence (LF): refers to the predominantly treated tumor after protocol therapy, corresponding to meeting both of the following two criteria: (1) an increase in tumor size of 20% of the above definition for local enhancement (LE); (2) measurable tumors meeting the criteria for LE should be of similar intensity uptake on positron emission tomography (PET) imaging as the pretreatment staging PET, or a biopsy of the measurable tumor should be taken to confirm that the cancer is viable.

[0331] Local control (LC): Absence of local recurrence.

[0332] The longest diameter (LD) of the irradiated target lesion calculated from the treatment planning CT scan was reported as the baseline LD using the appropriate tissue-specific window. The baseline LD was used as a reference to objectively characterize the tumor. For follow-up evaluation, a diagnostic CT scan performed using a 5 mm continuous reconstruction algorithm and lung window obtained as part of the scheduled protocol follow-up is the preferred method of response assessment. If a CT scan is not available, MRI or X-ray determination is acceptable as long as the target lesion is clearly visible.

[0333] result REGN2810 alone and in combination was safe and well tolerated by patients. Administration of REGN2810 alone or in combination with other treatment modalities inhibited tumor growth and / or promoted tumor regression in patients with advanced solid tumors. Overall response rates were favorable for combination therapy with radiation compared to monotherapy.

[0334] To date, 60 patients with advanced solid malignancies (47% had received 4 or more prior therapies) have been treated, including colorectal, head and neck, breast, soft tissue sarcoma, adrenal, anal, appendix, bladder, cervical, endometrial, esophageal, liver, non-small cell lung adenocarcinoma, ovarian, pancreatic, prostate, renal sarcomatoid, salivary gland, non-melanoma skin, Merkel cell carcinoma, squamous cell carcinoma, basal cell carcinoma, small intestine, thyroid, and uterine cancer.

[0335] Forty-two patients (70%) experienced one or more treatment-related adverse events (AEs). The most common treatment-related AEs were fatigue (28.3%), arthralgia (11.7%), and nausea (11.7%). Of the 60 patients evaluated for tumor response, 11 (18.3%) had an objective response (PR / CR), while 31 (51.7%) had disease control (CR / PR / SD). In the 36 patients who received combination therapy that included radiation therapy, objective responses were observed in 6 patients (16.7%) and disease control was observed in 19 patients (52.8%). For the 24 patients who did not receive radiation therapy, objective responses were observed in 5 patients (20.8%) and disease control was observed in 12 patients (50%). Table 12 shows a summary of responders.

[0336] [Table 12]

[0337] Among responders, the median time to response to monotherapy was 113 days (range, 52–226 days) and 59 days (range, 56–113 days) in patients who received radiation therapy. EXAMPLES

[0338] Case report of PD-1 blockade with monoclonal antibody REGN2810 achieving durable objective responses in metastatic non-melanoma skin cancer, basal cell carcinoma, and cutaneous squamous cell carcinoma Introduction Basal cell carcinoma (BCC) and cutaneous squamous cell carcinoma (CSCC) share exposure to UV light as a major risk factor, and therefore these tumors are highly mutated (Chalmers et al 2016, AACR Ann. Meeting, Abs 3576). In other malignancies, high mutational burden is associated with clinical benefit from treatment with antibodies against the PD-1 immune checkpoint [Le et al 2015, New Engl. J. Med. May 30 (epublished ahead of print)]. Hypermutated tumors are more likely to express immunogenic tumor neo-antigens that attract effector T cells that can be released by blockade of the PD-1 immune checkpoint (Mandal et al 2016). and Chan 2016, Cancer Discov. 6:1-12). This Example describes patients with metastatic BCC and metastatic CSCC treated with REGN2810, a fully human anti-PD-1 monoclonal antibody, in an ongoing Phase 1 clinical trial (NCT02383212; described herein in Example 7).

[0339] Case report 1 The patient was a 66-year-old woman diagnosed with stage 1 BCC arising on the left side of the jaw, which was removed by Mohs surgery. A local recurrence at the same site was identified 2 years later, and wide local excision revealed involvement of the left mandible and metastasis to 1 of 18 lymph nodes. The patient underwent adjuvant radiation and was in remission for 4 years, but surveillance lung imaging revealed a biopsy of an enlarging pulmonary nodule that confirmed the presence of metastatic BCC. The patient was then treated with vismodegib, a hedgehog pathway inhibitor (HHI), for 5 months. The patient initially responded but discontinued due to progression.

[0340] As the disease continued to progress slowly after 6 months of vismodegib therapy, the patient was enrolled in a Phase 1 trial of REGN2810 and placed in a cohort receiving 10 mg / kg IV every 2 weeks. The patient was randomized and received the initial dose. Two lung metastases were followed as target lesions. Response assessments at the end of 8 weeks (3% increase) and 16 weeks (10% increase) demonstrated stable disease by RECIST criteria. Response assessment at the end of 24 weeks demonstrated a 37% reduction in tumor measurements (Figure 14A), which was confirmed at 32 weeks. The patient tolerated the treatment well and is continuing treatment with REGN2810 for an additional 10 months.

[0341] Case report 2 The patient was a 52-year-old man diagnosed with cutaneous squamous cell carcinoma of the left cheek. The patient underwent Mohs surgery with securing margins. The patient experienced multiple recurrences and underwent at least nine additional Mohs surgeries. The patient underwent wide local excision of the entire left mandible 4 years later, followed by a left parotidectomy at 20 months. Adjuvant radiation therapy was similarly administered to the left cheek, left mandible, left neck (with concurrent cetuximab), and bilateral neck (with concurrent carboplatin). Other systemic treatments were capecitabine, and cisplatin plus docetaxel. Ten years after initial diagnosis, the patient underwent resection with securing margins for a 2.2 cm intrascar recurrence in the left neck. He subsequently required emergency decompressive surgery of the cervical spine with C4-C5 anterior fusion and C4-C6 posterior laminectomy due to an invasive CSCC at the C4-C5 vertebral bodies. The patient also developed muscle weakness in the lower extremities thought to be due to perineural involvement, necessitating the use of a walker for ambulation.

[0342] The patient was enrolled in the first cohort of the Phase 1 study and received REGN2810 at 1 mg / kg every 2 weeks. Within a few weeks of initiating treatment, the patient's lower extremity strength gradually recovered and he no longer required the use of a walker. Responses at 16 weeks are shown in Figure 14B. A radiographic complete response of the left cervical lesion was achieved at week 40. The patient completed the planned 48-week protocol treatment with REGN2810. The patient currently remains under active follow-up with his medical oncologist with no clinical or radiographic evidence of disease recurrence.

[0343] Consideration This example identifies for the first time partial responses in patients with metastatic BCC treated with a PD-1 inhibitor (REGN2810), as well as ongoing, durable complete responses in patients with metastatic CSCC. These heavily pretreated patients responded deeply and durable to anti-PD-1 monotherapy in this Phase 1 study, consistent with the hypothesis that the high mutational burden in BCC and CSCC induces antitumor cellular immunity that can be released by blockade of the PD-1 / PD-L1 checkpoint pathway.

[0344] This example supports the general principle that UV-associated skin cancers other than melanoma are sensitive to PD-1 blockade. Reductionist models predicted that UV-associated tumors with higher nonsynonymous mutation burdens would be more responsive to PD-1 blockade than tumors with lower mutation burdens. EXAMPLES

[0345] Safety and efficacy of anti-PD-1 antibodies in patients with unresectable locally advanced or metastatic cutaneous squamous cell carcinoma (CSCC) background There is no established standard treatment for unresectable locally advanced or metastatic CSCC. Most CSCC are frequently mutated due to UV-induced DNA damage. Therefore, these tumors may respond to PD-1 checkpoint blockade. This example describes a patient with locally advanced or metastatic CSCC treated with REGN2810, a fully human anti-PD-1 monoclonal antibody in an ongoing Phase 1 trial (NCT02383212; described in Example 7 herein).

[0346] method The expansion cohort (EC) of the phase 1 study of REGN2810 enrolled patients with distant metastatic CSCC (EC7) and locally advanced CSCC (EC8) (Table 9). All patients received REGN2810 3 mg / kg intravenously every 2 weeks for up to 48 weeks. Investigational biopsies were performed at baseline and day 29 (and at progression, if possible). Tumor measurements were performed every 8 weeks according to RECIST 1.1 to determine overall response rate.

[0347] result Twenty-five patients were enrolled (10 in EC7 and 15 in EC8): median age, 72.5 years (range, 56-88 years); median PS 1 (range, 0-1); 20 men and 5 women; median number of prior systemic treatment regimens 1 (range, 0-3). Median exposure to REGN2810 was 6 doses (range, 1-22). The most common treatment-related adverse events of any grade were fatigue (16.7%), nausea, arthralgia, and rash (8.3% each). Each of the following grade 3 or higher related adverse events (AEs) occurred once: AST elevated, ALT elevated, arthralgia, and rash.

[0348] The overall response rate (uPR+PR+CR) and disease control rate (ORR+SD) were 48% (11 of 23; 3 uPR, 5 PR, 2 CR, 1 uCR) and 70% (16 of 23 including 5 SD), respectively. Two patients were not yet evaluable. Median PFS and median OS were calculated, and only one patient experienced PD during REGN2810 treatment after initial response. Correlative scientific studies including whole-exome tumor DNA sequencing are ongoing.

[0349] conclusion REGN2810 demonstrated robust antitumor activity in patients with advanced CSCC. EXAMPLES

[0350] A Clinical Trial Comparing Anti-PD-1 Antibody Combined With Hypofractionated Radiation Therapy to Standard of Care in Patients Age 65 and Older With Newly Diagnosed Glioblastoma Introduction Glioblastoma is a fatal disease with a median survival of approximately 16 months in newly diagnosed patients (nGBM) and 9 months in the recurrent setting (rGBM) (Friedman et al, 2009, J. Clin. Oncol. 27:4733-4740). The current standard of care for newly diagnosed patients with glioblastoma is radiation (60 Gy over 6 weeks) with concurrent temozolomide (TMZ) followed by adjuvant temozolomide (Stupp et al, 2005, N. Engl. J. Med. 352:987-996), although subgroup analyses suggest that the addition of temozolomide may not improve efficacy in older patients (Laperriere et al, 2013, Cancer Treat. Rev. 39:350-357).

[0351] This example describes a phase 3 clinical trial to evaluate the efficacy of an anti-PD-1 antibody in combination with hypofractionated radiotherapy (hfRT) compared to standard of care (SoC) on overall survival in patients aged 65 years or older with nGBM.

[0352] The exemplary anti-PD-1 antibody used in this study was REGN2810 (also known as H4H7798N, as disclosed in U.S. Patent Application Publication No. 20150203579), a fully human monoclonal anti-PD-1 antibody comprising a heavy chain comprising the amino acid sequence of SEQ ID NO:9 and a light chain comprising the amino acid sequence of SEQ ID NO:10; an HCVR / LCVR amino acid sequence pair comprising SEQ ID NO:1 / 2; and heavy and light chain CDR sequences comprising SEQ ID NOs:3-8.

[0353] Test Purpose The primary objective of the study was to evaluate the efficacy of REGN2810 in combination with hfRT compared with standard of care in terms of overall survival (OS) for patients aged 65 years or older with nGBM.

[0354] A secondary objective of the study was to determine improvement in progression-free survival (PFS).

[0355] Other objectives of the study were: (i) improvement in objective response rate (ORR), durability of response, and sustained disease control; (ii) clinical assessment using the Neurologic Assessment in Neuro-Oncology (NANO) scale; (iii) safety; (iv) improvement in quality of life (QoL) and mental status; (v) changes in edema and steroid use; (vi) serum REGN2810 concentrations and anti-REGN2810 antibodies; and (vii) exploration of potential pharmacodynamic, predictive, or prognostic biomarkers.

[0356] Study design This is a phase 3 trial randomized 2:1 to REGN2810, a fully human antibody against PD-1, in combination with hypofractionated radiotherapy versus standard of care in patients aged 65 years or older with newly diagnosed glioblastoma. Patients were randomized 2:1 to REGN2810 in combination with hypofractionated radiotherapy versus standard of care, with stratification factors being methylation status (methylated vs. unmethylated vs. unknown) and extent of resection (subtotal vs. gross total resection). Efficacy was assessed by overall survival.

[0357] Patients with nGBM who were candidates for radiation therapy were randomized in a 2:1 ratio to receive one of the following treatments: Study treatment: REGN2810 3mg / kg IV (every 2 weeks) plus hypofractionated RT (6Gy x 5, week 2 only). Radiotherapy was provided in week 2 of cycle 1 but not in subsequent cycles. Comparator treatment: Standard RT (2 Gy fractionated radiation, 5 days per week) combined with standard TMZ (oral, 75 mg / m 2 , daily) followed by adjuvant TMZ (orally, 150 mg / m 2 ~200mg / m 2 Patients were randomly assigned to receive 6 cycles of chemotherapy (5 days over 28 days) with radiation therapy provided only in the first 6-week cycle.

[0358] Test period The study consisted of a 28-day screening period, after which eligible patients received up to twelve 56-day (8-week) treatment cycles for a total of up to 96 weeks of treatment. During the screening period (days -28 to -1), all eligible patients were required to have pre-treatment tumors that were resectable (subtotal or total resection), biopsy for pathology confirmation by a central laboratory, and determination and confirmation of MGMT methylation.

[0359] After Day 1 / baseline, patients visited on days 8±3, 15±3, 29±3, 43±3, and 56±3 during Cycle 1. For each subsequent 8-week cycle (Cycles 2-12), patients visited on days 1, 15±3, 29±3, 43±3, and 56±3. Tumor assessments (brain MRI, iRANO, and NANO assessments, MMSE, and EORTC QLQ-C30 / BN20 questionnaires) were performed on Day 1 / baseline and at the end of each treatment cycle. Extensive safety assessments were performed on Day 1 of each cycle, and routine safety assessments were performed at each visit. Samples for evaluation of biomarkers associated with REGN2810 treatment exposure (cellular or molecular markers as described herein), clinical activity, or underlying disease were also collected.

[0360] During the 24-week follow-up period, patients were randomized to receive their last treatment for the first follow-up visit. Patients were followed for follow-up visits 21-42 days later. Subsequent follow-up visits (follow-up visits 2 through 7) were conducted every 28 days ± 7 days. Tumor assessments (brain MRI, iRANO, and NANO assessments, MMSE, and EORTC QLQ-C30 / BN20 questionnaires) were conducted at follow-up visits 3, 5, and 7. Extensive safety assessments were conducted during the first follow-up visit, and routine safety assessments were conducted at subsequent follow-up visits. Samples were collected for evaluation of biomarkers associated with REGN2810 treatment exposure (cellular or molecular markers as described herein), clinical activity, or underlying disease.

[0361] Target Group The target population included patients aged 65 years and older with nGBM.

[0362] Inclusion Criteria: Patients must meet the following criteria to be eligible for study participation: (1) newly diagnosed, histologically confirmed primary glioblastoma ≤5 cm in longest dimension undergoing partial or complete surgical resection; (2) Eastern Cooperative Oncology Group (ECOG) performance status 0-2; (3) age ≥65 years; (4) liver function: (a) total bilirubin ≤1.5×upper limit of normal; (b) ALT and AST ≤3×ULN; (c) alkaline phosphatase (ALP) ≤2.5×ULN; (5) renal function: serum creatinine ≤1.5×ULN; (6) bone marrow function: hemoglobin ≥9.0 g / dL; absolute neutrophil count (ANC) ≥1.5×10 9 ≥ 75 × 10 9 (7) Able to read, understand, and willing to sign the ICF; and (8) Able and willing to consent to scheduled visits, treatment plans, laboratory tests, and other study-related procedures.

[0363] Exclusion Criteria: Patients who meet any of the following criteria will be excluded from the study: (1) any previous treatment for GBM (other than surgery); (2) have known contraindications to Gd-MRI; (3) evidence of ongoing or recent (within 5 years) significant autoimmune disease requiring treatment with systemic immunosuppressive measures that may indicate risk of immune-related adverse events (irAEs). The following are not excluded: vitiligo, childhood asthma in remission, residual hypothyroidism requiring only hormone replacement therapy, or psoriasis not requiring systemic treatment; (4) ongoing systemic corticosteroid treatment, excluding use of up to 10 mg / day prednisone or equivalent for other (non-oncology and non-immunosuppressive) indications; (5) any secondary active infection requiring treatment, including a primary tumor located in the brain stem, spinal cord, or known infection with human immunodeficiency virus, or active infection with hepatitis B or C virus; (6) history of pneumonia within the past 5 years; (7) any investigational drug or antitumor treatment within 30 days prior to the first dose of REGN2810; (8) any allergic reaction due to antibody treatment in general, or to any agent used in the study in particular. (9) inadequately controlled hypertension (defined as systolic pressure >150 mmHg and / or diastolic pressure >100 mmHg); (10) known allergy to doxycycline or tetracycline (precautions due to the presence of trace components in REGN2810); (11) history of hypertensive crisis or hypertensive encephalopathy; (12) history of invasive malignancy within the past 5 years other than the tumor being treated in this study, with the exception of excised / cauterized basal or squamous cell carcinoma of the skin or cervical intraepithelial neoplasia, or other localized tumors that were deemed cured by local treatment; (13) acute or chronic psychiatric problems that, as assessed by the investigator, would make the patient unfit to participate; (14) use of Novocure Tumor Treating Fields (Optune NovoTTF-100A device) at screening. Planned or anticipated use of Novocure Tumor Treating Fields during study participation; (15) previous treatment with carmustine wafers; (16) ongoing sexual activity in men who are not willing to practice adequate contraception during the study.

[0364] Test procedure Patients will receive one of the following treatment regimens:

[0365] Study drug treatment: REGN2810 3 mg / kg (administered as an IV infusion over 30 minutes every 2 weeks for up to 96 weeks) plus hfRT in week 2 of cycle 1.

[0366] Comparison treatment: Standard treatment: TMZ (oral, 75 mg / m 2 , daily) for 6 weeks, followed by adjuvant TMZ (oral, 150 mg / m 2 ~200mg / m 2 , 5 days out of 28 days) for 6 cycles. Radiotherapy was provided only in the first cycle.

[0367] REGN2810 was supplied as a liquid in sterile, single-use vials. Each vial contained sufficient volume to withdraw 10 mL of REGN2810 at a concentration of 25 mg / mL. REGN2810 was administered as a 30-minute IV infusion. Each patient's dose was dependent on their individual body weight. If body weight changed by 10% or more, the dose of REGN2810 must be adjusted each cycle.

[0368] Radiation therapy: Patients in the control group received standard radiation therapy (60 Gy over 6 weeks). Patients in the experimental treatment group received hfRT (6 Gy in daily fractions for 5 days) 1 week after the first dose of REGN2810.

[0369] REGN2810 Plus Radiation Therapy (Study Drug Treatment): REGN2810 was administered as an IV infusion over 30 minutes in combination with hfRT on days 8 through 12 every 14 days for 96 weeks.

[0370] Planned combination of REGN2810 and hfRT regimen: REGN2810 3 mg / kg infusion over 30 minutes every 14 days for 96 weeks plus radiation therapy (hfRT at 6 Gy x 5 daily fractions, preferably on consecutive days, given 1 week after the first dose of REGN2810).

[0371] Radiation Therapy Details: Patients received 6 Gy in 5 daily fractions of 30 Gy starting 1 week after the first dose of REGN2810.

[0372] Comparison treatment: Standard treatment: TMZ (oral, 75 mg / m 2 , daily) for 6 weeks, followed by adjuvant oral TMZ. The dose of TMZ was 150 mg / m for the first 5 days of the first adjuvant cycle. 2 and 200 mg / m with the start of the second cycle if there is no unacceptable hematologic toxicity in the first adjuvant cycle. 2 The treatment was increased to 5 days out of 28 days.

[0373] During the first adjuvant cycle, all observed non-hematologic toxicities were grade 2 or less (excluding alopecia, nausea, and vomiting) and platelet counts were ≥100 × 10 9 ≥ 1.5 x 10 9 If the number of cells / L is more than 1, the dose of TMZ is increased to dose level 1 (200 mg / m 2 ) and this dose should be used as the starting dose for subsequent cycles. If TMZ needs to be delayed after cycle 1 due to ongoing grade 2 or higher non-hematologic toxicity, it may not be escalated. If the dose is not escalated in the second cycle, it should not be escalated in subsequent cycles.

[0374] Treatment of CNS Edema: Any patient developing symptomatic intracranial edema during the study had REGN2810 and radiation therapy discontinued until the edema subsided.

[0375] For patients who developed intracranial edema, bevacizumab was administered IV as needed (PRN) at a reduced dose from the standard dose (suggested dose is 5 mg / kg twice weekly for a maximum of 3 doses, not to exceed 10 mg / kg per dose twice weekly) unless contraindicated (e.g., the patient had undergone surgery within the past 28 days).

[0376] If bevacizumab does not remit intracranial edema, systemic corticosteroids may be administered in addition to or as an alternative to bevacizumab at the lowest dose deemed appropriate for symptomatic control. For patients who are intolerant to bevacizumab, corticosteroids were used at doses deemed appropriate for symptomatic control.

[0377] Test variables The primary efficacy endpoint was overall survival (OS), defined as the time interval from the date of randomization to the date of death from any cause.

[0378] The key secondary endpoint was progression-free survival (PFS), defined as the time interval from the date of randomization to the date of first observed progression or death (from any cause), with progression determined by iRANO criteria.

[0379] Other secondary efficacy endpoints included:

[0380] Objective Response Rate (ORR): Defined as the proportion of patients with a confirmed complete response (CR) or a confirmed partial response (PR) as defined by the Immunotherapy Response Assessment in Neuro-Oncology (iRANO) criteria compared to the total number of patients in the analysis population.

[0381] Duration of Response: Determined for patients with a best overall response of CR or PR. Duration of response is measured from first meeting of metrics for CR / PR (whichever is first documented) to the first date of relapse or progression (by radiography), or death from any cause.

[0382] Duration of disease control: Determined for patients with a best overall response of SD, CR, or PR. Duration of disease control is measured from the start of treatment to the first day of recurrence or progression (by radiography), or death from any cause.

[0383] Quality of life and symptom control variables: Quality of life and symptom control variables were: The study will use the EORTC QLQ-C30 questionnaire, which includes five scales of function, three scales of symptoms, one scale of general health status, and six single-item scales assessing symptoms. During the study, the EORTC QLQ-BN20 questionnaire was used, with 4 scales and 7 single items. Clinical evaluation using NANO · Total MMSE score during the test Use of corticosteroids at baseline, cumulative use of corticosteroids during the study, and periods of steroid-free or low-dose steroid use during the progression-free period of the study Use of bevacizumab PRN at baseline, cumulative bevacizumab PRN during the study, and bevacizumab-free periods during progression-free time It was.

[0384] Exploratory biomarker variables: Other endpoints will be identified that are related to pharmacodynamics, prognosis, and clinical response. The study included predictive biomarkers, mechanism of action, and possible REGN2810-related AEs following treatment. Biomarker variables were: · Expression levels of immune checkpoint receptors PD-L1, GITR, and LAG3 and other potential biomarkers (e.g., EGFRvIII, Ki67, etc.) in tumor samples; Number and distribution of TILs in tumor samples · 1DH1 mutation status, microsatellite instability (MSI), and mutation burden in tumor samples; · Circulating biomarkers, including cytokines and angiogenic factors; · Cell subsets and expression levels of biomarkers of interest in PBMCs; MGMT promoter methylation status (also used for stratification) Includes:

[0385] Other variables included serum REGN2810 concentrations (a pharmacokinetic variable) and the occurrence of anti-REGN2810 antibodies.

[0386] Procedures and Evaluation After a screening period of up to 28 days, patients were offered up to twelve 56-day treatment cycles for a total of up to 96 weeks of treatment, followed by a 24-week follow-up period. Efficacy, safety, PK, ADA, and exploratory biomarker analyses were performed.

[0387] Efficacy Procedure MRI: MRI for tumor assessment was performed 72 hours after surgery, at the screening visit (within 28 days prior to infusion), at days 56 ± 3 of every cycle (approximately every 8 weeks), and if PD was suspected. Patients without disease progression underwent further tumor evaluation at follow-up visits 3, 5, and 7. Note: If PD is confirmed, no additional scans are required at follow-up visits. If pre- and post-operative MRIs were performed prior to study enrollment, those scans must also be submitted to the study to help determine tumor volume and tumor progression.

[0388] Tumor response assessments were performed according to iRANO and clinical neurological assessments were performed by NANO. Evaluations according to RANO were also performed as ancillary exploratory measures. However, primary determination of individual patient progression was performed according to iRANO.

[0389] The European Organization for Research and Treatment of Cancer Quality of Life Questionnaire (EORTC QLQ-C30) and EORTC Brain Cancer Module (EORTC QLQ-BN20) questionnaire: The EORTC QLQ-C30 is a 30-item questionnaire that assesses health-related quality of life (HRQoL) in cancer patients in 15 scales (single- or multi-item), each with a possible score ranging from 0 to 100. Of the 30 items, 24 are collated into 9 multi-item scales that represent various HRQoL dimensions: 5 scales of functioning (physical, role, emotional, cognitive, and social life), 3 scales of symptoms (fatigue, pain, and nausea), and 1 scale of general health status. The remaining 6 single-item scales assess symptoms: dyspnea, loss of appetite, insomnia, constipation and diarrhea, and perception of the economic impact of disease treatment. Higher scores indicate better HRQoL on global health measures and functional scales, and worse HRQoL on symptom scales.

[0390] The EORTC QLQ-BN20 is a 20-item QoL assessment specific to brain neoplasms, and is comparable to the EORTC QLQ-C3 when assessing health-related quality of life. 0. The EORTC QLQ-BN20 questionnaire assesses disease symptoms, treatment side effects, and some specific psychosocial issues important to brain cancer patients, using 4 scales (assessing future anxiety, visual impairment, motor dysfunction, and communication deficits) and 7 single items (assessing other disease symptoms [e.g., headaches and seizures] and toxic effects of treatment [e.g., hair loss]). Possible scores range from 0 to 100; higher scores indicate poorer HRQoL.

[0391] Mini Mental Status Assessment: The Mini Mental Status Examination (MMSE®) is a brief quantitative measurement of cognitive status in adults. It can be used to screen for cognitive impairment, estimate the severity of cognitive impairment at a given time point, and track the course of cognitive change in an individual over time. In this study, the MMSE score was part of the neurological examination carried out in the setting of disease evaluation.

[0392] MMSE was performed on Day 1 / baseline, at the end of each treatment cycle, and every 8 weeks during the follow-up period. MMSE assessments coincide with the disease assessment schedule, but they should be completed before patients are informed of the results of the radiographic evaluation. MMSE may be completed at the start of the next scheduled treatment administration. During the survival follow-up period, MMSE should continue to be performed at every other survival visit (every 8 weeks) unless the patient has yet to progress.

[0393] The MMSE total score has a possible range from 0 (worst) to 30 (best).

[0394] Safety Procedures Vital signs including temperature, resting blood pressure, pulse rate, and respiratory rate, along with body weight, were collected pre-infusion and approximately 15 minutes after the end of the infusion on Day 1 of Cycle 1 and all subsequent treatment days. A full physical examination and 12-lead ECG were performed at the start of each cycle.

[0395] Exploratory Tumor Biomarker Procedures Biomarkers of interest analyzed by immunohistochemistry (IHC) include, but are not limited to, EGFRvIII and cell proliferation biomarkers (e.g., Ki67). Expression levels (mRNA and / or protein) of PD-L1, GITR, and LAG-3, as well as lineage markers of tumor-infiltrating lymphocytes (CD4, CD8, CD25, FoxP3) were analyzed in tumor biopsy samples to explore the potential effects of REGN2810.

[0396] Tumor tissue samples may be used to extract tumor DNA and RNA and for subsequent analysis of putative genetic biomarkers associated with study treatment and glioblastoma. Blood samples were collected at Day 1 / baseline (pre-dose) or at any study visit if Day 1 / baseline collection was not possible to isolate germline DNA. Analysis of tumor DNA included (but was not limited to) MGMT promoter methylation status, 1DH1 mutation status, microsatellite instability (MSI), and tumor mutational burden, all of which may predict response to REGN2810 and other immunotherapeutic agents. Analysis of gene variants in tumor (somatic) DNA and germline DNA that may affect progression, drug response, and potential toxicity was performed. Germline DNA was also used for comparison with tumor DNA to explore potential novel gene variants underlying the malignant process.

[0397] result The combination of REGN2810 and hfRT was safe and well tolerated in patients with nGBM. Administration of REGN2810 in combination with hfRT inhibited tumor growth and / or promoted tumor regression in nGBM patients compared to standard of care. nGBM patients treated with REGN2810 and hfRT exhibited longer OS compared to standard of care. EXAMPLES

[0398] Clinical Trial of REGN2810 in Patients with Advanced Cutaneous Squamous Cell Carcinoma This example describes a Phase 2 study that was conducted to confirm the positive results seen in patients with advanced CSCC in a Phase 1 study (see Examples 7, 8 and 9 herein).

[0399] Test Purpose The primary objective of this study was to estimate the clinical benefit of REGN2810 monotherapy for patients with metastatic (nodal or distant) cutaneous squamous cell carcinoma (CSCC) (Arm 1) or unresectable locally advanced CSCC (Arm 2), as measured by overall response rate (ORR).

[0400] Secondary objectives of the study were: (i) to estimate the ORR; (ii) to estimate duration of response, progression-free survival (PFS), and overall survival (OS); (iii) to estimate the complete response (CR) rate; (iv) to evaluate the safety and tolerability of REGN2810; (v) to evaluate the pharmacokinetics (PK) of REGN2810; (vi) to evaluate the immunogenicity of REGN2810; and (vii) to evaluate the impact of REGN2810 on quality of life using the EORTC QLQ-C30.

[0401] Study design This was a phase 2, nonrandomized, two-arm, multicenter study of REGN2810 administered intravenously (IV) at a dose of 3 mg / kg every 2 weeks in patients with advanced CSCC. The study had two arms. Arm 1 was patients with metastatic CSCC. Arm 2 was patients with unresectable locally advanced CSCC. All patients underwent a screening procedure to determine eligibility within 28 days prior to their first dose of REGN2810.

[0402] After a screening period of up to 28 days, patients received up to twelve 56-day (8-week) treatment cycles for up to 96 weeks of treatment. Each patient received REGN2810 3 mg / kg IV on days 1, 15±3, 29±3, and 43±3 during each treatment cycle. Tumor evaluations were performed at the end of each treatment cycle. Extensive safety evaluations were performed on day 1 of each cycle, and routine safety evaluations were performed at each REGN2810 dosing visit.

[0403] Patients received treatment until completion of the 96-week treatment period or until progression, unacceptable toxicity, withdrawal of consent, or confirmed CR. Patients with confirmed CR after at least 48 weeks of treatment may choose to discontinue treatment and continue all relevant study evaluations (e.g., efficacy evaluations).

[0404] Test period Screening (up to 4 weeks), treatment up to 96 weeks, and follow-up up to 6 months.

[0405] Study population Patients with metastatic CSCC or unresectable locally advanced CSCC.

[0406] Test procedure REGN2810 3 mg / kg IV over 30 minutes every 14 days for 96 weeks.

[0407] Test variables The primary efficacy endpoint of the study was ORR over 12 treatment cycles. Overall response rate was assessed separately for patients with metastatic CSCC or locally advanced unresectable CSCC. In group 1, ORR was determined using the new Response Evaluation Criteria in Solid Tumors (RECIST) version 1.1. In group 2, ORR was determined using composite response criteria. In patients who achieved CR, tumor biopsies were used to determine final complete or partial response (PR).

[0408] Secondary efficacy endpoint measures were: duration of response; duration of disease control; PFS, OS, CR rate; change in patient-reported outcome score on the EORTC QLQ-C30; adverse events (AEs); serum concentrations of REGN2810; and anti-REGN2810 antibodies.

[0409] Procedures and Evaluation Tumor imaging (computed tomography [CT] or magnetic resonance imaging [MRI]) and digital medical images (externally visible lesions) were performed to measure tumor burden and to characterize the efficacy profile of study treatments using response criteria.

[0410] Physical examinations, laboratory tests, vital signs, electrocardiograms (ECGs), pregnancy tests for women of childbearing potential, and records of AEs and concomitant medications were performed to ensure patient safety and to characterize the safety profile of the study treatment.

[0411] Other assessments included: peripheral blood samples for PK; peripheral blood samples to evaluate anti-REGN2810 antibodies; tumor biopsies; and quality of life assessments.

[0412] result The trial has enrolled the planned number of patients and the results to date are consistent with the Phase 1 results (described herein in Examples 7, 8, and 9), in that patients demonstrated inhibition of tumor growth upon administration of REGN2810. Patients with metastatic CSCC who had been previously treated with therapy but who had not undergone surgery demonstrated complete responses, partial responses, or stable disease upon treatment with the anti-PD-1 antibody REGN2810. EXAMPLES

[0413] Clinical Trial of REGN2810 in Patients with Advanced Basal Cell Carcinoma A Phase 2 study was conducted to confirm the positive results seen in patients with advanced BCC in a Phase 1 study (see Examples 7 and 8 herein).

[0414] Test Purpose The primary objective of this study was to estimate the overall response rate (ORR) of metastatic basal cell carcinoma (BCC) (Group I) or unresectable locally advanced BCC (Group II) when treated with REGN2810 monotherapy in patients who had progressed on hedgehog pathway inhibitors (HHI) or were intolerant to previous HHI treatment.

[0415] The secondary objectives for both Arm I and Arm II are: (i) to estimate the ORR according to the investigator's survey; (ii) to estimate the duration of response, progression-free survival (PFS), and overall survival (OS); (iii) to estimate the complete response (CR) rate; (iv) to evaluate the safety and tolerability of REGN2810; (v) to evaluate the pharmacokinetics (PK) of REGN2810; (vi) to evaluate the immunogenicity of REGN2810; and (vii) the effect of REGN2810 on quality of life using the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire Core 30 (EORTC QLQ-C30) and Skindex-16. The objective of the study was to evaluate the impact of 0.

[0416] Study design The study was a phase 2, nonrandomized, two-arm, multicenter study of REGN2810 at a 350 mg dose administered intravenously (IV) every three weeks (Q3W) in patients with advanced BCC who had experienced disease progression during HHI treatment or were intolerant to previous HHI treatments. The study had two arms. Arm 1 was patients with metastatic BCC. Arm 2 was patients with unresectable locally advanced BCC. All patients underwent a screening procedure to determine eligibility within 28 days prior to the first dose of REGN2810. There was no randomization or placebo control.

[0417] After a screening period of up to 28 days, patients received treatment for up to 93 weeks. Each patient received a 350 mg dose of REGN2810 IV three times per week. Infusion time for REGN2810 was approximately 30 minutes (± 10 minutes). Tumor evaluations were performed at the end of each treatment cycle, at week 9 for the fifth treatment cycle, and then at week 12 for the fourth treatment cycle. Extensive safety evaluations were performed on day 1 of each cycle, and routine safety evaluations were performed at each REGN2810 dosing visit.

[0418] Patients received treatment until the completion of the 93-week treatment period or until progression (PD), unacceptable toxicity, withdrawal of consent, or confirmed CR. Patients with confirmed CR after at least 48 weeks of treatment may choose to discontinue treatment and continue all relevant study evaluations (e.g., efficacy evaluations). Patients who discontinued study treatment due to PD visited 30 days (range: 28 to 42 days) after their last study treatment to complete an end-of-study (EOS) evaluation. After the EOS visit, patients were followed for vital status until death, loss to follow-up, or end of study.

[0419] Test period After a screening period of up to 28 days, patients received treatment for up to 93 weeks. After the end-of-study visit, there was a follow-up period of 28 days. Patients were followed for vital status until death, loss to follow-up, or end of study.

[0420] Study population Patients with metastatic (group 1) or unresectable locally advanced (group 2) BCC who experienced disease progression during HHI treatment or were intolerant to previous HHI treatment.

[0421] Test procedure Study treatment involved REGN2810 350 mg IV over 30 minutes (± 10 minutes) every 3 weeks (q3w) for up to 93 weeks.

[0422] Evaluation items The primary efficacy endpoint of the study was ORR, which was assessed separately for patients with metastatic BCC (arm 1) or locally advanced, unresectable BCC (arm 2). For patients in group 1 (metastatic BCC), ORR was determined using the new Response Evaluation Criteria in Solid Tumors (RECIST) version 1.1. If not all metastatic disease was measurable by RECIST (as may occur in patients with bone-only metastases), clinical response criteria may be used for patients with externally visible target disease. For patients in group 2 (unresectable locally advanced BCC), ORR was determined using clinical criteria. For patients with disease measurable by both clinical response criteria and RECIST 1.1, combined response criteria were used.

[0423] Secondary endpoints were: (i) durability of response; (ii) PFS; (iii) OS; (iv) CR rate; (v) change in patient-reported outcome scores on EORTC QLQ-C30 and Skindex-16; (vi) adverse events (AEs); (vii) serum concentrations of REGN2810; and (viii) anti-REGN2810 antibodies.

[0424] Procedures and Evaluation Tumor imaging (computed tomography [CT] or magnetic resonance imaging [MRI]) and digital medical imaging (externally visible lesions) were performed to measure tumor burden and characterize the efficacy profile of the study treatment using efficacy criteria. Physical examinations, laboratory tests, vital signs, electrocardiograms (ECGs), pregnancy tests for women of childbearing potential, and records of AEs and concomitant medications were performed to ensure patient safety and characterize the safety profile of the study treatment. Other evaluations included blood samples for PK, blood samples to evaluate anti-REGN2810 antibodies, tumor biopsies, biomarkers, and quality of life assessments.

[0425] result Consistent with the results of the Phase 1 study (see Examples 7, 8, and 9 herein), administration of REGN2810 was expected to cause tumor regression in patients with advanced basal cell carcinoma who showed disease progression during treatment with a hedgehog pathway inhibitor (HHI) or who were intolerant to previous HHI therapy. Patients showed complete responses, partial responses, or stable disease with treatment with REGN2810.

[0426] The present invention is not limited in scope by the specific embodiments described herein. Indeed, various modifications of the present invention in addition to the embodiments described herein will become apparent to those skilled in the art from the foregoing description and accompanying drawings. Such modifications are intended to be included within the scope of the appended claims.

Claims

1. 1. A pharmaceutical composition comprising an isolated antibody or antigen-binding fragment thereof that specifically binds to Programmed Cell Death 1 (PD-1) for use in a method of treating or inhibiting the growth of a tumor, the method comprising: (a) selecting a patient with glioblastoma multiforme (GBM); and (b) administering to the patient a therapeutically effective amount of the antibody or antigen-binding fragment thereof in combination with radiation therapy; Including, The antibody or antigen-binding fragment thereof comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) of a heavy chain variable region (HCVR) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) of a light chain variable region (LCVR), wherein the HCDR1 comprises the amino acid sequence of SEQ ID NO:3; the HCDR2 comprises the amino acid sequence of SEQ ID NO:4; the HCDR3 comprises the amino acid sequence of SEQ ID NO:5; the LCDR1 comprises the amino acid sequence of SEQ ID NO:6; the LCDR2 comprises the amino acid sequence of SEQ ID NO:7; and the LCDR3 comprises the amino acid sequence of SEQ ID NO:

8. The pharmaceutical composition.

2. The pharmaceutical composition of claim 1 , wherein the GBM is newly diagnosed or recurrent GBM.

3. The pharmaceutical composition of claim 1 or 2, wherein the antibody or antigen-binding fragment thereof is for administration in one or more doses.

4. The pharmaceutical composition of claim 3, wherein each dose comprises 0.1 to 20 mg per kg of patient body weight.

5. 5. The pharmaceutical composition of claim 3 or 4, wherein each dose comprises 1, 3, or 10 mg / kg of patient body weight.

6. (a) each dose is to be administered 0.5 to 4 weeks after the immediately preceding dose; or (b) each dose is to be administered two weeks after the immediately preceding dose; The pharmaceutical composition according to any one of claims 3 to 5.

7. The pharmaceutical composition according to any one of claims 1 to 6, wherein the radiation therapy is hypofractionated radiation therapy.

8. The pharmaceutical composition of claim 7, wherein the radiation therapy is for administering to the patient 20-50 Gy in 2-20 fractions.

9. 9. The pharmaceutical composition of claim 7 or 8, wherein radiation therapy of 30 Gy is to be administered to the patient daily in 5 fractions.

10. The pharmaceutical composition according to any one of claims 1 to 9, wherein the radiotherapy is to be administered one week after the first dose of the antibody or antigen-binding fragment thereof.

11. 11. The pharmaceutical composition of any one of claims 1 to 10, wherein one or more doses of the antibody or antigen-binding fragment thereof are administered to the patient in one or more treatment cycles, each cycle comprising 1 to 6 doses of the antibody or antigen-binding fragment thereof.

12. 12. The pharmaceutical composition of claim 11, wherein each treatment cycle comprises four doses of the antibody or antigen-binding fragment thereof, each dose being to be administered to the patient two weeks after the immediately preceding dose.

13. 13. The pharmaceutical composition of any one of claims 1 to 12, wherein the method further comprises administering to the patient an anti-angiogenic agent if the patient develops intracranial edema following administration of the antibody or antigen-binding fragment thereof.

14. The pharmaceutical composition of claim 13, wherein the antiangiogenic agent is selected from a vascular endothelial growth factor (VEGF) inhibitor and an angiopoietin-2 (Ang-2) inhibitor.

15. 15. The pharmaceutical composition of claim 14, wherein the antiangiogenic agent is bevacizumab or aflibercept.

16. 16. The pharmaceutical composition of any one of claims 1 to 15, wherein said administration results in at least one effect selected from the group consisting of inhibition of tumor growth, tumor regression, reduction in tumor size, reduction in tumor cell count, slowing of tumor growth, abscopal effect, inhibition of tumor metastasis, reduction in metastatic lesions over time, reduction in the use of chemotherapeutic or cytotoxic agents, reduction in tumor burden, prolonged progression-free survival, prolonged overall survival, complete response, partial response, and stable disease.

17. further comprising administering to said patient an additional therapeutic agent or treatment; The additional therapeutic agent or treatment can be a hedgehog pathway inhibitor, surgery, a chemotherapy agent, a cancer vaccine, a programmed cell death ligand 1 (PD-L1) inhibitor, a lymphocyte activation gene 3 (LAG3) inhibitor, a cytotoxic T-lymphocyte associated protein 4 (CTLA-4) inhibitor, an anti-glucocorticoid-induced tumor necrosis factor receptor (GITR) antibody, a T-cell immunoglobulin and mucin domain-containing protein 3 (TIM3) inhibitor, a B and T lymphocyte T-cell immunoreceptor (BTLA) inhibitors, T-cell immunoreceptor with Ig and ITIM domains (TIGIT) inhibitors, CD47 inhibitors, indoleamine-2,3-dioxygenase (IDO) inhibitors, bispecific CD3 / CD20 antibodies, vascular endothelial growth factor (VEGF) inhibitors, F) selected from the group consisting of antagonists, angiopoietin-2 (Ang2) inhibitors, transforming growth factor beta (TGFβ) inhibitors, CD38 inhibitors, epidermal growth factor receptor (EGFR) inhibitors, granulocyte-macrophage colony stimulating factor (GM-CSF), cyclophosphamide, antibodies against tumor specific antigens, Bacillus Calmette-Guerin vaccines, cytotoxins, interleukin 6 receptor (IL-6R) inhibitors, interleukin 4 receptor (IL-4R) inhibitors, IL-10 inhibitors, IL-2, IL-7, IL-21, IL-15, antibody-drug conjugates, anti-inflammatory drugs, and dietary supplements; The pharmaceutical composition according to any one of claims 1 to 16.

18. The pharmaceutical composition of any one of claims 1 to 17, wherein the antibody or antigen-binding fragment thereof is for administration intravenously, subcutaneously, or intraperitoneally.

19. The pharmaceutical composition of any one of claims 1 to 18, wherein the HCVR comprises the amino acid sequence of SEQ ID NO:1 and the LCVR comprises the amino acid sequence of SEQ ID NO:

2.

20. The pharmaceutical composition according to any one of claims 1 to 19, wherein the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:9 and a light chain comprising the amino acid sequence of SEQ ID NO:

10.

21. The pharmaceutical composition of any one of claims 1 to 18, wherein the antibody or antigen-binding fragment thereof comprises an HCVR having 90% sequence identity with SEQ ID NO:1 and / or an LCVR having 90% sequence identity with SEQ ID NO:2.

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