Treatment method for urothelial carcinoma using anti-PD-1 antibodies

Abstract

The present invention provides a method for treating urothelial carcinoma or cancer derived therefrom. [Solution] The present invention provides a method for treating a subject suffering from urothelial carcinoma or a cancer derived therefrom, comprising administering an antibody or its antigen-binding moiety that specifically binds to the programmed cell death-1 (PD-1) receptor and inhibits PD-1 activity, or a method for treating a subject suffering from urothelial carcinoma or a cancer derived therefrom, comprising administering a combination of (a) an antibody or its antigen-binding moiety that specifically binds to the PD-1 receptor and inhibits PD-1 activity, and (b) an antibody or its antigen-binding moiety that specifically binds to cytotoxic T lymphocyte antigen-4 (CTLA-4) and inhibits CTLA-4 activity.

Description

[Technical Field]

[0001] Cross-reference with related applications This application claims the interests of US Provisional Application Serial No. 62 / 414,287, filed on 28 October 2016, which is incorporated herein by reference in its entirety.

[0002] Field of Invention The present invention relates to a method for treating urothelial carcinoma or cancer derived therefrom in a subject, comprising administering to the subject an anti-programmed cell death-1 (PD-1) antibody or a combination of an anti-PD-1 antibody and an anti-cytotoxic T lymphocyte antigen-4 (CTLA-4) antibody. [Background technology]

[0003] Background of the Invention Human cancers possess numerous genetic and epigenetic changes that generate nascent antigens potentially recognizable by the immune system (Sjoblom et al. (2006) Science 314:268-74). The adaptive immune system, composed of T and B lymphocytes, has potent anti-cancer capabilities and possesses broad ability and excellent specificity to respond to a variety of tumor antigens. Furthermore, the immune system exhibits considerable plasticity and memory components. Successfully utilizing all these characteristics of the adaptive immune system will make immunotherapy unique among all cancer treatment methods.

[0004] PD-1 is a key immune checkpoint receptor expressed by activated T and B cells that mediates immunosuppression. PD-1 is a member of the CD28 family of receptors, which includes CD28, CTLA-4, ICOS, PD-1, and BTLA. Two cell surface glycoprotein ligands for PD-1, programmed cell death ligand-1 (PD-L1) and programmed cell death ligand-2 (PD-L2), have been identified. These are expressed in antigen-presenting cells and many human cancers, and have been shown to downregulate T cell activation and cytokine secretion upon binding to PD-1.

[0005] Nivolumab (formerly known as 5C4, BMS-936558, MDX-1106, or ONO-4538) is a fully human IgG4(S228P) PD-1 immune checkpoint inhibitor antibody that selectively prevents interaction with PD-1 ligands (PD-L1 and PD-L2). This thereby blocks the downregulation of antitumor T cell function (US Patent No. 8, 008, 449; Wang et al., 2014 Cancer Immunol Res. 2(9):846-56).

[0006] Ipilimumab (YERVOY®) is a fully human IgG1 monoclonal antibody that blocks the binding of CTLA-4 to its B7 ligand, thereby stimulating T cell activation and improving overall survival (OS) in patients with advanced melanoma (Hodi et al. (2010) N Engl J Med 363:711-23). ​​In a phase I clinical trial, combination therapy with nivolumab and ipilimumab resulted in rapid and deep tumor regression in a significant proportion of patients with advanced melanoma, and was significantly more effective than either antibody alone (Wolchok et al. (2013) N Engl J Med 369(2)-122-33;WO2013 / 173223).

[0007] Urothelial carcinoma (UC) includes cancers of the bladder, ureters, and renal pelvis, with the majority presenting as bladder cancer. Bladder cancer accounts for approximately 5% of all new cancers in the United States, with an estimated 76,960 new cases diagnosed and approximately 16,390 deaths attributed to the disease in 2016 ("Key statistics for bladder cancer," cancer.org, May 23, 2016). Although considered a chemotherapy-sensitive disease, most patients with advanced or metastatic urothelial carcinoma relapse after first-line treatment with cisplatin (see, e.g., Oing et al., J. Urology 195(2):254-63 (2016)). Many patients respond initially to standard care, but in most cases, the disease progresses on average within about 8 months. At the time of relapse, there are few treatment options. The commonly used drugs, namely paclitaxel, carboplatin, and / or gemcitabine, are not approved by the FDA (Food and Drug Administration) for second-line systemic treatment, and most patients receive palliative care upon relapse (ibid.). Therefore, there is still a need for effective treatment methods for UC, particularly second-line therapies for patients who relapse after initial treatment. [Overview of the project]

[0008] Summary of the Invention This disclosure relates to a method for treating subjects suffering from urothelial carcinoma (UC) or a cancer derived therefrom, comprising administering to the subject an antibody ("anti-PD-1 antibody") or its antigen-binding moiety that specifically binds to the programmed cell death-1 (PD-1) receptor and inhibits PD-1 activity. In some embodiments, the method further comprises administering to the subject an antibody ("anti-CTLA-4 antibody") or its antigen-binding moiety that specifically binds to cytotoxic T lymphocyte antigen-4 (CTLA-4) and inhibits CTLA-4 activity.

[0009] Other embodiments of the present disclosure relate to a method for treating a subject with ulcerative colitis or cancer derived therefrom, comprising administering a combination of (a) an anti-PD-1 antibody and (b) an anti-CTLA-4 antibody to the subject.

[0010] In some embodiments, UC includes bladder cancer. In other embodiments, UC includes ureteral cancer. In other embodiments, UC includes renal pelvis carcinoma. In some embodiments, UC includes transitional cell carcinoma. In some embodiments, UC includes squamous cell carcinoma. In some embodiments, UC includes adenocarcinoma. In some embodiments, UC is recurrent UC. In some embodiments, UC is locally progressive. In certain embodiments, UC is metastatic.

[0011] In some embodiments, the subject received at least one, at least two, at least three, at least four, or at least five prior treatment lines to treat UC. In some embodiments, the prior treatment line includes chemotherapy. In some embodiments, the chemotherapy includes platinum-based therapy. In some embodiments, the platinum-based therapy includes a platinum-based antineoplastic agent selected from the group consisting of cisplatin, carboplatin, oxaliplatin, nedaplatin, triplatin tetranitrate, phenantriplatin, picoplatin, satraplatin, and any combination thereof. In certain embodiments, the platinum-based therapy includes cisplatin. In a particular embodiment, the platinum-based therapy includes carboplatin.

[0012] In some embodiments, the anti-PD-1 antibody cross-competes with nivolumab for binding to human PD-1. In some embodiments, the anti-PD-1 antibody binds to the same epitope as nivolumab. In some embodiments, the anti-PD-1 antibody is a chimeric, humanized, or human monoclonal antibody or a part thereof. In other embodiments, the anti-PD-1 antibody contains a heavy chain constant region of a human IgG1 or IgG4 isotype. In certain embodiments, the anti-PD-1 antibody is nivolumab. In one embodiment, the anti-PD-1 antibody is pembrolizumab.

[0013] In some embodiments, the anti-CTLA-4 antibody is a chimeric, humanized, or human monoclonal antibody or a part thereof. In some embodiments, the anti-CTLA-4 antibody contains the heavy chain constant region of a human IgG1 isotype. In certain embodiments, the anti-CTLA-4 antibody is ipilimumab. In other embodiments, the anti-CTLA-4 antibody is tremelimumab. In some embodiments, the anti-CTLA-4 antibody cross-competes with ipilimumab for binding to human CTLA-4.

[0014] In some embodiments, the anti-PD-1 antibody is administered once every 1, 2, 3, or 4 weeks in a dose ranging from at least about 0.1 mg / kg body weight to at least about 10.0 mg / kg body weight. In some embodiments, the anti-PD-1 antibody is administered in a dose of about 1 mg / kg body weight or about 3 mg / kg body weight. In some embodiments, the anti-PD-1 antibody is administered in a constant dose. In some embodiments, the anti-PD-1 antibody is administered in a constant dose of at least about 200 mg, at least about 220 mg, at least about 240 mg, at least about 260 mg, at least about 280 mg, at least about 300 mg, at least about 320 mg, at least about 340 mg, at least about 360 mg, at least about 380 mg, at least about 400 mg, at least about 420 mg, at least about 440 mg, at least about 460 mg, at least about 480 mg, at least about 500 mg, or at least about 550 mg. In some embodiments, the anti-PD-1 antibody is administered in a constant dose approximately once every 1, 2, 3, or 4 weeks. In some embodiments, the anti-PD-1 antibody is administered approximately once every 2 weeks. In some embodiments, the anti-PD-1 antibody is administered approximately once every 3 weeks. In some embodiments, the anti-PD-1 antibody is administered as long as a clinical benefit is observed or until uncontrollable toxicity or disease progression occurs.

[0015] In some embodiments, the anti-CTLA-4 antibody is administered at a dose ranging from at least about 0.1 mg / kg body weight to at least about 10.0 mg / kg body weight, once about every 1, 2, 3, or 4 weeks. In some embodiments, the anti-CTLA-4 antibody is administered at a dose of about 1 mg / kg body weight or about 3 mg / kg body weight. In some embodiments, the anti-PD-1 antibody is administered at a fixed dose. In some embodiments, the anti-CTLA-4 antibody is administered once about every 2 weeks. In some embodiments, the anti-CTLA-4 antibody is administered once about every 3 weeks.

[0016] In some embodiments, the anti-PD-1 antibody is administered at a dose of about 3 mg / kg body weight once about every 3 weeks, and the anti-CTLA-4 antibody is administered at a dose of about 1 mg / kg body weight once about every 3 weeks. In some embodiments, the anti-PD-1 antibody is administered at a dose of about 1 mg / kg body weight once about every 3 weeks, and the anti-CTLA-4 antibody is administered at a dose of about 3 mg / kg body weight once about every 3 weeks.

[0017] In certain embodiments, a subject treated by the disclosed method exhibits progression-free survival of at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 7 months, at least about 8 months, at least about 9 months, at least about 10 months, at least about 11 months, at least about 1 year, at least about 18 months, at least about 2 years, at least about 3 years, at least about 4 years, or at least about 5 years after the first administration.

[0018] In some embodiments, the subject has a tumor with PD-L1 expression of 1% or more. In other embodiments, the subject has a tumor with PD-L1 expression of 5% or more. In certain embodiments, the anti-PD-1 antibody, or the combination of an anti-PD-1 antibody and an anti-CTLA-4 antibody, is administered for as long as a clinical benefit is observed, or until disease progression or unmanageable toxicity occurs. In one embodiment, the anti-PD-1 and / or anti-CTLA-4 antibody is formulated for intravenous administration. In certain embodiments, the anti-PD-1 antibody and the anti-CTLA-4 antibody are administered sequentially to the subject. In some embodiments, the anti-PD-1 antibody and the anti-CTLA-4 antibody are administered to each other within 30 minutes. In one embodiment, the anti-PD-1 antibody is administered before the anti-CTLA-4 antibody. In another embodiment, the anti-CTLA-4 antibody is administered before the anti-PD-1 antibody. In some embodiments, the anti-PD-1 antibody and the anti-CTLA-4 antibody are administered simultaneously in separate compositions. In certain embodiments, the anti-PD-1 antibody and the anti-CTLA-4 antibody are administered simultaneously as a single composition.

[0019] In one embodiment, the anti-PD-1 antibody is administered at a sub-therapeutic dose. In certain embodiments, the anti-CTLA-4 antibody is administered at a sub-therapeutic dose. In some embodiments, the anti-PD-1 antibody and the anti-CTLA-4 antibody are each administered at a sub-therapeutic dose.

[0020] The present disclosure further relates to a kit for treating a subject suffering from UC or cancer derived therefrom, comprising: (a) an anti-PD-1 antibody in an amount ranging from about 4 mg to about 500 mg; and (b) instructions for using the anti-PD-1 antibody in any of the disclosed methods.

[0021] The disclosure further relates to a kit for treating subjects suffering from UC or cancer derived therefrom, comprising (a) an amount of anti-PD-1 antibody ranging from about 4 mg to about 500 mg; (b) an amount of anti-CTLA-4 antibody ranging from about 4 mg to about 500 mg; and (c) instructions for using the anti-PD-1 antibody and the anti-CTLA-4 antibody in any disclosed manner.

[0022] Other features and advantages of the present invention will become apparent from the following detailed description and examples, but should not be construed as limitations. The contents of all cited literature, including scientific papers, newspaper articles, GenBank entries, patents and patent applications, cited throughout this specification are expressly incorporated herein by reference. [Brief explanation of the drawing]

[0023] [Figure 1] Figure 1 shows a schematic diagram of a study design for the treatment of locally advanced or metastatic urothelial carcinoma (UC) previously treated with platinum-based therapy using an anti-PD-1 antibody or a combination of an anti-PD-1 antibody and an anti-CTLA-4 antibody.

[0024] Detailed description of the invention The present invention relates to a method for treating ulcerative colitis (UC) or cancer derived therefrom in a patient, comprising administering to the patient an anti-PD-1 antibody or a combination of an anti-PD-1 antibody and an anti-CTLA-4 antibody.

[0025] word To facilitate understanding of this disclosure, certain terms are defined first. Where used herein, unless otherwise expressly provided herein, each of the following terms shall have the meaning set forth below. Additional definitions are provided throughout this application.

[0026] "Administer" means physically introducing a composition containing a therapeutic agent into a target using any of the various methods and delivery systems known to those skilled in the art. The routes of administration of the antibodies of this application (e.g., anti-PD-1 antibody and / or anti-CTLA-4 antibody) include intravenous, intramuscular, subcutaneous, intraperitoneal, spinal, or other parenteral administration routes, for example, by injection or infusion. As used herein, the term "parenteral administration" means a mode of administration other than intestinal and topical administration, usually by injection, and includes, but is not limited to, intravenous, intramuscular, intra-arterial, intrathecal, intralymphatic, intrafocal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subepidermal, intra-articular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal injections and infusions, as well as in vivo electroporation. In some embodiments, the combination is administered via parenteral routes, and in some embodiments, it is administered orally. Other parenteral routes include topical, epidermal, or mucosal administration routes, such as intranasal, intravaginal, intrarectal, sublingual, or topical administration routes. Administration can also be carried out, for example, once, multiple times, and / or over one or more extended periods. In some embodiments, anti-PD-1 antibodies and anti-CTLA-4 antibodies can be administered simultaneously or sequentially. In some embodiments, the anti-PD-1 antibody is administered before the anti-CTLA-4 antibody. In other embodiments, the anti-CTLA-4 antibody is administered before the anti-PD-1 antibody.

[0027] As used herein, “adverse event” (AE) is any undesirable, generally unintended, or unwanted sign (including abnormal laboratory findings), symptom, or illness associated with the use of a medical procedure. For example, an adverse event may be associated with the activation of the immune system or the expansion of immune system cells (e.g., T cells) in response to a procedure. A medical procedure may have one or more associated AEs, and each AE may have the same or different levels of severity. References to methods that can “modify an adverse event” mean a treatment plan that reduces the incidence and / or severity of one or more AEs associated with the use of a different treatment plan.

[0028] An "antibody" (Ab) should include a glycoprotein immunoglobulin, or its antigen-binding moiety, which specifically binds to an antigen and comprises at least two heavy (H) chains and two light (L) chains interconnected by disulfide bonds. Each H chain has a heavy chain variable region (V in this specification). H It includes a heavy chain constant region (abbreviated as CH1, CH2, and CH3). Each light chain has a light chain variable region (V in this specification). L (Abbreviated as ) and includes a light chain constant region. The light chain constant region includes one constant domain, CL. V H and V L The region can be further subdivided into a highly variable region called the Complementarity Determination Region (CDR), which contains more conserved regions called the Framework Region (FR). H and V L It contains three CDRs and four FRs arranged in the order FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4 from the amino terminus to the carboxyl terminus. The variable regions of the heavy and light chains contain binding domains that interact with the antigen. The constant region of the antibody can mediate the binding of the immunoglobulin to various cells of the immune system (e.g., effector cells) and host tissues or factors including the first component (C1q) of the classical complement system.

[0029] Immunoglobulins may be derived from any of the commonly known isotypes, including but not limited to IgA, secretory IgA, IgG, and IgM. IgG subclasses are also well known to those skilled in the art, including but not limited to human IgG1, IgG2, IgG3, and IgG4. “Isotype” refers to an antibody class or subclass (e.g., IgM or IgG1) encoded by a heavy chain constant region gene. The term “antibody” includes, by example, both naturally occurring and non-naturally occurring antibodies; monoclonal and polyclonal antibodies; chimeric and humanized antibodies; human or non-human antibodies; fully synthetic antibodies; and single-chain antibodies. Non-human antibodies can be humanized by recombinant methods to reduce their immunogenicity in humans. Unless expressly stated and the context otherwise indicates, the term “antibody” also includes antigen-binding fragments or antigen-binding moieties of any of the aforementioned immunoglobulins, and monovalent and bivalent fragments or moieties, and single-chain antibodies.

[0030] "Isolated antibody" refers to an antibody that substantially does not contain other antibodies with different antigen specificities (for example, an isolated antibody that specifically binds to PD-1 substantially does not contain antibodies that specifically bind to antigens other than PD-1). However, an isolated antibody that specifically binds to PD-1 may cross-reactive to other antigens, such as PD-1 molecules from different species. Furthermore, an isolated antibody may not substantially contain other cellular material and / or chemicals.

[0031] The term "monoclonal antibody" (mAb) refers to a preparation of an antibody molecule that does not exist in nature, having a single molecular composition; that is, an antibody molecule whose primary sequence is essentially identical and which exhibits a single binding specificity and affinity for a particular epitope. Monoclonal antibodies are an example of isolated antibodies. Monoclonal antibodies can be produced by hybridoma, recombinant, transgenic, or other techniques known to those skilled in the art.

[0032] A “human” antibody (HuMAb) refers to an antibody having a variable region in which both the framework region and the CDR region are derived from a human germline immunoglobulin sequence. Furthermore, if the antibody contains a constant region, that constant region is also derived from a human germline immunoglobulin sequence. The human antibodies of this disclosure may contain amino acid residues not encoded by a human germline immunoglobulin sequence (e.g., mutations introduced by random or site-directed mutagenesis in vitro, or by somatic mutation in vivo). However, as used herein, the term “human antibody” is not intended to include antibodies in which a CDR sequence derived from the germline of another mammalian species, such as mouse, has been transplanted into a human framework sequence. The terms “human” antibody and “fully human” antibody are used synonymously.

[0033] A "humanized antibody" refers to an antibody in which some, most, or all of the amino acids outside the CDR domain of a non-human antibody are replaced with corresponding amino acids derived from human immunoglobulins. In one embodiment of a humanized antibody, some, almost, or all of the amino acids outside the CDR domain are replaced with amino acids derived from human immunoglobulins, while some, almost, or all of the amino acids within one or more CDR regions remain unchanged. Small additions, deletions, insertions, substitutions, or modifications of amino acids are acceptable as long as they do not invalidate the antibody's ability to bind to a particular antigen. "Humanized" antibodies retain similar antigen specificity to the original antibody.

[0034] A "chimeric antibody" refers to an antibody in which the variable region originates from one species and the constant region originates from another species; for example, an antibody in which the variable region originates from a mouse antibody and the constant region originates from a human antibody.

[0035] An "anti-antigen" antibody refers to an antibody that specifically binds to an antigen. For example, an anti-PD-1 antibody specifically binds to PD-1, and an anti-CTLA-4 antibody specifically binds to CTLA-4.

[0036] The "antigen-binding portion" (also called the "antigen-binding fragment") of an antibody refers to one or more fragments of the antibody that possess the ability to specifically bind to the antigen to which the entire antibody has bound.

[0037] As used herein, if both a primary and secondary antibody can bind to the same antigen, and the binding of the primary antibody to the antigen inhibits the binding of the secondary antibody to the antigen, the primary antibody is said to “cross-compete” with the secondary antibody. In some embodiments, the primary and secondary antibodies bind to the same epitope on the antigen. In some embodiments, the primary and secondary antibodies bind to epitopes that overlap or are adjacent to each other. In other embodiments, the primary and secondary antibodies bind to different epitopes distal to each other on the antigen. In some embodiments, the primary antibody physically blocks the binding of the secondary antibody to the epitope by masking the epitope. In other embodiments, the binding of the antigen by the primary antibody causes a conformational change in the three-dimensional structure of the antigen, inhibiting its ability to bind to the antigen by the secondary antibody. Cross-competition can be determined using any technique known in the art for measuring antibody-antigen interactions, including but not limited to BIACORE® analysis, ELISA assays, immunohistochemistry, and flow cytometry. The ability of a test antibody to inhibit the binding of a reference antibody demonstrates that the test antibody can compete with the reference antibody.

[0038] "Cancer" refers to a range of diseases characterized by the uncontrolled proliferation of abnormal cells within the body. "Cancer" or "cancer tissue" may include tumors. Uncontrolled cell division and proliferation can lead to the formation of cancer (e.g., malignant tumors) that invade adjacent tissues or lymph nodes (referred to herein as "locally advanced") and can also metastasize to distant parts of the body via the lymphatic system or bloodstream (referred to herein as "metastatic"). Locally advanced cancer can "originate" from an original pre-invasive cancer or tumor. After metastasis, cancer (e.g., distal tumors) can also "originate" from an original pre-metastatic cancer or tumor. For example, "cancer originating from UC" includes cancer or tumors that are the result of locally advanced or metastatic urothelial carcinoma.

[0039] Cytotoxic T lymphocyte antigen-4 (CTLA-4) refers to an immunosuppressive receptor belonging to the CD28 family. CTLA-4 is expressed exclusively on T cells in vivo and binds to two ligands, CD80 and CD86 (also known as B7-1 and B7-2, respectively). As used herein, the term "CTLA-4" includes human CTLA-4 (hCTLA-4), variants, isoforms, and species homologs of hCTLA-4, as well as analogs having at least one common epitope with hCTLA-4. The complete hCTLA-4 sequence can be found under GenBank registry number AAB59385.

[0040] The term "immunotherapy" refers to the treatment of a person who is suffering from a disease or is at risk of suffering from or relapsing from a disease, by means of inducing, enhancing, suppressing, or otherwise modifying the immune response.

[0041] The terms “treatment,” “to treat,” or “therapy” of the subject refer to any type of intervention or process, or administration of an active agent, performed on the subject for the purpose of reversing, reducing, improving, inhibiting, slowing, or preventing the onset, progression, development, severity, or recurrence of symptoms, complications, conditions, or biochemical signs associated with the disease. As used herein, “progression-free survival” refers to the period of time during which the subject is alive without progression of the disease (e.g., UC or cancer derived therefrom).

[0042] As used herein, “PD-L1 positive” can be used interchangeably with “at least about 1% PD-L1 expression.” In one embodiment, PD-L1 expression can be used by any method known in the art. In another embodiment, PD-L1 expression is measured by automated IHC. Thus, a PD-L1 positive tumor may have at least about 1%, at least about 2%, at least about 5%, at least about 10%, or at least about 20% of tumor cells expressing PD-L1 when measured by automated IHC. In certain embodiments, “PD-L1 positive” means that there are at least 100 cells expressing PD-L1 on the surface of the cells.

[0043] "Programmed cell death-1 (PD-1)" refers to an immunosuppressive receptor belonging to the CD28 family. PD-1 is primarily expressed on previously activated T cells in vivo and binds to two ligands, PD-L1 and PD-L2. As used herein, the term "PD-1" includes human PD-1 (hPD-1), variants, isoforms, and species homologs of hPD-1, as well as analogs having at least one common epitope with hPD-1. The complete hPD-1 sequence can be found under GenBank registration number U64863.

[0044] "Programmed cell death ligand-1 (PD-L1)" is one of two cell surface glycoprotein ligands to PD-1 (the other being PD-L2) that, upon binding to PD-1, downregulate T cell activation and cytokine secretion. As used herein, the term "PD-L1" includes human PD-L1 (hPD-L1), variants, isoforms, and species homologs of hPD-L1, as well as analogs having at least one common epitope with hPD-L1. The complete hPD-L1 sequence can be found under GenBank registry number Q9NZQ7.

[0045] "Subject" includes any human or non-human animal. The term "non-human animal" includes, but is not limited to, non-human primates, vertebrates such as sheep and dogs, and rodents such as mice, rats, and guinea pigs. In some embodiments, the subject is human. The terms "subject" and "patient" are used interchangeably herein.

[0046] As used herein, if a subject has a disease or disorder (e.g., UC or cancer derived therefrom), the subject is said to be “suffering” from the disease or disorder. In some embodiments, the subject has been diagnosed with UC or cancer derived therefrom. In some embodiments, the UC is recurrent UC after one or more prior treatments. In some embodiments, the subject is undergoing treatment for UC or cancer derived therefrom. In certain embodiments, the subject has been diagnosed with UC but does not exhibit any signs or symptoms generally associated with UC. In some embodiments, the subject previously had UC but is no longer detectable after one or more prior treatments.

[0047] The “therapeutically effective dose” or “therapeutically effective dosage” of a drug or therapeutic agent is any amount of the drug, when used alone or in combination with other therapeutic agents, that protects a subject from disease onset or promotes disease regression, as demonstrated by a reduction in the severity of disease symptoms, an increase in the frequency and duration of asymptomatic periods, or the prevention of functional impairment or disability due to the distress of the disease. The ability of a therapeutic agent to promote disease regression can be evaluated using various methods known to those skilled in the art, such as in human subjects during clinical trials, in animal model systems to predict efficacy in humans, or by assaying the activity of the drug in in vitro assays.

[0048] As used herein, “sub-therapeutic dose” means a dose of a therapeutic compound (e.g., an antibody) that is lower than the usual or typical dose of the therapeutic compound when administered alone for the treatment of a hyperproliferative disease (e.g., cancer).

[0049] For example, an "anti-cancer agent" promotes the regression of cancer in a subject or prevents further tumor growth. In certain embodiments, a therapeutically effective dose of the drug promotes cancer regression to the point of eliminating the cancer. "Promoting cancer regression" means that administration of an effective dose of the drug alone or in combination with an anti-neoplastic agent results in a reduction in tumor growth or size, tumor necrosis, a decrease in the severity of at least one disease symptom, an increase in the frequency and duration of asymptomatic periods, or prevention of functional impairment or disability due to the distress of the disease. Furthermore, the terms "effective" and "effective" in relation to a treatment include both pharmacological efficacy and physiological safety. Pharmacological efficacy refers to the ability of a drug to promote cancer regression in a patient. Physiological safety refers to the level of toxicity resulting from the administration of the drug, or other harmful physiological effects (adverse effects) at the cellular, organ, and / or biological levels.

[0050] As an example for tumor treatment, a therapeutically effective dose of an anticancer agent can inhibit cell proliferation or tumor growth by at least about 20%, at least about 40%, at least about 60%, or at least about 80% compared to an untreated subject. In other embodiments of this disclosure, tumor regression may be observed and continue for at least about 20 days, at least about 40 days, or at least about 60 days. Despite these final measurements of therapeutic efficacy, the evaluation of immunotherapy drugs must also take into account “immune-related” response patterns.

[0051] The “immune-associated” response pattern refers to a clinical response pattern frequently observed in cancer patients treated with immunotherapeutic agents that produce antitumor effects by inducing cancer-specific immune responses or by modifying innate immune processes. This response pattern is characterized by beneficial therapeutic effects followed by an initial increase in tumor volume or the appearance of new lesions, which would be classified as disease progression in the evaluation of traditional chemotherapy agents and would be synonymous with drug failure. Therefore, proper evaluation of immunotherapeutic agents may require long-term monitoring of their effects on the target disease.

[0052] A therapeutically effective dose of a drug includes a “prophylactic effective dose,” which is any amount of the drug that inhibits the development or recurrence of cancer when administered alone or in combination with an antineoplastic agent to a subject at risk of developing cancer (e.g., a subject with a pre-malignant condition) or a subject at risk of cancer recurrence. In certain embodiments, a prophylactic effective dose completely prevents the development or recurrence of cancer. To “inhibit” the development or recurrence of cancer means to reduce the likelihood of cancer developing or recurring, or to completely prevent cancer developing or recurring.

[0053] "Relapsing" UC refers to UC that has progressed after one or more previous lines of treatment. In some embodiments, UC progression occurred after a partial or complete response to one or more previous lines of treatment.

[0054] As used herein, “prior treatment line” refers to any therapy administered to a subject for the treatment of cancer in or arising from UC that occurred prior to or concurrently with a recurrence of UC. In certain embodiments, the prior treatment line includes chemotherapy. In some embodiments, chemotherapy includes platinum-based therapy. In certain embodiments, platinum-based therapy includes a platinum-based antineoplastic agent selected from the group consisting of cisplatin, carboplatin, oxaliplatin, nedaplatin, triplatin tetranitrate, phenanthriplatin, picoplatin, satraplatin, and any combination thereof. In one particular embodiment, platinum-based therapy includes cisplatin (e.g., cisplatin alone; in combination with methotrexate, vinblastine, and / or doxorubicin ("MVAC"); or in combination with paclitaxel and / or gemcitabine ("PGC")). In other embodiments, chemotherapy includes paclitaxel, docetaxel, gemcitabine, or any combination thereof. In some embodiments, the prior treatment line includes radiotherapy.

[0055] The use of substitutes (e.g., "or") should be understood to mean one, both, or any combination thereof of the substitutes. Where used herein, the indefinite articles "a" or "an" should be understood to refer to "one or more" of any recited or enumerated components.

[0056] The terms “approximately” or “essentially including” refer to a value or composition that falls within an acceptable margin of error for a particular value or composition as determined by those skilled in the art, and this will depend in part on how the value or composition is measured or determined, i.e., on the limits of the measuring system. For example, “approximately” or “essentially including” may mean within one or more standard deviations per run in the art. Alternatively, “approximately” or “essentially including” may mean a range of up to 10% or 20% (i.e., ±10% or ±20%). For example, approximately 3 mg may include any number between 2.7 mg and 3.3 mg (in the case of 10%) or 2.4 mg and 3.6 mg (in the case of 20%). Furthermore, particularly with respect to biological systems or processes, the term may mean up to one order of magnitude or up to five times the value. Where a particular value or composition is provided in this application and claims, unless otherwise specified, the meaning of “approximately” or “essentially including” should be considered to be within the acceptable margin of error for that particular value or composition.

[0057] The terms “about once a week,” “about once every two weeks,” or any other similar terms for dosing intervals used herein mean approximate numbers. “About once a week” may include once every 7 ± 1 days, i.e., once every 6 days through once every 8 days. “About once every two weeks” may include once every 14 ± 3 days, i.e., once every 11 days through once every 17 days. Similar approximations apply, for example, to once every 3 weeks, once every 4 weeks, once every 5 weeks, once every 6 weeks, and once every 12 weeks. In some embodiments, dosing intervals of about 6 weeks or about 12 weeks mean that the first dose can be administered on any day in the first week, and then the next dose can be administered on any day in the 6th or 12th week, respectively. In other embodiments, an interval of approximately 6 weeks or approximately 12 weeks means that the first dose is administered on a specific day in the first week (e.g., Monday), and then the next dose is administered on the same day in the 6th or 12th week (i.e., Monday), respectively.

[0058] Where described herein, any concentration range, percentage range, ratio range, or integer range is understood to include any integer value within the listed range, and, where appropriate, fractions thereof (e.g., 1 / 10 and 1 / 100 of an integer), unless otherwise specified.

[0059] Various embodiments of the present invention are described in further detail in the following subsections.

[0060] Method of the present invention This disclosure provides a method for treating a subject with ulcerative colitis (UC) or a cancer derived therefrom, comprising administering to a subject an antibody ("anti-PD-1 antibody") or its antigen-binding moiety that specifically binds to the programmed cell death-1 (PD-1) receptor and inhibits PD-1 activity. This disclosure further provides a method for treating a subject with UC or a cancer derived therefrom, comprising administering to a subject a combination of (a) an anti-PD-1 antibody and (b) an antibody ("anti-CTLA-4 antibody") or its antigen-binding moiety that specifically binds to cytotoxic T lymphocyte antigen-4 (CTLA-4) and inhibits CTLA-4 activity. In some embodiments, the subject is a human patient.

[0061] In certain embodiments, the subject is a patient who is naive to chemotherapy (e.g., a patient who has never received chemotherapy before). In other embodiments, the subject is receiving other cancer therapies (e.g., chemotherapy) but is resistant or refractory to such other cancer therapies. In a particular embodiment, the UC is relapsed UC. In some embodiments, the subject has received at least one, at least two, at least three, at least four, or at least five prior treatment lines to treat the tumor. In one embodiment, the subject has received one prior treatment line to treat the tumor. In another embodiment, the subject has received two prior treatment lines to treat the tumor. In another embodiment, the subject has received three prior treatment lines to treat the tumor. In another embodiment, the subject has received four prior treatment lines to treat the tumor. In another embodiment, the subject has received five prior treatment lines to treat the tumor. In another embodiment, the subject has received more than five prior treatment lines to treat the tumor.

[0062] In certain embodiments, the prior treatment line included chemotherapy. In some embodiments, the chemotherapy included platinum-based therapy. In certain embodiments, the platinum-based therapy included a platinum-based antineoplastic agent selected from the group consisting of cisplatin, carboplatin, oxaliplatin, nedaplatin, triplatin tetranitrate, phenantriplatin, picoplatin, satraplatin, and any combination thereof. In one particular embodiment, the platinum-based therapy included cisplatin (e.g., cisplatin alone; in combination with methotrexate, vinblastine, and / or doxorubicin ("MVAC"); or in combination with paclitaxel and / or gemcitabine ("PGC")). In other embodiments, the chemotherapy included paclitaxel, docetaxel, gemcitabine, or any combination thereof. In some embodiments, the subject had received prior radiotherapy.

[0063] In certain embodiments, the subject has cancer cells expressing a variant of the EGFR or KRAS gene. In certain embodiments, the subject has PD-L1-positive cancer cells. In certain embodiments, the subject has PD-L1-negative cancer cells. In some embodiments, the subject never smoked. In certain embodiments, the subject smoked in the past. In one embodiment, the subject smokes now. In some embodiments, the subject never ingested or abused one or more analgesics. In certain embodiments, the subject abused analgesics in the past. In one embodiment, the subject abuses one or more analgesics. In some embodiments, the analgesic contains phenacetin. In other embodiments, the analgesic does not contain phenacetin. In certain embodiments, analgesic abuse is characterized by ingestion of more than 100 mg, more than 1.0 g, more than 10 g, more than 100 g, more than 1 kg, more than 2 kg, more than 3 kg, more than 4 kg, more than 5 kg, or more than 10 kg. In one embodiment, analgesic abuse is characterized by the intake of more than 5 kg over the course of a patient's life.

[0064] In certain embodiments, UC includes bladder cancer. In other embodiments, UC includes ureteral cancer. In yet another embodiment, UC includes renal pelvis carcinoma. In certain embodiments, UC includes any one or more carcinomas of the bladder, ureter, and renal pelvis.

[0065] In some embodiments, UC includes transitional cell carcinoma. Transitional cell carcinoma arises from urothelial cells lining the inside of the bladder, ureters, and renal pelvis.

[0066] In some embodiments, UC includes squamous cell carcinoma. For example, squamous cell carcinoma of the bladder arises from bladder urothelium having a pure squamous epithelial phenotype.

[0067] In some embodiments, UC includes adenocarcinoma. For example, adenocarcinoma of the bladder is defined as a tumor consisting entirely of malignant glandular epithelium.

[0068] In certain embodiments, UC or cancer derived therefrom includes bladder cancer, ureteral cancer, renal pelvis cancer, transitional cell carcinoma, squamous cell carcinoma, adenocarcinoma, or any combination thereof.

[0069] In some embodiments, the method comprises administering an effective amount of anti-PD-1 antibody, or administering an effective amount of anti-PD-L1 antibody and an effective amount of anti-CTLA-4 antibody. The effective amount of anti-PD-1 antibody and / or anti-CTLA-4 antibody may be a constant dose or a body weight-based dose.

[0070] In embodiments, the present invention includes a method for treating cancer or a subject affected by cancer, comprising treating cancer by administering an anti-PD-1 antagonist in combination with an anti-CTLA-4 antibody. “Anti-PD-1 antagonist” as used herein includes any molecule that inhibits the interaction between PD-1 (receptor) and PD-L1 (ligand) such that the PD-1 / PD-L1 signaling pathway is blocked. In other embodiments, the anti-PD-1 antagonist is a PD-1-Fc fusion protein. In specific embodiments, the anti-PD-1 antagonist includes an anti-PD-1 fusion protein, antisense molecule, small molecule, ribozyme, or nanobody that inhibits or prevents the interaction between PD-1 and PD-L1.

[0071] In certain embodiments, the therapies of the present invention (e.g., administration of an anti-PD-1 antibody, or administration of an anti-PD-1 antibody and an anti-CTLA-4 antibody) effectively increase the duration of survival in the subject. In some embodiments, the anti-PD-1 antibody of the present invention, or combination therapy of an anti-PD-1 antibody and an anti-CTLA-4 antibody, extends progression-free survival in the subject. In certain embodiments, the anti-PD-1 antibody of the present invention, or combination therapy of an anti-PD-1 antibody and an anti-CTLA-4 antibody, increases progression-free survival in the subject compared to standard care therapy. In some embodiments, combination therapy of an anti-PD-1 antibody and an anti-CTLA-4 antibody of the present invention extends progression-free survival in the subject compared to an anti-PD-1 antibody alone. In some embodiments, combination therapy of an anti-PD-1 antibody and an anti-CTLA-4 antibody of the present invention extends progression-free survival in the subject compared to other combinations of anti-PD-1 antibodies.

[0072] In some embodiments, after administration of an anti-PD-1 antibody, or after administration of an anti-PD-1 antibody and an anti-CTLA-4 antibody, subjects with UC or cancer derived therefrom may demonstrate overall survival of at least about 2 months, at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 7 months, at least about 8 months, at least about 9 months, at least about 10 months, at least about 11 months, at least about 12 months, at least about 13 months, at least about 14 months, at least about 15 months, at least about 16 months, at least about 17 months, at least about 18 months, at least about 19 months, at least about 20 months, at least about 21 months, at least about 22 months, at least about 23 months, at least about 2 years, at least about 3 years, at least about 4 years, or at least about 5 years.

[0073] In other embodiments, after administration of the therapies disclosed herein (e.g., anti-PD-1 antibody therapy, or anti-PD-1 antibody and anti-CTLA-4 antibody therapy), the survival or overall survival period of subjects is increased by at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 6 months, at least about 1 year, at least about 2 years, at least about 3 years, at least about 4 years, at least about 5 years, at least about 10 years, or at least about 15 years, compared to other subjects treated only with standard care therapy (e.g., platinum-based chemotherapy) or different dosing schedules of said therapy. For example, the survival or overall survival of subjects treated with the anti-PD-1 antibodies disclosed herein is increased by at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 75%, at least about 100%, at least about 200%, at least about 300%, or at least about 500% compared to other subjects treated with standard care (e.g., platinum-based chemotherapy) or different dosing schedules of anti-PD-1 antibody therapy alone.

[0074] In other embodiments, after administration of combination therapy comprising an anti-PD-1 antibody and an anti-CTLA-4 antibody, the survival or overall survival of subjects increases by at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 6 months, at least about 1 year, at least about 2 years, at least about 3 years, at least about 4 years, at least about 5 years, at least about 10 years, or at least about 15 years compared to other subjects treated with standard care (e.g., platinum-based chemotherapy), anti-PD-1 antibody alone, or only different dosing schedules of the combination therapy. For example, the survival or overall survival of subjects treated with the combination therapy of anti-PD-1 antibody and anti-CTLA-4 antibody disclosed herein is increased by at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 75%, at least about 100%, at least about 200%, at least about 300%, or at least about 500% compared to other subjects treated with standard care (e.g., platinum-based chemotherapy), anti-PD-1 antibody alone, or only different dosing schedules of the combination therapy.

[0075] In certain embodiments, the treatment of the present invention effectively increases the period of progression-free survival for subjects. In some embodiments, subjects exhibit progression-free survival of at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 7 months, at least about 8 months, at least about 9 months, at least about 10 months, at least about 11 months, at least about 1 year, at least about 18 months, at least about 2 years, at least about 3 years, at least about 4 years, at least about 5 years, at least about 10 years, and at least about 15 years.

[0076] In some embodiments, anti-PD-1 antibodies and anti-CTLA-4 antibodies are formulated for intravenous administration. In certain embodiments, anti-PD-1 antibodies and anti-CTLA-4 antibodies are administered sequentially. In embodiments, anti-PD-1 antibodies and anti-CTLA-4 antibodies are administered within 30 minutes of each other. In one embodiment, the anti-PD-1 antibody is administered before the anti-CTLA-4 antibody. In another embodiment, the anti-CTLA-4 antibody is administered before the anti-PD-1 antibody. In yet another embodiment, anti-PD-1 antibodies and anti-CTLA-4 antibodies are administered simultaneously in separate compositions. In further embodiments, anti-PD-1 antibodies and anti-CTLA-4 antibodies are mixed as a single composition for simultaneous administration.

[0077] In some embodiments, the anti-PD-1 antibody and the anti-CTLA-4 antibody are administered in fixed doses.

[0078] In some embodiments, the PD-L1 status of the tumor in the subject is measured before administering any composition or before utilizing any method disclosed herein. In one embodiment, the PD-L1 expression level of the tumor is at least about 1%, at least about 2%, at least about 3%, at least about 4%, at least about 5%, at least about 6%, at least about 7%, at least about 8%, at least about 9%, at least about 10%, at least about 11%, at least about 12%, at least about 13%, at least about 14%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, or at least about 95%. In another embodiment, the PD-L1 status of the tumor is at least about 1%. In other embodiments, the PD-L1 status of the subject is at least about 5%. In other embodiments, the PD-L1 status of the tumor is at least about 10%. In other embodiments, the PD-L1 status of the tumor is at least about 20%. In other embodiments, the PD-L1 status of the tumor is at least about 30%. In other embodiments, the PD-L1 status of the tumor is at least about 40%. In other embodiments, the PD-L1 status of the tumor is at least about 50%. In other embodiments, the PD-L1 status of the tumor is at least about 60%. In other embodiments, the PD-L1 status of the tumor is at least about 70%. In other embodiments, the PD-L1 status of the tumor is at least about 80%. In other embodiments, the PD-L1 status of the tumor is at least about 90%.

[0079] In some embodiments, the median progression-free survival for subjects with tumors expressing 1% or more PD-L1 is at least about 1 week, at least about 2 weeks, at least about 3 weeks, at least about 4 weeks, at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 7 months, at least about 8 months, at least about 9 months, at least about 10 months, at least about 11 months, or at least about 1 year longer than the median progression-free survival for subjects with tumors expressing less than 1% PD-L1. In some embodiments, the progression-free survival for subjects with tumors expressing 1% or more PD-L1 is at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 7 months, at least about 8 months, at least about 9 months, at least about 10 months, at least about 11 months, at least about 1 year, at least about 18 months, at least about 2 years, at least about 3 years, at least about 4 years, or at least about 5 years.

[0080] In one embodiment, to evaluate PD-L1 expression, test tissue samples can be obtained from patients in need of treatment. In another embodiment, evaluation of PD-L1 expression can be achieved without taking test tissue samples. In some embodiments, selecting an appropriate patient includes (i) optionally providing test tissue samples obtained from patients with tissue cancer, wherein the test tissue samples contain tumor cells and / or tumor-infiltrating inflammatory cells; and (ii) evaluating the percentage of cells in the test tissue samples that express PD-L1 on the cell surface, based on the evaluation that the percentage of cells in the test tissue samples that express PD-L1 on the cell surface is higher than a predetermined threshold level.

[0081] However, it should be understood that in any method involving the measurement of PD-L1 expression in a test tissue sample, the step of providing a test tissue sample obtained from a patient is an optional step. It should also be understood that in certain embodiments, the “measurement” or “evaluation” step for identifying or determining the number or percentage of cells in a test tissue sample that express PD-L1 on the cell surface may be performed by a transformation method that assays for PD-L1 expression, for example by performing a reverse transcriptase-polymerase chain reaction (RT-PCR) assay or an IHC assay. In other certain embodiments, the transformation step is not involved, and PD-L1 expression is evaluated, for example, by reviewing a report of test results from a laboratory. In certain embodiments, the steps of a method that evaluates PD-L1 expression and a method that includes evaluating PD-L1 expression provide intermediate results that may be provided to a physician or other healthcare provider for use in selecting a suitable candidate for an anti-PD-1 antibody or anti-PD-L1 antibody therapy. In certain embodiments, the step of providing intermediate results is performed by a physician or a person acting under the direction of a physician. In other embodiments, these steps are performed by an independent laboratory or by an independent person such as a laboratory technician.

[0082] In any particular embodiment of this method, the percentage of cells expressing PD-L1 is assessed by performing an assay to determine the presence of PD-L1 RNA. In further embodiments, the presence of PD-L1 RNA is determined by RT-PCR, in situ hybridization, or RNase protection. In other embodiments, the percentage of cells expressing PD-L1 is assessed by performing an assay to determine the presence of PD-L1 polypeptide. In further embodiments, the presence of PD-L1 polypeptide is determined by immunohistochemistry (IHC), enzyme-linked immunosorbent assay (ELISA), in vivo imaging, or flow cytometry. In some embodiments, PD-L1 expression is assayed by IHC. In all other embodiments of these methods, cell surface expression of PD-L1 is assayed using, for example, IHC or in vivo imaging.

[0083] Imaging technologies have provided crucial tools in cancer research and treatment. Recent developments in molecular imaging systems, including positron emission tomography (PET), single-photon emission computed tomography (SPECT), fluorescence reflection imaging (FRI), fluorescence-mediated tomography (FMT), bioluminescence imaging (BLI), laser scanning confocal microscopy (LSCM), and multiphoton microscopy (MPM), suggest further use of these technologies in cancer research. Some of these molecular imaging systems not only allow clinicians to know where tumors are located in the body, but also visualize the expression and activity of specific molecules, cells, and biological processes that influence tumor behavior and responsiveness to therapeutic agents (Condeelis and Weissleder, "In vivo imaging in cancer," Cold Spring Harb. Perspect. Biol. 2(12):a003848 (2010)). Combined with the sensitivity and resolution of PET, the specificity of antibodies makes immunoPET imaging particularly attractive for monitoring and assaying antigen expression in tissue samples (McCabe and Wu, "Positive progress in immunoPET—not just a coincidence," Cancer Biother. Radiopharm. 25(3):253-61 (2010); Olafsen et al., "ImmunoPET imaging of B-cell lymphoma using 124I-anti-CD20 scFv dimers (diabodies)," Protein Eng. Des. Sel. 23(4):243-9 (2010)). In any specific embodiment of this method, PD-L1 expression is assayed by immunoPET imaging. In any specific embodiment of this method, the percentage of cells in a test tissue sample expressing PD-L1 is assessed by performing an assay to determine the presence of PD-L1 polypeptides on the cell surface in the test tissue sample. In a specific embodiment, the test tissue sample is an FFPE tissue sample.In other embodiments, the presence of PD-L1 polypeptide is determined by an IHC assay. In further embodiments, the IHC assay is performed using an automated process. In some embodiments, the IHC assay is performed using an anti-PD-L1 monoclonal antibody to bind to the PD-L1 polypeptide.

[0084] In one embodiment of this method, an automated IHC method is used to assay the expression of PD-L1 on the cell surface in FFPE tissue specimens. This disclosure provides a method for detecting the presence of human PD-L1 antigen in a test tissue specimen or quantifying the level or percentage of human PD-L1 antigen in a specimen expressing the antigen, comprising contacting a test specimen and a negative control specimen with a monoclonal antibody that specifically binds to human PD-L1 under conditions that allow for the formation of a complex between the antibody or a portion thereof and human PD-L1. In a particular embodiment, the test and control tissue specimens are FFPE specimens. Complex formation is then detected, where the difference in complex formation between the test specimen and the negative control specimen indicates the presence of human PD-L1 antigen in the specimen. Various methods are used to quantify PD-L1 expression.

[0085] In certain embodiments, the automated IHC method includes (a) deparaffinizing and rehydrating the loaded tissue sections in an automated staining machine; (b) recovering the antigen using a DeLouk chamber and pH 6 buffer heated to 110°C for 10 minutes; (c) loading the reagents onto the automated staining machine; and (d) operating the automated staining machine to neutralize endogenous peroxidase in the tissue specimen; blocking nonspecific protein binding sites on the slide; incubating the slide with a primary antibody; incubating with a post-primary blocking agent; incubating with NovoLink Polymer; developing with a dye matrix; and counterstaining with hematoxylin.

[0086] To assess PD-L1 expression in tumor tissue samples, pathologists examine the number of membrane PD-L1+ tumor cells in each field of view under a microscope, estimate the percentage of positive cells mentally, and then average them to obtain the final percentage. Different staining intensities are defined as 0 / negative, 1+ / weak, 2+ / moderate, and 3+ / strong. Typically, percentage values ​​are assigned first to the 0 and 3+ buckets, then the intermediate 1+ and 2+ intensities are considered. In the case of highly heterogeneous tissue, the specimen is divided into multiple zones, each zone is scored separately, and then combined into a single set of percentage values. Percentages of negative and positive cells for different staining intensities are determined from each region, and the median is given to each zone. Final percentage values ​​of negative, 1+, 2+, and 3+ are given to the tissue for each staining intensity category. The sum of all staining intensities must be 100%. In one embodiment, the threshold number of cells that need to be PD-L1 positive is at least about 100, at least about 125, at least about 150, at least about 175, or at least about 200 cells. In a particular embodiment, the threshold number of cells that need to be PD-L1 positive is at least about 100 cells.

[0087] Staining is also evaluated in tumor-infiltrating inflammatory cells such as macrophages and lymphocytes. Since staining is observed in most macrophages, macrophages usually serve as internal positive controls. While staining with 3+ intensity is not required, any technical failure should be ruled out considering the absence of macrophage staining. Macrophages and lymphocytes are evaluated for plasma membrane staining and recorded for all samples as positive or negative for each cell category. Staining is also characterized according to the designation of extraneous / internal tumor immune cells. "Internal" means that immune cells are located within tumor tissue and / or on the boundary of the tumor region without being physically inserted into tumor cells. "External" means that immune cells are found in the periphery, associated with connective tissue or any related adjacent tissue, without physical association with the tumor.

[0088] In certain embodiments of these scoring methods, the sample is scored by two independently operating pathologists, and the scores are subsequently combined. In other specific embodiments, the identification of positive and negative cells is scored using appropriate software.

[0089] Histological scores are used as a more quantitative measure of IHC data. Histological scores are calculated as follows: Hist score = [(% tumor × 1 (low intensity)) + (% tumor × 2 (medium intensity)) + (% tumor × 3 (high intensity)]

[0090] Pathologists estimate the percentage of stained cells in each intensity category within a specimen to determine the histscore. Since the expression of most biomarkers is heterogeneous, the histscore more accurately represents overall expression. The final histscore ranges from 0 (no expression) to 300 (maximum expression).

[0091] An alternative method for quantifying PD-L1 expression in IHC test tissue samples is to determine the adjusted inflammation score (AIS) score, which is defined as the ratio of PD-L1 expression by tumor-infiltrating inflammatory cells to the inflammation density (Taube et al., "Colocalization of inflammatory response with B7-h1 expression in human melanocytic lesions supports an adaptive resistance mechanism of immune escape," Sci. Transl. Med. 4(127):127ra37 (2012)).

[0092] This method can treat UC or cancers derived therefrom at any stage. There are at least six stages used for UC (incorporated herein by reference as a whole, Edge et al. (eds), AJCC Cancer Staging Manual, 7th Edition, New York, Springer, See 2010): Stage 0a (Ta, N0, and M0), Stage 0is (carcinoma at original site; Tis, N0, and M0), Stage I (T1, N0, and M0), Stage II (T2a, N0, M0; or T2b, N0, and M0), Stage III (T3a, N0, and M0; T3b, N0, and M0; or T4a, N0, and M0), and Stage IV (T1-T3, N2, and M0; T3, N1, and M0; or T4, N0-N1, and M0), Stage IIIB (any T, N3, M0; or T4, N2, and M0), and Stage IV (T4b, N0, and M0; any T, N1-N3, and M0; or any T, any N, and M1).

[0093] In one embodiment, this method treats stage I ulcerative colitis (UC). In stage I, the tumor has infiltrated, for example, the subepithelial convective tissue (lamina propria).

[0094] In another embodiment, the method of the present invention treats stage II UC. Stage II UC can be characterized, for example, by either tumor invasion of the superficial muscularis propria (internal half) or invasion of the deep muscularis propria (lateral half).

[0095] In other embodiments, any method of the present invention treats stage III UC. Stage III can be characterized, for example, by (1) microscopic invasion of the periorbital tissue, (2) macroscopic invasion of the periorbital tissue, or (3) invasion of the prostatic matrix, seminal vesicles, uterus, vagina, pelvic wall, abdominal wall, or any combination thereof.

[0096] In some embodiments, the methods of the present invention treat stage IV UC. Stage IV UC can be characterized, for example, by (1) infiltration of the pelvic wall and / or abdominal wall; (2) local metastasis combined with metastasis to one or more lymph nodes; or (3) any local metastasis combined with or uncombined with metastasis to one or more lymph nodes and distant metastasis.

[0097] Anti-PD-1 antibody An anti-PD-1 antibody suitable for use in the disclosed methods is an antibody that binds to PD-1 with high specificity and affinity and inhibits PD-1 activity (e.g., by blocking the binding of PD-1 to PD-L1 and inhibiting the immunosuppressive effect of the PD-1 signaling pathway). In any of the therapeutic methods disclosed herein, the anti-PD-1 or anti-PD-L1 "antibody" includes an antigen-binding moiety that binds to the PD-1 or PD-L1 receptor, respectively, and exhibits functional properties similar to those of the whole antibody in inhibiting ligand binding and upregulating the immune system. In certain embodiments, the anti-PD-1 antibody cross-competes with nivolumab for binding to human PD-1. In other embodiments, the anti-PD-L1 antibody competes with BMS-936559, MPDL3280A, MEDI4736, or MSB0010718C for binding to human PD-L1.

[0098] In other embodiments, the anti-PD-1 antibody or anti-PD-L1 antibody is a chimeric, humanized, or human monoclonal antibody or a part thereof. In certain embodiments for treating human subjects, the antibody is a humanized antibody. In other embodiments for treating human subjects, the antibody is a human antibody. Antibodies of the IgG1, IgG2, IgG3, or IgG4 isotype can be used.

[0099] In certain embodiments, the anti-PD-1 antibody or anti-PD-L1 antibody contains a heavy chain constant region of a human IgG1 or IgG4 isotype. In other specific embodiments, the sequence of the IgG4 heavy chain constant region of the anti-PD-1 antibody or anti-PD-L1 antibody contains the S228P mutation, which replaces a serine residue in the hinge region with a proline residue typically found at the corresponding position in the IgG1 isotype antibody. The mutation present in nivolumab prevents Fab arm exchange with endogenous IgG4 antibody while maintaining a low affinity for activating the Fc receptor associated with wild-type IgG4 antibody (Wang et al. In vitro characterization of the anti-PD-1 antibody nivolumab, BMS-936558, and in vivo toxicology in non-human primates, Cancer Imm Res, 2(9):846-56 (2014)). In yet another embodiment, the antibody contains a light chain constant region which is a human kappa or lambda constant region. In other embodiments, the anti-PD-1 antibody, or the anti-PD-L1 antibody, or their antigen-binding portion, is a monoclonal antibody or their antigen-binding portion.

[0100] Any anti-PD-1 antibody known in the art can be used in the currently described method. In particular, various human monoclonal antibodies that bind specifically to PD-1 with high affinity are disclosed in US Patent No. 8,008,449. Other anti-PD-1 monoclonal antibodies are described, for example, in US Patent Nos. 6,808,710, 7,488,802, 8,168,757 and 8,354,509, and PCT Publication No. WO2012 / 145493. Each anti-PD-1 human monoclonal antibody disclosed in US Patent No. 8,008,449 has the following characteristics: (a) When determined by surface plasmon resonance using a Biacore biosensor system, it has an affinity of 1 × 10⁻¹⁶ -7It has been demonstrated that an antibody has one or more of the following characteristics: (b) binding to human PD-1 with a KD of M or less; (c) substantially not binding to human CD28, CTLA-4, or ICOS; (d) increasing T cell proliferation in a mixed lymphocyte reaction (MLR) assay; (e) increasing interferon-γ production in an MLR assay; (f) binding to human PD-1 and cynomolgus monkey PD-1; (g) inhibiting the binding of PD-L1 and / or PD-L2 to PD-1; (h) stimulating antigen-specific memory reactions; (i) stimulating antibody reactions; and (j) inhibiting tumor cell proliferation in vivo. Anti-PD-1 antibodies available in this invention include monoclonal antibodies that specifically bind to human PD-1 and exhibit at least one, and in some embodiments, at least five of the aforementioned characteristics. In some embodiments, the anti-PD-1 antibody is nivolumab. In one embodiment, the anti-PD-1 antibody is pembrolizumab.

[0101] Other anti-PD-1 monoclonal antibodies include, for example, US Patent Nos. 6,808,710, 7,488,802, 8,168,757 and 8,354,509, US Publication No. 2016 / 0272708, and PCT Publication. No.WO2012 / 145493, WO2008 / 156712, WO2015 / 112900, WO2012 / 145493, WO2015 / 112800, WO2014 / 206107, WO2 015 / 35606, WO2015 / 085847, WO2014 / 179664, WO2017 / 020291, WO2017 / 020858, WO2016 / 197367, WO2017 / 024 The following are described in 515, WO2017 / 025051, WO2017 / 123557, WO2016 / 106159, WO2014 / 194302, WO2017 / 040790, WO2017 / 133540, WO2017 / 132827, WO2017 / 024465, WO2017 / 025016, and WO2017 / 106061, each of which is incorporated herein by reference in whole.

[0102] In some embodiments, the anti-PD-1 antibody is nivolumab (also known as "OPDIVO®"; formerly also called 5C4, BMS-936558, MDX-1106, or ONO-4538), pembrolizumab (Merck, also known as "KEYTRUDA®", lambrolizumab, and MK-3475; see WO2008156712A1), PDR001 (Novartis; see WO2015 / 112900), MEDI-0680 (AstraZeneca; see AMP-514; see WO2012 / 145493), REGN-2810 (Regeneron; see WO2015 / 112800), JS001 (TAIZHOU JUNSHI PHARMA; Si-Yang Liu et al., J. Hematol. Oncol. 10:136 (2017)), BGB-A317 (Beigene; see WO2015 / 35606 and US2015 / 0079109), INCSHR1210 (SHR-1210; Jiangsu Hengrui Medicine; see WO2015 / 085847; Si-Yang Liu et al., J. Hematol. Oncol. 10:136 (2017)), TSR-042 (ANB011; Tesaro Biopharmaceutical; see WO2014 / 179664), GLS-010 (WBP3055; Wuxi / Harbin Gloria Pharmaceuticals; Si-Yang Liu et al., J. Hematol. Oncol. 10:136 The group is selected from (see 2017), AM-0001 (Armo), STI-1110 (Sorrento Therapeutics; see WO2014 / 194302), AGEN2034 (Agenus; see WO2017 / 040790), and MGD013 (Macrogenics).

[0103] In one embodiment, the anti-PD-1 antibody is nivolumab. Nivolumab is a fully human IgG4(S228P)PD-1 immune checkpoint inhibitor antibody that selectively prevents interaction with PD-1 ligands (PD-L1 and PD-L2), thereby blocking the downregulation of antitumor T cell function (USPatent No. 8,008,449; Wang et al., Cancer Imm Res, 2(9):846-56 (2014)).

[0104] In another embodiment, the anti-PD-1 antibody is pembrolizumab. Pembrolizumab is a humanized monoclonal IgG4 antibody against the human cell surface receptor PD-1 (programmed death-1). Pembrolizumab is described, for example, in US Patent Nos. 8,354,509 and 8,900,587; see also www.cancer.gov / drugdictionary?cdrid=695789 (last accessed December 14, 2014). Pembrolizumab is approved by the FDA for the treatment of relapsed or refractory melanoma.

[0105] The anti-PD-1 antibodies available in the disclosed methods also include isolated antibodies that specifically bind to human PD-1 and cross-compete with any anti-PD-1 antibodies disclosed herein for binding to human PD-1, such as nivolumab (see, e.g., US Patent No. 8,008,449 and 8,779,105; WO2013 / 173223). In some embodiments, the anti-PD-1 antibody binds to the same epitope as any of the anti-PD-1 antibodies described herein, such as nivolumab. The ability of antibodies to cross-compete for binding to the antigen indicates that these monoclonal antibodies bind to the same epitope region of the antigen and sterically prevent the binding of other cross-competitive antibodies to that particular epitope region. These cross-competitive antibodies are expected to have very similar functional properties to the reference antibody, such as nivolumab, for binding to the same epitope region of PD-1. Cross-competitive antibodies can be readily identified based on their ability to cross-compete with nivolumab in standard PD-1 binding assays such as Biacore analysis, ELISA assays, or flow cytometry (see, for example, WO2013 / 173223).

[0106] In certain embodiments, an antibody that cross-competes with nivolumab, a human PD-1 antibody, for binding to human PD-1, or that binds to the same epitope region of human PD-1, is a monoclonal antibody. For administration to human subjects, these cross-competitive antibodies are chimeric antibodies, genetically engineered antibodies, or humanized or human antibodies. Such chimeric, engineered, humanized, or human monoclonal antibodies can be prepared and isolated by methods well known in the art.

[0107] The anti-PD-1 antibody usable in the disclosed method of the invention also includes the antigen-binding portion of the antibody described above. It has been well demonstrated that the antigen-binding function of the antibody can be carried out by a fragment of the full-length antibody.

[0108] An anti-PD-1 antibody suitable for use in the disclosed methods or compositions is an antibody that binds to PD-1 with high specificity and affinity, blocks the binding of PD-L1 and / or PD-L2, and inhibits the immunosuppressive effect of the PD-1 signaling pathway. In any of the compositions or methods disclosed herein, the anti-PD-1 "antibody" comprises an antigen-binding moiety or fragment that binds to the PD-1 receptor and exhibits functional properties similar to those of the whole antibody in inhibiting ligand binding and upregulating the immune system. In certain embodiments, the anti-PD-1 antibody cross-competes with nivolumab for binding to human PD-1.

[0109] Anti-PD-L1 antibody Any anti-PD-L1 antibody can be used in the methods of this disclosure. Examples of anti-PD-L1 antibodies useful in the methods of this disclosure include the antibodies disclosed in US Patent No. 9,580,507. Each anti-PD-L1 human monoclonal antibody disclosed in US Patent No. 9,580,507 has the following characteristics: (a) When determined by surface plasmon resonance using a Biacore biosensor system, it has a frequency of 1 × 10⁻¹⁶ -7 Antibodies have been demonstrated to bind to human PD-L1 with a KD of M or less; (b) increase T cell proliferation in mixed lymphocyte reaction (MLR) assays; (c) increase interferon-γ production in MLR assays; (d) increase IL-2 secretion in MLR assays; (e) stimulate antibody responses; and (f) reverse the effect of T regulatory cells on T cell effector cells and / or dendritic cells. Anti-PD-L1 antibodies usable in the present invention include monoclonal antibodies that specifically bind to human PD-L1 and exhibit at least one, and in some embodiments, at least five of the aforementioned features.

[0110] In certain embodiments, the anti-PD-L1 antibody is selected from BMS-936559 (formerly 12A4 or MDX-1105; see, for example, U.S. Patent No. 7,943,743 and WO2013 / 173223), MPDL3280A (also known as RG7446, atezolizumab, and TECENTRIQ®; see also US8,217,149; Herbst et al. (2013) J Clin Oncol 31(suppl):3000), durvalumab (IMFINZI TM ; MEDI-4736; AstraZeneca; see WO2011 / 066389), avelumab (Pfizer; MSB-0010718C; BAVENCIO®; see WO2013 / 079174), STI-1014 (Sorrento; see WO2013 / 181634), CX-072 (Cytomx; see WO2016 / 149201), KN035 (3D Med / Alphamab; see Zhang et al., Cell Discov. 7:3 (March 2017)), LY3300054 (Eli Lilly Co.; see, for example, WO2017 / 034916), and CK-301 (Checkpoint Therapeutics; see Gorelik et al., AACR:Abstract 4606 (Apr 2016)).

[0111] In certain embodiments, the PD-L1 antibody is atezolizumab (TECENTRIQ®). Atezolizumab is a fully humanized IgG1 monoclonal anti-PD-L1 antibody.

[0112] In certain embodiments, the PD-L1 antibody is durvalumab (IMFINZI™). Durvalumab is a human IgG1 kappa monoclonal anti-PD-L1 antibody.

[0113] In certain embodiments, the PD-L1 antibody is avelumab (BAVENCIO®). Avelumab is a human IgG1 lambda monoclonal anti-PD-L1 antibody.

[0114] In other embodiments, the anti-PD-L1 monoclonal antibody is selected from the group consisting of 28-8, 28-1, 28-12, 29-8, 5H1, and any combination thereof.

[0115] The anti-PD-L1 antibodies available in the disclosed methods also include isolated antibodies that specifically bind to human PD-L1 and cross-compete with any anti-PD-L1 antibodies disclosed herein for binding to human PD-L1, such as atezolizumab and / or avelumab. In some embodiments, the anti-PD-L1 antibody binds to the same epitope as any of the anti-PD-L1 antibodies described herein, such as atezolizumab and / or avelumab. The ability of antibodies to cross-compete for binding to an antigen indicates that these antibodies bind to the same epitope region of the antigen and sterically prevent the binding of other cross-competitive antibodies to that particular epitope region. These cross-competitive antibodies are expected to have functional properties very similar to the reference antibody, such as atezolizumab and / or avelumab, by binding to the same epitope region of PD-L1. Cross-competitive antibodies can be readily identified based on their ability to cross-compete with atezolizumab and / or avelumab in standard PD-L1 binding assays such as Biacore analysis, ELISA assays, or flow cytometry (see, e.g., WO2013 / 173223).

[0116] In certain embodiments, antibodies that cross-compete with human PD-L1 antibodies, such as atezolizumab and / or avelumab, for binding to human PD-L1, or that bind to the same epitope region of human PD-L1, are monoclonal antibodies. For administration to human subjects, these cross-competing antibodies are chimeric antibodies, genetically engineered antibodies, or humanized or human antibodies. Such chimeric, engineered, humanized, or human monoclonal antibodies can be prepared and isolated by methods well known in the art.

[0117] The anti-PD-L1 antibody usable in the disclosed method of the invention also includes the antigen-binding portion of the antibody described above. It has been well demonstrated that the antigen-binding function of the antibody can be carried out by fragments of the full-length antibody.

[0118] An anti-PD-L1 antibody suitable for use in the disclosed methods or compositions is an antibody that binds to PD-L1 with high specificity and affinity, blocks PD-1 binding, and inhibits the immunosuppressive effect of the PD-1 signaling pathway. In any of the compositions or methods disclosed herein, the anti-PD-L1 "antibody" comprises an antigen-binding moiety or fragment that exhibits functional properties similar to those of the whole antibody in binding to PD-L1, inhibiting receptor binding, and upregulating the immune system. In certain embodiments, the anti-PD-L1 antibody cross-competes with atezolizumab and / or avelumab for binding to human PD-L1.

[0119] Combination therapy with anti-PD-1 or anti-PD-L1 antibodies In certain embodiments, an anti-PD-1 antibody or an anti-PD-L1 antibody is administered in combination with one or more other anticancer agents. In certain embodiments, one or more anticancer agents are administered to the target before administration of the anti-PD-1 or anti-PD-L1 antibody, or before combination with the anti-PD-1 or anti-PD-1 antibody. In certain embodiments, one or more anticancer agents were not effective in treating cancer. In some embodiments, the other anticancer agent is any anticancer agent described herein or known in the art. In one embodiment, the other anticancer agent is an anti-CTLA-4 antibody. In one embodiment, the other anticancer agent is chemotherapy or platinum-based doublet chemotherapy (PT-DC). In certain embodiments, the other anticancer agent is an EGFR-targeted tyrosine kinase inhibitor (TKI). In one embodiment, the other anticancer agent is an anti-VEGF antibody. In other embodiments, anticancer agents include platinum-based agents (e.g., cisplatin, carboplatin, oxaliplatin, and satraplatin), mitotic inhibitors (e.g., paclitaxel, albumin-bound paclitaxel, docetaxel, taxotere, docecad, vinblastine, doxorubicin, and eribulin), fluorinated Vinca alkaloids (e.g., vinflunin and jabrol), PI3K / AKT / mTOR inhibitors (e.g., sirolimus, temsirolimus, and everolimus), epidermal growth factor receptor (EGFR) inhibitors (e.g., gefitinib, cetuximab, erlotinib, and panitumumab), HER2 inhibitors (e.g., trastuzumab and These include lapatinib, fibroblast growth factor receptor (FGFR) inhibitors (e.g., dovitinib), vascular endothelial growth factor (VEGF) inhibitors (e.g., bevacizumab, aflibercept, ramucirumab, and sunitinib), MET / hepatocyte growth factor 1 (HGF1) inhibitors (e.g., cabozantinib), vinorelbine, vinblastine, etoposide, pemetrexed, gemcitabine, cabazitaxel, fluorouracil, topotecan, pazopanib, pyrazoloacridine, pralatrexate, pyretrexime, trimethrexate, ixabepyrone, irinotecan, ifosfamide, interleukin-2, irinotecan, arsenic trioxide, or any combination thereof.In one embodiment, the other anticancer agent is 5-fluorouracil (5-FU). In a particular embodiment, the other anticancer agent is any other anticancer agent known in the art. In some embodiments, two or more additional anticancer agents are administered in combination with an anti-PD-1 or anti-PD-L1 antibody. In some embodiments, the PD-1 or PD-L1 antibody is combined with surgical resection and / or radiotherapy.

[0120] Anti-CTLA-4 antibody Any anti-CTLA-4 antibody known in the art can be used in the methods of this disclosure. The anti-CTLA-4 antibody of the present invention binds to human CTLA-4 in such a way as to disrupt the interaction of CTLA-4 with the human B7 receptor. Since the interaction of CTLA-4 with B7 transmits a signal that leads to the inactivation of T cells having a CTLA-4 receptor, disruption of the interaction effectively induces, enhances or prolongs the activation of such T cells, thereby inducing, enhancing or prolonging an immune response.

[0121] Human monoclonal antibodies that specifically bind to CTLA-4 with high affinity are disclosed in US Patent Nos. 6,984,720 and 7,605,238. Other anti-CTLA-4 monoclonal antibodies are described, for example, in US Patent Nos. 5,977,318, 6,051,227, 6,682,736, and 7,034,121, and in International Publication Nos. WO2012 / 122444, WO2007 / 113648, WO2016 / 196237, and WO2000 / 037504, each of which is incorporated herein by reference in its entirety. The anti-CTLA-4 human monoclonal antibodies disclosed in US Patent Nos. 6,984,720 and 7,605,238 have the following characteristics: (a) When determined by Biacore analysis, they bind at least about 10 7 M -1 , or about 10 9 M -1 , or about 10 10 M -1 ~10 11 M-1 (b) Binds specifically to human CTLA-4 with a binding affinity reflected by an equilibrium association constant (Ka) greater than or equal to that; (b) at least about 10 3 , about 10 4 , or about 10 5 m -1 s -1 The dynamic association constant (ka); (c) at least about 10 3 , about 10 4 , or about 10 5 m -1 s -1 It has been demonstrated that the dynamic dissociation constant (kd) of (d) inhibits the binding of CTLA-4 to B7-1 (CD80) and B7-2 (CD86). Anti-CTLA-4 antibodies useful in the present invention include monoclonal antibodies that specifically bind to human CTLA-4 and exhibit at least one, at least two, or at least three of the aforementioned features.

[0122] In certain embodiments, the CTLA-4 antibody is selected from the group consisting of ipilimumab (YERVOY®; US Patent No. 6,984,720), MK-1308 (Merck), AGEN-1884 (Agenus Inc.; WO2016 / 196237), and tremelimumab (formerly tisilimmab, CP-675,206; AstraZeneca; see, e.g., WO2000 / 037504 and Ribas, Update Cancer Ther. 2(3): 133-39 (2007)). In certain embodiments, the anti-CTLA-4 antibody is ipilimumab.

[0123] In certain embodiments, the anti-CTLA-4 antibody is a human monoclonal antibody 10D1 (now known as ipilimumab and marketed as YERVOY®), as disclosed in US Patent No. 6,984,720. Ipilimumab is an anti-CTLA-4 antibody for use in the methods disclosed herein. Ipilimumab is a fully human IgG1 monoclonal antibody that blocks the binding of CTLA-4 to its B7 ligand, thereby promoting T cell activation and improving overall survival (OS) in patients with progressive melanoma.

[0124] In certain embodiments, the anti-CTLA-4 is tremelimumab (also known as CP-675,206). Tremelimumab is a human IgG2 monoclonal anti-CTLA-4 antibody. Tremelimumab is described in WO / 2012 / 122444, USPubl.No.2012 / 263677, or WO Publ.No.2007 / 113648A2.

[0125] In certain embodiments, the CTLA-4 antibody is MK-1308, which is an anti-CTLA-4 antibody under development by Merck.

[0126] In certain embodiments, the CTLA-4 antibody is AGEN-1884, a recombinant human monoclonal antibody against human CTLA-4 developed by Agenus Inc.

[0127] The anti-CTLA-4 antibodies available in the disclosed methods also include isolated antibodies that specifically bind to human CTLA-4 and cross-compete with any anti-CTLA-4 antibodies disclosed herein for binding to human CTLA-4, e.g., ipilimumab and / or tremelimumab. In some embodiments, the anti-CTLA-4 antibody binds to the same epitope as any of the anti-CTLA-4 antibodies described herein, e.g., ipilimumab and / or tremelimumab. The ability of antibodies to cross-compete for binding to the antigen indicates that these antibodies bind to the same epitope region of the antigen and sterically prevent the binding of other cross-competitive antibodies to that particular epitope region. These cross-competitive antibodies are expected to have functional properties very similar to the reference antibody, e.g., ipilimumab and / or tremelimumab, due to their binding to the same epitope region of CTLA-4. Cross-competitive antibodies can be readily identified based on their ability to cross-compete with ipilimumab and / or tremelimumab in standard CTLA-4 conjugation assays such as Biacore analysis, ELISA assay, or flow cytometry (see, e.g., WO2013 / 173223).

[0128] In certain embodiments, antibodies that cross-compete with ipilimumab and / or tremelimumab for binding to human CTLA-4, or that bind to the same epitope region of human CTLA-4, are monoclonal antibodies. For administration to human subjects, these cross-competing antibodies are chimeric antibodies, genetically modified antibodies, or humanized or human antibodies. Such chimeric, modified, humanized, or human monoclonal antibodies can be prepared and isolated by methods well known in the art.

[0129] The anti-CTLA-4 antibody usable in the method of the disclosed invention also includes the antigen-binding portion of the antibody described above. It has been well demonstrated that the antigen-binding function of the antibody can be carried out by fragments of the full-length antibody.

[0130] An anti-CTLA-4 antibody suitable for use in the disclosed methods or compositions is an antibody that binds to CTLA-4 with high specificity and affinity, blocks the activity of CTLA-4, and disrupts the interaction of CTLA-4 with the human B7 receptor. In any of the compositions or methods disclosed herein, the anti-CTLA-4 "antibody" comprises an antigen-binding moiety or fragment that binds to CTLA-4 and exhibits functional properties similar to those of the whole antibody in inhibiting the interaction of CTLA-4 with the human B7 receptor and upregulating the immune system. In certain embodiments, the anti-CTLA-4 antibody cross-competes with ipilimumab and / or tremelimumab for binding to human CTLA-4.

[0131] Combination of anti-PD-1 antibody with anti-CTLA-4 antibody to treat UC This disclosure provides a combination therapy method for treating ulcerative colitis (UC) or cancer derived therefrom, in which an anti-PD-1 antibody is combined with another anticancer agent, which is an antibody or its antigen-binding portion that specifically binds to CTLA-4 and inhibits CTLA-4 activity. The combination of nivolumab, an anti-PD-1 antibody, and ipilimumab, an anti-CTLA-4 antibody, has been demonstrated herein to produce early and sustained antitumor activity in UC patients, particularly with a specific dosing schedule (see Example 1). Therefore, in certain embodiments, the anti-CTLA-4 antibody used in combination with the anti-PD-1 antibody is ipilimumab. In embodiments, the anti-CTLA-4 antibody is tremelimumab. In other embodiments, the anti-CTLA-4 antibody is an antibody or a portion thereof that cross-competes with ipilimumab for binding to human CTLA-4. In yet another embodiment, the anti-CTLA-4 antibody is a chimeric, humanized, or human monoclonal antibody or a portion thereof. In yet another embodiment, the anti-CTLA-4 antibody comprises a heavy chain constant region of a human IgG1 or IgG4 isotype. In some embodiments, the anti-CTLA-4 antibody contains a heavy chain constant region of a human IgG1 isotype.

[0132] Due to the sustained clinical efficacy previously demonstrated with immunotherapy by inhibiting immune checkpoints (see, for example, WO2013 / 173223), in alternative embodiments, the combination therapy may include a finite number of doses, e.g., about 1 to 10 doses, or a single dose at long intervals, e.g., once every 3 to 6 months or once every 1 to 2 years or longer.

[0133] In certain embodiments of this method, the anti-PD-1 antibody is nivolumab. In other embodiments, it is pembrolizumab. In yet another embodiment, the anti-CTLA-4 antibody is ipilimumab. In yet another embodiment, the anti-CTLA-4 antibody is tremelimumab. Typically, the anti-PD-1 antibody and the anti-CTLA-4 antibody are formulated for intravenous administration. In certain embodiments, when the anti-PD-1 antibody and the anti-CTLA-4 antibody are administered together, they are administered within 30 minutes of each other. Either antibody can be administered first; that is, in some embodiments, the anti-PD-1 antibody is administered before the anti-CTLA-4 antibody, while in other embodiments, the anti-CTLA-4 antibody is administered before the anti-PD-1 antibody. Typically, each antibody is administered by intravenous infusion over 60 minutes. In certain embodiments, the anti-PD-1 antibody and the anti-CTLA-4 antibody are mixed and administered simultaneously as a single composition in a pharmaceutically acceptable formulation for co-administration, or simultaneously as separate compositions having each antibody in a pharmaceutically acceptable formulation.

[0134] In certain embodiments, the anti-PD-1 antibody is administered in a sub-therapeutic dose. In other embodiments, the anti-CTLA-4 antibody is administered in a sub-therapeutic dose. In further embodiments, both the anti-PD-1 antibody and the anti-CTLA-4 antibody are administered in sub-therapeutic doses, respectively.

[0135] Standard care for ulcerative colitis (UC) Standard care therapies for various types of cancer are well known to those skilled in the art. For example, the National Comprehensive Cancer Network (NCCN), an alliance of 21 major cancer centers in the United States, publishes the NCCN Oncology Guidelines (NCCN GUIDELINES®), which provide detailed and up-to-date information on standard care therapies for a wide variety of cancers (see NCCN GUIDELINES® (2014), available at http: / / www.nccn.org / professionals / physician_gls / f_guidelines.asp, last accessed June 2, 2016).

[0136] Chemotherapy, surgery, and radiotherapy (RT) are the three most commonly used treatments for ulcerative colitis (UC). The most commonly used initial chemotherapy regimens are methotrexate, vinblastine, doxorubicin, and cisplatin (MVAC) and gemcitabine and cisplatin (GC).

[0137] While many UC patients respond to initial treatment, including chemotherapy and / or surgery, patients with relapsed UC have increasingly unreliable treatment options, and many receive palliative care. Therefore, there is a particularly unmet need among patients with relapsed UC due to the lack of effective therapies after first-line therapy.

[0138] Pharmaceutical composition and dosage The therapeutic agent of the present invention may be composed in a pharmaceutical composition containing, for example, an antibody and a pharmaceutically acceptable carrier. As used herein, “pharmaceutically acceptable carrier” includes any physiologically compatible solvent, dispersion medium, coating, antimicrobial and antifungal agents, isotonic agents and absorption retarders, etc. In one embodiment, the carrier for an antibody-containing composition is suitable for intravenous, intramuscular, subcutaneous, parenteral, spinal, or epidermal administration (e.g., by injection or infusion). The pharmaceutical composition of the present invention may contain one or more pharmaceutically acceptable salts, antioxidants, aqueous and non-aqueous carriers, and / or adjuvants such as preservatives, wetting agents, emulsifiers and dispersants.

[0139] The dosage regimen is adjusted to provide the optimal desired response, e.g., maximum therapeutic response and / or minimum adverse effects. When an anti-PD-1 antibody is administered as monotherapy or in combination with other anticancer agents (e.g., in combination with an anti-CTLA-4 antibody), the dosage can range from approximately 0.01 to 20 mg / kg, 0.1 to 10 mg / kg, 0.1 to 5 mg / kg, 1 to 5 mg / kg, 2 to 5 mg / kg, 7.5 to 12.5 mg / kg, or 0.1 to 30 mg / kg of the subject's body weight. For example, the dosage could be approximately 0.1, 0.3, 1, 2, 3, 5, or 10 mg / kg of body weight, or approximately 0.3, 1, 2, 3, or 5 mg / kg of body weight. Typically, the dosing schedule is designed to achieve exposure that results in sustained receptor occupancy (RO) based on the typical pharmacokinetic properties of the antibody. Exemplary treatment plans include administrations approximately once a week, once every two weeks, once every three weeks, once every four weeks, once a month, and once every three to six months or more. In certain embodiments, an anti-PD-1 antibody, such as nivolumab, is administered to the subject approximately every two weeks. In other embodiments, the antibody is administered approximately every three weeks. Dosage and schedule may change during the course of treatment. For example, a dosing schedule for anti-PD-1 monotherapy may include (i) every two weeks in an approximately six-week cycle; (ii) every four weeks for approximately six doses, then every three months; (iii) every three weeks; and (iv) administration of the antibody at approximately 3 to 10 mg / kg once, followed by approximately 1 mg / kg every two to three weeks. Typically, considering that IgG4 antibodies have a half-life of 2–3 weeks, the administration regimen for the anti-PD-1 antibody of the present invention comprises intravenous administration of at least about 0.3–at least about 10 mg / kg body weight, at least about 1–at least about 5 mg / kg body weight, or at least about 1–at least about 3 mg / kg body weight, with the antibody administered every 14–21 days in cycles of up to about 6 weeks or about 12 weeks until a complete response or confirmed progressive disease. In certain embodiments, anti-PD-1 monotherapy is administered at 3 mg / kg once every two weeks until progressive disease or unacceptable toxicity occurs.In some embodiments, the antibody treatment disclosed herein, or any combination treatment, is continued for at least about 1 month, at least about 3 months, at least about 6 months, at least about 9 months, at least about 1 year, at least about 18 months, at least about 24 months, at least about 3 years, at least about 5 years, or at least about 10 years.

[0140] When used in combination with other anticancer agents (for example, in combination with an anti-CTLA-4 antibody), the dose of the anti-PD-1 antibody can be lower compared to the monotherapy dose. A dose of nivolumab that is lower than the typical 3 mg / kg but greater than or equal to 0.001 mg / kg is a sub-therapeutic dose. Sub-therapeutic doses of anti-PD-1 antibodies used in the methods described herein are higher than 0.001 mg / kg and lower than 3 mg / kg. In some embodiments, sub-therapeutic doses are about 0.001 mg / kg to about 1 mg / kg, about 0.01 mg / kg to about 1 mg / kg, about 0.1 mg / kg to about 1 mg / kg, or about 0.001 mg / kg to about 0.1 mg / kg body weight. In some embodiments, sub-therapeutic doses are at least about 0.001 mg / kg, at least about 0.005 mg / kg, at least about 0.01 mg / kg, at least about 0.05 mg / kg, at least about 0.1 mg / kg, at least about 0.5 mg / kg, or at least about 1.0 mg / kg body weight. Receptor occupancy data from 15 subjects administered nivolumab at doses of 0.3 mg / kg to 10 mg / kg indicate that PD-1 occupancy is dose-independent within this dose range. Across all doses, the mean occupancy was 85% (range, 70% to 97%) and the mean plateau occupancy was 72% (range, 59% to 81%). In some embodiments, a dose of 0.3 mg / kg may allow sufficient exposure to produce maximum biological activity. Receptor occupancy data from 15 subjects administered nivolumab at doses of 0.3 mg / kg to 10 mg / kg indicate that PD-1 occupancy is dose-independent within this dose range. Across all doses, the mean occupancy rate was 85% (range, 70% to 97%), and the mean plateau occupancy rate was 72% (range, 59% to 81%) (Brahmer et al. (2010) J Clin Oncol 28:3167-75). Therefore, a dose of 0.3 mg / kg can allow sufficient exposure to produce maximum biological activity.

[0141] Although higher nivolumab monotherapy doses up to approximately 10 mg / kg every two weeks were achieved without reaching the maximum tolerable dose (MTD), significant toxicity reported in other trials of checkpoint inhibitors + anti-angiogenic therapy (see, e.g., Johnson et al. (2013) Cancer Immunol Res 1:373-77; Rini et al. (2011) Cancer 117:758-67) supports the selection of nivolumab doses less than 10 mg / kg.

[0142] In certain embodiments, the dose of anti-PD-1 antibody (or anti-PD-L1 antibody) and / or anti-CTLA-4 antibody is a fixed dose in the pharmaceutical composition. In other embodiments, the method of the present invention can be used in a constant dose (a dose given to the patient regardless of the patient's weight). For example, a constant dose of nivolumab may be about 240 mg. For example, a constant dose of pembrolizumab may be about 200 mg. In embodiments, the anti-PD-1 antibody is administered in a dose of about 240 mg. In embodiments, the anti-PD-1 antibody is administered in a dose of about 360 mg. In embodiments, the anti-PD-1 antibody is administered in a dose of about 480 mg. In one embodiment, 360 mg of anti-PD-1 antibody or antigen-binding fragment is administered once every three weeks. In another embodiment, 480 mg of anti-PD-1 antibody or antigen-binding fragment is administered once every four weeks.

[0143] Ipilimumab (YERVOY®) is approved for the treatment of melanoma, administered intravenously at a dose of 3 mg / kg once every three weeks for four doses. Therefore, in some embodiments, approximately 3 mg / kg is the maximum dose of ipilimumab used in combination with an anti-PD-1 antibody, but in certain embodiments, an anti-CTLA-4 antibody such as ipilimumab, when combined with nivolumab, may be administered at a dose of approximately 0.3 to approximately 10 mg / kg, approximately 0.5 to approximately 10 mg / kg, approximately 0.5 to approximately 5 mg / kg, or approximately 1 to approximately 5 mg / kg body weight every two or three weeks. In other embodiments, ipilimumab is administered on a different dosing schedule than nivolumab. In some embodiments, ipilimumab is administered approximately every 1 week, every 2 weeks, every 3 weeks, every 4 weeks, every 5 weeks, every 6 weeks, every 7 weeks, every 8 weeks, every 9 weeks, every 10 weeks, every 11 weeks, every 12 weeks, or every 15 weeks. A dose of ipilimumab every 3 weeks that is lower than a typical 3 mg / kg but greater than or equal to 0.001 mg / kg is a sub-therapeutic dose. Sub-therapeutic doses of anti-CTLA-4 antibodies used in the methods herein are higher than 0.001 mg / kg and lower than 3 mg / kg. In some embodiments, sub-therapeutic doses are approximately 0.001 mg / kg to approximately 1 mg / kg, approximately 0.01 mg / kg to approximately 1 mg / kg, approximately 0.1 mg / kg to approximately 1 mg / kg, or approximately 0.001 mg / kg to approximately 0.1 mg / kg body weight. In some embodiments, sub-therapeutic doses are at least about 0.001 mg / kg, at least about 0.005 mg / kg, at least about 0.01 mg / kg, at least about 0.05 mg / kg, at least about 0.1 mg / kg, at least about 0.5 mg / kg, or at least about 1.0 mg / kg body weight. While the combination of 3 mg / kg nivolumab and 3 mg / kg ipilimumab exceeds the MTD in the melanoma population, the combination of 1 mg / kg nivolumab + 3 mg / kg ipilimumab or 3 mg / kg nivolumab + 1 mg / kg ipilimumab has been found to be tolerable in melanoma patients (Wolchok et al., N Engl J Med 369(2):122-33(2013)).Therefore, while nivolumab is tolerated up to a maximum of 10 mg / kg when administered intravenously every two weeks, in certain embodiments, the dose of the anti-PD-1 antibody does not exceed approximately 3 mg / kg when combined with ipilimumab. In certain embodiments, based on risk-benefit and PK-PD assessments, the doses used include combinations of approximately 1 mg / kg nivolumab + approximately 3 mg / kg ipilimumab, approximately 3 mg / kg nivolumab + approximately 1 mg / kg ipilimumab, or approximately 3 mg / kg nivolumab + approximately 3 mg / kg ipilimumab, each administered approximately once every two to four weeks, or in certain embodiments, once every two weeks or once every three weeks. In certain other embodiments, nivolumab is administered in combination with ipilimumab, which is administered at doses of approximately 0.1, approximately 0.3, approximately 1, approximately 2, approximately 3, or approximately 5 mg / kg,

[0144] In certain embodiments, the combination of anti-PD-1 antibody and anti-CTLA-4 antibody is administered intravenously to the subject during the induction phase in 1, 2, 3, or 4 doses, approximately every 2 or 3 weeks. In certain embodiments, the combination of nivolumab and ipilimumab is administered intravenously during the induction phase in approximately 4 doses, approximately every 2 or 3 weeks. The induction phase is followed by a maintenance phase, during which the anti-PD-1 antibody alone is administered to the subject at doses of approximately 0.1, 0.3, 1, 2, 3, 5, or 10 mg / kg approximately every 2 or 3 weeks, for as long as the treatment proves effective or until uncontrollable toxicity or disease progression occurs. In certain embodiments, nivolumab is administered during the maintenance phase at a dose of approximately 3 mg / kg body weight approximately every 2 weeks.

[0145] In certain embodiments, the anti-PD-1 antibody and the anti-CTLA-4 antibody are formulated as a single composition, where the doses of the anti-PD-1 antibody and the anti-CTLA-4 antibody are combined in ratios of 1:50, 1:40, 1:30, 1:20, 1:10, 1:5, 1:3, 1:1, 3:1, 5:1, 10:1, 20:1, 30:1, 40:1, or 50:1. In other embodiments, the dose of the anti-CTLA-4 antibody is a fixed dose. In certain embodiments, the dose of the anti-CTLA-4 antibody is a constant dose, which is given to the patient regardless of body weight. In certain embodiments, the constant dose of the anti-CTLA-4 antibody is approximately 80 mg.

[0146] For combination therapy with nivolumab and other anticancer agents, these agents are administered at their approved doses. Treatment continues as long as a clinical benefit is observed or until unacceptable toxicity or disease progression occurs. Nevertheless, in certain embodiments, the dose of these anticancer agents administered is significantly lower than the approved dose, i.e., a sub-therapeutic dose of the agent, administered in combination with an anti-PD-1 antibody. The anti-PD-1 antibody can be administered at the dose that has shown to produce the highest efficacy as monotherapy in clinical trials, e.g., approximately 3 mg / kg of nivolumab administered approximately every three weeks (Topalian et al., N Engl J Med 366:2443-54 (2012); Topalian et al., Curr Opin Immunol 24:207-12 (2012)), or at a significantly lower dose, i.e., a sub-therapeutic dose. In certain embodiments, the anti-PD-1 antibody is administered approximately every three weeks at approximately 3 mg / kg.

[0147] Dosage and frequency vary depending on the half-life of the antibody in the subject. Generally, human antibodies have the longest half-lives, followed by humanized antibodies, chimeric antibodies, and non-human antibodies. Dosage and frequency may also vary depending on whether the treatment is prophylactic or therapeutic. In prophylactic use, typically, relatively low doses are administered relatively infrequently over a long period. Some patients continue treatment for the rest of their lives. In therapeutic use, relatively high doses are sometimes required at relatively short intervals until the progression of the disease decreases or ends, or until the patient shows partial or complete improvement of the disease symptoms. After that, the patient may be given a prophylactic regimen.

[0148] The actual dose levels of the active ingredient or multiple components in the pharmaceutical composition of the present invention can be varied for a particular patient, composition, and mode of administration to obtain an amount of the active ingredient that is not excessively toxic to the patient and is effective in achieving the desired therapeutic response. The selected dose level depends on a variety of pharmacokinetic factors, including the activity of the particular composition of the present invention used, the route of administration, the time of administration, the excretion rate of the particular compound used, the duration of treatment, other drugs, compounds and / or substances used in combination with the particular composition used, the age, sex, weight, condition, general health status and prior medical history of the patient being treated, and similar factors well known in the medical field. The compositions of the present invention can be administered by one or more routes of administration using one or more different methods well known in the art. As will be understood by those skilled in the art, the route and / or mode of administration will vary depending on the desired outcome.

[0149] Since anti-PD-1 and anti-PD-L1 target the same signaling pathway and have been shown in clinical trials to exhibit similar levels of efficacy in a variety of cancers, including renal cell carcinoma (see Brahmer et al. (2012) N Engl J Med 366:2455-65; Topalian et al. (2012a) N Engl J Med 366:2443-54; WO2013 / 173223), an anti-PD-L1 antibody may be used instead of an anti-PD-1 antibody in any of the therapeutic methods disclosed herein. In certain embodiments, the anti-PD-L1 antibody is BMS-936559 (formerly 12A4 or MDX-1105) (see, for example, USPatent No. 7,943,743; WO2013 / 173223). In other embodiments, the anti-PD-L1 antibody is MPDL3280A (also known as RG7446) (see, e.g., Herbst et al. (2013) J Clin Oncol 31(suppl):3000. Abstract; USPatent No.8,217,149) or MEDI4736 (Khleif (2013) In: Proceedings from the European Cancer Congress 2013; September 27-October 1, 2013; Amsterdam, The Netherlands. Abstract 802). In certain embodiments, the antibody that cross-competes with the above PD-L1 antibody for binding to human PD-L1, or the antibody that binds to the same epitope region of the above PD-L1 antibody and human PD-L1, is a monoclonal antibody. For administration to human subjects, these cross-competing antibodies may be chimeric antibodies or humanized or human antibodies. Humanized or human monoclonal antibodies of such chimeras can be prepared and isolated by methods well known in the art.

[0150] kit Kits containing anti-PD-1 antibodies and other anticancer agents for therapeutic use are also within the scope of the present invention. Typically, a kit includes a label indicating the intended use and instructions for use of the kit's contents. The term "label" includes any documents or recorded materials on or supplied with the kit, or otherwise accompanying the kit. Accordingly, this disclosure provides a kit for treating subjects suffering from UC or cancer derived therefrom, which includes (a) an amount of anti-PD-1 antibody in the range of about 4 mg to about 500 mg; and (b) instructions for using the anti-PD-1 antibody in any manner disclosed herein. This disclosure further provides a kit for treating subjects suffering from UC or cancer derived therefrom, which includes (a) an amount of anti-PD-1 antibody in the range of about 4 mg to about 500 mg; (b) an amount of CTLA-4 antibody in the range of about 4 mg to about 500 mg; and (c) instructions for using the anti-PD-1 antibody and CTLA-4 antibody in any manner disclosed herein. In some embodiments, the kit contains an anti-PD-1 antibody and a CTLA-4 antibody as a separation composition. In some embodiments, the kit contains the anti-PD-1 antibody or its antigen-binding moiety and the CTLA-4 antibody as a single composition. In certain embodiments, the anti-PD-1 antibody and the anti-CTLA-4 antibody can be packaged together in a unit dosage form. In certain embodiments for treating human patients, the kit contains an anti-human PD-1 antibody disclosed herein, for example, nivolumab or pembrolizumab. In other embodiments, the kit contains an anti-human CTLA-4 antibody disclosed herein, for example, ipilimumab or tremelimumab.

[0151] The present invention will be further illustrated by the following embodiments, which should not be construed as further limitations. The contents of all documents cited throughout this application are expressly incorporated herein by reference.

[0152] Embodiment E1. A method for treating a subject suffering from urothelial carcinoma (UC) or a cancer derived therefrom, comprising administering to the subject an antibody ("anti-PD-1 antibody") or an antigen-binding moiety thereof that specifically binds to the programmed cell death-1 (PD-1) receptor and inhibits PD-1 activity.

[0153] E2. The method of E1, further comprising administering an antibody ("anti-CTLA-4 antibody") that specifically binds to cytotoxic T lymphocyte antigen-4 (CTLA-4) and inhibits CTLA-4 activity, or its antigen-binding portion.

[0154] E3.UC is a method of E1 or E2, including bladder cancer.

[0155] E4.UC is an E1 or E2 method, including ureteral cancer.

[0156] E5.UC includes renal pelvis carcinoma, E1 or E2 method.

[0157] E6.UC includes transitional cell carcinoma, and is defined by any of the E1-E5 methods.

[0158] E7.UC includes squamous cell carcinoma, and is defined by any of the methods from E1 to E5.

[0159] E8.UC includes adenocarcinoma, and is one of the methods E1-E3.

[0160] E9.UC is relapsed UC, and one of the methods from E1 to E8 is used.

[0161] E10.UC is locally progressive, and one of the methods E1-E9 is used.

[0162] E11.UC is metastatic, and one of the methods E1-E9 applies.

[0163] E12. The subject has received at least one, at least two, at least three, at least four, or at least five prior treatment lines to treat UC, by any of the methods in E1 to E11.

[0164] E13. Method E12, where the previous therapy included chemotherapy.

[0165] E14. Chemotherapy including platinum-based therapy, as in E13.

[0166] E15. The method of E14, wherein the platinum-based therapy comprises a platinum-based antineoplastic agent selected from the group consisting of cisplatin, carboplatin, oxaliplatin, nedaplatin, triplatin tetranitrate, phenantriplatin, picoplatin, satraplatin, and any combination thereof.

[0167] E16. Platinum-based therapy including cisplatin, methods E14 or E15.

[0168] E17. Platinum-based therapy including carboplatin, methods E14 or E15.

[0169] E18. The anti-PD-1 antibody cross-competes with nivolumab for binding to human PD-1, as described in one of the methods E1-E17.

[0170] E19. An anti-PD-1 antibody binds to the same epitope as nivolumab, using one of the methods E1-E18.

[0171] E20. The anti-PD-1 antibody is a chimeric, humanized, or human monoclonal antibody or a portion thereof, as described in any of the methods from E1 to E19.

[0172] E21. Any method from E1 to E20, wherein the anti-PD-1 antibody comprises a heavy chain constant region that is a human IgG1 or IgG4 isotype.

[0173] E22. The anti-PD-1 antibody is nivolumab, using one of the methods from E1 to E21.

[0174] E23. The anti-PD-1 antibody is pembrolizumab, using one of the methods from E1 to E21.

[0175] E24. The anti-CTLA-4 antibody is a chimeric, humanized, or human monoclonal antibody or a portion thereof, as described in any of the methods in E2-E23.

[0176] E25. An anti-CTLA-4 antibody containing the heavy chain constant region of a human IgG1 isotype, according to any of the methods E2-E24.

[0177] E26. The anti-CTLA-4 antibody is ipilimumab, using one of the methods from E2 to E25.

[0178] E27. The anti-CTLA-4 antibody is tremelimumab, using one of the methods from E2 to E25.

[0179] E28. The anti-CTLA-4 antibody cross-competes with ipilimumab for binding to human CTLA-4, as described in one of the methods E2-E27.

[0180] E29. One of the methods E1-E28, in which an anti-PD-1 antibody is administered approximately once every 1, 2, 3, or 4 weeks at a dose ranging from at least approximately 0.1 mg / kg to at least approximately 10.0 mg / kg body weight.

[0181] E30. One of the methods E1-E29, in which an anti-PD-1 antibody is administered at a dose of approximately 1 mg / kg body weight or approximately 3 mg / kg body weight.

[0182] E31. One of the methods E1 to E28, in which an anti-PD-1 antibody or its antigen-binding portion is administered in a fixed dose.

[0183] E32. Any of the methods E1-E28 and E31, wherein an anti-PD-1 antibody or its antigen-binding moiety is administered in a constant dose of at least about 200, at least about 220, at least about 240, at least about 260, at least about 280, at least about 300, at least about 320, at least about 340, at least about 360, at least about 380, at least about 400, at least about 420, at least about 440, at least about 460, at least about 480, at least about 500, or at least about 550 mg.

[0184] E33. An anti-PD-1 antibody or its antigen-binding portion is administered in a fixed dose approximately once every 1, 2, 3, or 4 weeks, using one of the methods E1-E28, E31, and E32.

[0185] E34. One of the methods E1-E33, in which an anti-PD-1 antibody is administered approximately once every two weeks.

[0186] E35. One of the methods E1-E33, in which an anti-PD-1 antibody is administered approximately once every three weeks.

[0187] E36. Administer anti-PD-1 antibodies as described in any of E1-E35, for as long as clinical benefit is observed or until uncontrollable toxicity or disease progression occurs.

[0188] E37. One of the methods E2-E36, in which an anti-CTLA-4 antibody is administered approximately once every 1, 2, 3, or 4 weeks at a dose ranging from at least approximately 0.1 mg / kg body weight to at least approximately 10.0 mg / kg body weight.

[0189] E38. Anti-CTLA-4 is administered at a dose of approximately 1 mg / kg body weight or approximately 3 mg / kg body weight, using one of the methods described in E2-E37.

[0190] E39. One of the methods E2 to E38, in which an anti-PD-1 antibody or its antigen-binding portion is administered in a fixed dose.

[0191] E40. One of the methods E2-E39, in which an anti-CTLA-4 antibody is administered approximately once every two weeks.

[0192] E41. One of the methods E2-E40, in which an anti-CTLA-4 antibody is administered approximately once every three weeks.

[0193] E42. One of the methods E2 to E41, in which an anti-PD-1 antibody is administered approximately once every three weeks at a dose of approximately 3 mg / kg body weight, and an anti-CTLA-4 antibody or its antigen-binding portion is administered approximately once every three weeks at a dose of approximately 1 mg / kg body weight.

[0194] E43. One of the methods E2 to E41, in which an anti-PD-1 antibody is administered approximately once every three weeks at a dose of approximately 1 mg / kg body weight, and an anti-CTLA-4 antibody or its antigen-binding portion is administered approximately once every three weeks at a dose of approximately 3 mg / kg body weight.

[0195] E44. Any of the methods E1 to E43, in which the subject demonstrates progression-free survival for at least approximately 1 month, at least approximately 2 months, at least approximately 3 months, at least approximately 4 months, at least approximately 5 months, at least approximately 6 months, at least approximately 7 months, at least approximately 8 months, at least approximately 9 months, at least approximately 10 months, at least approximately 11 months, at least approximately 1 year, at least approximately 18 months, at least approximately 2 years, at least approximately 3 years, at least approximately 4 years, or at least approximately 5 years after the initial dose.

[0196] E45. Any of the methods E1 to E44, where the subject has a tumor with 1% or more PD-L1 expression.

[0197] E46. Any of the methods E1 to E45, where the subject has a tumor with 5% or more PD-L1 expression.

[0198] E47. The combination is administered by any of the methods E2-E46, as long as a clinical benefit is observed or until disease progression or uncontrollable toxicity occurs.

[0199] E48. Anti-PD-1 antibody prescribed for intravenous administration using one of the methods E1-E47.

[0200] E49. Anti-CTLA-4 antibody is prescribed for intravenous administration using one of the methods E2-E48.

[0201] E50. One of the methods E2-E49, in which anti-PD-1 antibody and anti-CTLA-4 antibody are administered sequentially to the target.

[0202] E51. Any of the methods E2-E50, in which the anti-PD-1 antibody and the anti-CTLA-4 antibody are administered to each other within 30 minutes of each other.

[0203] E52. Any method from E2 to E51, wherein the anti-PD-1 antibody or its antigen-binding portion is administered before the anti-CTLA-4 antibody or its antigen-binding portion.

[0204] E53. Any method from E2 to E51, wherein the anti-CTLA-4 antibody or its antigen-binding portion is administered before the anti-PD-1 antibody or its antigen-binding portion.

[0205] E54. Any method from E2 to E49, wherein an anti-PD-1 antibody or its antigen-binding portion and an anti-CTLA-4 antibody or its antigen-binding portion are administered simultaneously in separate compositions.

[0206] E55. Any method from E2 to E49, wherein an anti-PD-1 antibody or its antigen-binding portion and an anti-CTLA-4 antibody or its antigen-binding portion are administered simultaneously as a single composition.

[0207] E56. Any of the methods E1 to E55, in which an anti-PD-1 antibody or its antigen-binding moiety is administered in a subtherapeutic dose.

[0208] E57. Any of the methods E2 to E56, in which an anti-CTLA-4 antibody or its antigen-binding moiety is administered in a subtherapeutic dose.

[0209] E58. Any of the methods E2 to E57, wherein an anti-PD-1 antibody or its antigen-binding portion and an anti-CTLA-4 antibody or its antigen-binding portion are each administered in subtherapeutic doses.

[0210] A kit for treating subjects suffering from E59.UC or cancer derived therefrom, (a) an anti-PD-1 antibody or its antigen-binding moiety in the range of approximately 4 mg to approximately 500 mg; and (b) Instructions for using PD-1 antibodies in any of the methods E1-58 A kit that includes this.

[0211] A kit for treating subjects suffering from E60.UC or cancer derived therefrom, (a) an anti-PD-1 antibody or its antigen-binding moiety in the range of approximately 4 mg to 500 mg; (b) CTLA-4 antibody or its antigen-binding moiety ranging from approximately 4 mg to approximately 500 mg; and (c) Instructions for using PD-1 antibody and CTLA-4 antibody in any of the methods E2-58. A kit that includes this.

[0212] Example 1 Example 1 The initial efficacy and safety results of combined nivolumab plus ipilimumab, given in two different dosing schedules in an open-label, multicenter Phase I / II trial of patients with locally advanced or metastatic UC that had progressed after prior platinum-based therapy, are reported herein.

[0213] material and method Patients with locally advanced or metastatic UC previously treated with platinum-based therapy were included in this study (Figure 1). Patients were treated with either (1) 3 mg / kg ipilimumab in combination with 1 mg / kg nivolumab ("N1I3") or 1 mg / kg ipilimumab in combination with 3 mg / kg nivolumab ("N3I1") administered every 3 weeks for 4 cycles, followed by 3 mg / kg nivolumab every 2 weeks, or (2) 3 mg / kg nivolumab monotherapy (N3) administered every 2 weeks. All patients were treated until disease progression or unacceptable toxicity. The primary endpoint was the investigator-assessed objective response rate (ORR) according to RECIST v1.1. Secondary endpoints included safety and duration of response (DoR).

[0214] Results and Conclusions The minimum follow-up period was 3.9 months for the N1I3 (n=26) group, 14.5 months for the N3I1 (n=104) group, and 13.8 months for the N3 (n=78) group. The objective response rates were 38.5%, 26.0%, and 25.6% for the N1I3, N3I1, and N3 groups, respectively (Table 1). At the time of data collection, the median duration of response (DoR) had not been reached in any treatment group. The frequency of drug-related grade 3-4 adverse events was 30.8% in N1I3, 31.7% in N3I1, and 23.1% in N3. Treatment-related adverse events led to discontinuation in 7.7% (N1I3), 13.5% (N3I1), and 3.8% (N3) of patients. One death was reported in group N3I1 (pneumonia), and two deaths were reported in group N3 (pneumonia and thrombocytopenia). [Table 1]

[0215] Second-line treatment with 3 mg / kg ipilimumab (N1I3) combined with 1 mg / kg nivolumab may offer the most favorable benefit-risk ratio among the regimens tested. This study is ongoing.

[0216] The above description of the specific embodiments sufficiently reveals the general nature of the invention, so others can readily modify and / or adapt such specific embodiments for various applications without departing from the general concept of the invention and without excessive experimentation, by applying the knowledge of those skilled in the art. Accordingly, such adaptations and modifications are intended to be within the meaning and scope of equivalents of the disclosed embodiments, based on the teachings and guidance presented herein. It should be understood that the words or technical terms used herein are intended to be descriptive, not restrictive, so that they may be interpreted by those skilled in the art in light of the teachings and guidance.

[0217] Other embodiments of the present invention will be apparent to those skilled in the art from the discussion herein and the practice of the present invention disclosed herein. The specification and examples are intended to be for illustrative purposes only, and the true scope and spirit of the invention are shown in the claims.

Claims

1. A method for treating a subject suffering from urothelial carcinoma (UC) or a cancer derived therefrom, comprising administering an antibody ("anti-PD-1 antibody") or its antigen-binding moiety specifically bound to the programmed cell death-1 (PD-1) receptor and inhibiting PD-1 activity.

2. The method according to claim 1, further comprising administering an antibody ("anti-CTLA-4 antibody") or its antigen-binding portion that specifically binds to cytotoxic T lymphocyte antigen-4 (CTLA-4) and inhibits CTLA-4 activity.

3. The method according to claim 1 or 2, wherein the UC includes bladder cancer, ureteral cancer, renal pelvis cancer, or any combination thereof.

4. The method according to any one of claims 1 to 3, wherein the UC includes transitional cell carcinoma, squamous cell carcinoma, adenocarcinoma, or any combination thereof.

5. The method according to any one of claims 1 to 4, wherein the ulcerative colitis (UC) is recurrent UC.

6. The method according to any one of claims 1 to 5, wherein the UC is locally progressive or metastatic.

7. The method according to any one of claims 1 to 6, wherein the subject has received at least one, at least two, at least three, at least four, or at least five prior treatment lines to treat UC.

8. The method according to any one of claims 1 to 7, wherein the anti-PD-1 antibody is nivolumab.

9. The method according to any one of claims 2 to 8, wherein the anti-CTLA-4 antibody is ipilimumab.

10. The method according to any one of claims 1 to 9, wherein an anti-PD-1 antibody is administered once every 1, 2, 3, or 4 weeks in a dose ranging from at least about 0.1 mg / kg body weight to at least about 10.0 mg / kg body weight.

11. The method according to any one of claims 2 to 10, wherein an anti-CTLA-4 antibody is administered once every 1, 2, 3, or 4 weeks in a dose ranging from at least about 0.1 mg / kg body weight to at least about 10.0 mg / kg body weight.

12. A method according to any one of claims 2 to 11, (a) The anti-PD-1 antibody is administered at a dose of approximately 3 mg / kg body weight once every three weeks, and the anti-CTLA-4 antibody or its antigen-binding moiety is administered at a dose of approximately 1 mg / kg body weight once every three weeks; or (b) The anti-PD-1 antibody is administered approximately once every three weeks at a dose of approximately 1 mg / kg body weight, and the anti-CTLA-4 antibody or its antigen-binding portion is administered approximately once every three weeks at a dose of approximately 3 mg / kg body weight. method.

13. The method according to any one of claims 1 to 12, wherein the subject has a tumor having 1% or more PD-L1 expression.

14. A kit for treating subjects suffering from UC or cancer derived therefrom, (a) an amount of anti-PD-1 antibody or its antigen-binding moiety ranging from approximately 4 mg to approximately 500 mg; and (b) Instructions for using the PD-1 antibody in the method according to any one of claims 1 to 58. A kit that includes this.

15. A kit for treating subjects suffering from UC or cancer derived therefrom, (a) an amount of anti-PD-1 antibody or its antigen-binding moiety ranging from approximately 4 mg to approximately 500 mg; (b) an amount ranging from approximately 4 mg to approximately 500 mg of CTLA-4 antibody or its antigen-binding moiety; and (c) Instructions for using the PD-1 antibody and the CTLA-4 antibody in the method according to any one of claims 2 to 58. A kit that includes this.

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