Use of Anti-PD-1 antibody in treatment of patient having colorectal cancer

Abstract

To provide methods for treating a tumor derived from a colorectal cancer exhibiting a high degree of microsatellite instability in a subject.SOLUTION: The methods comprise administering an anti-PD-1 antibody to a subject, and further comprise administering an anti-CTLA-4 antibody. The colorectal cancer is rectal cancer, colon cancer, or any combination thereof.SELECTED DRAWING: None

Description

[Technical Field]

[0001] FIELD OF THE INVENTION The present invention relates to a method for treating a tumor derived from a colorectal cancer exhibiting high microsatellite instability (MSI-H) in a subject in need thereof, comprising administering an anti-programmed death-1 (PD-1) antibody to the subject. In one embodiment, the anti-PD-1 antibody is administered in combination with an anti-cytotoxic T-lymphocyte antigen-4 (CTLA-4) antibody. In one embodiment, the colorectal cancer is rectal cancer, colon cancer, or any combination thereof. [Background technology]

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

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

[0004] Nivolumab (formerly named 5C4, BMS-936558, MDX-1106, or ONO-4538) is a fully human IgG4(S228P) PD-1 immune checkpoint inhibitor antibody that selectively inhibits interaction with PD-1 ligands (PD-L1 and PD-L2), thereby blocking downregulation of antitumor T cell function (U.S. Patent 8,008,449; Wang et al., (2014) Cancer Immunol Res 2:846-56). Nivolumab has shown activity in a variety of advanced solid tumors, including renal cell carcinoma (renal adenocarcinoma or adrenal neoplasm), melanoma, and non-small cell lung cancer (NSCLC) (Topalian et al., (2012) N Engl J Med 366:2443-54; Topalian et al., (2014) J Clin Oncol 32:1020-30; Drake et al., (2013) BJU Int 112:1-17; Ansell et al., (2015) Blood 126:583 [Abstract]; PCT Publication WO2013 / 173223).

[0005] Colorectal cancer is the third most common type of cancer in both men and women in the United States (see http: / / www.cancer.gov / types / colorectal; last accessed December 9, 2015). Most colorectal cancers are adenocarcinomas. Some colorectal cancers are associated with high levels of microsatellite instability (MSI-H), resulting from DNA mismatch repair defects. In particular, patients with DNA mismatch repair deficient / microsatellite instability-high (dMMR / MSI-H) metastatic colorectal cancer (mCRC) (approximately 5% of patients) are less likely to benefit from conventional chemotherapy than patients with pMMR mCRC. dMMR-MSI-H CRC, with its high mutational burden, is particularly sensitive to immune checkpoint inhibitor blockade. Summary of the Invention [Problem to be solved by the invention]

[0006] Targeting the multiple non-overlapping molecular pathways that control immune response can enhance anti-tumor immunotherapy. However, not all therapeutic agents are acceptable. Compared with monotherapy and other immunotherapy combinations, there remains a need for therapeutic agents that have acceptable safety profiles and high efficacy to enhance anti-tumor immune response. [Means for solving the problem]

[0007] Summary of the Invention The present invention relates to methods of treating a subject having a tumor derived from a colorectal cancer that exhibits high levels of microsatellite instability (MSI-H), comprising administering to the subject a therapeutically effective amount of an antibody or antigen-binding portion thereof that specifically binds to programmed death-1 receptor (PD-1) and inhibits PD-1 activity (an "anti-PD-1 antibody"), hi certain embodiments, the method further comprises administering a therapeutically effective amount of an antibody or antigen-binding portion thereof that specifically binds to cytotoxic T-lymphocyte antigen-4 (CTLA-4) and inhibits CTLA-4 activity (an "anti-CTLA-4 antibody").

[0008] In some embodiments, the colorectal cancer is rectal cancer, colon cancer, or any combination thereof. In some embodiments, the administration treats colorectal cancer.

[0009] 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 portion thereof. In some embodiments, the anti-PD-1 antibody comprises a heavy chain constant region of a human IgG1 or IgG4 isotype. In some embodiments, the anti-PD-1 antibody is nivolumab. In other embodiments, the anti-PD-1 antibody is pembrolizumab.

[0010] In certain embodiments, the anti-PD-1 antibody is administered at a dose ranging from at least about 0.1 mg / kg to at least about 10.0 mg / kg body weight approximately once every 1, 2, or 3 weeks. In certain embodiments, the anti-PD-1 antibody (e.g., nivolumab) is administered at a dose of at least about 1 mg / kg body weight approximately once every 3 weeks. In other embodiments, the anti-PD-1 antibody (e.g., nivolumab) is administered at a dose of at least about 3 mg / kg body weight approximately once every 3 weeks. In still other embodiments, the anti-PD-1 antibody (e.g., nivolumab) is administered at a dose of at least about 3 mg / kg body weight approximately once every 2 weeks. In other embodiments, the anti-PD-1 antibody (e.g., pembrolizumab) is administered at a dose of at least about 200 mg every 3 weeks or 2 mg / kg (maximum 200 mg) every 3 weeks. In certain embodiments, the anti-PD-1 antibody (e.g., avelumab) is administered at a dose of 10 mg / kg every 2 weeks. In some embodiments, the anti-PD-1 antibody is administered in a flat dose. In some embodiments, the anti-PD-1 antibody is administered in a flat 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, at least about 550 mg, at least about 600 mg, at least about 650 mg, at least 700 mg, at least 750 mg, or at least 800 mg. In some embodiments, the anti-PD-1 antibody is administered as long as clinical benefit is observed or until unmanageable toxicity or disease progression occurs.

[0011] In some embodiments, the anti-CTLA-4 antibody cross-competes with ipilimumab for binding to human CTLA-4. In some embodiments, the anti-CTLA-4 antibody binds to the same epitope as ipilimumab. In some embodiments, the anti-CTLA-4 antibody is a chimeric, humanized or human monoclonal antibody or a portion thereof. In some embodiments, the anti-CTLA-4 antibody comprises a heavy chain constant region of human IgG1 or IgG2 isotype. In some embodiments, the anti-CTLA-4 antibody is ipilimumab. In other embodiments, the anti-CTLA-4 antibody is tremelimumab.

[0012] In certain embodiments, the anti-CTLA-4 antibody is administered at a dose ranging from about 0.01 mg / kg to at least about 10 mg / kg body weight about once every 1, 2, or 3 weeks. In certain embodiments, the anti-CTLA-4 antibody is administered at a dose of at least about 1 mg / kg body weight about once every 3 weeks. In other embodiments, the anti-CTLA-4 antibody is administered at a dose of at least about 3 mg / kg body weight about once every 3 weeks. In certain embodiments, the anti-CTLA-4 antibody is administered as long as clinical benefit is observed or until unmanageable toxicity or disease progression occurs.

[0013] In some embodiments, the anti-PD-1 and anti-CTLA-4 antibodies are formulated for intravenous administration. In some embodiments, the anti-PD-1 and anti-CTLA-4 antibodies are administered sequentially. In some embodiments, the anti-PD-1 and anti-CTLA-4 antibodies are administered within 30 minutes of each other. 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. In some embodiments, the anti-PD-1 antibody and anti-CTLA-4 antibody are administered simultaneously in separate compositions. In other embodiments, the anti-PD-1 antibody and anti-CTLA-4 antibody are combined into a single composition for simultaneous administration.

[0014] In some embodiments, the anti-PD-1 antibody is administered at a subtherapeutic dose. In other embodiments, the anti-CTLA-4 antibody is administered at a subtherapeutic dose. In further embodiments, the anti-PD-1 antibody and the anti-CTLA-4 antibody are each administered at a subtherapeutic dose.

[0015] In certain embodiments, the subject has a tumor that expresses PD-L1, PD-L2, or both. In certain embodiments, the subject exhibits progression-free survival for 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 initiation of administration.

[0016] The present invention relates to kits for treating a subject with cancer, the kits including: (a) a single dose of an anti-PD-1 antibody in the range of about 0.1 mg / kg to about 10 mg / kg; (b) a single dose of an anti-CTLA-4 antibody in the range of about 0.1 mg / kg to about 10 mg / kg; and (c) instructions for using the anti-PD-1 antibody and the anti-CTLA-4 antibody in any of the methods disclosed herein.

[0017] In one embodiment, the subject has a microsatellite stable (MSS) tumor or a highly microsatellite instability (MSI-H) tumor. Some embodiments further comprise measuring the microsatellite status prior to administration of the anti-PD-1 antibody and / or anti-CTLA-4 antibody. In one embodiment, the tumor is an MSS tumor or an MSI-H tumor. In one embodiment, the subject has colorectal cancer. In another embodiment, the subject has colon cancer. In yet another embodiment, the subject has rectal cancer.

[0018] Implementation E1. A method of treating a subject having a tumor resulting from colorectal cancer, comprising administering to the subject a therapeutically effective amount of an antibody or antigen-binding portion thereof that specifically binds to the programmed death-1 receptor (PD-1) and inhibits PD-1 activity (an "anti-PD-1 antibody"), wherein the tumor exhibits high microsatellite instability ("MSI-H").

[0019] E2. The method of embodiment E1, wherein the tumor is colon cancer.

[0020] E3. The method of embodiment E1, wherein the tumor is a rectal cancer.

[0021] E4. The method of any of embodiments E1-E3, wherein the tumor exhibits one or more characteristics selected from the group consisting of: (a) the tumor contains germline alterations in at least two, at least three, at least four, or at least five DNA mismatch repair genes ("MMR genes"); (b) the tumor contains germline alterations in at least 30% of five or more MMR genes; (c) at least one protein encoded by a DNA MMR gene is not detectable in the tumor; and (d) any combination thereof.

[0022] E5. The method of embodiment E4, wherein the DNA MMR genes include MSH2, MLH1, MSH6, PMS2, PMS1, or any combination thereof.

[0023] E6. The method of embodiment E4, wherein the germline variation in (a) or (b) is determined by polymerase chain reaction.

[0024] E7. The method of embodiment E4, wherein at least one protein encoded by a DNA MMR gene is detected by immunohistochemistry.

[0025] E8. The method of any of embodiments E1-E7, wherein the subject 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, or at least about 12 months after administration of the treating agent.

[0026] E9. The method of any of embodiments E1-E8, wherein the subject exhibits an overall 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, or at least about 12 months after administration of the treating agent.

[0027] E10. The method of any of embodiments E1-E9, wherein the administering treats cancer.

[0028] E11. The method of any of embodiments E1-E10, wherein the anti-PD-1 antibody cross-competes with nivolumab for binding to human PD-1.

[0029] E12. The method of any of embodiments E1-E11, wherein the anti-PD-1 antibody binds to the same epitope as nivolumab.

[0030] E13. The method of any of embodiments E1-E12, wherein the anti-PD-1 antibody is a chimeric, humanized, or human monoclonal antibody or portion thereof.

[0031] E14. The method of any of embodiments E1-E13, wherein the anti-PD-1 antibody comprises a heavy chain constant region that is of the human IgG1 or IgG4 isotype.

[0032] E15. The method of any of embodiments E1-E14, wherein the anti-PD-1 antibody is nivolumab.

[0033] E16. The method of any of embodiments E1-E14, wherein the anti-PD-1 antibody is pembrolizumab.

[0034] E17. The method of any of embodiments E1-E16, wherein the anti-PD-1 antibody is administered at a dose ranging from at least about 0.1 mg / kg to at least about 10.0 mg / kg body weight approximately once every 1, 2, or 3 weeks.

[0035] E18. The method of embodiment E17, wherein the anti-PD-1 antibody is administered at a dose of about 3 mg / kg body weight about once every three weeks.

[0036] E19. The method of any of embodiments E1-E18, further comprising administering a therapeutically effective amount of an antibody or antigen-binding portion thereof that specifically binds to CTLA-4 and inhibits CTLA-4 activity (an "anti-CTLA-4 antibody").

[0037] E20. The method of embodiment E19, wherein the anti-CTLA-4 antibody cross-competes with ipilimumab for binding to human CTLA-4.

[0038] E21. The method of embodiment E19 or E20, wherein the anti-CTLA-4 antibody binds to the same epitope as ipilimumab.

[0039] E22. The method of any of embodiments E19-E21, wherein the anti-CTLA-4 antibody is a chimeric, humanized, or human monoclonal antibody or portion thereof.

[0040] E23. The method of any of embodiments E19-E22, wherein the anti-CTLA-4 antibody comprises a heavy chain constant region that is a human IgG1 or IgG2 isotype.

[0041] E24. The method of any of embodiments E19-E23, wherein the anti-CTLA-4 antibody is ipilimumab.

[0042] E25. The method of any of embodiments E19-E23, wherein the anti-CTLA-4 antibody is tremelimumab.

[0043] E26. The method of any of embodiments E19-E25, wherein the anti-CTLA-4 antibody is administered at a dose ranging from at least about 0.1 mg / kg to at least about 10.0 mg / kg body weight approximately once every 1, 2, or 3 weeks.

[0044] E27. The method of embodiment E26, wherein the anti-CTLA-4 antibody is administered at a dose of about 1 mg / kg body weight about once every three weeks.

[0045] E28. The method of any of embodiments E19-E25, wherein the anti-PD-1 antibody is administered at a dose of about 1 mg / kg body weight and the anti-CTLA-4 antibody is administered at a dose of about 1 mg / kg body weight.

[0046] E29. The method of any of embodiments E19-E25, wherein the anti-PD-1 antibody is administered at a dose of about 1 mg / kg body weight and the anti-CTLA-4 antibody is administered at a dose of about 3 mg / kg body weight.

[0047] E30. The method of any of embodiments E19-E25, wherein the anti-PD-1 antibody is administered at a dose of about 3 mg / kg body weight and the anti-CTLA-4 antibody is administered at a dose of about 1 mg / kg body weight.

[0048] E31. The method of any of embodiments E19-E25, wherein the anti-PD-1 antibody is administered at a dose of about 3 mg / kg body weight and the anti-CTLA-4 antibody is administered at a dose of about 3 mg / kg body weight.

[0049] E32. The method of any of embodiments E19-E31, wherein administration of the anti-PD-1 antibody and the anti-CTLA-4 antibody is followed by anti-PD-1 antibody monotherapy.

[0050] E33. The method of embodiment E32, wherein the anti-PD-1 antibody monotherapy comprises administering the anti-PD-1 antibody at a dose of about 1 mg / kg body weight.

[0051] E34. The method of embodiment E32, wherein the anti-PD-1 antibody monotherapy comprises administering the anti-PD-1 antibody at a dose of about 3 mg / kg body weight.

[0052] E35. The method of embodiment E32, wherein the anti-PD-1 antibody monotherapy comprises administering the anti-PD-1 antibody at a dose of about 4 mg / kg body weight.

[0053] E36. The method of any of embodiments E19-E35, wherein the anti-PD-1 antibody and anti-CTLA-4 antibody are administered about once every 1, 2, 3, or 4 weeks.

[0054] E37. The method of any of embodiments E32-E35, wherein the anti-PD-1 antibody monotherapy is administered about once every 1, 2, 3, or 4 weeks.

[0055] E38. The method of any of embodiments E1-E37, wherein the subject exhibits a complete or partial response to treatment at about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 9 weeks, about 10 weeks, about 11 weeks, about 12 weeks, about 18 weeks, about 24 weeks, about 30 weeks, or about 36 weeks after the first administration of the treating agent.

[0056] E39. The method of any of embodiments E19-E38, wherein the anti-PD-1 and anti-CTLA-4 antibodies are formulated for intravenous administration.

[0057] E40. The method of any of embodiments E19-E39, wherein the anti-PD-1 and anti-CTLA-4 antibodies are administered sequentially.

[0058] E41. The method of any of embodiments E19-E40, wherein the anti-PD-1 and anti-CTLA-4 antibodies are administered within 30 minutes of each other.

[0059] E42. The method of any of embodiments E19-E41, wherein the anti-PD-1 antibody is administered before the anti-CTLA-4 antibody.

[0060] E43. The method of any of embodiments E19-E41, wherein the anti-CTLA-4 antibody is administered before the anti-PD-1 antibody.

[0061] E44. The method of any of embodiments E1-E39, wherein the anti-PD-1 antibody and the anti-CTLA-4 antibody are administered simultaneously in separate compositions.

[0062] E45. The method of any of embodiments E1-E39, wherein the anti-PD-1 antibody and the anti-CTLA-4 antibody are combined in a single composition for simultaneous administration.

[0063] E46. A kit for treating a patient having a tumor resulting from colorectal cancer, wherein the tumor exhibits high microsatellite instability ("MSI-H"), the kit comprising: (a) a single dose of an anti-PD-1 antibody in the range of 0.1 mg / kg to 10 mg / kg of body weight; (b) a single dose of an anti-CTLA-4 antibody in the range of 0.1 mg / kg to 10 mg / kg of body weight; and (c) instructions for using the anti-PD-1 antibody and the anti-CTLA-4 antibody in the method of any of embodiments E1-E45. [Brief explanation of the drawings]

[0064] [Figure 1] Figure 1 shows a trial outline for patients with microsatellite stable (MSS) colorectal cancer receiving 1 mg / kg nivolumab in combination with 3 mg / kg ipilimumab or 3 mg / kg nivolumab in combination with 1 mg / kg ipilimumab.

[0065] [Figure 2] Figure 2 shows a trial outline for patients with microsatellite instability-high (MSI-H) colorectal cancer receiving 3 mg / kg nivolumab monotherapy or 3 mg / kg nivolumab in combination with 1 mg / kg ipilimumab.

[0066] [Figure 3] Figure 3 shows investigator-assessed objective response in patients with MSI-H colorectal cancer receiving 3 mg / kg nivolumab monotherapy.

[0067] [Figure 4] Figure 4 shows investigator-assessed objective response of patients with MSI-H colorectal cancer receiving 3 mg / kg nivolumab in combination with 1 mg / kg ipilimumab.

[0068] [Figure 5] Figures 5A-5B show the best reduction in target lesion size comparing MSI-H colorectal cancer patients receiving 3 mg / kg nivolumab monotherapy (Figure 5A) and MSI-H colorectal cancer patients receiving 3 mg / kg nivolumab in combination with 1 mg / kg ipilimumab (Figure 5B).

[0069] [Figure 6] Figure 6 shows investigator-assessed progression-free survival of patients with MSI-H colorectal cancer receiving 3 mg / kg nivolumab monotherapy or 3 mg / kg nivolumab in combination with 1 mg / kg ipilimumab.

[0070] [Figure 7] Figure 7 shows overall survival of patients with MSI-H colorectal cancer receiving 3 mg / kg nivolumab monotherapy or 3 mg / kg nivolumab in combination with 1 mg / kg ipilimumab.

[0071] [Figure 8] FIG. 8 shows a clinical trial outline for patients with DNA mismatch repair deficient / microsatellite instability-high (dMMR / MSI-H) metastatic colorectal cancer receiving 3 mg / kg nivolumab monotherapy or 3 mg / kg nivolumab + 1 mg / kg ipilimumab combination treatment.

[0072] [Figure 9] FIG. 9 shows the best reduction in target lesion size from baseline in patients with dMMR / MSI-H metastatic colorectal cancer receiving nivolumab monotherapy in an expanded clinical case study.

[0073] [Figure 10] FIG. 10 shows the best reduction in target lesion size from baseline in patients with dMMR / MSI-H metastatic colorectal cancer receiving nivolumab + ipilimumab combination treatment.

[0074] [Figure 11]FIG. 11 shows the time course of tumor burden in patients with dMMR / MSI-H metastatic colorectal cancer receiving nivolumab + ipilimumab combination treatment.

[0075] [Figure 12] Figures 12A-12B show investigator-assessed progression-free survival (PFS) (Figure 12A) and investigator-assessed overall survival (OS) (Figure 12B) for patients with dMMR / MSI-H metastatic colorectal cancer receiving nivolumab + ipilimumab combination treatment. DETAILED DESCRIPTION OF THE INVENTION

[0076] Detailed Description of the Invention The present invention relates to a method for treating colorectal cancer in a subject, comprising administering an anti-programmed death-1 (PD-1) antibody to the subject. In some embodiments, the colorectal cancer exhibits high microsatellite instability (MSI-H). In some embodiments, the method further comprises administering an anti-cytotoxic T-lymphocyte antigen-4 (CTLA-4) antibody. In some embodiments, the colorectal cancer is rectal cancer, colon cancer, or any combination thereof.

[0077] term In order that the present invention may be more readily understood, certain terms will first be defined. As used herein, unless expressly indicated otherwise herein, each of the following terms has the meaning set forth below. Additional definitions are set forth throughout the specification.

[0078] As used herein, the term "and / or" should be construed as specifically disclosing each of the two specified features or elements, with or without the other. Thus, the term "and / or" when used in a phrase such as "A and / or B" is intended to encompass "A and B," "A or B," "A" alone, and "B" alone. Similarly, the term "and / or" when used in a phrase such as "A, B, and / or C" is intended to encompass each of the following embodiments: A, B, and C; A, B, or C; A or C; A or B; B, or C; A and C; A and B; B and C; A alone; B alone; and C alone.

[0079] Where embodiments are described herein using the term "comprising," it is understood that otherwise similar embodiments described using the terms "consisting of" and / or "consisting essentially of" are also provided.

[0080] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this invention pertains. For example, Concise Dictionary of Biomedicine and Molecular Biology, Juo, Pei-Show, 2nd ed., 2002, CRC Press; The Dictionary of Cell and Molecular Biology, 3rd ed., 1999, Academic Press; and the Oxford Dictionary of Biochemistry and Molecular Biology, Revised, 2000, Oxford University Press provide those skilled in the art with a general dictionary of most of the terms used herein.

[0081] Units, prefixes, and symbols are shown in the form accepted by the International System of Units (SI). Numeric ranges are inclusive of the numbers defining the range. Headings provided herein do not limit aspects of the invention but should be read with reference to the specification as a whole. Accordingly, the definitions of the terms immediately below are more fully understood by reference to the specification in its entirety.

[0082] "Administration" refers to the physical introduction of a therapeutic agent into a subject using any of a variety of methods and delivery systems known to those skilled in the art. Examples of routes of administration for anti-PD-1 antibodies include intravenous, intramuscular, subcutaneous, intraperitoneal, spinal, or other parenteral routes, e.g., by injection or infusion. As used herein, the term "parenteral administration" refers to modes of administration other than enteral and topical administration, usually by injection, and includes, but is not limited to, intravenous, intramuscular, intraarterial, intrathecal, intralymphatic, intralesional, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal injection and infusion, and in vivo electroporation. Therapeutic agents may be administered by non-parenteral routes or orally. Other non-parenteral routes include topical, epithelial, or mucosal administration routes, e.g., intranasal, vaginal, rectal, sublingual, or topical. Administration can also be carried out, for example, once, multiple times and / or one or more times over an extended period of time.

[0083] As used herein, an "adverse event" (AE) is any untoward, generally unintended or undesired sign (including laboratory findings), symptom, or disease associated with the use of a medical treatment. A medical treatment may have one or more associated AEs, and each AE may have the same or different levels of severity. References to methods that can "alter an adverse event" refer to a treatment regimen that reduces the incidence and / or severity of one or more AEs associated with the use of a different treatment regimen.

[0084] An "antibody" (Ab) shall include, but is not limited to, a glycoprotein immunoglobulin or antigen-binding portion thereof that specifically binds to an antigen and comprises at least two heavy (H) chains and two light (L) chains interconnected by disulfide bonds. Each H chain comprises a heavy chain variable region (herein referred to as V H The heavy chain constant region comprises at least three constant domains: C H1 , C H2 and C H3 Each light chain comprises a light chain variable region (herein V L The light chain constant region contains one constant domain, C L Includes V H and V L The regions can be further subdivided into regions of hypervariability interspersed with more conserved regions called framework regions (FR). H and V L contains three CDRs and four FRs, arranged from the amino terminus to the carboxy terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. The variable regions of the heavy and light chains contain binding domains that interact with antigens. The constant regions of the antibody can mediate the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (C1q) of the classical complement pathway.

[0085] Immunoglobulins can be derived from any of the commonly known isotypes, including, but not limited to, IgA, secretory IgA, IgG, and IgM. IgG subclasses are also well known to those skilled in the art and include, but are not limited to, human IgG1, IgG2, IgG3, and IgG4. "Isotype" refers to the antibody class or subclass (e.g., IgM or IgG1) encoded by heavy chain constant region genes. The term "antibody" includes, by way of 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 may be humanized by recombinant methods to reduce immunogenicity in humans. Unless expressly stated and unless the context indicates otherwise, the term "antibody" also includes antigen-binding fragments or portions of any of the foregoing immunoglobulins, including monovalent and bivalent fragments or portions and single-chain antibodies.

[0086] An "isolated antibody" refers to an antibody that is substantially free of other antibodies having different antigenic specificities (e.g., an isolated antibody that specifically binds PD-1 is substantially free of antibodies that specifically bind to antigens other than PD-1). An isolated antibody that specifically binds PD-1 may, however, have cross-reactivity with other antigens, such as PD-1 molecules from different species. Moreover, an isolated antibody may be substantially free of other cellular material and / or chemicals.

[0087] The term "monoclonal antibody" ("mAb") refers to a preparation of non-naturally occurring antibody molecules of single molecular composition, i.e., antibody molecules which are essentially identical in primary sequence and which display a single binding specificity and affinity for a particular epitope. Monoclonal antibodies are examples of isolated antibodies. Monoclonal antibodies may be produced by hybridoma, recombinant, transgenic, or other techniques known to those skilled in the art.

[0088] A "human" antibody (HuMAb) refers to an antibody having variable regions in which both the framework and CDRs are derived from human germline immunoglobulin sequences. Furthermore, if the antibody contains a constant region, the constant region also is derived from human germline immunoglobulin sequences. The human antibodies of the invention may include amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutation in vivo). However, as used herein, the term "human antibody" is not intended to include antibodies in which CDR sequences from other mammalian species, such as mouse, have been grafted onto human framework sequences. The terms "human" antibody and "fully human" antibody are used interchangeably.

[0089] A "humanized antibody" refers to an antibody in which some, most, or all of the amino acids outside the CDRs of a non-human antibody have been replaced with corresponding amino acids from a human immunoglobulin. In some embodiments of humanized forms of antibodies, some, most, or all of the amino acids outside the CDRs have been replaced with those from a human immunoglobulin, while some, most, or all of the amino acids within one or more CDRs remain unchanged. Minor additions, deletions, insertions, substitutions, or modifications of amino acids are permissible as long as they do not abolish the antibody's ability to bind to a particular antigen. A "humanized" antibody maintains antigen specificity similar to that of the original antibody. In some embodiments, the CDRs of a humanized antibody comprise CDRs from a non-human mammalian antibody. In other embodiments, the CDRs of a humanized antibody comprise CDRs from an engineered synthetic antibody.

[0090] "Chimeric antibody" refers to an antibody in which the variable region is derived from one species and the constant region is derived from another species, such as when the variable region is derived from a mouse antibody and the constant region is derived from a human antibody.

[0091] 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.

[0092] An "antigen-binding portion" (also called an "antigen-binding fragment") of an antibody refers to one or more fragments of an antibody that retain the ability to specifically bind to the antigen bound by the whole antibody.

[0093] "Cancer" refers to a broad group of diverse diseases characterized by the uncontrolled growth of abnormal cells in the body. "Cancer" or "cancerous tissue" can include tumors. Uncontrolled cell division and growth leads to the formation of malignant tumors that can invade nearby tissues and even metastasize to distant sites via the lymphatic system or bloodstream. After metastasis, the distant tumor can be said to "originate" from the original pre-metastatic tumor. For example, a "tumor derived from" a colorectal cancer refers to a tumor that is the result of metastatic colorectal cancer. Because a distant tumor originates from a metastatic tumor, "originating from" a tumor can also include a pre-metastatic tumor; for example, a tumor derived from a colorectal cancer can include a colorectal cancer.

[0094] "Cytotoxic T-lymphocyte antigen-4" (CTLA-4) refers to an immunoinhibitory receptor belonging to the CD28 family. CTLA-4 is expressed exclusively on T cells in vivo and binds to two ligands, CD80 and CD86 (also known as B7-1 and B7-2, respectively). As used herein, the term "CTLA-4" includes human CTLA-4 (hCTLA-4), variants, isoforms, and species homologs of hCTLA-4, and analogs that share at least one epitope with hCTLA-4. The complete hCTLA-4 sequence can be found under GenBank Accession No. AAB59385.

[0095] The term "immunotherapy" refers to the treatment of a subject having, at risk of recurring, or having a disease by methods involving the induction, enhancement, suppression, or other modification of the immune response.

[0096] "Treatment" or "treatment" of a subject refers to any type of intervention or procedure or administration of an active agent to a subject with the intent of ameliorating, alleviating, ameliorating, arresting, delaying or preventing the onset, progression, occurrence, severity or recurrence of symptoms, complications, conditions or biochemical manifestations associated with a disease.

[0097] As used herein, "PD-L1 positive" or "PD-L2 positive" can be used interchangeably with "at least about 1% PD-L1 and / or PD-L2 expression." In certain embodiments, PD-L1 and / or PD-L2 expression can be measured by any method known in the art. In other embodiments, PD-L1 and / or PD-L2 expression is measured by automated in situ hybridization (IHC). A PD-L1- and / or PD-L2-positive tumor can have at least about 1%, at least about 2%, at least about 5%, at least about 10%, or at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100% of tumor cells expressing PD-L1 and / or PD-L2, i.e., as measured by automated IHC. In one embodiment, "PD-L1 positive" means that there are at least 100 cells that express PD-L1 on their surface. In another embodiment, "PD-L2 positive" means that there are at least 100 cells that express PD-L2 on their surface.

[0098] "Programmed death-1 (PD-1)" refers to an immunoinhibitory receptor belonging to the CD28 family. PD-1 is found primarily on 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, and analogs that share at least one epitope with hPD-1. The complete hPD-1 sequence can be found in GenBank Accession No. U64863.

[0099] "Programmed death-ligand-1" (PD-L1) is one of two cell surface glycoprotein ligands for PD-1 (the other being PD-L2) that downregulates T cell activation and cytokine secretion upon binding to PD-1. As used herein, the term "PD-L1" includes human PD-L1 (hPD-L1), variants, isoforms, and species homologs of hPD-L1, and analogs that share at least one epitope with hPD-L1. The complete hPD-L1 sequence can be found under GenBank Accession No. Q9NZQ7.

[0100] A "subject" includes any human or non-human animal. The term "non-human animal" includes, but is not limited to, vertebrates such as non-human primates, sheep, dogs, and rodents such as mice, rats, and guinea pigs. In some embodiments, the subject is a human. The terms "subject" and "patient" are used interchangeably herein.

[0101] A "therapeutically effective amount" or "therapeutically effective dose" of a drug or therapeutic agent is any amount of drug that, when used alone or in combination with other therapeutic agents, protects a subject against disease onset or promotes disease regression, as evidenced by a reduction in the severity of disease symptoms, an increase in the frequency and duration of disease-free periods, or prevention of functional impairment or disability due to disease morbidity. The ability of a therapeutic agent to promote disease regression can be assessed using a variety of methods known to those of skill in the art, such as by assaying the activity of the agent in human subjects during clinical trials, in animal model systems predictive of efficacy in humans, or in in vitro assays.

[0102] As used herein, a "sub-therapeutic dose" refers to a dose of a therapeutic compound (e.g., an antibody) that is lower than the usual or typical dose of the therapeutic compound when used alone to treat a hyperproliferative disease (e.g., cancer).

[0103] By way of example, an "anti-cancer agent" promotes cancer regression in a subject. In certain embodiments, a therapeutically effective amount of the drug promotes cancer regression to the point of eliminating the cancer. "Promoting cancer regression" means that administration of an effective amount of the drug, alone or in combination with an anti-cancer agent, results in a decrease 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 disease-free symptom periods, or prevention of functional impairment or disability due to disease morbidity. Furthermore, the terms "effective" and "efficacy" in relation to treatment include both pharmacological effectiveness and physiological safety. Pharmacological effectiveness refers to the ability of a drug to promote cancer regression in a patient. Physiological safety refers to the level of toxicity or other adverse physiological effects (adverse effects) at the cellular, organ, and / or organismal level resulting from drug administration.

[0104] As an example of tumor treatment, a therapeutically effective amount of an anti-cancer agent inhibits cell proliferation or tumor growth by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90% or about 100% compared to an untreated subject.

[0105] In other embodiments of the invention, tumor regression may be observed and sustained for a period of at least about 20 days, at least about 30 days, at least about 40 days, at least about 50 days, or at least about 60 days. Even with these ultimate assessments of therapeutic efficacy, evaluation of immunotherapeutic agents must also take into account "immune-related" response patterns.

[0106] The term "immune-related" response pattern refers to a clinical response pattern frequently observed in cancer patients treated with immunotherapeutic agents that produce antitumor effects by eliciting cancer-specific immune responses or by modulating innate immune processes. This response pattern, in the evaluation of conventional chemotherapeutic agents, is classified as disease progression, synonymous with drug failure, and characterized by an initial increase in tumor burden or the appearance of new lesions followed by a beneficial therapeutic effect. Therefore, proper evaluation of immunotherapeutic agents may require long-term monitoring of the effects of these agents on the target disease.

[0107] A therapeutically effective amount of a drug includes any amount of drug that, when administered alone or in combination with an anti-cancer drug to a subject at risk of developing cancer (e.g., a subject with a pre-malignant condition) or having a recurrence of cancer, prevents the onset or recurrence of cancer, i.e., a "prophylactically effective amount." In some embodiments, a prophylactically effective amount completely prevents the onset or recurrence of cancer. "Preventing" the onset or recurrence of cancer means reducing the likelihood of cancer onset or recurrence or completely preventing the onset or recurrence of cancer.

[0108] As used herein, the term "weight-based dose" means that the dose administered to a patient is calculated based on the patient's weight. For example, if a patient weighing 60 kg requires 3 mg / kg of anti-PD-1 antibody, the appropriate amount of anti-PD-1 antibody for administration (i.e., 180 mg) can be calculated and used.

[0109] The use of the term "fixed dose" in reference to the methods of the invention means that two or more different antibodies (e.g., an anti-PD-1 antibody and an anti-CTLA-4 antibody) in a single composition are present in the composition in a specific (fixed) ratio to each other. In certain embodiments, the fixed dose is based on the weight (e.g., mg) of the antibody. In certain embodiments, the fixed dose is based on the concentration (e.g., mg / ml) of the antibody. In certain embodiments, the ratio is at least about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, about 1:30, about 1:40, about 1:50, about 1:60, about 1:70, about 1:80, about 1:90, about 1:100, about 1:120, about 1:140, about 1:160, about 1:180, about 1:200, about 1:200, about 1:300, about 1:300, about 1:400, about 1:500, about 1:600, about 1:700, about 1:800, about 1:900, about 1:1000, about 1:1200, about 1:1400, about 1:1600, about 1:1800, about 1:1800, about 1:20 ... 1, about 160:1, about 140:1, about 120:1, about 100:1, about 90:1, about 80:1, about 70:1, about 60:1, about 50:1, about 40:1, about 30:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, or about 2:1 mg of first antibody (e.g., anti-PD-1 antibody) to mg of second antibody (e.g., anti-CTLA-4 antibody). For example, a 3:1 ratio of anti-PD-1 antibody to anti-CTLA-4 antibody can mean that a vial can contain about 240 mg of anti-PD-1 antibody and 80 mg of anti-CTLA-4 antibody, or about 3 mg / ml of anti-PD-1 antibody and 1 mg / ml of anti-CTLA-4 antibody.

[0110] The use of the term "flat dose" in reference to the methods and dosages of the present invention refers to a dose administered to a patient regardless of the patient's weight or body surface area (BSA). Thus, a flat dose is provided as an absolute amount of agent (e.g., an anti-CTLA-4 antibody and / or an anti-PD-1 antibody) rather than as a mg / kg dose. For example, a 60 kg human and a 100 kg human would receive the same dose of antibody (e.g., 240 mg of an anti-PD-1 antibody).

[0111] The use of the alternative (e.g., "or") should be construed to mean either one, both, or any combination thereof of the alternatives. As used herein, the terms "a," "an," and "the" should be construed to refer to "one or more" of any referenced or listed components.

[0112] The terms "about" or "essentially comprising" refer to a value or composition that is within an acceptable error range for a particular value or composition, as determined by one of ordinary skill in the art, which depends in part on how the value or composition is measured or determined, i.e., the limitations of the measurement system. For example, "about" or "essentially comprising" can mean within one or more standard deviations according to the practice in the art. Alternatively, "about" or "essentially comprising" can mean a range of up to 20%. Moreover, particularly with respect to biological systems or processes, the term can mean up to an order of magnitude or up to five times the value. When a particular value or composition is provided in the specification and claims, unless otherwise indicated, the meaning of "about" or "essentially comprising" should be assumed to be within an acceptable error range for that particular value or composition.

[0113] As used herein, the terms "about once a week," "about once every two weeks," or any other similar administration interval refer to approximate numbers. "About once a week" can include every 7 days ± 1 day, i.e., every 6 to 8 days. "About once every two weeks" can include every 14 days ± 3 days, i.e., every 11 to 17 days. Similar approximations apply, for example, to about once every 3 weeks, about once every 4 weeks, about once every 5 weeks, about once every 6 weeks, and about once every 12 weeks. In certain embodiments, an administration interval of about once every 6 weeks or about once every 12 weeks means that the first administration can be administered on any day of the first week, followed by the next administration on any day of the sixth or twelfth week, respectively. In other embodiments, a dosing interval of about once every 6 weeks or about once every 12 weeks means that the first dose is administered on a particular day of the week (e.g., Monday) in the first week, and then the next dose is administered on a particular day of the week (i.e., Monday) in the sixth or twelfth week, respectively.

[0114] Unless otherwise specified, any concentration range, percentage range, ratio range, or integer range described herein should be interpreted to include every integer value within the described range and fractions thereof, where appropriate (e.g., 1 / 10 and 1 / 100 of an integer).

[0115] Various aspects of the invention are described in further detail in the following subsections. [Table 1]

[0116] Methods of the Invention The present invention relates to methods for treating a subject having a tumor resulting from colorectal cancer, comprising administering to the subject a therapeutically effective amount of an antibody or antigen-binding portion thereof that specifically binds to the programmed death-1 receptor (PD-1) and inhibits PD-1 activity (an "anti-PD-1 antibody"), or an antibody or antigen-binding portion thereof that specifically binds to the programmed death-ligand 1 (PD-L1) and inhibits PD-L1 activity (an "anti-PD-L1 antibody").

[0117] In some embodiments, the treatment treats tumors derived from cancer that is colorectal cancer. In some embodiments, the colorectal cancer is colon cancer. In other embodiments, the colorectal cancer is rectal cancer. In some embodiments, the colorectal cancer has microsatellite instability (MSI) (see Pawlik et al., Dis. Markers 20(4-5): 199-206 (2004)). In other embodiments, the colorectal cancer has high microsatellite instability (MSI-H).

[0118] Colon cancer is classified into five stages: stage 0 (carcinoma in situ), stage I, stage II, stage III, and stage IV. Six types of standard treatments are used for colon cancer: 1) surgery, including local resection, colon resection and anastomosis, or colon resection and colostomy; 2) radiofrequency ablation; 3) cryosurgery; 4) chemotherapy; 5) radiation therapy; and 6) targeted therapy, including monoclonal antibodies and angiogenesis inhibitors. In some embodiments, the combination therapy of the present invention treats colon cancer in conjunction with standard of care therapeutic agents.

[0119] Rectal cancer is classified into five stages: stage 0 (intraepithelial carcinoma), stage I, stage II, stage III, and stage IV. Six types of standard treatments are used for rectal cancer: 1) surgery, including polypectomy, local excision, resection, radiofrequency ablation, cryosurgery, and pelvic exenteration; 2) radiation therapy; 3) chemotherapy; and 4) targeted therapy, including monoclonal antibody therapy. In some embodiments, the methods of the present invention treat rectal cancer in conjunction with standard treatment therapeutic agents.

[0120] In colorectal cancer, MSI-H is associated with increased immune infiltration and expression of immune checkpoint regulators. Therefore, methods of the invention include treating colorectal cancer that exhibits high levels of microsatellite instability (MSI-H). In one embodiment, an anti-PD-1 antibody is administered in combination with a therapeutically effective amount of an antibody or antigen-binding portion thereof that specifically binds to CTLA-4 (an "anti-CTLA-4 antibody").

[0121] Microsatellite instability is a state of genetic hypermutability resulting from DNA mismatch repair (MMR) defects. The presence of MSI represents phenotypic evidence that MMR does not function normally. In most cases, the genetic basis of instability in MSI tumors is inherited germline mutations in any one of five human MMR genes: MSH2, MLH1, MSH6, PMS2, and PMS1. In one embodiment, the subject undergoing tumor treatment has high microsatellite instability (MSI-H), with at least one mutation in the gene MSH2, MLH1, MSH6, PMS2, or PMS1. In another embodiment, the subject undergoing tumor treatment in the control group does not have microsatellite instability (MSS or MSI stable), and does not have a mutation in the gene MSH2, MLH1, MSH6, PMS2, or PMS1.

[0122] The present invention also relates to a method for treating a tumor, e.g., a colon tumor, comprising identifying a subject who responds to an anti-PD-1 antibody administered alone or in combination with an anti-CTLA-4 antibody, wherein the subject has an MSI-H tumor. In one embodiment, the present invention relates to a method for treating a tumor, e.g., a colon tumor, comprising (i) identifying a subject whose tumor is an MSI-H tumor, and (ii) administering to the subject an effective amount of a PD-1 antibody, alone or in combination with an effective amount of an anti-CTLA-4 antibody. As used herein, an MSI-H tumor refers to a tumor with at least about 30% or more unstable MSI biomarkers. In one embodiment, a colorectal cancer is MSI-H when germline alterations are detected in at least two, at least three, at least four, or at least five MMR genes. In another embodiment, a colorectal cancer is MSI-H when germline alterations are detected in at least 30% of five or more MMR genes. In one embodiment, germline alterations in MMR genes are measured by polymerase chain reaction. In other embodiments, colorectal cancer is MSI-H when at least one protein encoded by a DNA MMR gene is not detected in the tumor. In certain embodiments, at least one protein encoded by a DNA MMR gene is detected by immunohistochemistry. In certain embodiments, the present invention relates to methods of treating cancer, comprising 1) identifying the microsatellite status of the tumor, and 2) administering a therapeutic agent to the subject based on the microsatellite status. In certain embodiments, the subject has MSI-H.

[0123] In some embodiments, the colorectal cancer is rectal cancer, colon cancer, or any combination thereof. In some embodiments, the subject has undergone one, two, three, four, five, or more previous cancer treatments. In other embodiments, the subject is treatment-naive. In some embodiments, the subject has progressed with other cancer treatments. In some embodiments, the cancer has recurred. In some embodiments, the cancer is metastatic. In other embodiments, the cancer is not metastatic.

[0124] In some embodiments, colorectal cancer is histologically confirmed. In some embodiments, colorectal cancer is metastatic or recurrent. In some embodiments, the subject progresses during or after the last administration of standard treatment, or is intolerant after administration. In some embodiments, the subject has microsatellite instability. In other embodiments, the colorectal cancer has high microsatellite instability (MSI-H).

[0125] In other embodiments, the MSI-H colorectal cancer expresses PD-L1. In yet other embodiments, the MSI-H colorectal cancer expresses at least 1%, at least 2%, at least 3%, at least 4%, at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, or at least 50% membrane PD-L1.

[0126] In some embodiments, the method comprises administering an effective amount of an anti-PD-1 antibody. In other embodiments, the method comprises administering an effective amount of an anti-PD-1 antibody and an effective amount of an anti-CTLA-4 antibody. The effective amounts of the anti-PD-1 antibody and / or anti-CTLA-4 antibody can be a flat dose or a weight-based dose.

[0127] In some embodiments, the present invention includes a method of treating a subject with a tumor derived from colorectal cancer, comprising administering an anti-PD-1 antagonist in combination with an anti-CTLA-4 antibody to treat the cancer. As used herein, "anti-PD-1 antagonist" 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 some embodiments, the anti-PD-1 antagonist includes an anti-PD-1 fusion protein, an antisense molecule, a small molecule, a ribozyme, or a nanobody that inhibits or blocks the interaction between PD-1 and PD-L1.

[0128] In certain embodiments, a treatment of the invention (e.g., administration of an anti-PD-1 antibody alone or in combination with an anti-CTLA-4 antibody) effectively extends the subject's survival, e.g., by 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 or more.

[0129] In some embodiments, the treatment of the present invention effectively prolongs the progression-free survival period of the subject.For example, the progression-free survival period of the subject is prolonged by 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.In some embodiments, the treatment of the present invention effectively increases the response rate of the subject group. For example, the response rate of a subject group is increased by at least about 2%, at least about 3%, at least about 4%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 99% or about 100%.

[0130] In some embodiments, the anti-PD-1 antibody is formulated for intravenous administration. In some embodiments, the anti-PD-1 and anti-CTLA-4 antibodies are formulated for intravenous administration. In some embodiments, the anti-PD-1 and anti-CTLA-4 antibodies are administered sequentially. In some embodiments, the anti-PD-1 and anti-CTLA-4 antibodies are administered within 30 minutes of each other. In some embodiments, the anti-PD-1 antibody, or antigen-binding portion thereof, is administered before the anti-CTLA-4 antibody, or antigen-binding portion thereof. In other embodiments, the anti-CTLA-4 antibody, or antigen-binding portion thereof, is administered before the anti-PD-1 antibody, or antigen-binding portion thereof. In other embodiments, the anti-PD-1 antibody, or antigen-binding portion thereof, and the anti-CTLA-4 antibody, or antigen-binding portion thereof, are administered simultaneously in separate compositions. In a further embodiment, the anti-PD-1 antibody, or antigen-binding portion thereof, and the anti-CTLA-4 antibody, or antigen-binding portion thereof, are combined into a single composition for simultaneous administration.

[0131] In certain embodiments, the anti-PD-1 antibody and the anti-CTLA-4 antibody are administered as a fixed dose.

[0132] In some embodiments, the cancer is microsatellite stable (MSS) (or "MSI stable") and therefore does not have microsatellite instability. In other embodiments, the cancer has high microsatellite instability (MSI-H).

[0133] Anti-PD-1 and anti-PD-L1 antibodies Therapies of the present invention may utilize anti-PD-1 antibodies or antigen-binding portions thereof. PD-1 is an important immune checkpoint receptor expressed by activated T and B cells and mediates immunosuppression. PD-1 is a member of the CD28 family of receptors, which also includes CD28, CTLA-4, ICOS, PD-1, and BTLA. Two cell surface glycoprotein ligands for PD-1, programmed death-ligand-1 (PD-L1) and programmed death-ligand-2 (PD-L2), have been identified and are expressed on antigen-presenting cells and many human cancers. Binding to PD-1 has been shown to downregulate T cell activation and cytokine secretion. Inhibition of PD-1 / PD-L1 interaction mediates potent antitumor activity in preclinical models.

[0134] Anti-PD-1 antibodies suitable for use in the methods of the present invention are antibodies that bind to PD-1 with high specificity and affinity, block PD-L1 binding, and inhibit the immunosuppressive effects of the PD-1 signaling pathway. In any of the therapeutic methods described herein, an anti-PD-1 or anti-PD-L1 "antibody" comprises an antigen-binding portion that binds to the PD-1 or PD-L1 receptor, respectively, and exhibits functional properties similar to whole antibodies in blocking ligand binding and upregulating the immune system. In one embodiment, the anti-PD-1 antibody or antigen-binding portion thereof cross-competes with nivolumab for binding to human PD-1. In other embodiments, the anti-PD-L1 antibody or antigen-binding fragment thereof competes with BMS-936559, MPDL3280A, MEDI4736, or MSB0010718C for binding to human PD-L1.

[0135] In other embodiments, the anti-PD-1 antibody or anti-PD-L1 antibody, or antigen-binding portion thereof, is a chimeric, humanized, or human monoclonal antibody, or portion thereof. In certain embodiments for treating human subjects, the antibody is a humanized antibody. In other embodiments for treating human subjects, the antibody may be of the human IgG1, IgG2, IgG3, or IgG4 isotype.

[0136] In certain embodiments, the anti-PD-1 antibody or anti-PD-L1 antibody, or antigen-binding portion thereof, comprises a heavy chain constant region of the human IgG1 or IgG4 isotype. In certain other embodiments, the sequence of the IgG4 heavy chain constant region of the anti-PD-1 antibody or anti-PD-L1 antibody, or antigen-binding portion thereof, comprises an S228P mutation, replacing a serine residue in the hinge region with a proline residue normally found at the corresponding position in IgG1 isotype antibodies. This mutation, present in nivolumab, prevents Fab arm exchange with endogenous IgG4 antibodies while maintaining the low affinity for Fc receptor activation associated with wild-type IgG4 antibodies (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 other embodiments, the antibody comprises a light chain constant region that is a human kappa or lambda constant region. In other embodiments, the anti-PD-1 antibody or anti-PD-L1 antibody, or antigen-binding portion thereof, is a monoclonal antibody, or antigen-binding portion thereof.

[0137] Human monoclonal antibodies that specifically bind to PD-1 with high affinity are disclosed in U.S. Patent 8,008,449. Other anti-PD-1 monoclonal antibodies are disclosed, for example, in U.S. Patents 6,808,710, 7,488,802, 8,168,757, and 8,354,509 and PCT Publication WO 2012 / 145493. Each of the anti-PD-1 human monoclonal antibodies disclosed in U.S. Patent 8,008,449 has been shown to exhibit one or more of the following characteristics: (a) a specific binding affinity of 1×10 to human PD-1 as determined by surface plasmon resonance using a Biacore biosensor system; -7 K below M D(b) binds to human CD28, CTLA-4, or ICOS; (c) increases T-cell proliferation in a mixed lymphocyte reaction (MLR) assay; (d) increases interferon-γ production in an MLR assay; (e) increases IL-2 secretion in an MLR assay; (f) binds to human PD-1 and cynomolgus PD-1; (g) inhibits the binding of PD-L1 and / or PD-L2 to PD-1; (h) stimulates an antigen-specific memory response; (i) stimulates an antibody response; and / or (j) inhibits tumor cell growth in vivo. Anti-PD-1 antibodies that can be used in the present invention include monoclonal antibodies that specifically bind to human PD-1 and exhibit at least one, at least two, at least three, at least four, or at least five of the above characteristics. In one embodiment, the anti-PD-1 antibody comprises nivolumab. In one embodiment, the anti-PD-1 antibody comprises pembrolizumab.

[0138] In one embodiment, the anti-PD-1 antibody is nivolumab. (登録商標) Nivolumab, also known as nivolumab; formerly named 5C4, BMS-936558, MDX-1106, or ONO-4538, is a fully human IgG4(S228P) PD-1 immune checkpoint inhibitor antibody that selectively inhibits the interaction with PD-1 ligands (PD-L1 and PD-L2), thereby blocking the downregulation of antitumor T cell function (U.S. Patent 8,008,449; Wang et al., 2014 Cancer Immunol Res. 2(9):846-56). Nivolumab has shown activity in a variety of advanced solid tumors, including renal cell carcinoma (renal adenocarcinoma or adrenal nephroma), melanoma, and non-small cell lung cancer (NSCLC) (Topalian et al., 2012a; Topalian et al., 2014; Drake et al., 2013; WO2013 / 173223). In other embodiments, the anti-PD-1 antibody (or antigen-binding portion thereof) cross-competes with nivolumab. In certain embodiments, the anti-PD-1 antibody binds to the same epitope as nivolumab. In certain embodiments, the anti-PD-1 antibody has the same CDRs as nivolumab.

[0139] In other embodiments, the anti-PD-1 antibody (or antigen-binding portion thereof) cross-competes with pembrolizumab. In certain embodiments, the anti-PD-1 antibody binds to the same epitope as pembrolizumab. In certain embodiments, the anti-PD-1 antibody has the same CDRs as pembrolizumab. In other embodiments, the anti-PD-1 antibody is pembrolizumab. Pembrolizumab ("Keytruda") (登録商標) ", also known as lambrolizumab and MK-3475) is a humanized monoclonal IgG4 antibody directed against the human cell surface receptor PD-1 (programmed death-1 or programmed cell death-1). Pembrolizumab is described, e.g., in U.S. Patent 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 recurrent or refractory melanoma and advanced NSCLC.

[0140] In other embodiments, the anti-PD-1 antibody (or antigen-binding portion thereof) cross-competes with MEDI0680. In certain embodiments, the anti-PD-1 antibody binds to the same epitope as MEDI0680. In certain embodiments, the anti-PD-1 antibody has the same CDRs as MEDI0680. In other embodiments, the anti-PD-1 antibody is MEDI0680 (formerly AMP-514), a monoclonal antibody against the PD-1 receptor. MEDI0680 is described, for example, in U.S. Patent 8,609,089 B2 or www.cancer.gov / drugdictionary?cdrid=756047 (last accessed: December 14, 2014).

[0141] In one embodiment, the immune checkpoint inhibitor is AMP-224, a B7-DC Fc fusion protein. AMP-224 is described in U.S. Publication 2013 / 0017199 or http: / / www.cancer.gov / publications / dictionaries / cancer-drug?cdrid=700595 (last accessed July 8, 2015).

[0142] In other embodiments, the anti-PD-1 antibody (or antigen-binding portion thereof) cross-competes with BGB-A317. In certain embodiments, the anti-PD-1 antibody binds to the same epitope as BGB-A317. In certain embodiments, the anti-PD-1 antibody has the same CDRs as BGB-A317. In certain embodiments, the anti-PD-1 antibody is the humanized monoclonal antibody BGB-A317. BGB-A317 is described in U.S. Publication No. 2015 / 0079109.

[0143] In other embodiments, the anti-PD-1 antibody (or antigen-binding portion thereof) cross-competes with INCSHR1210 (SHR-1210). In certain embodiments, the anti-PD-1 antibody binds to the same epitope as INCSHR1210 (SHR-1210). In certain embodiments, the anti-PD-1 antibody has the same CDRs as INCSHR1210 (SHR-1210). In certain embodiments, the anti-PD-1 antibody is the human monoclonal antibody INCSHR1210 (SHR-1210). INCSHR1210 (SHR-1210) is described in WO2015 / 085847.

[0144] In other embodiments, the anti-PD-1 antibody (or antigen-binding portion thereof) cross-competes with REGN-2810. In certain embodiments, the anti-PD-1 antibody binds to the same epitope as REGN-2810. In certain embodiments, the anti-PD-1 antibody has the same CDRs as REGN-2810. In certain embodiments, the anti-PD-1 antibody is the human monoclonal antibody REGN-2810. REGN-2810 is described in WO2015 / 112800.

[0145] In other embodiments, the anti-PD-1 antibody (or antigen-binding portion thereof) cross-competes with PDR001. In certain embodiments, the anti-PD-1 antibody binds to the same epitope as PDR001. In certain embodiments, the anti-PD-1 antibody has the same CDRs as PDR001. In certain embodiments, the anti-PD-1 antibody is PDR001, a humanized monoclonal antibody. PDR001 is described in WO2015 / 112900.

[0146] In other embodiments, the anti-PD-1 antibody (or antigen-binding portion thereof) cross-competes with TSR-042 (ANB011). In certain embodiments, the anti-PD-1 antibody binds to the same epitope as TSR-042 (ANB011). In certain embodiments, the anti-PD-1 antibody has the same CDRs as TSR-042 (ANB011). In certain embodiments, the anti-PD-1 antibody is the humanized monoclonal antibody TSR-042 (ANB011). TSR-042 (ANB011) is described in WO2014 / 179664.

[0147] In other embodiments, the anti-PD-1 antibody (or antigen-binding portion thereof) cross-competes with STI-1110. In certain embodiments, the anti-PD-1 antibody binds to the same epitope as STI-1110. In certain embodiments, the anti-PD-1 antibody has the same CDRs as STI-1110. In certain embodiments, the anti-PD-1 antibody is the human monoclonal antibody STI-1110. STI-1110 is described in WO2014 / 194302.

[0148] Anti-PD-1 antibodies that can be used in the methods of the present invention also include isolated antibodies that specifically bind to human PD-1 and cross-compete with nivolumab for binding to human PD-1 (see, e.g., U.S. Patent No. 8,008,449; WO2013 / 173223). The ability of antibodies to cross-compete for binding to an antigen indicates that these antibodies bind to the same epitope region of the antigen and sterically interfere with the binding of other cross-competing antibodies to that particular epitope region. These cross-competing antibodies are expected to have functional properties very similar to nivolumab by binding to the same epitope region of PD-1. Cross-competing antibodies can be readily identified based on their ability to cross-compete with nivolumab in standard PD-1 binding assays, such as Biacore analysis, ELISA assays, or flow cytometry (see, e.g., WO2013 / 173223).

[0149] In some embodiments, the antibody that cross-competes with the binding of nivolumab to human PD-1 or binds to the same epitope region of human PD-1 is a monoclonal antibody. For administration to human subjects, these cross-competing antibodies can be chimeric, humanized, or human antibodies. Such chimeric, humanized, or human monoclonal antibodies can be produced and isolated by methods well known in the art.

[0150] Anti-PD-1 antibodies useful in the methods of the present invention also include antigen-binding portions of such antibodies. It is well established that the antigen-binding function of an antibody can be performed by fragments of a full-length antibody. Examples of binding fragments encompassed by the term "antigen-binding portion" of an antibody include: (i) V L , V H , C L and C H1 (ii) a F(ab')2 fragment, a bivalent fragment containing two Fab fragments linked by disulfide bridges in the hinge region; and (iii) a V H and C H1 (iv) a single-arm V fragment of an antibody; L and V Han Fv fragment consisting of a domain; or any combination thereof.

[0151] In certain embodiments, the anti-PD-1 antibody, or antigen-binding portion thereof, comprises a heavy chain constant region of a human IgG1 or IgG4 isotype. In certain other embodiments, the sequence of the IgG4 heavy chain constant region of the anti-PD-1 antibody, or antigen-binding portion thereof, contains a S228P mutation, replacing a serine residue in the hinge region with a proline residue normally found at the corresponding position in IgG1 isotype antibodies. This mutation, present in nivolumab, prevents Fab arm exchange with endogenous IgG4 antibodies while maintaining the low affinity for Fc receptor activation associated with wild-type IgG4 antibodies (Wang et al., 2014). In yet other embodiments, the antibody comprises a light chain constant region that is a human kappa or lambda constant region. In other embodiments, the anti-PD-1 antibody, or antigen-binding portion thereof, is a monoclonal antibody or antigen-binding portion thereof. In certain embodiments of any of the methods of treatment described herein that comprise administration of an anti-PD-1 antibody, the anti-PD-1 antibody is nivolumab. In other embodiments, the anti-PD-1 antibody is pembrolizumab. In other embodiments, the anti-PD-1 antibody is selected from human antibodies 17D8, 2D3, 4H1, 4A11, 7D3, and 5F4, described in U.S. Patent 8,008,449. In yet other embodiments, the anti-PD-1 antibody is MEDI0680 (formerly AMP-514), AMP-224, or BGB-A317.

[0152] In other embodiments, the anti-PD-1 antibody, or antigen-binding portion thereof, is a chimeric, humanized, or human monoclonal antibody, or portion thereof. In certain embodiments for treating a human subject, the antibody is a humanized antibody. In other embodiments for treating a human subject, the antibody is a human antibody. Antibodies of the IgG1, IgG2, IgG3, or IgG4 isotype may be used.

[0153] In certain embodiments, the anti-PD-1 antibody used in the methods can be replaced with another PD-1 or anti-PD-L1 antagonist. For example, an anti-PD-L1 antibody can be substituted for the use of an anti-PD-1 antibody in the methods disclosed herein, because anti-PD-L1 antibodies block the interaction between PD-1 and PD-L1, thereby exerting a similar effect on the PD-1 signaling pathway. Thus, in certain embodiments, the present invention relates to a method for treating a subject with a tumor derived from colorectal cancer, comprising administering to the subject a therapeutically effective amount of an anti-cancer agent that is an antibody or antigen-binding portion thereof (anti-PD-L1 antibody) that specifically binds to the programmed death-ligand 1 (PD-L1) receptor and inhibits PD-L1 activity. In other embodiments, the present invention relates to a method for treating a subject with a tumor derived from colorectal cancer, comprising administering to the subject a therapeutically effective amount of an anti-PD-L1 antibody in combination with an anti-CTLA-4 antibody. In some embodiments, the anti-PD-L1 Ab is BMS-936559 (formerly 12A4 or MDX-1105) (see, e.g., U.S. Patent 7,943,743; WO2013 / 173223). In other embodiments, the anti-PD-L1 Ab is MPDL3280A (also known as RG7446 or atezolizumab) (see, e.g., Herbst; U.S. Patent 8,217,149), MEDI4736 (also known as durvalumab; see Khleif, 2013; U.S. Patent 8,779,108; or US2014 / 0356353, filed May 6, 2014), or MSB0010718C (also known as avelumab; see US2014 / 0341917). In other embodiments, the anti-PD-L1 antibody is CX-072 (also known as CytomX; see WO2016 / 149201).

[0154] Anti-CTLA-4 antibody In one embodiment, the method of the present invention can be a combination therapy of an anti-PD-1 antibody or anti-PD-L1 antibody and an anti-CTLA-4 antibody. The anti-CTLA-4 antibody of the present invention binds to human CTLA-4 in a manner that disrupts the interaction between CTLA-4 and human B7 receptor. Because the interaction between CTLA-4 and human B7 transmits a signal that leads to the inactivation of CTLA-4 receptor-bearing T cells, disrupting this interaction effectively induces, enhances, or prolongs the activation of such T cells, thereby inducing, enhances, or prolongs an immune response.

[0155] Human monoclonal antibodies that specifically bind to CTLA-4 with high affinity are disclosed in U.S. Patents 6,984,720 and 7,605,238. Other anti-PD-1 monoclonal antibodies are described, for example, in U.S. Patents 5,977,318, 6,051,227, 6,682,736, and 7,034,121. The anti-PD-1 human monoclonal antibodies disclosed in U.S. Patents 6,984,720 and 7,605,238 have been shown to exhibit one or more of the following properties: (a) a cytoplasmic affinity of at least about 10 as determined by Biacore analysis; 7 M -1 or about 10 9 M -1 or about 10 10 M -1 ~10 11 M -1 or higher equilibrium binding constant (K a ), (b) specifically binds human CTLA-4 with a binding affinity reflected by at least about 10 3 , about 10 4 or about 10 5 m -1 s -1 The kinetic binding constant (k a );(c) at least about 10 3 , about 10 4 or about 10 5 m -1 s -1 The kinetic dissociation constant (k dand (d) inhibit 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 above properties.

[0156] An example of a clinical anti-CTLA-4 antibody is the human monoclonal antibody 10D1 (now known as ipilimumab and Yervoy), disclosed in U.S. Patent 6,984,720. (登録商標) Ipilimumab is an anti-CTLA-4 antibody for use in the methods described herein. Ipilimumab is a fully human, IgG1 monoclonal antibody that blocks the binding of CTLA-4 to its B7 ligand, thereby stimulating T cell activation and improving overall survival (OS) in patients with advanced melanoma.

[0157] Another anti-CTLA-4 antibody useful for the present method is tremelimumab (also known as CP-675,206). Tremelimumab is a human IgG2 monoclonal anti-CTLA-4 antibody. Tremelimumab is described in WO / 2012 / 122444, U.S. Publication No. 2012 / 263677, or WO Publication No. 2007 / 113648A2.

[0158] Anti-CTLA-4 antibodies useful in the compositions of the present invention also include isolated antibodies that specifically bind to human CTLA-4 and cross-compete with ipilimumab or tremelimumab for binding to human CTLA-4 or bind to the same epitope region of human CTLA-4 as ipilimumab or tremelimumab. In some embodiments, antibodies that cross-compete with ipilimumab or tremelimumab for binding to human CTLA-4 or bind to the same epitope region of human CTLA-4 are antibodies containing a heavy chain of human IgG1 isotype. For administration to human subjects, these cross-competing antibodies are chimeric antibodies or humanized or human antibodies. Useful anti-CTLA-4 antibodies also include antigen-binding portions of the above antibodies, such as Fab, F(ab')2, Fd, or Fv fragments.

[0159] Cancer and standard of care treatments In certain embodiments, the methods described herein are used in place of standard of care treatments. In certain embodiments, standard of care treatments are used in combination with any of the methods disclosed herein. Standard of care treatments for various types of cancer are well known to those of skill in the art. For example, the National Comprehensive Cancer Network (NCCN), an alliance of 21 major cancer centers in the United States, has published the NCCN Clinical Practice Guidelines in Oncology (NCCN GUIDELINES), which provide detailed, up-to-date information on standard of care treatments for a variety of cancers. (登録商標) ) has been published (NCCN GUIDELINES (登録商標) , 2014).

[0160] Pharmaceutical Compositions and Dosages Therapeutic agents of the present invention may comprise compositions, e.g., pharmaceutical compositions, containing an antibody and a pharmaceutically acceptable carrier. As used herein, "pharmaceutically acceptable carrier" includes any and all physiologically compatible solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like. In some embodiments, the carrier for the antibody-containing composition is suitable for intravenous, intramuscular, subcutaneous, parenteral, spinal, or epidermal administration (e.g., by injection or infusion). Pharmaceutical compositions of the present invention may include one or more pharmaceutically acceptable salts, antioxidants, aqueous and non-aqueous carriers, and / or adjuvants such as preservatives, wetting agents, emulsifying agents, and dispersing agents.

[0161] The dosage regimen is adjusted to provide the optimal desired response, e.g., maximal therapeutic response and / or minimal adverse effects. In certain embodiments, the anti-PD-1 antibody is administered in a weight-based dose. For anti-PD-1 antibody administration, the dosage can range from at least about 0.01 to at least about 20 mg / kg, or at least about 0.1 to at least about 10 mg / kg of the subject's body weight. For example, the dosage can be at least about 0.1 mg / kg, at least about 0.3 mg / kg, at least about 1 mg / kg, at least about 2 mg / kg, at least about 3 mg / kg, at least about 5 mg / kg, or at least about 10 mg / kg of body weight. In certain embodiments, the dosage of the anti-PD-1 antibody is 3 mg / kg body weight. In certain embodiments, the anti-PD-1 antibody is administered in a flat dose. In some embodiments, the flat dose of the anti-PD-1 antibody is at least about 100-300 mg, e.g., at least about 200-300 mg, at least about 220-260 mg, at least about 230-250 mg, or at least about 240 mg, e.g., at least about 60 mg, at least about 80 mg, at least about 100 mg, at least about 120 mg, at least about 140 mg, at least about 160 mg, at least about 180 mg, 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, or at least about 300 mg (e.g., a flat dose). In some embodiments, the anti-PD-1 antibody is administered in a fixed dose with the anti-CTLA-4 antibody.In certain embodiments, the ratio is at least about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, about 1:30, about 1:40, about 1:50, about 1:60, about 1:70, about 1:80, about 1:90, about 1:100, about 1:120, about 1:140, about 1:160, about 1:180, about 1:200, about 200:1, about 180:1, about 160:1, about 140:1, about 120:1, about 100:1, about 90:1, about 80:1, about 70:1, about 60:1, about 50:1, about 40:1, about 30:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1 or about 2:1 mg anti-PD-1 antibody to mg anti-CTLA-4 antibody.

[0162] The administration schedule is generally based on the typical pharmacokinetics of the antibody and is designed to achieve an exposure that results in sustained receptor occupancy (RO). Exemplary treatment regimens include administration once a week, about once every two weeks, about once every three weeks, about once every four weeks, about once a month, about once every three to six months, or at longer intervals. In one embodiment, an anti-PD-1 antibody such as is administered to a subject about once every two weeks. In another embodiment, the antibody is administered about once every three weeks. Dosage and schedule may vary during the course of treatment.

[0163] When used in combination with other anti-cancer drugs, the dosage of the anti-PD-1 antibody may be reduced compared to the monotherapy dose. A typical dose of nivolumab lower than 3 mg / kg but not less than 0.001 mg / kg is a subtherapeutic dose. A subtherapeutic dose of the anti-PD-1 antibody used in the methods herein is greater than 0.001 mg / kg and less than 3 mg / kg. In some embodiments, the subtherapeutic dose is about 0.001 mg / kg to about 1 mg / kg, about 0.01 mg / kg to about 1 mg / kg, about 0.1 mg / kg to about 1 mg / kg, or about 0.001 mg / kg to about 0.1 mg / kg of body weight. In certain embodiments, the subtherapeutic dose is at least about 0.001 mg / kg, at least about 0.005 mg / kg, at least about 0.01 mg / kg, at least about 0.05 mg / kg, at least about 0.1 mg / kg, at least about 0.5 mg / kg, or at least about 1.0 mg / kg body weight. Receptor occupancy data from 15 subjects receiving nivolumab at doses of 0.3 mg / kg to 10 mg / kg indicate that PD-1 occupancy appears to be dose-independent within this dose range. Across all doses, the mean occupancy was 85% (range, 70%-97%), and the mean plateau occupancy was 72% (range, 59%-81%). In certain embodiments, a 0.3 mg / kg dose may allow sufficient exposure to achieve maximal biological activity.

[0164] In certain embodiments, the anti-CTLA-4 antibody is administered at a weight-based dose. For administration of the anti-CTLA-4 antibody, the dosage can range from about 0.01 to about 20 mg / kg, about 0.05 to about 20 mg / kg, about 0.1 to about 20 mg / kg, about 0.1 to about 15 mg / kg, about 0.1 to about 10 mg / kg, about 0.1 to about 5 mg / kg, and about 1 to about 10 mg / kg of patient body weight. For example, the dosage can be about 0.05 mg / kg, about 0.1 mg / kg, about 0.5 mg / kg, about 1 mg / kg, about 2 mg / kg, about 3 mg / kg, about 4 mg / kg, about 5 mg / kg, about 6 mg / kg, about 7 mg / kg, about 8 mg / kg, about 9 mg / kg, about 10 mg / kg, about 11 mg / kg, about 12 mg / kg, about 13 mg / kg, about 14 mg / kg, about 15 mg / kg, or about 20 mg / kg of patient body weight. In one embodiment, the dosage of the anti-CTLA-4 antibody is 0.1 mg / kg body weight. In another embodiment, the dosage of the anti-CTLA-4 antibody is 1 mg / kg body weight. In a further embodiment, the dosage of the anti-CTLA-4 antibody is 10 mg / kg body weight. In one embodiment, the anti-CTLA-4 antibody is administered in a flat dose.In certain embodiments, the uniform dose of anti-CTLA-4 is at least about 60 to 1500 mg, e.g., at least about 100 to 1400 mg, at least about 100 to 1000 mg, at least about 200 to 1000 mg, or at least about 200 to 500 mg, e.g., at least about 60 mg, at least about 80 mg, at least about 100 mg, at least about 120 mg, at least about 140 mg, at least about 160 mg, at least about 180 mg, 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 400 mg, at least about 500 mg, at least about 600 mg, at least about 800 mg, at least about 100 mg, at least about 120 mg, at least about 140 mg, at least about 160 mg, at least about 180 mg, 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 400 mg, at least about 500 mg, at least about 600 mg, at least about 700 mg, at least about 800 mg, at least about 900 mg, at least about 900 mg, at least about 1000 mg, at least about 1200 mg, at least about 140 mg, at least about 160 mg, at least about 180 mg, 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 The dose (e.g., a flat dose) is 0 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, at least about 600 mg, at least about 700 mg, at least about 800 mg, at least about 900 mg, at least about 1000 mg, at least about 1100 mg, at least about 1200 mg, at least about 1300 mg, at least about 1400 mg, or at least about 1500 mg.

[0165] Exemplary treatment regimens include administration once a week, about once every two weeks, about once every three weeks, about once every four weeks, about once a month, about once every three to six months, or longer. In certain embodiments, the anti-CTLA-4 antibody is administered about once every three weeks.

[0166] In some embodiments, a subtherapeutic dose of an anti-CTLA-4 antibody is used in the methods herein. The subtherapeutic dose of an anti-CTLA-4 antibody used in the methods herein is greater than 0.001 mg / kg and less than 10 mg / kg. In some embodiments, the subtherapeutic dose is about 0.001 mg / kg to about 10 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 of body weight. In some embodiments, the subtherapeutic dose is at least about 0.001 mg / kg, at least about 0.005 mg / kg, at least about 0.01 mg / kg, at least about 0.05 mg / kg, at least about 0.1 mg / kg, at least about 0.5 mg / kg, at least about 1.0 mg / kg, at least about 2 mg / kg, at least about 3 mg / kg, at least about 4 mg / kg, at least about 5 mg / kg, at least about 6 mg / kg, at least about 7 mg / kg, at least about 8 mg / kg, at least about 9 mg / kg, or at least about 10 mg / kg body weight. In some embodiments, the subtherapeutic dose is about 10 mg / kg, about 5 mg / kg, about 2 mg / kg, about 1 mg / kg, about 0.1 mg / kg, or about 0.01 mg / kg body weight.

[0167] In one embodiment, at least about 0.1 to about 10 mg / kg of anti-CTLA-4 antibody and at least about 0.1 to about 10 mg / kg of anti-PD-1 antibody are administered to a subject approximately once every three weeks. In one embodiment, at least about 1 mg / kg of anti-CTLA-4 antibody and at least about 1 mg / kg of anti-PD-1 antibody are administered to a subject approximately once every three weeks. In one embodiment, at least about 1 mg / kg of anti-CTLA-4 antibody and at least about 3 mg / kg of anti-PD-1 antibody are administered to a subject approximately once every three weeks. In one embodiment, at least about 3 mg / kg of anti-CTLA-4 antibody and at least about 1 mg / kg of anti-PD-1 antibody are administered to a subject approximately every three weeks. In one embodiment, at least about 3 mg / kg of anti-CTLA-4 antibody and at least about 3 mg / kg of anti-PD-1 antibody are administered to a subject approximately every three weeks. In one embodiment, at least about 3 mg / kg of anti-CTLA-4 antibody and at least about 3 mg / kg of anti-PD-1 antibody are administered to a subject approximately every three weeks. In one embodiment, the anti-CTLA-4 antibody is ipilimumab. In one embodiment, the anti-PD-1 antibody is nivolumab.

[0168] In one embodiment, the combination of an anti-PD-1 antibody and an anti-CTLA-4 antibody is intravenously administered to the subject approximately once every three weeks for a total of 12 weeks. In one embodiment, the 12-week cycle is repeated three or four times. In one embodiment, the subject is treated with the combination of an anti-PD-1 antibody and an anti-CTLA-4 antibody every three weeks for a total of 12 weeks, with three 12-week cycles being performed. In one embodiment, the subject is treated with the combination of an anti-PD-1 antibody and an anti-CTLA-4 antibody every three weeks for a total of 12 weeks, with four 12-week cycles being performed. In one embodiment, the subject is treated with the anti-PD-1 antibody for twelve 12-week cycles.

[0169] In some embodiments, administration of an anti-PD-1 antibody and an anti-CTLA-4 antibody is followed by anti-PD-1 antibody monotherapy. In some embodiments, the antibody monotherapy after the combination treatment with an anti-PD-1 antibody and an anti-CTLA-4 antibody comprises administering the anti-PD-1 antibody at a dose of about 1 mg / kg body weight, about 2 mg / kg body weight, about 3 mg / kg body weight, about 4 mg / kg body weight, about 5 mg / kg body weight, about 6 mg / kg body weight, about 7 mg / kg body weight, about 8 mg / kg body weight, about 9 mg / kg body weight, or about 10 mg / kg body weight. In some embodiments, the anti-PD-1 monotherapy provided after the combination treatment with an anti-PD-1 antibody and an anti-CTLA-4 antibody is administered about once every 1, 2, 3, or 4 weeks. In some embodiments, the anti-PD-1 antibody monotherapy provided after the combination treatment with an anti-PD-1 antibody and an anti-CTLA-4 antibody is administered at a dose of about 3 mg / kg body weight once every two weeks.

[0170] In other embodiments, the anti-PD-1 antibody is administered at 3 mg / kg once every three weeks, and the anti-CTLA-4 antibody is administered at 1 mg / kg once every three weeks. In certain embodiments, the anti-PD-1 antibody and the anti-CTLA-4 antibody are administered in combination once, twice, three times, four times, five times, six times, seven times, eight times, nine times, or ten times. In certain embodiments, the anti-PD-1 antibody and the anti-CTLA-4 antibody are administered in combination four times. In certain embodiments, the subject is administered an anti-PD-1 antibody at a dose of 3 mg / kg once every two weeks before and / or after the combination treatment with the anti-PD-1 antibody and the anti-CTLA-4 antibody. In certain embodiments, subjects with MSI-H colorectal cancer are treated in an initial stage (e.g., anti-PD-1 antibody monotherapy, e.g., nivolumab at a dose of 3 mg / kg once every two weeks), a combination stage (e.g., anti-PD-1 antibody and anti-CTLA-4 antibody combination, e.g., nivolumab at a dose of 3 mg / kg once every three weeks and ipilimumab at a dose of 1 mg / kg once every week), and an end-stage (e.g., anti-PD-1 antibody monotherapy, e.g., nivolumab at a dose of 3 mg / kg once every two weeks).

[0171] Treatment may continue as long as clinical benefit is observed or until unacceptable toxicity or disease progression occurs. In certain embodiments, the anti-PD-1 antibody can be administered at a dose shown in clinical trials to produce maximal efficacy as monotherapy, e.g., about 3 mg / kg nivolumab administered about once every 3 weeks (Topalian et al., 2012 N Engl J Med 366:2443-54; Topalian et al., 2012 Curr Opin Immunol 24:207-12), or at a significantly lower dose, i.e., a subtherapeutic dose.

[0172] The actual dosage level of the active ingredient in the pharmaceutical composition of the present invention can be varied to obtain an amount of active ingredient that is effective to achieve the desired therapeutic effect for a particular patient, composition, and administration method without causing excessive toxicity to the patient. The selected dosage depends on various 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, and previous medical history of the patient being treated, and similar factors well known in the medical field. The composition of the present invention can be administered by one or more routes of administration using one or more of a variety of methods well known in the art. As known to those skilled in the art, the route and / or mode of administration will vary depending on the desired results.

[0173] kit Also within the scope of the present invention are kits containing an anti-PD-1 antibody and another anti-cancer agent for therapeutic use. The kit generally includes a label indicating the intended use and instructions for use of the contents of the kit. The term label includes any writing or recorded material on or with the kit or that otherwise accompanies the kit. Thus, the present invention provides kits for treating a subject having a tumor resulting from colorectal cancer, the kit including: (a) a single dose of an anti-PD-1 antibody, or antigen-binding portion thereof, in the range of about 0.1 mg / kg to about 10 mg / kg; and (b) instructions for using the anti-PD-1 antibody in any of the monotherapy methods described herein. In another embodiment, a kit for treating a subject with a tumor derived from colorectal cancer comprises: (a) a single dose of an anti-PD-1 antibody, or antigen-binding portion thereof, in the range of about 0.1 mg / kg to about 10 mg / kg; and (b) a single dose of an anti-CTLA-4 antibody, or antigen-binding portion thereof, in the range of about 0.1 mg / kg to about 10 mg / kg; and (c) instructions for using the anti-PD-1 antibody and the anti-CTLA-4 antibody in any of the combination treatment methods described herein. In some embodiments, the anti-PD-1 antibody and the anti-CTLA-4 antibody may be co-packaged in a unit dosage form. In some embodiments for treating a human patient, the kit comprises an anti-human PD-1 antibody disclosed herein, e.g., nivolumab, pembrolizumab, MEDI0608 (formerly AMP-514), AMP-224, or BGB-A317. In other embodiments, the kit comprises an anti-human CTLA-4 antibody described herein, e.g., ipilimumab or tremelimumab.

[0174] The present invention is further illustrated by the following examples, which should not be construed as further limiting. The contents of all references cited throughout this specification are expressly incorporated herein by reference.

[0175] CROSS-REFERENCE TO PRIOR-FILED APPLICATIONS: This invention claims the benefit of U.S. Provisional Application No. 62 / 345,662, filed June 3, 2017, which is incorporated herein by reference in its entirety. [Example]

[0176] Example 1 Clinical Study of Nivolumab ± Ipilimumab in the Treatment of Patients with Metastatic Colorectal Cancer With or Without High Microsatellite Instability Patients were eligible for inclusion in this trial based on the following criteria: 1) histologically confirmed colorectal cancer; 2) recurrent or metastatic disease measurable by Response Evaluation Criteria in Solid Tumors (RECIST) 1.1; 3) patients were adults at least 18 years of age and had an Eastern Cooperative Oncology Group (ECOG) score of 0 to 1; and 4) disease progression after ≥1 prior treatment regimen (for microsatellite instability-high patients) or most recent treatment (for all patients) or chemotherapy intolerance or refusal. Patients were excluded from the trial based on the following criteria: 1) central nervous system complications; 2) history of malignancy within 3 years; 3) active or history of autoimmune disease; 4) need for treatment with immunosuppressive regimens, including corticosteroids; or 5) prior treatment targeting T-cell costimulation or immune checkpoint pathways.

[0177] Patients with colorectal cancer were defined as microsatellite instability-high (MSI-H) by polymerase chain reaction (PCR) if they had instability at two or more markers out of five loci tested or at least 30% of the markers out of five or more loci tested. Patients with colorectal cancer were also defined as MSI-H by immunohistochemistry (IHC) if they had loss of one or more markers.

[0178] Patients with non-MSI-H, stage 3L or later colon cancer and an ECOG activity index of 0-1 were selected for inclusion in the microsatellite stable (MSS) cohort. Selected MSS patients received 1 mg / kg nivolumab in combination with 1 mg / kg ipilimumab every 3 weeks for a total of four doses, followed by 3 mg / kg nivolumab every 2 weeks. After 6 weeks or more of this initial phase of treatment, MSS patients who tolerated the initial treatment regimen were divided into two treatment groups. In the first treatment group, 10 MSS patients received 1 mg / kg nivolumab in combination with 3 mg / kg ipilimumab every 3 weeks for a total of four doses, followed by 3 mg / kg nivolumab every 2 weeks. In the second treatment arm, 10 patients with MSS received 3 mg / kg nivolumab in combination with 1 mg / kg ipilimumab every 3 weeks for a total of four doses, followed by 3 mg / kg nivolumab every 2 weeks. Together, these treatment arms served as a separate safety arm in patients with MSS and informed the dose of nivolumab in combination with ipilimumab for use in patients with MSI-H. Figure 1 shows the overall trial design for patients in the MSS cohort.

[0179] Patients with second-line MSI-H colon cancer who had received at least one prior treatment for metastatic disease, had at least one target lesion, and had an ECOG activity index of 0 to 1 were selected for inclusion in the MSI-H cohort of patients. Nineteen MSI-H patients received nivolumab at 3 mg / kg every two weeks in the initial monotherapy stage of treatment (mStage 1). Some MSI-H patients who demonstrated a positive response were selected for inclusion in the combination therapy stage of treatment (cStage 1). These 19 MSI-H patients then received nivolumab at 3 mg / kg every three weeks for a total of four doses in cStage 1, followed by nivolumab at 3 mg / kg every two weeks. The majority of patients with positive responses from cStage 1 are currently enrolled in the second stage of combination therapy (cStage 2). In cStage 2, 48 patients (19 patients from cStage 1 + 29 additional patients) received 3 mg / kg nivolumab in combination with 1 mg / kg ipilimumab every 3 weeks for a total of 4 doses, followed by 3 mg / kg nivolumab every 2 weeks. Another group of patients who showed a positive response in mStage 1 were selected for inclusion in the second stage (mStage 2) of monotherapy. In mStage 2, 48 patients (19 patients from mStage 1 + 29 additional patients) received 3 mg / kg nivolumab every 2 weeks. Figure 2 shows the overall trial design for patients in the MSI-H cohort.

[0180] The primary endpoint of this trial was determination of investigator-assessed objective response rate (ORR) using RECIST 1.1 in patients with MSI-H. The secondary endpoint of this trial was assessment of independent radiological review committee-assessed ORR in patients with MSS. Exploratory endpoints of this trial included safety and tolerability, progression-free survival (PFS), overall survival (OS), investigator-assessed ORR, and biomarkers in patients with MSS.

[0181] Patient demographics collected for this trial included median age, number of patients under 65 years of age, number of male patients, number of patients of a particular race, and number of patients with an ECOG score of 0 or 1. Table 1 shows a comparison of the demographics of MSI-H patients who received 3 mg / kg nivolumab monotherapy and MSI-H patients who received 3 mg / kg nivolumab in combination with 1 mg / kg ipilimumab. Table 2 also shows a comparison of the demographics of MSS patients who received 1 mg / kg nivolumab in combination with 3 mg / kg ipilimumab and MSS patients who received 3 mg / kg nivolumab in combination with 1 mg / kg ipilimumab. [Table 2]

[0182] Data collected on patient profiles and prior treatment included the number of patients with specific disease stages at diagnosis, the number of patients with specific mutational status, the number of patients receiving prior treatment, the number of patients undergoing prior surgery, and the number of patients receiving prior radiation therapy. Table 3 shows the disease characteristics and prior treatments of MSI-H patients receiving 3 mg / kg nivolumab monotherapy compared with MSI-H patients receiving 3 mg / kg nivolumab in combination with 1 mg / kg ipilimumab. Table 3 also shows the disease characteristics and prior treatments of MSS patients receiving 1 mg / kg nivolumab in combination with 3 mg / kg ipilimumab compared with MSS patients receiving 3 mg / kg nivolumab in combination with 1 mg / kg ipilimumab. [Table 3]

[0183] Data on the disposition of MSI-H patients collected in this trial included the number of patients who continued treatment, the number of patients who did not continue treatment, and the number of patients who did not continue treatment due to disease progression, study drug toxicity, consent withdrawal, or other or unreported reasons. Table 4 shows the disposition of MSI-H patients who received 3 mg / kg nivolumab monotherapy compared with MSI-H patients who received 3 mg / kg nivolumab in combination with 1 mg / kg ipilimumab. [Table 4]

[0184] Investigator-assessed best overall response in MSI-H patients receiving 3 mg / kg nivolumab monotherapy was determined by objective response rate, median time to response, and median duration of response. As shown in Table 5, 12 of 47 patients (25.5%) demonstrated an objective response to 3 mg / kg nivolumab monotherapy, with a median time to response after treatment of 2.12 months. Figure 3 shows the percent change from baseline for individual patients receiving 3 mg / kg nivolumab monotherapy. [Table 5]

[0185] Investigator-assessed best overall response in MSI-H patients receiving 3 mg / kg nivolumab in combination with 1 mg / kg ipilimumab was determined by objective response rate, median time to response, and median duration of response. As shown in Table 6, 9 of 27 patients (33.3%) demonstrated an objective response with treatment with 3 mg / kg nivolumab in combination with 1 mg / kg ipilimumab, with a median time to response of 2.73 months after treatment. Figure 4 shows the percent change from baseline for individual patients receiving 3 mg / kg nivolumab in combination with 1 mg / kg ipilimumab. [Table 6]

[0186] The best reduction in target lesion size in MSI-H patients who received 3 mg / kg nivolumab monotherapy was compared to the best reduction in target lesion size in MSI-H patients who received 3 mg / kg nivolumab in combination with 1 mg / kg ipilimumab. As shown in Figure 5, 56% of patients who received 3 mg / kg nivolumab monotherapy showed a reduction in target lesion size from baseline, while 81% of patients who received 3 mg / kg nivolumab in combination with 1 mg / kg ipilimumab showed a reduction in target lesion size from baseline.

[0187] Investigator-assessed PFS was measured at 6, 9, and 12 months for MSI-H patients receiving 3 mg / kg nivolumab monotherapy and at 6 months for MSI-H patients receiving 3 mg / kg nivolumab in combination with 1 mg / kg ipilimumab. As shown in Table 7, 45.9% of patients receiving 3 mg / kg nivolumab monotherapy demonstrated progression-free survival at 6 months, while 66.6% of patients receiving 3 mg / kg nivolumab in combination with 1 mg / kg ipilimumab demonstrated progression-free survival at 6 months. Figure 6 compares the percentage of progression-free survival for patients receiving 3 mg / kg nivolumab monotherapy and 3 mg / kg nivolumab in combination with 1 mg / kg ipilimumab. [Table 7]

[0188] OS was measured at 6, 9, and 12 months for MSI-H patients receiving 3 mg / kg nivolumab monotherapy and at 6 and 9 months for MSI-H patients receiving 3 mg / kg nivolumab in combination with 1 mg / kg ipilimumab. As shown in Table 8, 75.0% of patients receiving 3 mg / kg nivolumab monotherapy demonstrated progression-free survival at 6 months, while 85.1% of patients receiving 3 mg / kg nivolumab in combination with 1 mg / kg ipilimumab demonstrated progression-free survival at 6 months. At 9 months, 65.6% of patients receiving 3 mg / kg nivolumab monotherapy demonstrated progression-free survival, while 85.1% of patients receiving 3 mg / kg nivolumab in combination with 1 mg / kg ipilimumab demonstrated progression-free survival. Figure 7 shows a comparison of the percentages showing overall survival for patients receiving 3 mg / kg nivolumab monotherapy and patients receiving 3 mg / kg nivolumab in combination with 1 mg / kg ipilimumab. [Table 8]

[0189] Efficacy in treating patients with MSS colorectal cancer was measured by ORR, PFS, and OS. Table 9 summarizes the efficacy data comparing MSS patients who received 1 mg / kg nivolumab in combination with 3 mg / kg ipilimumab with those who received 3 mg / kg nivolumab in combination with 1 mg / kg ipilimumab. [Table 9]

[0190] Treatment-related adverse events seen in patients with ≥15% MSI-H colorectal cancer included fatigue, diarrhea, pruritus, nausea, fever, and vomiting. Table 10 shows treatment-related adverse events in patients with ≥15% MSI-H comparing MSI-H patients receiving 3 mg / kg nivolumab with MSI-H patients receiving 3 mg / kg nivolumab in combination with 1 mg / kg ipilimumab. [Table 10]

[0191] Treatment-related adverse events observed in ≥15% of MSS colorectal cancer patients were diarrhea, asthenia, nausea, fever, vomiting, fatigue, dry skin, and cough.

[0192] The results of this trial demonstrated encouraging activity of nivolumab monotherapy in patients with MSI-H status. The combination of nivolumab and ipilimumab also showed promising preliminary activity. The results of this trial demonstrated durable responses to nivolumab monotherapy and nivolumab plus ipilimumab combination therapy. Nivolumab and the combination of nivolumab and ipilimumab demonstrated acceptable safety profiles consistent with clinical benefit, consistent with those observed in other solid tumors. These encouraging results support continued evaluation of nivolumab monotherapy and nivolumab plus ipilimumab combination therapy in patients with MSI-H metastatic colorectal cancer and other tumors with underlying mismatch repair deficiencies.

[0193] Example 2 Expanded clinical study of nivolumab plus ipilimumab in the treatment of patients with deficient DNA mismatch repair / microsatellite instability-high metastatic colorectal cancer Following the clinical trial described in Example 1, the efficacy and safety of nivolumab in combination with ipilimumab was investigated in an expanded population of patients with DNA mismatch repair deficient / microsatellite instability-high (dMMR / MSI-H) metastatic colorectal cancer (mCRC) who had received their first treatment dose at least 6 months prior to the data cutoff. Patients were eligible for inclusion in this trial based on the following criteria: 1) histologically confirmed metastatic / recurrent colorectal cancer; 2) dMMR / MSI-H as documented by a local testing laboratory; and 3) at least one prior line of therapy.

[0194] Figure 8 shows the overall trial design for patients included in the monotherapy or combination therapy arms of treatment. Specifically, patients included in the monotherapy arm received 3 mg / kg nivolumab every two weeks in Stage 1 treatment. If at least seven responders out of 19 patients were confirmed, enrollment continued for the second stage of treatment. In the second stage of treatment, patients received 3 mg / kg nivolumab every two weeks. Patients included in the other combination arm received 3 mg / kg nivolumab combined with 1 mg / kg ipilimumab every three weeks for four doses in Stage 1 treatment, followed by 3 mg / kg nivolumab every two weeks. If at least seven responders out of 19 patients were confirmed, enrollment continued for the second stage of treatment. In stage 2 treatment, patients received 3 mg / kg nivolumab in combination with 1 mg / kg ipilimumab every 3 weeks for 4 doses, followed by 3 mg / kg nivolumab every 2 weeks. Further details regarding the trial design are described in Overman M et al., J Clin Oncol. 2017; 35: (suppl 4S; abstract 519) and Overman M et al., Ann Oncol. 2016; 27 (6): 149-206 (abstract 479P).

[0195] The primary endpoint of this trial was the determination of investigator-assessed objective response rate (ORR) using RECIST 1.1. Other key endpoints included determination of ORR by blinded independent central review (BICR) and assessment of progression-free survival (PFS), overall survival (OS), and safety. Tumor imaging assessments were performed every 6 weeks for 24 weeks, then every 12 weeks until disease progression or discontinuation. Post-progression treatment was permitted if the patient was determined by the investigator to benefit from and tolerate the study treatment and if patient consent was obtained.

[0196] Investigator-assessed response to nivolumab monotherapy was evaluated in 74 patients. Nivolumab monotherapy demonstrated durable responses, sustained disease control, and a 74% 12-month overall survival (OS) rate. The objective response rate (ORR) for patients receiving nivolumab monotherapy was 31%, with a median time to response (TTR) of 2.8 months. The median duration of response (DOR) was not reached in patients receiving nivolumab monotherapy, and 83% (19 / 23) had an ongoing response. As shown in Figure 9, 62% of patients receiving nivolumab monotherapy demonstrated a reduction in tumor burden from baseline. Further details regarding nivolumab monotherapy are provided in Overman M et al., J Clin Oncol. 2017;35:(suppl 4S; abstract 519).

[0197] The following patient demographic profiles for this trial included median age, number of patients under 65 years of age, number of male patients, number of patients of a particular race, number of patients with an ECOG score of 0 or 1, number of patients with a particular disease stage at initial diagnosis, number of patients with a particular history of Lynch syndrome, number of patients with a particular BRAF or KRAS mutation status, number of patients with tumor PD-L1 expression of ≥ 1% or < 1% at baseline, number of patients who had received a particular number of prior lines of therapy, and number of patients who had received prior radiation therapy. Table 11 shows the baseline patient demographic profile and disease status of 84 patients with dMMR / MSI-H metastatic colorectal cancer who received the combination of 3 mg / kg nivolumab and 1 mg / kg ipilimumab. [Table 11]

[0198] Data on the disposition of dMMR / MSI-H metastatic colorectal cancer patients collected in this trial included the number of doses received, the number of patients continuing treatment, the number of patients who discontinued treatment, and the number of patients who discontinued treatment for specific reasons. The median time from first dose to data cutoff was 8.6 months, with a range of 6.3 to 19.4 months. Table 12 shows the patient disposition of 84 dMMR / MSI-H metastatic colorectal cancer patients who received the combination of 3 mg / kg nivolumab and 1 mg / kg ipilimumab. [Table 12]

[0199] Investigator-assessed objective response rate (ORR), best overall response, number of patients with disease control of ≥ 12 weeks, median time to response (TTR), and median duration of response (DOR) were measured in patients with dMMR / MSI-H metastatic colorectal cancer treated with nivolumab plus ipilimumab combination therapy. Best overall response was determined by measuring the number of patients with complete response, partial response, stable disease, or progressive disease. As shown in Table 13, investigator-assessed ORR was achieved in 55% of patients, and the disease control rate (DCR) was 79%. [Table 13]

[0200] The best reduction in target lesion size and change in tumor burden over time was determined in patients with dMMR / MSI-H metastatic colorectal cancer treated with nivolumab + ipilimumab combination therapy. As shown in Figure 10, 80% of patients demonstrated a reduction in tumor burden from baseline. The change in tumor burden over time by patient is shown in Figure 11. At the database cutoff, 85% (39 / 46) of responses were ongoing.

[0201] Progression-free survival (PFS) and overall survival (OS) were evaluated for patients with dMMR / MSI-H metastatic colorectal cancer treated with the nivolumab plus ipilimumab combination. The median time from first dose to death or last known alive date was 8.7 months for these patients, ranging from 0.1 to 20.1 months. As shown in Figure 12A, the 9-month PFS rate was 77%, while the 6-month and 12-month PFS rates were 77% (66.5%, 95% CI 85.1%), respectively. The median PFS in months was not reached at the time of data cutoff (11.47, 95% CI not estimable). As shown in Figure 12B, the 9-month OS rate was 88%, while the 6-month OS rate was 89% (80.2%, 95% CI 94.2%) and the 12-month OS rate was 88% (78.1%, 95% CI 93.1%). Median OS in months was not reached at the time of data cutoff (not estimable, 95% CI not estimable).

[0202] Treatment-related adverse events (TRAEs) observed in patients with dMMR / MSI-H metastatic colorectal cancer treated with the nivolumab plus ipilimumab combination included diarrhea, fatigue, elevated aspartate aminotransferase, fever, pruritus (itching), elevated alanine aminotransferase, nausea, hyperthyroidism, and hypothyroidism. All adverse events were manageable, with grade 3 / 4 TRAEs reported in 29% of patients. No treatment-related deaths were reported.

[0203] The results of this trial demonstrated that the combination of nivolumab and ipilimumab induced durable responses, sustained disease control, and favorable survival in previously treated patients with dMMR / MSI-H metastatic colorectal cancer. The combination also demonstrated a manageable safety profile. These encouraging results support continued evaluation of the combination of nivolumab and ipilimumab in patients with dMMR / MSI-H metastatic colorectal cancer and potentially other tumors.

Claims

1. An antibody or antigen-binding portion thereof ("anti-PD-1 antibody") that specifically binds to the programmed death-1 receptor (PD-1) and inhibits PD-1 activity, for use in a method of treating a subject having a tumor derived from colorectal cancer, wherein the tumor exhibits high-level microsatellite instability ("MSI-H").

2. The anti-PD-1 antibody for use according to claim 1, wherein the tumor is colon cancer or rectal cancer.

3. The tumor (a) the tumor contains germline alterations in at least two, at least three, at least four, or at least five DNA mismatch repair genes (“MMR genes”); (b) the tumor contains germline alterations in at least 30% of 5 or more MMR genes; (c) at least one protein encoded by a DNA MMR gene is not detectable in the tumor; and (d) Any combination of these The anti-PD-1 antibody for use according to claim 1 or 2, which exhibits one or more properties selected from the group consisting of:

4. 4. The anti-PD-1 antibody for use according to claim 3, wherein the DNA MMR genes include MSH2, MLH1, MSH6, PMS2, PMS1, or any combination thereof.

5. 5. The anti-PD-1 antibody for use according to any of claims 1 to 4, wherein (i) the subject exhibits progression-free survival for 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 12 months after administration; or (ii) the subject exhibits overall survival for 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 12 months after administration.

6. The anti-PD-1 antibody for use according to any one of claims 1 to 5, wherein the anti-PD-1 antibody cross-competes with nivolumab for binding to human PD-1.

7. The anti-PD-1 antibody for use according to any one of claims 1 to 6, wherein the anti-PD-1 antibody is nivolumab or pembrolizumab.

8. 8. The anti-PD-1 antibody for use according to any of claims 1 to 7, wherein the subject is further administered a therapeutically effective amount of an antibody or antigen-binding portion thereof that specifically binds to CTLA-4 and inhibits CTLA-4 activity (an "anti-CTLA-4 antibody").

9. 9. The anti-PD-1 antibody for use according to claim 8, wherein the anti-CTLA-4 antibody cross-competes with ipilimumab for binding to human CTLA-4.

10. The anti-PD-1 antibody for use according to claim 8 or 9, wherein the anti-CTLA-4 antibody is ipilimumab or tremelimumab.

11. The anti-PD-1 antibody for use according to any one of claims 8 to 10, wherein the anti-PD-1 antibody and / or anti-CTLA-4 antibody is administered at a dose ranging from at least about 0.1 mg / kg to at least about 10.0 mg / kg of body weight, approximately once every one, two, or three weeks.

12. 12. The anti-PD-1 antibody for use according to any of claims 8 to 11, wherein: (i) the anti-PD-1 antibody is administered at a dose of about 1 mg / kg body weight, and the anti-CTLA-4 antibody is administered at a dose of about 1 mg / kg body weight; (ii) the anti-PD-1 antibody is administered at a dose of about 1 mg / kg body weight, and the anti-CTLA-4 antibody is administered at a dose of about 3 mg / kg body weight; (iii) the anti-PD-1 antibody is administered at a dose of about 3 mg / kg body weight, and the anti-CTLA-4 antibody is administered at a dose of about 1 mg / kg body weight; or (iv) the anti-PD-1 antibody is administered at a dose of about 3 mg / kg body weight, and the anti-CTLA-4 antibody is administered at a dose of about 3 mg / kg body weight.

13. The anti-PD-1 antibody for use according to any one of claims 8 to 12, wherein administration of the anti-PD-1 antibody and the anti-CTLA-4 antibody is followed by anti-PD-1 antibody monotherapy.

14. The anti-PD-1 antibody for use according to any one of claims 8 to 13, wherein the anti-PD-1 antibody and the anti-CTLA-4 antibody are administered approximately once every 1 week, 2 weeks, 3 weeks, or 4 weeks.

15. 1. A kit for treating a patient having a tumor derived from colorectal cancer, wherein the tumor exhibits high microsatellite instability ("MSI-H"); (a) one dose of an anti-PD-1 antibody in the range of 0.1 mg / kg to 10 mg / kg of body weight; (b) one dose of an anti-CTLA-4 antibody in the range of 0.1 mg / kg to 10 mg / kg of body weight; and (c) Instructions for using the anti-PD-1 antibody and anti-CTLA-4 antibody according to any one of claims 8 to 14. Includes a kit.

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