Treatment of renal cancer using combination of Anti-PD-1 antibody and another Anti-cancer agent

Abstract

To provide a method for treating a subject afflicted with renal cancer.SOLUTION: The method comprises administering to the subject therapeutically effective amounts of: (a) an anti-cancer agent which is an antibody or an antigen-binding portion thereof that specifically binds to a Programmed Death-1 (PD-1) receptor and inhibits PD-1 activity; and (b) another anti-cancer agent. The other anti-cancer agent may be an anti-angiogenic tyrosine kinase inhibitor or an anti-Cytotoxic T-Lymphocyte Antigen-4 (CTLA-4) antibody. The disclosure also provides a kit for treating a subject afflicted with a renal cancer, the kit comprising a dosage of an anti-PD-1 antibody, a dosage of another anti-cancer agent which is an anti-angiogenic tyrosine kinase inhibitor or an anti-CTLA-4 antibody, and instructions for using the anti-PD-1 antibody and the other anti-cancer agent in any of the disclosed methods for treating a renal cancer.SELECTED DRAWING: Figure 2

Description

[Technical field]

[0001] Throughout this specification, various publications are referenced by author name and date in parentheses or patent or patent publication number. Full citations for these publications appear at the end of the specification. The descriptions of these publications are incorporated herein by reference in their entireties in order to more fully describe the state of the art at the time of writing. However, citation of these publications herein should not be construed as an admission that such publications are prior art to the present invention.

[0002] FIELD OF THEINVENTION The present invention relates to a method of treating renal cancer in a subject, comprising administering a combination of an anti-cancer agent that is an anti-programmed cell death (PD-1) antibody and another anti-cancer agent. In some embodiments, the other anti-cancer agent is an anti-cytotoxic T-lymphocyte antigen-4 (CTLA-4) antibody, an anti-angiogenic tyrosine kinase inhibitor, or a combination thereof. [Background technology]

[0003] 2. Background of the Invention Human cancers possess numerous genetic and epigenetic alterations that produce novel antigens potentially recognizable by the immune system (Sjoblom et al., 2006). The adaptive immune system, consisting of T and B lymphocytes, has a strong anticancer potential with a broad capacity and exquisite specificity to respond to a wide variety of tumor antigens. Furthermore, the immune system exhibits considerable plasticity and memory components. The successful exploitation of all these properties of the adaptive immune system makes immunotherapy unique among all cancer treatment approaches.

[0004] Until recently, cancer immunotherapy has focused substantially on approaches to enhance antitumor immune responses through adoptive transfer of activated effector cells, immunization against appropriate antigens, or provision of nonspecific immune stimulants such as cytokines. In the last decade, however, intensive efforts to develop specific immune checkpoint pathway inhibitors have led to the development of an antibody (Ab) that binds to and blocks cytotoxic T-lymphocyte antigen-4 (CTLA-4), ipilimumab (Yervoy), for the treatment of patients with advanced melanoma. (登録商標) ) (Hodi et al., 2010) and the development of Abs such as nivolumab and pembrolizumab (formerly lambrolizumab; USAN Council Statement, 2013) that specifically bind to the programmed cell death (PD-1) receptor and block the PD-1 / PD-1 inhibitory ligand pathway (Brahmer et al., 2010; Topalian et al., 2012a; Topalian et al., 2014; Hamid et al., 2013; Brahmer et al., 2012; Flies et al., 2011; Pardoll, 2012; Hamid and Carvajal, 2013).

[0005] PD-1 is an important immune checkpoint receptor expressed by activated T and B cells and mediates immune suppression. PD-1 is a member of the CD28 family of receptors that 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 as well as many human cancers and have been shown to downregulate T cell activation and cytokine secretion upon binding to PD-1. Inhibition of the PD-1 / PD-L1 interaction mediates potent antitumor activity in preclinical models (U.S. Patents Nos. 8,008,449 and 7,943,743), and Ab inhibitors of the PD-1 / PD-L1 interaction for the treatment of cancer have entered clinical trials (Brahmer et al., 2010; Topalian et al., 2012a; Topalian et al., 2014; Hamid et al., 2013; Brahmer et al., 2012; Flies et al., 2011; Pardoll, 2012; Hamid and Carvajal, 2013).

[0006] Nivolumab (previously called 5C4, BMS-936558, MDX-1106 or ONO-4538) is a fully human IgG4(S228P)PD-1 immune checkpoint inhibitor Ab that selectively blocks interaction with PD-1 ligands (PD-L1 and PD-L2), thereby blocking downregulation of antitumor T cell function (US Patent No. 8,008,449; Wang et al., 2014). Nivolumab has shown activity in a variety of advanced solid tumors, including renal cell carcinoma (RCC; renal adenocarcinoma or adrenal tumor), melanoma and non-small cell lung cancer (Topalian et al., 2012a; Topalian et al., 2014; Drake et al., 2013; WO2013 / 173223).

[0007] Ipilimumab (Yervoy®) is a fully human, IgG1 monoclonal Ab that blocks the binding of CTLA-4 to its B7 ligand, thereby stimulating T cell activation and improving overall survival (OS) in patients with advanced melanoma (Hodi et al., 2010). Current treatment with nivolumab and ipilimumab in phase 1 clinical trials has resulted in rapid and robust tumor regression in a significant proportion of patients with advanced melanoma and was significantly more effective than either Ab alone (Wolchok et al., 2013; WO2013 / 173223). However, it is not known to date whether this combination of immunomodulatory Abs is similarly effective in other tumor types.

[0008] Renal cancer (also known as kidney cancer) is cancer that originates in the kidney. The most common type of renal cancer is renal cell carcinoma (RCC). If RCC is diagnosed and treated while localized in the kidney and immediately surrounding tissues (Stage I), it can often be cured by surgical resection, and total resection is permitted and often the curative treatment in stage II and stage III RCC. In contrast, when distant metastases are present (Stage IV), disease-free survival is poor. Furthermore, the prognosis of any treated RCC patients with progressive, recurrent or recurrent disease is also poor, regardless of cell type or stage. Approximately 25%-30% of RCC patients have metastatic disease at the time of diagnosis, and the median survival time for metastatic RCC is only about 24 months (Gupta et al., 2008; NCCN GUIDELINES®, Version 3.2014 - Kidney Cancer; Heng et al., 2009). Responses to cytotoxic chemotherapy agents generally do not exceed 10% for any regimen tested in an adequate number of patients. However, an improved understanding of the biology of RCC has led to the development and Food and Drug Administration (FDA) approval of seven novel agents targeting specific proliferation pathways. Two of the commercialized targeted therapeutic agents, temsirolimus and everolimus, block the mammalian target of rapamycin (mTOR), a serine / threonine protein kinase that controls cell growth, division, and survival. Several agents have been developed that target the vascular endothelial growth factor (VEGF)-mediated proangiogenic pathway, including five oral tyrosine kinase inhibitors (TKIs), pazopanib, sorafenib, sunitinib, axitinib, and tivozanib, and the anti-VEGF monoclonal antibody (mAb), bevacizumab. These agents show clinical efficacy and improved progression-free survival (PFS), but durable responses are rare. VEGF TKIs have been shown to suppress immune-inhibitory regulatory T cells (Tregs) and myeloid-derived suppressor cells (MDSCs), making the immune environment more conducive to T cell-mediated antitumor activity. Furthermore, targeting the VEGF axis may attenuate RCC-induced immunosuppression, making tumors more responsive to immune modulation when used in combination, thereby resulting in greater and more durable therapeutic effects. [Prior art documents] [Patent documents]

[0009] [Patent Document 1] International Publication No. WO 2012 / 145493 [Patent Document 2] International Publication No. 2013 / 173223 [Patent Document 3] U.S. Pat. No. 5,977,318 [Patent Document 4] U.S. Patent No. 6,051,227 [Patent Document 5] U.S. Patent No. 6,682,736 [Patent Document 6] U.S. Patent No. 6,808,710 [Patent Document 7] U.S. Patent No. 6,984,720 [Patent Document 8] U.S. Patent No. 7,034,121 [Patent Document 9] U.S. Patent No. 7,605,238 [Patent Document 10] U.S. Patent No. 7,488,802 [Patent Document 11] U.S. Pat. No. 7,943,743 [Patent Document 12] U.S. Pat. No. 8,008,449 [Patent Document 13] U.S. Pat. No. 8,168,757 [Patent Document 14] U.S. Pat. No. 8,217,149 [Patent Document 15] U.S. Pat. No. 8,354,509 [Non-patent literature]

[0010] [Non-Patent Document 1] Amin et al. (2014) Nivolumab (anti-PD-1; BMS-936558, ONO-4538) in combination with sunitinib or pazopanib in patients (pts) with metastatic renal cell carcinoma (mRCC). J Clin Oncol 32:5s, (suppl; Abstract 5010, Clinical Science Symposium).

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[0011] As described herein, the activity of nivolumab in combination with two anti-angiogenic (anti-VEGF) TKIs, sunitinib (Sutent®) or sorafenib (Nexavar®), has been tested in preclinical RCC mouse models. Clinical data for the combination of nivolumab with sunitinib, pazopanib (Votrient®) or ipilimumab is also provided herein. [Means for solving the problem]

[0012] Summary of the Invention The present invention provides a method for treating a subject suffering from renal cancer, comprising administering to the subject a therapeutically effective amount of a combination of (a) an Ab or antigen-binding portion thereof that specifically binds and inhibits PD-1; and (b) another anti-cancer agent. In some embodiments, the other anti-cancer agent is (i) an Ab or antigen-binding portion thereof that specifically binds and inhibits CTLA-4, (ii) an anti-angiogenic TKI, or (iii) a combination thereof. In some embodiments, the renal cancer is RCC. In some embodiments of any of the therapeutic methods disclosed herein, the anti-PD-1 Ab is nivolumab. In other embodiments, the anti-PD-1 Ab is pembrolizumab. In some other embodiments of any of the therapeutic methods disclosed herein, the anti-CTLA-4 Ab is ipilimumab. In other embodiments, the anti-CTLA-4 Ab is tremelimumab. In still other embodiments, the TKI is sunitinib, pazopanib, sorafenib, axitinib, or tivozanib.

[0013] In some embodiments, the subject is pretreated for renal cancer. In other embodiments, the renal cancer is advanced, metastatic and / or refractory cancer. In some embodiments, administration of the combination of Ab or its antigen-binding portion with TKI induces a durable clinical response in the subject.

[0014] In some embodiments, administration of a combination of an anti-PD-1 Ab, or antigen-binding portion thereof, and another anti-cancer agent results in increased T cell infiltration of renal cancer tissue or tumor compared to untreated subjects or subjects treated with an anti-PD-1 Ab, or antigen-binding portion thereof, or another anti-cancer agent alone. In other embodiments, the increased T cell infiltration is due to increased CD4 T cell proliferation in renal cancer tissue or tumor. + T cells, CD8 + In yet another embodiment, administration of the anti-PD-1 Ab, or antigen-binding portion thereof, in combination with another anti-cancer agent results in increased T cell proliferation. In another embodiment, administration of the anti-PD-1 Ab, or antigen-binding portion thereof, in combination with another anti-cancer agent results in a reduction in T regulatory cells compared to untreated subjects or subjects treated with the anti-PD-1 Ab, or antigen-binding portion thereof, or another anti-cancer agent alone.

[0015] In some embodiments, administration of the combination of an anti-PD-1 Ab, or antigen-binding portion thereof, and another anti-cancer agent reduces the number of monocytic myeloid-derived suppressor cells. In some embodiments, administration of the combination of an anti-PD-1 Ab, or antigen-binding portion thereof, and another anti-cancer agent increases the number of granulocytic myeloid cells compared to untreated subjects or subjects treated with an anti-PD-1 Ab, or antigen-binding portion thereof, or another anti-cancer agent alone. In some embodiments, the monocytic myeloid-derived suppressor cells are CD11b + / Ly6C hi / Ly6G - Expression of CD11b + / Ly6C low / Ly6G - In other embodiments, the granulocytic myeloid cells are characterized by expression of CD11b + / Ly6C - / Ly6G + It is characterized by the expression of

[0016] In some embodiments of the methods involving the use of a combination of an anti-PD-1 Ab or antigen-binding portion thereof and a TKI, a therapeutically effective amount of the anti-PD-1 Ab or antigen-binding portion thereof ranges from about 0.1 to about 10.0 mg / kg body weight administered by intravenous infusion about once every 2 to 4 weeks, hi some embodiments, the anti-PD-1 Ab or antigen-binding portion thereof is administered at a dose of about 2 mg / kg or about 5 mg / kg once every 3 weeks.

[0017] For combinations of anti-PD-1 Ab and anti-CTLA-4 Ab, in some embodiments, the method comprises (a) an induction phase comprising four doses of anti-PD-1 Ab and anti-CTLA-4 Ab administered three weeks apart at the following doses: (i) about 1 mg / kg anti-PD-1 Ab and about 3 mg / kg anti-CTLA-4 Ab; (ii) about 3 mg / kg anti-PD-1 Ab and about 3 mg / kg anti-CTLA-4 Ab; or (iii) about 3 mg / kg anti-PD-1 Ab and about 1 mg / kg anti-CTLA-4 Ab; and (b) a maintenance phase comprising repeated administration of anti-PD-1 Ab at a dose of about 3 mg / kg every two weeks.

[0018] The invention also provides kits for treating a subject suffering from renal cancer, the kit comprising: (a) a dose of an Ab or antigen-binding portion thereof that specifically binds and inhibits PD-1; (b) a dose of one of (i) an Ab or antigen-binding portion thereof that specifically binds and inhibits CTLA-4, or (ii) an anti-angiogenic TKI, e.g., sunitinib or pazopanib; and (c) instructions for using the anti-PD-1 Ab and anti-CTLA-4 Ab or TKI to treat the subject.

[0019] Other features and advantages of the present invention will be apparent from the following detailed description and examples, which should not be construed as limiting. The contents of all references cited throughout this application, including scientific articles, journal articles, GenBank entries, patents, and patent applications, are expressly incorporated herein by reference. [Brief description of the drawings]

[0020] [Figure 1] FIG. 1 shows the effect of treatment with the following therapeutic agents on mean tumor volume increase: control (filled circles), sunitinib (open circles), sorafenib (filled triangles), anti-PD-1 (filled inverted triangles), sunitinib + anti-PD-1 (open squares) and sorafenib + anti-PD-1 (filled diamonds).

[0021] [Diagram 2] FIG. 2 shows the time for tumors treated with control (closed circles), sunitinib (open circles), sorafenib (closed triangles), anti-PD-1 (closed inverted triangles), sunitinib + anti-PD-1 (open squares) and sorafenib + anti-PD-1 (closed diamonds) to reach a volume of 500 mm3.

[0022] [Diagram 3] FIG. 3 shows the effect of treatment with the following therapeutic agents on percentage change in body weight: control (filled circles), sunitinib (open circles), sorafenib (filled triangles), anti-PD-1 (filled inverted triangles), sunitinib + anti-PD-1 (open squares) and sorafenib + anti-PD-1 (filled diamonds).

[0023] [Figure 4]FIG. 4 shows CD4+ and CD8+ T cell infiltration into Renca tumors following treatment with control, anti-PD-1 alone, sunitinib alone, or anti-PD-1 in combination with sunitinib.

[0024] [Diagram 5] FIG. 5 shows assessment of the composition of tumor-infiltrating CD4+ immune cells by flow cytometry following treatment with sunitinib, sorafenib, or anti-PD-1 Ab alone or in combination with sunitinib or sorafenib in the Renca tumor model.

[0025] [Figure 6] FIG. 6 shows assessment of the composition of tumor-infiltrating CD8+ immune cells by flow cytometry following treatment with sunitinib, sorafenib, or anti-PD-1 Ab alone or in combination with sunitinib or sorafenib in the Renca tumor model.

[0026] [Figure 7] Figure 7 shows assessment of bone marrow derived tumor infiltrating cell subsets (CD11b+ / Ly6Chi / Ly6G-, CD11b+ / Ly6Clow / Ly6G- or CD11b+ / Ly6C- / Ly6G+) following treatment with sunitinib, sorafenib or anti-PD-1 Ab alone or in combination with sunitinib or sorafenib in the Renca tumor model. Gating: cells→singlets→Cd45+ / viable→CD11b+.

[0027] [Figure 8] Figure 8 shows the effect of anti-PD-1 mAb and sunitinib combination on peptide-specific cytotoxicity response in CT26 tumor model. X-axis shows percentage of cell death. Each bar shows from top to bottom: (i) control, (ii) anti-PD-1 10mg / kg, Q3Dx2 IP, (iii) sunitinib 120mg / kg, QDx5, PO and (iv) anti-PD-1 + sunitinib.

[0028] [Figure 9]Figure 9 shows the effect of anti-PD-1 mAb and sunitinib combination on tumor volume in the CT26 tumor model. Tumor volume (mm3) is shown on the y-axis for control, anti-PD-1 10 mg / kg Q3D x IP, sunitinib 120 mg / kg QD x 5 PO and anti-PD-1 + sunitinib treatments.

[0029] [Figure 10A]Figures 10A and 10B show the Phase 1 study design. Figures 10A and 10B show the study design of the combination of nivolumab and sunitinib or pazopanib (A) or nivolumab and ipilimumab (B) in subjects with advanced or metastatic RCC (mRCC), respectively (NCT01472081). *For advanced / metastatic RCC, subjects who had received at least one systemic treatment regimen but had not previously received pazopanib or sunitinib were assigned to two dose-escalation arms in an alternating fashion when both arms were open. If only one arm was open, these subjects were assigned to the open arm. **Until disease progression (PD) or drug-related toxicity led to discontinuation or until the subject withdrew consent. ***Subjects who had received at least one prior systemic treatment but were not qualified in the escalation arms S and / or P when they were open were assigned to the two ipilimumab combination arms (I-1 and I-3) in an alternating fashion when both were open. If only one arm was open, these subjects were assigned to the open arm. #Untreated subjects were randomly assigned in a 1:1:1:1 ratio to one of the four arms (expansion arms S and P and I-1 and I-3) if all arms were open, until full enrollment was completed. ##Until disease progression (PD) or drug-related toxicity led to discontinuation or the subject withdrew consent. Treatment beyond the first investigator-assessed RECIST 1.1-defined progression was considered in subjects who experienced investigator-assessed clinical benefit and tolerated the investigational agent. Such subjects were required to discontinue treatment when further progression was reported. ###For the I-1 and I-3 cohort expansion and IN-3 dose accrual cohorts, one prior adjuvant or neoadjuvant therapy for localized or locally advanced RCC was allowed as long as recurrence occurred ≥6 months after the last dose of adjuvant or neoadjuvant therapy. Prior cytokine-based treatment for RCC [e.g., interferon-alpha (IFN-α) or interleukin-2 (IL-2)] as prior therapy was allowed.These minimally treated (including treatment-naïve) subjects were randomly assigned to one of three arms (I-1, I-3, and IN-3) in a 1:1:1 ratio if all arms were open until full enrollment was completed. [Figure 10B]Figures 10A and 10B show the Phase 1 study design. Figures 10A and 10B show the study design of the combination of nivolumab and sunitinib or pazopanib (A) or nivolumab and ipilimumab (B) in subjects with advanced or metastatic RCC (mRCC), respectively (NCT01472081). *For advanced / metastatic RCC, subjects who had received at least one systemic treatment regimen but had not previously received pazopanib or sunitinib were assigned to two dose-escalation arms in an alternating fashion when both arms were open. If only one arm was open, these subjects were assigned to the open arm. **Until disease progression (PD) or drug-related toxicity led to discontinuation or until the subject withdrew consent. ***Subjects who had received at least one prior systemic treatment but were not qualified in the escalation arms S and / or P when they were open were assigned to the two ipilimumab combination arms (I-1 and I-3) in an alternating fashion when both were open. If only one arm was open, these subjects were assigned to the open arm. #Untreated subjects were randomly assigned in a 1:1:1:1 ratio to one of the four arms (expansion arms S and P and I-1 and I-3) if all arms were open, until full enrollment was completed. ##Until disease progression (PD) or drug-related toxicity led to discontinuation or the subject withdrew consent. Treatment beyond the first investigator-assessed RECIST 1.1-defined progression was considered in subjects who experienced investigator-assessed clinical benefit and tolerated the investigational agent. Such subjects were required to discontinue treatment when further progression was reported. ###For the I-1 and I-3 cohort expansion and IN-3 dose accrual cohorts, one prior adjuvant or neoadjuvant therapy for localized or locally advanced RCC was allowed as long as recurrence occurred ≥6 months after the last dose of adjuvant or neoadjuvant therapy. Prior cytokine-based treatment for RCC [e.g., interferon-alpha (IFN-α) or interleukin-2 (IL-2)] as prior therapy was allowed.These minimally treated (including untreated) subjects were randomly assigned to one of three arms (I-1, I-3, and IN-3) in a 1:1:1 ratio if all arms were open until full enrollment was completed.

[0030] [Figure 11] Figure 11 shows the dose escalation performed in the S and P arms of the nivolumab combination trial. The S arm consisted of 7 pretreated subjects who received S+N2; 7 pretreated subjects who received S+N5; and 19 untreated patients who entered the S+N5 expansion. The P arm consisted of 20 pretreated subjects who received P+N2. There was no expansion of the P arm.

[0031] [Figure 12A] Figures 12A-D show clinical activity of RCC subjects treated with a combination of nivolumab and sunitinib or pazopanib. Radar charts show the percentage change from baseline in tumor burden, measured as a two-way sum of products of all target lesions, in subjects receiving various regimens. Figure 12A shows a regimen of sunitinib and 2 mg / kg nivolumab. Figure 12B shows a regimen of sunitinib and 5 mg / kg nivolumab. Figure 12C shows a regimen of pazopanib and 2 mg / kg nivolumab. "+" indicates the first occurrence of new lesions. Figure 12D is a waterfall plot showing the maximum response percentage in baseline target lesions of subjects in the S and P arms. Subjects with baseline target lesions and at least one post-baseline target lesion assessment are shown. A positive change in tumor burden indicates tumor growth; a negative change in tumor burden indicates tumor shrinkage. In the radar charts and waterfall plots, horizontal lines indicate a 30% decrease (RECIST 1.1 threshold for PR) and a 20% increase (RECIST 1.1 threshold for PD). Not all decreases of 30% or more from baseline are PR. [Figure 12B]Figures 12A-D show clinical activity of RCC subjects treated with a combination of nivolumab and sunitinib or pazopanib. Radar charts show the percentage change from baseline in tumor burden, measured as a two-way sum of products of all target lesions, in subjects receiving various regimens. Figure 12A shows a regimen of sunitinib and 2 mg / kg nivolumab. Figure 12B shows a regimen of sunitinib and 5 mg / kg nivolumab. Figure 12C shows a regimen of pazopanib and 2 mg / kg nivolumab. "+" indicates the first occurrence of new lesions. Figure 12D is a waterfall plot showing the maximum response percentage in baseline target lesions of subjects in the S and P arms. Subjects with baseline target lesions and at least one post-baseline target lesion assessment are shown. A positive change in tumor burden indicates tumor growth; a negative change in tumor burden indicates tumor shrinkage. In the radar charts and waterfall plots, horizontal lines indicate a 30% decrease (RECIST 1.1 threshold for PR) and a 20% increase (RECIST 1.1 threshold for PD). Not all decreases of 30% or more from baseline are PR. [Figure 12C]Figures 12A-D show clinical activity of RCC subjects treated with a combination of nivolumab and sunitinib or pazopanib. Radar charts show the percentage change from baseline in tumor burden, measured as a two-way sum of products of all target lesions, in subjects receiving various regimens. Figure 12A shows a regimen of sunitinib and 2 mg / kg nivolumab. Figure 12B shows a regimen of sunitinib and 5 mg / kg nivolumab. Figure 12C shows a regimen of pazopanib and 2 mg / kg nivolumab. "+" indicates the first occurrence of new lesions. Figure 12D is a waterfall plot showing the maximum response percentage in baseline target lesions of subjects in the S and P arms. Subjects with baseline target lesions and at least one post-baseline target lesion assessment are shown. A positive change in tumor burden indicates tumor growth; a negative change in tumor burden indicates tumor shrinkage. In the radar charts and waterfall plots, horizontal lines indicate a 30% decrease (RECIST 1.1 threshold for PR) and a 20% increase (RECIST 1.1 threshold for PD). Not all decreases of 30% or more from baseline are PR. [Figure 12D]Figures 12A-D show clinical activity of RCC subjects treated with a combination of nivolumab and sunitinib or pazopanib. Radar charts show the percentage change from baseline in tumor burden, measured as a two-way sum of products of all target lesions, in subjects receiving various regimens. Figure 12A shows a regimen of sunitinib and 2 mg / kg nivolumab. Figure 12B shows a regimen of sunitinib and 5 mg / kg nivolumab. Figure 12C shows a regimen of pazopanib and 2 mg / kg nivolumab. "+" indicates the first occurrence of new lesions. Figure 12D is a waterfall plot showing the maximum response percentage in baseline target lesions of subjects in the S and P arms. Subjects with baseline target lesions and at least one post-baseline target lesion assessment are shown. A positive change in tumor burden indicates tumor growth; a negative change in tumor burden indicates tumor shrinkage. In the radar charts and waterfall plots, horizontal lines indicate a 30% decrease (RECIST 1.1 threshold for PR) and a 20% increase (RECIST 1.1 threshold for PD). Not all decreases of 30% or more from baseline are PR.

[0032] [Figure 13] Figure 13 shows the progression-free survival (PFS) rate plotted against time for subjects treated with nivolumab and sunitinib (S+N) or nivolumab and pazopanib (P+N). Symbols represent censored observations. The number of patients at risk listed is the number at risk before entering the period. Tx: treatment.

[0033] [Figure 14A]Figures 14A-14D show clinical activity of RCC subjects treated with a combination of ipilimumab and nivolumab. Radar charts show the percentage change from baseline in tumor burden, measured as a two-way sum of products of all target lesions, in subjects receiving various regimens. Figure 14A shows a regimen of 3 mg / kg ipilimumab and 1 mg / kg nivolumab. Figure 14B shows a regimen of 1 mg / kg ipilimumab and 3 mg / kg nivolumab. Figure 14C shows a regimen of 3 mg / kg ipilimumab and 3 mg / kg nivolumab. "+" indicates the first occurrence of new lesions. Figure 14D is a waterfall plot showing the maximum response percentage in baseline target lesions of subjects in the I-1, I-3 and IN-3 arms. Subjects with baseline target lesions and at least one post-baseline target lesion assessment are shown. A positive change in tumor burden indicates tumor growth; a negative change in tumor burden indicates tumor shrinkage. In the radar charts and waterfall plots, horizontal lines indicate a 30% decrease (RECIST 1.1 threshold for PR) and a 20% increase (RECIST 1.1 threshold for PD). Not all decreases of 30% or more from baseline are PR. [Figure 14B]Figures 14A-14D show clinical activity of RCC subjects treated with a combination of ipilimumab and nivolumab. Radar charts show the percentage change from baseline in tumor burden, measured as a two-way sum of products of all target lesions, in subjects receiving various regimens. Figure 14A shows a regimen of 3 mg / kg ipilimumab and 1 mg / kg nivolumab. Figure 14B shows a regimen of 1 mg / kg ipilimumab and 3 mg / kg nivolumab. Figure 14C shows a regimen of 3 mg / kg ipilimumab and 3 mg / kg nivolumab. "+" indicates the first occurrence of new lesions. Figure 14D is a waterfall plot showing the maximum response percentage in baseline target lesions of subjects in the I-1, I-3 and IN-3 arms. Subjects with baseline target lesions and at least one post-baseline target lesion assessment are shown. A positive change in tumor burden indicates tumor growth; a negative change in tumor burden indicates tumor shrinkage. In the radar charts and waterfall plots, horizontal lines indicate a 30% decrease (RECIST 1.1 threshold for PR) and a 20% increase (RECIST 1.1 threshold for PD). Not all decreases of 30% or more from baseline are PR. [Figure 14C]Figures 14A-14D show clinical activity of RCC subjects treated with a combination of ipilimumab and nivolumab. Radar charts show the percentage change from baseline in tumor burden, measured as a two-way sum of products of all target lesions, in subjects receiving various regimens. Figure 14A shows a regimen of 3 mg / kg ipilimumab and 1 mg / kg nivolumab. Figure 14B shows a regimen of 1 mg / kg ipilimumab and 3 mg / kg nivolumab. Figure 14C shows a regimen of 3 mg / kg ipilimumab and 3 mg / kg nivolumab. "+" indicates the first occurrence of new lesions. Figure 14D is a waterfall plot showing the maximum response percentage in baseline target lesions of subjects in the I-1, I-3 and IN-3 arms. Subjects with baseline target lesions and at least one post-baseline target lesion assessment are shown. A positive change in tumor burden indicates tumor growth; a negative change in tumor burden indicates tumor shrinkage. In the radar charts and waterfall plots, horizontal lines indicate a 30% decrease (RECIST 1.1 threshold for PR) and a 20% increase (RECIST 1.1 threshold for PD). Not all decreases of 30% or more from baseline are PR. [Figure 14D]Figures 14A-14D show clinical activity of RCC subjects treated with a combination of ipilimumab and nivolumab. Radar charts show the percentage change from baseline in tumor burden, measured as a two-way sum of products of all target lesions, in subjects receiving various regimens. Figure 14A shows a regimen of 3 mg / kg ipilimumab and 1 mg / kg nivolumab. Figure 14B shows a regimen of 1 mg / kg ipilimumab and 3 mg / kg nivolumab. Figure 14C shows a regimen of 3 mg / kg ipilimumab and 3 mg / kg nivolumab. "+" indicates the first occurrence of new lesions. Figure 14D is a waterfall plot showing the maximum response percentage in baseline target lesions of subjects in the I-1, I-3 and IN-3 arms. Subjects with baseline target lesions and at least one post-baseline target lesion assessment are shown. A positive change in tumor burden indicates tumor growth; a negative change in tumor burden indicates tumor shrinkage. In the radar charts and waterfall plots, horizontal lines indicate a 30% decrease (RECIST 1.1 threshold for PR) and a 20% increase (RECIST 1.1 threshold for PD). Not all decreases of 30% or more from baseline are PR.

[0034] [Figure 15] Figure 15 shows a comparison of waterfall plots of subjects in S, P and I arms. Subjects with baseline target lesions and at least one post-baseline target lesion assessment are shown. A positive change in tumor burden indicates tumor growth; a negative change in tumor burden indicates tumor shrinkage. Horizontal lines indicate a 30% decrease (RECIST 1.1 threshold for PR) and a 20% increase (RECIST 1.1 threshold for PD). Not all decreases of 30% or more from baseline are PRs.

[0035] [Figure 16]Figure 16 shows the proportion of PFS plotted against time (weeks) from first dose for subjects treated with 1 mg / kg ipilimumab + 3 mg / kg nivolumab (IPI1 + INV3) (stars), 3 mg / kg ipilimumab + 1 mg / kg nivolumab (IPI3 + NIV1) (circles), and 3 mg / kg nivolumab + 3 mg / kg ipilimumab (IPI3 + NIV3) (triangles). Symbols represent censored observations. The number of patients at risk listed is the number at risk prior to entering the time period. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0036] Detailed Description of the Invention The present invention relates to a method of treating a patient with renal cancer, comprising administering to the patient a combination of an anti-PD-1 Ab or antigen-binding fragment thereof and another anti-cancer agent, in one embodiment, the other anti-cancer agent is an anti-CTLA-4 Ab, an anti-angiogenic TKI, or any combination thereof.

[0037] term So that the present invention may be more readily understood, certain terms are first defined. As used herein, unless expressly provided otherwise herein, each of the following terms has the meaning set forth below. Additional definitions are set forth throughout the specification.

[0038] "Administering" refers to the physical introduction of a composition containing a therapeutic agent into a subject using any of a variety of methods and delivery systems known to those of skill in the art. Preferred routes of administration for anti-PD-1 Ab include intravenous, intramuscular, subcutaneous, intraperitoneal, spinal or other parenteral routes of administration, for example by injection or infusion. The term "parenteral administration" as used herein generally refers to modes of administration other than enteral and topical administration by injection, including, but 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. TKIs are generally administered by a non-parenteral route, preferably orally. Other non-parenteral routes include topical, epithelial or mucosal routes of administration, for example, intranasal, vaginal, rectal, sublingual or topical. Administration may be, for example, once, multiple times and / or over one or more extended periods.

[0039] As used herein, an "adverse event" (AE) is any unfavourable and generally unintended or undesirable sign (including laboratory findings), symptom or disease associated with the use of a medical treatment. For example, an adverse event may be associated with activation of the immune system or expansion of immune system cells (e.g., T cells) in response to the treatment. A medical treatment may be associated with one or more AEs, and the severity level of each AE may be the same or different. Reference to a method that can "modify an adverse event" refers to a treatment regimen that reduces the incidence and / or severity of one or more AEs associated with the use of a different treatment regimen.

[0040] "Antibody" (Ab) refers to a glycoprotein immunoglobulin or antigen-binding portion thereof that specifically binds 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 includes three constant domains, C H1 , C H2 and C H3 Each light chain comprises a light chain variable region (herein V LThe light chain constant region comprises one constant domain, C L Includes: V H and V L The regions can be further subdivided into regions of hypervariability called complementarity determining regions (CDRs) interspersed with more conserved regions called framework regions (FRs). H and V L contains 3 CDRs and 4 FRs, arranged from amino terminus to carboxy terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable regions of the heavy and light chains contain binding domains that interact with antigens. The constant regions of the Abs may mediate the binding of immunoglobulins to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (C1q) of the classical complement system.

[0041] Immunoglobulins may be derived from any of the commonly known isotypes, including, but not limited to, IgA, secretory IgA, IgG, and IgM. IgG subclasses are also well known to those skilled in the art, including, but not limited to, human IgG1, IgG2, IgG3, and IgG4. "Isotype" refers to the Ab class or subclass (e.g., IgM or IgG1) encoded by the heavy chain constant region genes. The term "antibody" includes, by way of example, both naturally occurring and non-naturally occurring Abs; monoclonal and polyclonal Abs; chimeric and humanized Abs; human or non-human Abs; fully synthetic Abs; and single-chain Abs. Non-human Abs may be recombinantly humanized 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 above immunoglobulins, including monovalent and bivalent fragments or portions and single-chain Abs.

[0042] An "isolated antibody" refers to an Ab that is substantially free of other Abs having different antigen specificities (e.g., an isolated Ab that specifically binds PD-1 is substantially free of Abs that specifically bind antigens other than PD-1). An isolated Ab that specifically binds PD-1 may, however, have cross-reactivity with other antigens, such as PD-1 molecules from different species. Moreover, an isolated Ab may be substantially free of other cellular material and / or chemicals.

[0043] The term "monoclonal antibody" ("mAb") refers to a non-naturally occurring preparation of Ab molecules of single molecular composition, i.e., Ab molecules which are essentially identical in primary sequence and have a single binding specificity and affinity for a particular epitope. A mAb is an example of an isolated Ab. mAbs may be produced by hybridoma, recombinant, transgenic, or other techniques known to those of skill in the art.

[0044] A "human" antibody (HuMAb) refers to an Ab having variable regions in which both the framework and CDR regions are derived from human germline immunoglobulin sequences. Additionally, if the Ab contains a constant region, the constant region is also derived from human germline immunoglobulin sequences. Human Abs 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, the term "human antibody" as used herein is not intended to include Abs in which CDR sequences derived from the germline of another mammalian species, such as mouse, have been grafted onto human framework sequences. The terms "human" Ab and "fully human" Ab are used interchangeably.

[0045] "Humanized antibody" refers to an Ab in which some, most or all of the amino acids outside the CDR domains of a non-human Ab are replaced with the corresponding amino acids from a human immunoglobulin. In some embodiments of a humanized form of an Ab, some, most or all of the amino acids outside the CDR domains are replaced with amino acids from a human immunoglobulin, and some, most or all of the amino acids within one or more CDR regions remain unchanged. Small additions, deletions, insertions, substitutions or modifications of amino acids are permissible as long as they do not abrogate the ability of the Ab to bind a particular antigen. A "humanized" Ab retains antigen specificity similar to the original Ab.

[0046] "Chimeric antibody" refers to an Ab whose variable region is derived from one species and whose constant region is derived from another species, such as an Ab whose variable region is derived from a murine Ab and whose constant region is derived from a human Ab.

[0047] "Anti-antigen" Ab refers to an Ab that specifically binds to an antigen. For example, anti-PD-1 Ab specifically binds to PD-1, and anti-CTLA-4 Ab specifically binds to CTLA-4.

[0048] An "antigen-binding portion" of an Ab (also called an "antigen-binding fragment") refers to one or more fragments of an Ab that retain the ability to specifically bind to the antigen bound by the whole Ab.

[0049] "Cancer" refers to a broad group of various diseases characterized by the unregulated growth of abnormal cells in the body. Unregulated cell division and proliferation leads to the formation of malignant tumors that can invade adjacent tissues and even metastasize to distant parts of the body via the lymphatic system or bloodstream. "Cancer" or "cancerous tissue" can include tumors.

[0050] "Cytotoxic T-lymphocyte antigen-4 (CTLA-4)" refers to an immune inhibitory receptor belonging to the CD28 family. CTLA-4 is expressed exclusively on T cells in vivo and binds two ligands, CD80 and CD86 (also called 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 common epitope with hCTLA-4. The complete hCTLA-4 sequence can be found under GenBank Accession No. AAB59385.

[0051] The term "immunotherapy" refers to the treatment of a subject having a disease or at risk of developing or relapsing with a disease by methods that induce, enhance, suppress or otherwise modify the immune response.

[0052] "Treatment" or "treatment" of a subject refers to any type of intervention or process or administration of an active agent performed on a subject for the purpose of reversing, alleviating, ameliorating, arresting, slowing or preventing the onset, progression, development, severity or recurrence of a symptom, complication or biochemical manifestation associated with a condition or disease.

[0053] "Programmed cell death (PD-1)" is an immune inhibitory receptor belonging to the CD28 family. PD-1 is expressed predominantly on preactivated T cells in vivo and binds 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 common epitope with hPD-1. The complete hPD-1 sequence can be found under GenBank Accession No. U64863.

[0054] "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 in common with hPD-L1. The complete hPD-L1 sequence can be found under GenBank Accession No. Q9NZQ7.

[0055] 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.

[0056] A "therapeutically effective amount" or "therapeutically effective dose" of a drug or therapeutic agent is any amount of drug that, when used alone or in combination with other therapeutic agents, protects a subject from developing a disease 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 or disability impairment 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 in human subjects during clinical trials, assays of the agent's activity in animal model systems predictive of efficacy in humans, or in in vitro assays.

[0057] As used herein, the term "subtherapeutic dose" or "subtherapeutically effective amount" refers to a dose of a therapeutic compound (e.g., an antibody) that is lower than the usual or typical dose of the therapeutic compound when administered alone for the treatment of a hyperproliferative disease (e.g., cancer). For example, a subtherapeutic dose of an anti-PD1 antibody (nivolumab) is a single dose of less than about 3 mg / kg of the antibody.

[0058] The use of the alternative (e.g., "or") should be understood to mean one, both, or any combination of the alternatives. As used herein, the terms "a," "an," and "the" should be understood to mean "one or more" of the cited or listed elements.

[0059] An "anti-cancer agent" promotes, for example, cancer regression in a subject. In other 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-neoplastic 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 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 cell, organ and / or organism level resulting from administration of the drug.

[0060] As an example of tumor treatment, a therapeutically effective amount of an anti-cancer agent preferably inhibits cell proliferation or tumor growth by at least about 20%, more preferably at least about 40%, even more preferably at least about 60%, and even more preferably at least about 80% compared to untreated subjects. In other embodiments of the invention, tumor regression can be observed and continued for a period of at least about 20 days, more preferably at least about 40 days, or even more preferably at least about 60 days. Regardless of these ultimate measures of therapeutic efficacy, evaluation of immunotherapeutic agents must also take into account "immune-related" response patterns.

[0061] The "immune-related" response pattern is a clinical response pattern frequently observed in cancer patients treated with immunotherapeutic agents that produce antitumor effects by inducing a cancer-specific immune response or by modifying natural immune processes. This response pattern is characterized by a favorable therapeutic effect after an initial increase in tumor burden or the appearance of new lesions, which in the evaluation of traditional chemotherapeutic agents would be classified as disease progression and is synonymous with drug ineffectiveness. Therefore, proper evaluation of immunotherapeutic agents may require long-term monitoring of the effect of these agents on the target disease.

[0062] A therapeutically effective amount of a drug includes a "prophylactically effective amount," which is any amount of drug that prevents the onset or recurrence of cancer when administered alone or in combination with an anti-neoplastic agent to a subject at risk of developing cancer (e.g., a subject with a pre-malignant condition) or a subject at risk of recurrence of cancer. In some embodiments, a prophylactically effective amount prevents the onset or recurrence of cancer entirely. "Preventing" the onset or recurrence of cancer means reducing the likelihood of the onset or recurrence of cancer or preventing the onset or recurrence of cancer entirely.

[0063] The use of the alternative (e.g., "or") should be understood to mean one, both, or any combination of the alternatives. As used herein, the terms "a," "an," and "the" should be understood to mean "one or more" of the cited or listed elements.

[0064] The term "about" or "essentially comprising" refers 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 practice in the art. Alternatively, "about" or "essentially comprising" can mean a range of up to 10% or 20% (i.e., ±10% or ±20%). For example, about 3 mg / kg can include any number between 2.7 mg / kg and 3.3 mg / kg (at 10%) or 2.4 mg / kg and 3.6 mg / kg (at 20%). Furthermore, particularly with respect to biological systems or processes, the term can mean within an order of magnitude or up to 5-fold of the value. When a particular value or composition is described in the present 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.

[0065] As described herein, any concentration range, percentage range, ratio range, or integer range should be understood to include any integer value within the recited range, and, where appropriate, fractions thereof (e.g., 1 / 10 and 1 / 100 of an integer), unless otherwise specified.

[0066] Various aspects of the invention are described in further detail in the following subsections.

[0067] Anti-PD-1 antibody PD-1 is a key immune checkpoint receptor expressed by activated T and B cells and mediates immune suppression. PD-1 is a member of the CD28 family of receptors that 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 that are expressed on antigen-presenting cells as well as many human cancers and have been shown to downregulate T cell activation and cytokine secretion upon binding to PD-1. Inhibition of PD-1 / PD-L1 interaction mediates potent antitumor activity in preclinical models.

[0068] HuMAbs that specifically bind PD-1 with high affinity are disclosed in U.S. Patents 8,008,449 and 8,779,105. Other anti-PD-1 mAbs 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 HuMAbs disclosed in U.S. Patent 8,008,449 exhibits one or more of the following characteristics: (a) a 1×10 mAb specific binding affinity determined by surface plasmon resonance using a Biacore biosensor system; -7 M or lower K D and binds to human PD-1; (b) does not substantially bind to human CD28, CTLA-4, or ICOS; (c) does not increase 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 Ab response; and (j) blocks tumor cell proliferation in vivo. Anti-PD-1 Abs that may be used in the present invention are mAbs that specifically bind to human PD-1 and exhibit at least one, preferably at least five, of the above characteristics.

[0069] In one embodiment, the anti-PD-1 Ab is nivolumab. (登録商標) " and formerly referred to as 5C4, BMS-936558, MDX-1106, or ONO-4538) is a fully human IgG4 (S228P) PD-1 immune checkpoint inhibitor Ab that selectively blocks interaction with PD-1 ligands (PD-L1 and PD-L2), thereby blocking downregulation of antitumor T cell function (U.S. Patent No. 8,008,449; Wang et al., 2014 Cancer Immunol Res. 2(9):846-56).

[0070] In other embodiments, the anti-PD-1 Ab is pembrolizumab. (登録商標) ", also known as lambrolizumab and MK-3475), is a humanized monoclonal IgG4 antibody against the human cell surface receptor PD-1 (programmed death-1 or programmed cell death-1). Pembrolizumab is described, e.g., in U.S. Patents Nos. 8,354,509 and 8,900,587, see also http: / / www.cancer.gov / drugdictionary?cdrid=695789 (last accessed: December 14, 2014). Pembrolizumab is approved by the FDA for the treatment of recurrent or refractory melanoma.

[0071] In other embodiments, the anti-PD-1 Ab is MEDI0608 (formerly AMP-514), which is a monoclonal antibody. MEDI0608 is described, for example, in U.S. Patent No. 8,609,089B2 or http: / / www.cancer.gov / drugdictionary?cdrid=756047 (last accessed December 14, 2014).

[0072] In one embodiment, the anti-PD-1 antibody is pidilizumab (CT-011), which is a humanized monoclonal antibody. Pidilizumab is described in U.S. Patent No. 8,686,119 B2 or WO 2013 / 014668 A1.

[0073] Anti-PD-1 Abs that may be used in the methods of the present disclosure also include isolated Abs that specifically bind human PD-1 and cross-compete with nivolumab for binding to human PD-1 (see, e.g., U.S. Patents Nos. 8,008,449 and 8,779,105; WO2013 / 173223). The ability of Abs to cross-compete for binding to an antigen indicates that these Abs bind to the same epitope region of the antigen and sterically hinder other cross-competing Abs from binding to that particular epitope region. These cross-competing Abs are expected to have functional properties very similar to nivolumab due to the fact that they bind to the same epitope region of PD-1. Cross-competing Abs can be easily identified based on their ability to cross-compete against nivolumab in standard PD-1 binding assays such as Biacore analysis, ELISA assays, or flow cytometry (see, e.g., WO2013 / 173223).

[0074] In some embodiments, the Ab that cross-competes with nivolumab for binding to human PD-1 or binds to the same epitope region of human PD-1 as nivolumab is a mAb.For administration to human subjects, these cross-competing Abs are preferably chimeric Abs or more preferably humanized or human Abs.Such chimeric, humanized or human mAbs can be produced and isolated by methods well known in the art.

[0075] Anti-PD-1 Abs that may be used in the methods of the invention also include antigen-binding portions of the Abs. It has been well established that the antigen-binding function of an Ab can be performed by fragments of a full-length Ab. Binding fragments encompassed by the term "antigen-binding portion" of an Ab include: (i) V L , V H , C L and C H1 (ii) a F(ab')2 fragment, which is a bivalent fragment containing two Fab fragments linked by a disulfide bridge at the hinge region; (iii) a V H and C H1(iv) an Fd fragment consisting of the V domain of a single arm of an Ab; and L and V H It contains an Fv fragment consisting of domains.

[0076] Anti-CTLA-4 antibody The anti-CTLA-4 antibodies of the invention bind to human CTLA-4 and disrupt the interaction of CTLA-4 with the human B7 receptor. Because the interaction of CTLA-4 with B7 transmits a signal that inactivates CTLA-4 receptor-bearing T cells, disruption of the interaction effectively induces, enhances or prolongs the activation of such T cells, thereby inducing, enhancing or prolonging an immune response.

[0077] HuMAbs 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 mAbs are disclosed, for example, in U.S. Patents 5,977,318, 6,051,227, 6,682,736, and 7,034,121. The anti-PD-1 HuMAbs disclosed in U.S. Patents 6,984,720 and 7,605,238 have been demonstrated to exhibit one or more of the following characteristics: (a) a specific binding 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 or 10 11 M -1 or higher equilibrium binding constant (K a (b) specifically binds human CTLA-4 with a binding affinity of at least about 10 3 , about 10 4 or about 10 5 m -1 s -1 The dynamic binding constant (k a );(c) at least about 10 3 , about 10 4 or about 10 5 m -1 s -1 The dynamic dissociation constant (k dand (d) inhibit the binding of CTLA-4 to B7-1 (CD80) and B7-2 (CD86). Anti-CTLA-4 Abs that may be used in the present invention are mAbs that specifically bind to human CTLA-4 and exhibit at least one, and preferably at least three, of the above characteristics.

[0078] An example of an anti-CTLA-4 Ab in clinical use is the human mAb 10D1 (also known as ipilimumab and Yervoy), disclosed in U.S. Patent No. 6,984,720. (登録商標) Ipilimumab is an anti-CTLA-4 Ab for use in the methods disclosed herein. Ipilimumab is a fully human, IgG1 monoclonal Ab that blocks the binding of CTLA-4 to the B7 ligand, thereby stimulating T cell activation and improving overall survival (OS) in patients with advanced melanoma.

[0079] Another anti-CTLA-4 Ab that can be used in 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.

[0080] The anti-CTLA-4 that can be used in the methods disclosed herein is an isolated Ab that specifically binds to human PD-1 and cross-competes with ipilimumab or tremelimumab for binding to human CTLA-4 or binds to the same epitope region of human CTLA-4 as ipilimumab or tremelimumab. In some embodiments, the Ab that cross-competes with ipilimumab or tremelimumab for binding to human CTLA-4 or binds to the same epitope region of human CTLA-4 as ipilimumab or tremelimumab is an Ab that comprises a heavy chain of human IgG1 isotype. For administration to human subjects, these cross-competing Abs are preferably chimeric Abs or more preferably humanized or human Abs. Useful anti-CTLA-4 Abs also include antigen-binding portions of the above Abs, such as Fab, F(ab')2, Fd or Fv fragments.

[0081] Standard treatment for kidney cancer The present invention also includes a combination therapy of anti-PD-1 antibody and standard therapy for the treatment of renal cancer. Standard therapy can be administered to the subject at any time before, during or after anti-PD1 antibody treatment. In some examples, standard therapy and PD-1 antibody treatment are also combined with additional treatment. Standard therapy for various types of cancer is well known to those skilled in the art. For example, the National Comprehensive Cancer Network (NCCN), a federation of 21 major cancer centers in the United States, has published NCCN Clinical Practice Guidelines in Oncology (NCCN GUIDELINES®), which provides cutting-edge information on detailed standard therapy treatment for a wide range of cancers (see NCCN GUIDELINES®, 2014).

[0082] RCC is the most common type of renal cancer in adults, accounting for approximately 90% of kidney tumors, with 80-90% of these having clear cell histology (see NCCN GUIDELINES® Version 3.2014 - Kidney Cancer). RCC accounts for approximately 3% of all cancers in the United States, with an estimated 63,920 patients diagnosed with renal cancer in the United States in 2014 and 13,860 deaths due to the disease (Siegel et al., 2014). Metastases are seen in approximately 30% of subjects at the time of diagnosis.

[0083] For clinically localized RCC (stages IA and IB), surgical resection, including radical nephrectomy and nephron-preserving surgery, is an effective treatment. Partial nephrectomy is generally not suitable for patients with locally advanced tumors (stages II and III), in which case radical nephrectomy is preferred. When the tumor is confined to the renal parenchyma, the 5-year survival rate is 60-70%, but this drops considerably in stage IV disease with widespread metastases. Stage IV RCC is fairly resistant to RT and chemotherapy, although patients may benefit from surgery, and in patients with a surgically resectable primary and resectable metastases, cytoreductive nephrectomy is recommended prior to systemic treatment.

[0084] Until recently, the cytokines IL-2 and IFNα, e.g., IFN-2b and PegIFN-2b, were the only active systemic treatments for advanced or metastatic RCC (mRCC), both of which were reported to provide objective response rates (ORRs) of 5–27%. IL-2 was approved based on a durable response rate of 15%. However, due to significant toxicities, including hypotension (71%), diarrhea (67%), dyspnea (43%), rash (42%), supraventricular tachycardia (12%), and a variety of metabolic and hematological disorders (Fyfe et al., 1995), IL-2 is administered only to relatively young, exceptionally eligible subjects. Furthermore, due to its severe acute toxicity, its administration is limited to intensive care settings in tertiary care centers. IFN-α is not approved for the treatment of RCC, and due to the limited clinical benefit and significant toxicity profiles of IL-2 and IFN-α, newer targeted agents have largely replaced cytokines in the treatment of advanced or metastatic renal cell carcinoma.

[0085] Recognition of the importance of hypoxia-inducible factor alpha (HIFα) signaling in the pathogenesis of clear cell RCC has sparked extensive investigation of two classes of targeted therapeutic agents, antiangiogenic TKIs and mTOR inhibitors, in 1L and 2L treatment (Mulders, 2009). Targeting angiogenesis is rational because constitutive HIFα activation leads to upregulation or activation of several proteins, including vascular endothelial growth factor (VEGF), which can subsequently lead to tumor growth and vasculature formation. Furthermore, blocking VEGF activity may modulate the immune environment and stimulate antitumor responses. Targeting the mTOR pathway is important because activation of the upstream PI3K / Akt / mTOR signaling pathway is one means by which constitutive HIFα activation or upregulation occurs (Mulders, 2009).

[0086] Agents targeting angiogenesis include VEGF receptor (VEGFr) TKIs (e.g., sorafenib, sunitinib, pazopanib, axitinib, and tivozanib) and VEGF-binding mAbs (e.g., bevacizumab), while agents targeting the mTOR pathway include mTOR inhibitors (e.g., everolimus and temsirolimus) (Mulders, 2009; NCCN GUIDELINES®, Version 3.2014 - Kidney Cancer). However, durable responses are rare as most patients develop resistant, ultimately progressive disease, and improved OS has only been shown in one Phase 3 trial in high-risk patients: temsirolimus (Torisel®) showed a statistically significant benefit in OS for patients with advanced RCC compared to IFNα (10.9 months vs. 7.3 months) (Hudes et al., 2007). Everolimus (Afinitor®) showed a 2.1 month improvement in median progression-free survival (PFS) versus placebo, but no OS improvement (Motzer et al., 2008). Of the five approved antiangiogenic agents (sorafenib, sunitinib, pazopanib, axitinib, and bevacizumab) and the two approved mTOR inhibitors (temsirolimus, everolimus), only everolimus is specifically approved for use after treatment failure with antiangiogenic therapy. In the United States, everolimus is indicated for the treatment of advanced RCC after failure of first-line treatment with sunitinib or sorafenib, and in the EU, everolimus is more broadly indicated for patients with advanced RCC whose disease has progressed on or after VEGF-targeted therapy. No recommendations exist for patients who have progressed on mTOR inhibitors.

[0087] Kidney Cancer Immunotherapy There is a clear need for agents that are effective in patients progressing on multiple lines of targeted therapy, as well as therapies that extend survival for longer periods beyond the current standard of care. Newer approaches to immunotherapy other than IL-2 and IFN-α have recently shown promise. Immunotherapies were first tested in patients with mRCC because traditional chemotherapy and radiation therapy have proven disappointing, kidney tumors are known to be able to elicit immune responses, and RCC is one of the tumors with the most common spontaneous regression (Elhilali et al., 2000;Inman et al., 2013). Currently, a variety of immunotherapeutic strategies, including T cell modulation, immune priming, and innate immunity, are in clinical development for the treatment of RCC (Rosenblatt and McDermott, 2011; Inman et al., 2013). Reversing T cell immunosuppression using Abs targeting the PD-1 receptor or its PD-L1 ligand has also proven very promising for the treatment of RCC (Topalian et al., 2012a; Brahmer et al., 2012; WO2013 / 173223).

[0088] Combination immunotherapy for renal cancer Immunotherapeutic approaches, including combinations of drugs such as anti-PD-1 and anti-CTLA-4 Ab, have proven highly effective in the treatment of melanoma (Wolchok et al., 2013; WO2013 / 173223). By analogy, RCC patients may benefit from combinations of different immunotherapeutic drugs or combinations of such drugs with targeted drugs or other treatments including surgery, radiation, standard cancer chemotherapy or vaccines. However, surprising and unexpected complications may be observed when immunotherapy is combined with other anticancer drugs. For example, first-line treatment of two melanoma patients with BRAF V600E mutations with anti-PD-1 agents (nivolumab and MK-3475, respectively) did not result in significant toxicity, but treatment with vemurafenib (Zelboraf®) at the time of disease progression led to severe hypersensitivity drug rash and multi-organ injury early in the course of vemurafenib treatment (Johnson et al., 2013). One patient subsequently developed acute inflammatory demyelinating polyneuropathy, and the other developed anaphylaxis upon rechallenge with low-dose vemurafenib.

[0089] With regard to RCC specifically, all drugs approved to date for the treatment of mRCC have been approved as monotherapy, with the exception of bevacizumab (Avastin®), which is approved only in combination with IFN-α. This combination was approved based on improved PFS combined with an acceptable safety profile, although it did not result in a significant improvement in median OS and resulted in a higher incidence of grade ≥3 hypertension and proteinuria compared to patients receiving IFN-α alone (Escudier et al., 2010; Rini et al., 2010). Many other combinations of antiangiogenic therapy plus immunotherapy are being evaluated in clinical trials, including trials of sunitinib plus IFN-α (Motzer et al., 2009), sunitinib plus IL-21 (Grunwald et al., 2011), sunitinib plus tremelimumab (Rini et al., 2011), sorafenib plus IFN-α (Escudier et al., 2007; Ryan et al., 2007; Gollob et al., 2007) and sorafenib plus IL-2 (Procopio et al., 2011), but significant toxicity is often observed.

[0090] In a phase 1 study testing the use of sunitinib + IFN-α in treatment-naïve subjects with mRCC (n=25), all subjects experienced grade 3 / 4 treatment-emergent AEs, most commonly neutropenia, fatigue, and thrombocytopenia (Motzer et al., 2009). The low starting dose was well tolerated, but clinical activity was reduced. The 12% response rate seen with sunitinib + IFN-α was lower than the historical response rate of 27.5% seen with sunitinib alone. Sunitinib + IL-21 was also tested in a phase 1 study in treatment-naïve subjects (n=9) (Procopio et al., 2011). The combination of maximum dose sunitinib with 10 mcg / kg IL-21 was considered too toxic based on the hematologic dose-limiting toxicities (DLTs) that occurred, while the combination with low dose IL-21 was not considered adequate for treatment. Sunitinib plus tremelimumab (anti-CTLA-4 Ab; Ribas, 2010; US Patent 6,682,736) was tested in a Phase 1 dose-escalation study in 28 subjects with mRCC who had received ≤1 prior systemic therapy (Rini et al., 2011). Unexpected toxicity of rapidly developing renal failure was observed in 4 subjects receiving sunitinib 37.5 mg daily in combination with tremelimumab 10 mg / kg or 15 mg / kg every 12 weeks. A 43% partial response rate was observed, but toxicity of the combination at the MTD (sunitinib 37.5 mg daily + tremelimumab 10 mg / kg every 12 weeks) was deemed unacceptable.

[0091] Combinations with sorafenib also proved too toxic or less effective than sorafenib monotherapy. A phase 1 study of sorafenib + IFN-α in a population including 12 mRCC subjects found that a maximum dose of sorafenib + IFN 9MIU was well tolerated, with only one patient experiencing a grade 3 DLT of asthenia (Escudier et al., 2007), and two subsequent phase 2 studies found increased toxicity when using this combination (Ryan et al., 2007;Gollob et al., 2007). In a phase 2 study of sorafenib + IFN-α in 62 subjects with untreated mRCC, the confirmed response rate with the combination (19%) was higher than the historical response rate with either agent alone, but increased toxicity, including grade 3 or worse toxicity in 77% of subjects, limited further development (Ryan et al., 2007). In a phase 2 trial of sorafenib plus IFN-α in 40 first- or second-line subjects, the response rate was 33%, including two complete responses (Gollop et al., 2007). However, the combination was more toxic than either drug alone, and treatment was only tolerable with treatment interruptions and dose reductions (65% of subjects). Sorafenib plus subcutaneous IL-2 versus sorafenib alone was tested in a randomized phase 2 trial of 128 subjects with untreated mRCC (Procopio et al., 2011). After treatment in 40 subjects, the IL-2 dose was reduced from 4.5 MIU to 3 MIU due to toxicity. Response rates were higher in the combination arm than in the sorafenib monotherapy arm (27% vs. 15%, respectively), but median PFS was not significantly different in the two treatment arms. The authors concluded that efficacy was not improved by the use of the sorafenib plus IL-2 combination.

[0092] Therefore, combination regimens including antiangiogenic TKIs have not been able to demonstrate improved efficacy in RCC without increased and generally unacceptable toxicity compared to monotherapy of either drug. The above examples in melanoma and RCC clearly show that the combination of immunotherapy (including immune checkpoint inhibitors such as anti-CTLA-4 or anti-PD-1 Ab) with targeted anticancer therapy (e.g., antiangiogenic TKIs) is unpredictable and must be carefully evaluated for safety and efficacy in clinical trials. Although the combination of nivolumab (anti-PD-1) and ipilimumab (anti-CTLA-4) is highly effective in the treatment of melanoma with manageable toxicity (Wolchok et al., 2013), it is not known to date whether this combination is significantly more effective in human subjects in the treatment of RCC and other cancers than the individual agents.

[0093] Pharmaceutical Compositions and Dosages The therapeutic agent of the present invention may comprise a composition, e.g., a pharmaceutical composition comprising an Ab or TKI and a pharmaceutically acceptable carrier. As used herein, a "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. Preferably, the carrier for a composition comprising an Ab is suitable for intravenous, intramuscular, subcutaneous, parenteral, spinal or epidermal administration (e.g., by injection or infusion), whereas the carrier for a composition comprising a TKI is suitable for non-parenteral, e.g., oral, administration. The pharmaceutical composition 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.

[0094] Dosing regimens are adjusted to provide the optimal desired response, e.g., maximum therapeutic response and / or minimal adverse effects. For administration of anti-PD-1 Ab, including in combination with other anti-cancer agents, the dose can range from about 0.01 to about 20 mg / kg, about 0.1 to about 10 mg / kg of the subject's body weight. For example, the dose can be about 0.1 mg / kg body weight, about 0.3 mg / kg body weight, about 1 mg / kg body weight, about 2 mg / kg body weight, about 3 mg / kg body weight, about 5 mg / kg body weight, or about 10 mg / kg body weight, and about 0.3 mg / kg body weight, about 1 mg / kg body weight, about 2 mg / kg body weight, about 3 mg / kg body weight, or about 5 mg / kg body weight. Dosing schedules are generally based on the typical pharmacokinetic properties of the Ab and are designed to achieve an exposure that results in sustained receptor occupancy (RO). Representative treatment regimes include once a week, about once every 2 weeks, about once every 3 weeks, about once every 4 weeks, about once a month, about once every 3-6 months, or longer. In some embodiments, an anti-PD-1 Ab, such as nivolumab, is administered to a subject about once every 2 weeks. In other embodiments, the Ab is administered about once every 3 weeks. Dosage and scheduling may vary over the course of treatment. For example, dosing schedules for anti-PD-1 monotherapy may include administration of Ab: (i) every 2 weeks for a 6-week cycle; (ii) 6 times every 4 weeks, then every 3 months; (iii) every 3 weeks; (iv) 3-10 mg / kg once, followed by 1 mg / kg every 2-3 weeks. Considering that IgG4 Abs generally have a half-life of 2-3 weeks, dosing regimens for the anti-PD-1 Abs of the invention include about 0.3 to about 10 mg / kg body weight, preferably about 1 to about 5 mg / kg body weight, more preferably about 1 to about 3 mg / kg body weight, administered intravenously, with the Ab given every 14-21 days for up to 6 weeks or in 12 week cycles until complete remission or disease progression is confirmed.

[0095] When used in combination with other cancer drugs, the dose of anti-PD-1 Ab may be lower compared to monotherapy doses. For example, a dose of nivolumab significantly lower than the typical dose of about 3 mg / kg every 3 weeks, such as about 0.1 mg / kg or less every 3 or 4 weeks, is considered a subtherapeutic dose. In some embodiments, a subtherapeutic dose is about 2 mg / kg, about 1 mg / kg, about 0.1 mg / kg, or about 0.01 mg / kg. Receptor-occupancy data from 15 subjects receiving 0.3 mg / kg to 10 mg / kg doses of nivolumab indicates that PD-1 occupancy appears to be dose-independent in this dose range. Across all doses, the mean occupancy was 85% (range, 70%-97%), with a mean plateau occupancy of 72% (range, 59%-81%) (Brahmer et al., 2010). Therefore, a dose of about 0.3 mg / kg may allow sufficient exposure to produce maximum bioactivity.

[0096] Although higher nivolumab monotherapy dosing up to 10 mg / kg every 2 weeks has been achieved without reaching the maximum tolerated dose (MTD), significant toxicity has been reported in other trials of checkpoint inhibitor plus anti-angiogenic therapy (see, e.g., Johnson et al., 2013; Rini et al., 2011), supporting the selection of a lower nivolumab dose than 10 mg / kg.

[0097] Ipilimumab (Yervoy®) is approved for the treatment of melanoma at 3 mg / kg intravenously once every three weeks for four doses. Thus, in some embodiments, about 3 mg / kg is the maximum dose of ipilimumab used in combination with an anti-PD-1 Ab, but in some embodiments, an anti-CTLA-4 Ab such as ipilimumab, when combined with nivolumab, can be dosed within the range of about 0.3-10 mg / kg body weight about every two or three weeks. Doses of ipilimumab significantly lower than the approved 3 mg / kg every three weeks, for example about 0.3 mg / kg or less about every three or four weeks, are considered subtherapeutic doses. In some embodiments, a subtherapeutic dose is about 2 mg / kg, about 1 mg / kg, about 0.1 mg / kg, or about 0.01 mg / kg.

[0098] Although combined dosing of nivolumab 3 mg / kg and ipilimumab 3 mg / kg has been shown to exceed the maximum tolerated dose (MTD) in melanoma populations, combinations of nivolumab 1 mg / kg + ipilimumab 3 mg / kg or nivolumab 3 mg / kg + ipilimumab 1 mg / kg have been found to be tolerable in melanoma patients (Wolchok et al., 2013). Thus, nivolumab is tolerated at up to 10 mg / kg intravenously every 2 weeks, but in some embodiments, the dose of anti-PD-1 Ab does not exceed about 3 mg / kg when combined with ipilimumab. In certain embodiments, based on the risk-benefit and PK-PD assessment, the doses used are about 1 mg / kg nivolumab + about 3 mg / kg ipilimumab, about 3 mg / kg nivolumab + about 1 mg / kg ipilimumab, or about 3 mg / kg nivolumab + about 3 mg / kg ipilimumab, each administered at a dosing frequency of about once every 2-4 weeks or about once every 3 weeks. In certain other embodiments, nivolumab is administered at a dose of about 0.1 mg / kg, about 0.3 mg / kg, about 1 mg / kg, about 2 mg / kg, about 3 mg / kg, or about 5 mg / kg, in combination with ipilimumab administered at a dose of about 0.1 mg / kg, about 0.3 mg / kg, about 1 mg / kg, about 2 mg / kg, about 3 mg / kg, or about 5 mg / kg once every 2 weeks, once every 3 weeks, or once every 4 weeks. In some embodiments, the combination of anti-PD-1 Ab and anti-CTLA-4 Ab is administered intravenously to the subject about every 2 or 3 weeks for 2, 3 or 4 times during the induction phase. In some embodiments, the combination of nivolumab and ipilimumab is administered intravenously about every 3 weeks for 4 times during the induction phase. The induction phase is followed by a maintenance phase during which anti-PD-1 Ab alone is administered to the subject at a dose of about 0.1 mg / kg, about 0.3 mg / kg, about 1 mg / kg, about 2 mg / kg, about 3 mg / kg, about 5 mg / kg or about 10 mg / kg every 2 or 3 weeks, as long as treatment is proven effective or until uncontrollable toxicity or disease progression occurs. In some embodiments, nivolumab is administered at a dose of about 3 mg / kg body weight about every 2 weeks during the maintenance phase.

[0099] For the combination of nivolumab and other anti-cancer drugs, in some embodiments, these drugs can be administered at their approved doses. Treatment continues as long as clinical benefit is observed or until unacceptable toxicity or disease progression occurs. Nevertheless, in some embodiments, the doses of these anti-cancer drugs administered are significantly lower than their approved doses, i.e., subtherapeutic doses of the drugs are administered in combination with anti-PD-1 Ab. Anti-PD-1 Ab can be administered at the doses shown in clinical trials to produce the best efficacy as monotherapy, for example, about 3mg / kg nivolumab is administered about once every 3 weeks or at significantly lower doses, i.e., subtherapeutic doses.

[0100] The dosing schedule of TKI varies with different TKI.For example, the usual dose of sunitinib includes a fixed dose of 50mg orally administered to adults once a day with or without food, and dose adjustment is recommended at about 12.5mg up or down based on individual patient safety and tolerability.Sunitinib is administered on a schedule of 4 weeks treatment followed by 2 weeks off.In some embodiments, sunitinib is administered at a dose of about 12.5mg, about 25mg, about 37.5mg, about 50mg, about 62.5mg, about 75mg, about 87.5mg or about 90mg once a day.Sorafenib is administered orally to adult patients at a dose of 400mg twice a day, at least 1 hour before or 2 hours after a meal. In some embodiments, sorafenib is administered at a dose of about 100 mg, about 200 mg, about 300 mg, about 400 mg, about 500 mg, about 600 mg or about 800 mg twice a day. Pazopanib is administered orally to patients once a day at a dose of 800 mg without food (at least 1 hour before or 2 hours after a meal). In some embodiments, pazopanib is administered at a dose of about 200 mg, about 400 mg, about 600 mg, about 700 mg, about 800 mg, about 900 mg or about 1000 mg once a day. Axitinib (Inlyta®) is administered orally to patients at a dose of about 5 mg twice a day about 12 hours apart with or without food, but can be titrated up to about 2 mg, about 3 mg, about 7 or about 10 mg twice a day as needed. Tivozanib has not yet been approved by the FDA, but a daily oral dose of about 1 mg or 1.5 mg, preferably 1.5 mg, for 28 days followed by 14 days off (Eskens et al., 2011) or 21 days on medication followed by 7 days off (Nosov et al., 2012). In some embodiments, tivozanib is administered at a dose of about 0.5 mg, about 0.75 mg, about 1 mg, about 1.25 mg, about 1.5 mg, about 1.75 mg, or about 2 mg once daily. With regard to the combination treatment of TKI and anti-PD-1 Ab, in some embodiments, the TKI is administered at its approved or recommended dose. Treatment is continued as long as clinical benefit is observed or until unacceptable toxicity or disease progression occurs.

[0101] In some embodiments, the dose of TKI administered is the approved or recommended dose.In other embodiments, the TKI is administered in combination with anti-PD-1 Ab at a dose significantly lower than the approved dose, i.e., subtherapeutic dose.Anti-PD-1 Ab may be administered at a dose that has been shown in clinical trials to produce the best efficacy as monotherapy, for example, about 3mg / kg nivolumab once every 3 weeks (Topalian et al., 2012a; Topalian et al., 2012) or at a low dose, i.e., subtherapeutic dose. In some embodiments, the anti-PD-1 Ab is administered at a dose of about 0.1 mg / kg, about 0.3 mg / kg, about 1 mg / kg, about 2 mg / kg, about 3 mg / kg, or about 5 mg / kg, and the TKI is administered at a dose of about 0.1 mg, about 0.5, about 1, about 2, about 3, about 5, about 7.5, about 10, about 12.5, about 15, about 20, about 30, about 40, about 50, or about 75 mg / kg.

[0102] Dose and frequency vary according to the half-life of Ab in the subject. In general, human Ab has the longest half-life, followed by humanized Ab, chimeric Ab and non-human Ab. Dose and frequency of administration can vary according to whether the treatment is preventive or therapeutic. In preventive applications, relatively low doses are generally administered relatively infrequently for a long period of time. Some patients are treated for the rest of their lives. In therapeutic applications, relatively high doses at relatively short intervals may be required until the progression of the disease slows or stops, preferably until the patient shows partial or complete improvement of disease symptoms. The patient can then be administered a prophylactic regime.

[0103] The actual dosage level of the active ingredient in the pharmaceutical composition of the present invention may vary in order to obtain an amount of the active ingredient that is effective for achieving the desired therapeutic response for a particular patient, composition and administration method without excessive toxicity to the patient.The selected dosage level depends on the activity of the particular composition of the present invention used, the route of administration, the timing of administration, the excretion rate of the particular compound used, the duration of treatment, the variation of various pharmacokinetic factors, including 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 may be administered by one or more routes of administration using one or more of a variety of methods well known in the art.As will be recognized by those skilled in the art, the route and / or method of administration will vary depending on the desired result.

[0104] Methods of the Invention The present invention provides a method for treating a subject having renal cancer. In some embodiments, the present invention includes a method for treating a subject having renal cancer or renal cancer, comprising administering a combination therapy. In some embodiments, the method comprises administering to the subject a therapeutically effective amount of: (a) an anti-cancer agent that is an Ab or antigen-binding portion thereof that specifically binds to the PD-1 receptor and inhibits PD-1 activity; and (b) a combination of other anti-cancer therapeutic agents. In some embodiments, the present invention provides a method for treating a subject having renal cancer or renal cancer in a subject in need of treatment, comprising administering to the subject a therapeutically effective amount of: (a) an anti-PD-1 antibody or antigen-binding portion thereof; and (b) a combination of other anti-cancer agents. In other embodiments, the present invention relates to a method for treating a subject having renal cancer or renal cancer, comprising administering to the subject a therapeutically effective amount of: (a) an anti-PD-1 antibody or antigen-binding portion thereof; and (b) a standard of care therapy as disclosed herein. In some embodiments, the renal cancer is RCC, since RCC constitutes approximately 90% of kidney tumors. In some other embodiments, the subject is a human patient.

[0105] In one embodiment, the present invention relates to a method of treating renal cancer or a subject having renal cancer in a subject in need of treatment, the method comprising administering to the subject an anti-PD-1 antibody or an antigen-binding portion thereof, wherein the subject is concurrently treated with other anti-cancer agents. In another embodiment, the present invention relates to a method of treating renal cancer or a subject having renal cancer in a subject in need of treatment, the method comprising administering to the subject an anti-PD-1 antibody or an antigen-binding portion thereof, wherein the subject is concurrently treated with a standard of care therapy as disclosed herein. In one embodiment, the renal cancer is RCC, as RCC constitutes approximately 90% of kidney tumors. In another embodiment, the subject is a human patient.

[0106] In some embodiments, the PD-1 antibody, or antigen-binding portion thereof, and / or the anti-CTLA-4 antibody, or antigen-binding portion thereof, is administered in combination with a VEGF receptor (VEGFr) TKI (e.g., sorafenib, sunitinib, pazopanib, axitinib, and tivozanib), a VEGF binding mAb and inhibitor (e.g., bevacizumab, aflibercept, and ziv-aflibercept), an mTOR inhibitor (e.g., everolimus and temsirolimus), a cytotoxic T cell inhibitor (e.g., cytotoxic T cell inhibitor ... The anti-PD-1 antibody is administered in combination with one or more additional anti-cancer agents selected from the group consisting of anticancer agents (e.g., IFN-α, IFN-b2, pegylated IFNb2 (PegIFN-b2), and IL-2) and mitotic inhibitors (e.g., paclitaxel, docetaxel, vincristine, eribulin, estramustine, etoposide, ixabepilone, cabazitaxel, vincristine liposomes, vinorelbine, vincristine, vinblastine, and teniposide). In some embodiments, the anti-cancer agent is any other agent described herein. In some embodiments, the treatment of the invention (e.g., administration of an anti-PD-1 antibody or administration of an anti-PD-1 antibody and another anti-cancer treatment) effectively prolongs the survival of the subject. For example, the subject's survival is extended 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 when compared to other subjects treated with either the other therapy alone (e.g., bevacizumab or temozolomide) or, in the case of a combination therapy, with one of the two members of the combination therapy alone (e.g., an anti-PD-1 antibody alone). In some embodiments, survival is extended by at least about 2 months. In some embodiments, the treatment of the invention effectively extends the subject's progression-free survival.For example, the progression-free survival of the subject is extended 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, when compared to an untreated subject or a subject treated with either the other treatment alone (e.g., bevacizumab or temozolomide) or, in the case of a combination treatment, one of the two members of the combination treatment alone (e.g., an anti-PD-1 antibody alone). In some embodiments, the progression-free survival is extended by at least about 2 months. In some embodiments, the treatment of the present invention effectively increases the response rate in a group of subjects. For example, the response rate in a group of subjects 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 at least about 100%, when compared to another group of subjects treated with either the other treatment alone (e.g., bevacizumab or temozolomide) or, in the case of a combination treatment, with one of the two members of the combination treatment alone (e.g., an anti-PD-1 antibody alone), i.e., monotherapy.

[0107] Anti-PD-1 and Anti-PD-L1 Antibodies Suitable for Use in the Methods of the Invention Anti-PD-1 Abs suitable for use in the methods of the invention are Abs that bind PD-1 with high specificity and affinity, block binding of PD-L1 and / or PD-L2, and inhibit the immunosuppressive effects of the PD-1 signaling pathway. In any of the therapeutic methods disclosed herein, the anti-PD-1 or anti-CTLA-4 "antibody" is an antigen-binding portion or fragment that binds to the PD-1 or CTLA-4 receptor, respectively, inhibits ligand binding, and exhibits functional properties similar to full Abs that upregulate the immune system. In some embodiments, the anti-PD-1 Ab or antigen-binding portion thereof cross-competes with nivolumab for binding to human PD-1. In other embodiments, the anti-PD-1 Ab or antigen-binding portion thereof is a chimeric, humanized, or human monoclonal Ab or portion thereof. In some embodiments for treating a human subject, the Ab is a humanized Ab. In other embodiments for treating a human subject, the Ab is a human Ab. Abs of IgG1, IgG2, IgG3, or IgG4 isotypes may be used.

[0108] In some embodiments, the anti-PD-1 Ab, or antigen-binding portion thereof, comprises a heavy chain constant region that is 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 Ab, or antigen-binding portion thereof, comprises a S228P mutation that replaces a serine residue in the hinge region with a proline residue normally found at the corresponding position in IgG1 isotype antibodies. This mutation, present in nivolumab, prevents Fab arm exchange with endogenous IgG4 antibodies while retaining the low affinity for activating Fc receptors associated with wild-type IgG4 antibodies (Wang et al., 2014). In yet other embodiments, the Ab comprises a light chain constant region that is a human kappa or lambda constant region. In other embodiments, the anti-PD-1 Ab, or antigen-binding portion thereof, is a mAb, or antigen-binding portion thereof. In certain embodiments of any of the therapeutic methods described herein that include administration of an anti-PD-1 Ab, the anti-PD-1 Ab is nivolumab. In other embodiments, the anti-PD-1 Ab is pembrolizumab. In other embodiments, the anti-PD-1 Ab is selected from human antibodies 17D8, 2D3, 4H1, 4A11, 7D3, and 5F4, described in U.S. Patent No. 8,008,449. In yet other embodiments, the anti-PD-1 Ab is MEDI0608 (formerly AMP-514), AMP-224, or pidilizumab (CT-011).

[0109] In another embodiment, the present invention includes a method of treating renal cancer or a subject with renal cancer in a subject in need of treatment, comprising administering an anti-PD-1 antagonist or an anti-PD-1 antagonist in combination with one or more anti-cancer agents described herein to treat the cancer. In one embodiment, the renal cancer is RCC. As used herein, an "anti-PD-1 antagonist" includes any molecule that inhibits the interaction of PD-1 (receptor) with PD-L1 (ligand), such that the PD-1 / PD-L1 signaling pathway is blocked. PD-L1 and PD-L2 are two identified ligands for PD-1. In one embodiment, the anti-PD-1 antagonist is an anti-PD-L1 antibody. PD-L1 and PD-L2 have been shown to downregulate T cell activation by binding to PD-1. Both PD-L1 and PD-L2 are B7 homologs that bind to PD-1 but not other CD28 family members. PD-L1 has been shown to be abundant in a variety of human cancers. The interaction of PD-1 and PD-L1 results in a reduction in tumor-infiltrating lymphocytes, a decrease in T cell receptor-mediated proliferation, and immune evasion by cancerous cells. Immune suppression can be restored by inhibition of the local interaction of PD-1 and PD-L1, and this effect is additive when the interaction of PD-1 and PD-L2 is similarly blocked. In some embodiments, an anti-PD-L1 Ab can be substituted for an anti-PD-1 Ab in any of the therapeutic methods disclosed herein. In some embodiments, the anti-PD-L1 Ab is BMS-936559 (formerly 12A4 or MDX-1105) (see, e.g., U.S. Patent No. 7,943,743; WO2013 / 173223). In other embodiments, the anti-PD-L1 Ab is MPDL3280A (also known as RG7446) (see, e.g., Herbst et al. 2013; U.S. Patent No. 8,217,149) or MEDI4736 (Khleif, 2013).

[0110] In other embodiments, the anti-PD-1 antagonist is a soluble PD-1 protein. In other embodiments, the anti-PD-1 antagonist is a PD-1-Fc fusion protein. In some embodiments, the PD-1-Fc fusion protein is the result of a fusion of any of the PD-1 antibodies described herein with an Ig Fc domain. In some embodiments, the Ig Fc domain is an IgG, IgA, or IgM domain. In some embodiments, fusion proteins can be produced by chemical synthesis or by generating a polynucleotide encoding the desired fusion protein.

[0111] In some embodiments, the anti-PD-1 antagonist is an anti-PD-1 fusion protein, antisense molecule, small molecule, ribozyme, or nanobody that inhibits or blocks the interaction of PD-1 and PD-L1.

[0112] Combination of anti-PD-1 Ab and anti-angiogenic TKI for treating RCC In some embodiments of the present invention for treating RCC, the other anti-cancer agent, i.e., the anti-cancer agent other than anti-PD-1 Ab used in combination with anti-PD-1 Ab, is an anti-angiogenic TKI. Various TKIs approved for treating RCC or proven effective in clinical trials, including sorafenib, sunitinib, pazopanib, axitinib and tivozanib, may be used in combination therapy with anti-PD-1 Ab. However, until the efficacy and safety of these combinations are evaluated in preclinical or clinical trials, there may be unexpected toxicity or no significant enhanced efficacy over monotherapy. Preclinical data disclosed herein in Renca mouse RCC model shows that anti-PD-1 Ab acts synergistically with sunitinib in inhibiting RCC tumor growth, but the combination of anti-PD-1 Ab and sorafenib does not significantly enhance the low-level anti-tumor activity of sorafenib in this mouse RCC model (Figures 1 and 2). To further demonstrate that the combination of anti-PD-1 Ab and sunitinib is highly effective in treating RCC in mouse models, the efficacy and safety of this combination as well as the combination of nivolumab and pazopanib in human RCC patients are being confirmed in clinical trials.

[0113] Example 5 provides interim results of an ongoing Phase 1 study (NCT01472081; see Example 4) of nivolumab in combination with sunitinib or pazopanib in patients with mRCC. The data supports that nivolumab in combination with either of these TKIs shows significantly higher antitumor activity and a manageable safety profile in these patients (see FIG. 12). Thus, in one embodiment of this combination therapy, the antiangiogenic TKI is sunitinib. In another embodiment, the TKI is pazopanib. In a further embodiment, the TKI is axitinib. Axitinib is a highly selective, potent inhibitor of VEGF receptors 1, 2 and 3 that has a favorable toxicity profile (Sonpavde et al., 2008) and is FDA approved for the treatment of advanced RCC after failure of one prior systemic therapy. In another embodiment, the TKI is sorafenib. In yet another embodiment, the TKI is tivozanib. Tivozanib is a potent, selective, long half-life inhibitor of VEGFR-1, -2 and -3, which is being tested in clinical trials for the treatment of RCC.Axitinib and tivozanib have not been tested in preclinical or clinical trials here.However, based on the highly selective inhibition of all three VEGF receptors, which overlaps with the activity of sunitinib, pazopanib and sorafenib to inhibit VEGFR while minimizing off-target toxicity, both axitinib and tivozanib are excellent candidates to be combined with anti-PD-1 Ab in the therapeutic method disclosed herein.

[0114] In some embodiments, administration of an anti-PD-1 Ab or antigen-binding portion thereof and an anti-angiogenic TKI results in increased infiltration of cancer tissues by T cells compared to untreated subjects or subjects treated with either an anti-PD-1 antibody or other anti-cancer agent monotherapy. As described in Example 2, combination therapy can be used to, for example, increase the infiltration of CD4 + and CD8 +This can result in increased localization of T cells to kidney tumors. This increase in T cell infiltration is 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%, or at least about 90% greater than the T cell infiltration levels in untreated subjects or subjects treated with either anti-PD-1 antibodies or other anti-cancer agents monotherapy. The methods disclosed herein can further increase T cell proliferation compared to untreated subjects or subjects treated with either anti-PD-1 antibodies or other anti-cancer agents monotherapy. T cell proliferation is increased 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%, or at least about 90%. The methods disclosed herein may also, in some embodiments, result in a decrease in T regulatory cells compared to untreated subjects or subjects treated with a monotherapy of either an anti-PD-1 antibody or another anti-cancer agent.

[0115] In other embodiments, the methods for treating RCC decrease the number of monocytic myeloid-derived suppressor cells or increase the number of granulocytic myeloid cells compared to untreated subjects or subjects treated with either an anti-PD-1 antibody or other anti-cancer agent monotherapy. + / Ly6C hi / Ly6G - Expression or CD11b + / Ly6C low / Ly6G - In other embodiments, the granulocytic myeloid cells are characterized by expression of CD11b + / Ly6C - / Ly6G +The number of monocytic myeloid derived suppressor cells can be decreased by at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90%. The number of granulocytic myeloid cells can be increased by at least about 5%, 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%, or at least about 90%.

[0116] Doses of anti-PD-1 Ab and anti-angiogenic TKI used in combination therapy In certain embodiments of the combination therapy, a therapeutically effective amount of an anti-PD-1 Ab or antigen-binding portion thereof in combination with an anti-angiogenic TKI includes a dose range of about 0.1 to about 10.0 mg / kg body weight administered by intravenous infusion about once a week, about once every two weeks, about once every three weeks, or about once a month. In certain embodiments, the anti-PD-1 Ab is administered at a dose of 2 mg / kg once every three weeks. In other embodiments, the anti-PD-1 Ab is administered at a dose of about 5 mg / kg once every three weeks.

[0117] The dose of sunitinib may range, for example, from about 25 to about 50 mg, including in one embodiment 50 mg administered to adult RCC patients in a 42-day cycle with 28 days of daily administration and 14 days off, and repeating the cycle as long as clinical benefit is observed or until uncontrollable toxicity or disease progression occurs. Alternatively, the 28-day dose of 25-50 mg and 14 days off may be followed by daily administration of 37.5 mg of sunitinib as long as clinical benefit is observed or until uncontrollable toxicity or disease progression occurs. In some embodiments, sunitinib is administered at a dose of about 5 mg / kg body weight, about 10 mg / kg body weight, about 15 mg / kg body weight, about 20 mg / kg body weight, about 25 mg / kg body weight, about 30 mg / kg body weight, about 35 mg / kg body weight, about 40 mg / kg body weight, about 45 mg / kg body weight, about 50 mg / kg body weight, about 55 mg / kg body weight, about 60 mg / kg body weight, about 65 mg / kg body weight, about 70 mg / kg body weight or about 75 mg / kg body weight.

[0118] Pazopanib in combination with anti-PD-1 Ab, orally once daily at about 50 mg, about 100 mg, about 200 mg, about 300 mg, about 400 mg, about 500 mg, about 600 mg, or about 700 mg Or at a dose of about 800 mg. In some embodiments, pazopanib is administered at a recommended dose of 800 mg once daily without food.

[0119] The dose of TKIs other than sunitinib and pazopanib that have not been clinically tested in combination with anti-PD-1 Ab is based on known therapeutic doses. Thus, in some embodiments, sorafenib is administered orally at its recommended dose of 400 mg twice daily. In some embodiments, axitinib (Inlyta®) is administered orally to patients at its recommended dose of 5 mg twice daily, approximately 12 hours apart. In some embodiments, tivozanib is administered orally at its recommended daily dose of 1.5 mg for 28 days, followed by 14 days off (Eskens et al., 2011) or 21 days, followed by 7 days off (Nosov et al., 2012).

[0120] In some embodiments, nivolumab is administered by intravenous infusion at about 2 or about 5 mg / kg on days 1 and 22 of each 6-week (42-day) cycle, and is combined with a TKI, such as sunitinib administered orally at about 50 mg on days 1-28 of each 42-day cycle, or pazopanib administered orally at about 800 mg on days 1-42 of each 42-day cycle. Other TKIs used in the method are administered according to their approved or recommended dosing regimens. Treatment is continued as long as clinical benefit is observed or until unacceptable toxicity or disease progression occurs.

[0121] The synergistic interactions observed in preclinical models, for example, between anti-PD-1 and sunitinib, suggest that one or both of these therapeutic agents may be administered to patients at subtherapeutic doses, i.e., at doses of the therapeutic agents that are significantly lower than the usual or FDA approved doses when administered as monotherapy for the treatment of cancer. In some embodiments of the invention, the anti-PD-1 Ab or antigen-binding portion thereof is administered to RCC patients at subtherapeutic doses. In other embodiments, the TKI is administered at subtherapeutic doses as described herein. In further embodiments, the anti-PD-1 Ab or antigen-binding portion thereof and the TKI are each administered at subtherapeutic doses as described herein.

[0122] The combination treatment of the anti-PD-1 antibody, or antigen-binding portion thereof, and the other anti-cancer agent can be administered simultaneously or sequentially (e.g., the anti-PD-1 antibody, or antigen-binding portion thereof, is administered first and the other anti-cancer agent is administered second, or the other anti-cancer agent is administered first and the anti-PD-1 antibody, or antigen-binding portion thereof, is administered second).

[0123] Combination of anti-PD-1 and anti-CTLA-4 Abs for the treatment of RCC The present disclosure also provides combination therapy for the treatment of RCC, in which anti-PD-1 Ab is combined with another anti-cancer agent that is an Ab or antigen-binding portion thereof that specifically binds to CTLA-4 and inhibits CTLA-4 activity. It is shown herein that the combination of anti-PD-1 Ab, nivolumab, and anti-CTLA-4 Ab, ipilimumab, produces a significantly higher level of clinical activity, as measured by ORR, than that observed with nivolumab or ipilimumab alone (see Example 6). Thus, in some embodiments, the anti-CTLA-4 Ab used in combination with anti-PD-1 Ab is ipilimumab. In some embodiments, the anti-CTLA-4 Ab is tremelimumab. In other embodiments, the anti-CTLA-4 Ab or antigen-binding portion thereof is an Ab or portion thereof that cross-competes with ipilimumab for binding to human CTLA-4. In some other embodiments, the anti-CTLA-4 Ab or antigen-binding portion thereof is a chimeric, humanized, or human mAb or portion thereof. In yet other embodiments, the anti-CTLA-4 Ab, or antigen-binding portion thereof, comprises a heavy chain constant region that is of the human IgG1 or IgG4 isotype. In some embodiments, the anti-CTLA-4 Ab comprises a heavy chain constant region that is of the human IgG1 isotype.

[0124] For a combination of anti-PD-1 and anti-CTLA-4 Ab, the dosing regimen includes an induction period (also referred to herein as the induction phase) during which one or more, preferably about four, combined doses of anti-PD-1 and anti-CTLA-4 Ab are administered to the patient, followed by a maintenance period or phase in which the anti-PD-1 Ab is administered alone, i.e., without the anti-CTLA-4 Ab. In certain embodiments, the method includes (a) an induction phase in which 2, 4, 6, 8, or 10 doses of anti-PD-1 and anti-CTLA-4 antibodies, or antigen-binding portions thereof, are administered in combination, each dose ranging from about 0.1 to about 10.0 mg / kg body weight, administered at least about once every two weeks, about once every three weeks, or about once every four weeks, and (b) a maintenance phase in which no anti-CTLA-4 antibody, or antigen-binding portion thereof, is administered, and the anti-PD-1 antibody, or antigen-binding portion thereof, is administered at a dose of about 0.1 to about 10 mg / kg, repeatedly, at least about once every two weeks, about once every three weeks, or about once every four weeks.

[0125] In some embodiments, the anti-PD-1 Ab, or antigen-binding portion thereof, is administered at a sub-therapeutic dose. In certain other embodiments, the anti-CTLA-4 Ab, or antigen-binding portion thereof, is administered at a sub-therapeutic dose. In further embodiments, both the anti-PD-1 Ab, or antigen-binding portion thereof, and the anti-CTLA-4 Ab, or antigen-binding portion thereof, are each administered at a sub-therapeutic dose. In some embodiments, the anti-PD-1 Ab, or antigen-binding portion thereof, and / or the anti-CTLA-4 Ab, or antigen-binding portion thereof, are administered at a therapeutic dose.

[0126] In some embodiments, (a) the induction phase comprises at least four administrations, three weeks apart, in which the anti-PD-1 and anti-CTLA-4 Abs are administered at the following doses: (i) 0.1 mg / kg anti-PD-1 Ab and 3 mg / kg anti-CTLA-4 Ab; (ii) 0.3 mg / kg anti-PD-1 Ab and 3 mg / kg anti-CTLA-4 Ab; (iii) 1 mg / kg anti-PD-1 Ab and 3 mg / kg anti-CTLA-4 Ab; (iv) 3 mg / kg anti-PD-1 Ab and 3 mg / kg anti-CTLA-4 Ab; (v) 5 mg / kg anti-PD-1 Ab and 3 mg / kg anti-CTLA-4 Ab; (vi) 10 mg / kg anti-PD-1 Ab and 3 mg / kg anti-CTLA-4 Ab; (vii) 0.1 mg / kg anti-PD-1 Ab and 1 mg / kg anti-CTLA-4 Ab; (viii) 0.3 mg / kg anti-PD-1 Ab and 1 mg / kg anti-CTLA-4 Ab; (ix) 1 mg / kg anti-PD-1 Ab and 1 mg / kg anti-CTLA-4 Ab; (x) 3 mg / kg anti-PD-1 Ab and 1 mg / kg anti-CTLA-4 Ab; (xi) 5 mg / kg anti-PD-1 Ab and 1 mg / kg anti-CTLA-4 Ab; or (xii) 10 mg / kg anti-PD-1 Ab and 1 mg / kg anti-CTLA-4 Ab and (b) the maintenance phase comprises repeated administration of anti-PD-1 Ab at a dose of 3 mg / kg every 2 weeks.

[0127] Due to the durability of clinical efficacy previously demonstrated with immunotherapy by inhibition of immune checkpoints (see, e.g., WO2013 / 173223), the maintenance phase may, in another embodiment, include a finite number of doses, e.g., 1-10 doses, or may include dosing at longer intervals, e.g., about once every 3-6 months or about once every 1-2 years or longer. The maintenance phase may continue as long as clinical benefit is observed or until uncontrollable toxicity or disease progression occurs.

[0128] Given the uncertainty of the contribution of ipilimumab administered the previous 12 weeks to clinical benefit in melanoma and the Food and Drug Administration (FDA) and European Medicines Agency (EMA) approved schedule for Yervoy® is every 3 weeks for a total of 4 doses, in one embodiment, the anti-CTLA-4 Ab is administered once every 3 weeks for a total of 4 doses during the induction phase. Thus, in some embodiments, the methods consist of (a) administration of four combined doses at three week intervals, where (i) an anti-PD-1 antibody, or antigen-binding portion thereof, is administered at 3 mg / kg body weight and an anti-CTLA-4 antibody, or antigen-binding portion thereof, is administered at 1 mg / kg body weight; (ii) an induction phase where an anti-PD-1 antibody, or antigen-binding portion thereof, is administered at 1 mg / kg body weight and an anti-CTLA-4 antibody, or antigen-binding portion thereof, is administered at 3 mg / kg body weight; or (iii) an anti-PD-1 antibody, or antigen-binding portion thereof, is administered at 3 mg / kg body weight and an anti-CTLA-4 antibody, or antigen-binding portion thereof, is administered at 3 mg / kg body weight; and (b) a maintenance phase comprising repeated administration of an anti-PD-1 antibody, or antigen-binding portion thereof, at a dose of 3 mg / kg every two weeks. In further embodiments of these methods, the maintenance phase continues as long as clinical benefit is observed or until uncontrollable toxicity or disease progression occurs.

[0129] In some embodiments of the method, the anti-PD-1 Ab is nivolumab. In other embodiments, it is pembrolizumab. In yet other embodiments, the anti-CTLA-4 Ab is ipilimumab. In a further embodiment, the anti-CTLA-4 Ab is tremelimumab. In general, the anti-PD-1 and anti-CTLA-4 Abs are formulated for intravenous administration. In some embodiments, the combination of the anti-PD-1 antibody or antigen-binding portion thereof and the anti-CTLA-4 antibody or antigen-binding portion thereof is administered simultaneously as a single composition or as separate compositions. In some embodiments, the anti-PD-1 and anti-CTLA-4 Abs are administered sequentially. In some embodiments, the anti-PD-1 and anti-CTLA-4 Abs are administered sequentially during the induction phase. In some embodiments, when the anti-PD-1 and anti-CTLA-4 Abs are administered in combination, they are administered within 30 minutes of each other. Either Ab may be administered first, i.e., in some embodiments, the anti-PD-1 Ab is administered before the anti-CTLA-4 Ab, while in other embodiments, the anti-CTLA-4 Ab is administered before the anti-PD-1 Ab. Generally, each Ab is administered by intravenous infusion over 60 minutes. In some embodiments, the anti-PD-1 and anti-CTLA-4 Abs are administered simultaneously, either mixed as one composition in a pharma- ceutically acceptable formulation for simultaneous administration, or as separate compositions with each Ab in a pharma- ceutically acceptable formulation.

[0130] Certain embodiments of the method include (a) an induction phase consisting of administration of nivolumab by intravenous infusion for four combined doses every three weeks, followed by administration of ipilimumab by intravenous infusion, and (b) followed by maintenance dosing with nivolumab administered by intravenous infusion every two weeks, beginning three weeks after the fourth dose of induction therapy or on or after day 113 if the fourth dose of induction therapy was not administered due to a treatment delay.

[0131] In some embodiments, anti-PD-1 Ab or its antigen-binding portion, anti-angiogenic TKI and anti-CTLA-4 Ab or its antigen-binding portion can be administered to a subject for the treatment of RCC.In some embodiments, anti-angiogenic TKI is sorafenib, sunitinib, pazopanib, axitinib or tivozanib.In some embodiments, anti-angiogenic TKI is sunitinib or pazopanib.In other embodiments, anti-CTLA-4 Ab is ipilimumab or tremelimumab. In certain embodiments, the combination of anti-PD-1 Ab or antigen-binding portion thereof, anti-angiogenic TKI and anti-CTLA-4 Ab or antigen-binding portion thereof is administered at: (i) 0.1 mg / kg anti-PD-1 Ab and 3 mg / kg anti-CTLA-4 Ab; (ii) 0.3 mg / kg anti-PD-1 Ab and 3 mg / kg anti-CTLA-4 Ab; (iii) 1 mg / kg anti-PD-1 Ab and 3 mg / kg anti-CTLA-4 Ab; (iv) 3 mg / kg anti-PD-1 Ab and 3 mg / kg anti-CTLA-4 Ab; (v) 5 mg / kg anti-PD-1 Ab and 3 mg / kg anti-CTLA-4 Ab; (vi) 10 mg / kg anti-PD-1 Ab and 3 mg / kg anti-CTLA-4 Ab; (vii) 0.1 mg / kg anti-PD-1 Ab and 1 mg / kg anti-CTLA-4 Ab; (viii) 0.3 mg / kg anti-PD-1 Ab and 1 mg / kg anti-CTLA-4 Ab. Ab; (ix) 1 mg / kg anti-PD-1 Ab and 1 mg / kg anti-CTLA-4 Ab; (x) 3 mg / kg anti-PD-1 Ab and 1 mg / kg anti-CTLA-4 Ab; (xi) 5 mg / kg anti-PD-1 Ab and 1 mg / kg anti-CTLA-4 Ab; or (xii) 10 mg / kg anti-PD-1 Ab and 1 mg / kg anti-CTLA-4 Ab, where the TKI is administered at about its recommended dose. For example, sunitinib can be administered at a dose ranging from about 25 to about 50 mg, in one embodiment at a dose of 50 mg; pazopanib can be administered at a dose of about 200 mg, about 400 mg, or about 800 mg; sorafenib can be administered at a dose of about 400 mg twice a day; and tivozanib can be administered at its recommended oral daily dose of about 1.5 mg per day.

[0132] Combination of anti-PD-1 Ab with other known anticancer drugs for the treatment of RCC Those skilled in the art will recognize that the described method of treating RCC with anti-PD-1 antibody or its antigen-binding fragment can be combined with other known treatments for RCC. For example, anti-PD-1 Ab or its antigen-binding fragment can be combined with mitotic inhibitors such as paclitaxel, docetaxel, vincristine, eribulin, estramustine, etoposide, ixabepilone, cabazitaxel, vincristine liposome, vinorelbine, vincristine, vinblastine or teniposide. As mentioned above, various cytokines are also known to be useful in treating cancer, including IFN-α or IL-2. Anti-PD-1 Ab or its antigen-binding fragment can be combined with cytokines such as IFN-α, IFN-b2, pegylated IFNb2 (PegIFN-b2) or IL-2. Other small molecule treatments known in the art, including but not limited to vinflunine, may also be suitable. In some embodiments, the anti-PD-1 antibodies described herein, including combination treatments of anti-PD-1 antibodies and TKIs or anti-CTLA-4 antibodies, are used in combination with any other treatments described herein, including SOC treatments.

[0133] The anti-PD-1 antibodies can be administered in combination with one or more other known anti-cancer agents described herein in dosing that is standard for the particular anti-cancer agent. For example, pazopanib can be administered at a dose of about 400 to about 800 mg; axitinib can be administered at a dose of about 1 mg, about 2 mg, about 3 mg, about 4 mg, about 5 mg, about 6 mg, about 7 mg, about 8 mg, about 9 mg, or at least about 10 mg; paclitaxel and / or docetaxel can be administered at a dose of about 10 mg / m 2 , about 25mg / m 2 , about 30mg / m 2 , about 35mg / m 2 , about 40mg / m 2 , about 45mg / m 2 , about 50mg / m 2 , about 75mg / m 2 , about 100mg / m 2 , about 125mg / m 2 , about 150mg / m 2, about 175mg / m 2 , about 200mg / m 2 , about 225mg / m 2 , about 250mg / m 2 , about 275mg / m 2 or at least about 300 mg / m 2 Vinflunine can be administered at a dose of approximately 50 mg / m 2 , about 75mg / m 2 , about 100mg / m 2 , about 125mg / m 2 , about 150mg / m 2 , about 175mg / m 2 , about 200mg / m 2 , about 225mg / m 2 , about 250mg / m 2 , about 275mg / m 2 , about 300mg / m 2 , about 325mg / m 2 , about 350mg / m 2 , about 375mg / m 2 , about 400mg / m 2 , about 425mg / m 2 , about 450mg / m 2 , about 475mg / m 2 or at least about 500 mg / m 2 can be administered at a dose of

[0134] kit Kits containing anti-PD-1 Ab and TKI for therapeutic use are also within the scope of the present invention. The kits generally include a label for the intended use and instructions for use of the contents of the kit. The term label includes any writing or recording medium on or delivered with the kit or which otherwise accompanies the kit. Thus, the present invention provides kits for treating a subject suffering from renal cancer, the kits comprising: (a) an anti-cancer agent, which is an Ab or antigen-binding portion thereof that specifically binds to the PD-1 receptor and inhibits PD-1 activity, at a dose in the range of 0.1-10 mg / kg body weight; (b) a fixed dose of another anti-cancer agent, which is (i) an anti-angiogenic TKI, e.g., sunitinib at a dose of 12.5 mg, 25 mg or 50 mg once daily, pazopanib at a dose of 200 mg, 400 mg or 800 mg once daily, or sorafenib at a dose of 400 mg once or twice daily, on a schedule of 4 weeks on followed by 2 weeks off; or (ii) an antibody or antigen-binding portion thereof that specifically binds and inhibits CTLA-4, at a dose range of 0.1-10 mg / kg body weight; and (c) instructions for using the anti-PD-1 Ab and the other anti-cancer agent in any of the combination therapies disclosed herein. In some embodiments, the anti-PD-1, anti-CTLA-4 Ab and / or TKI can be co-packaged in a unit dose form. In some embodiments for the treatment of human patients, the kit includes an anti-human PD-1 Ab disclosed herein, such as nivolumab or pembrolizumab. In other embodiments, the kit includes an anti-human CTLA-4 Ab disclosed herein, such as ipilimumab or tremelimumab.

[0135] This invention is further illustrated by the following examples which should not be construed as further limiting. All references cited throughout this specification are expressly incorporated herein by reference. EXAMPLES

[0136] Example 1 Treating renal cancer with anti-PD-1 and TKIs in mouse models material and method animal Female 8-12 week old Balb / c and C57 / BL6 mice (Harlan Laboratories, Federick, MD) were used in all studies. Mice were housed in sterile microisolator cages unless otherwise noted and were provided with sterile food and water ad libitum.

[0137] reagent Sunitinib (Sutent®) is an orally bioavailable, small molecule, multikinase inhibitor that targets several receptor tyrosine kinases, including VEGFR-1, VEGFR-2 and VEGFR-3, PDGFR-α and -β, c-Kit, FLT-3, CSF-1R and RET, that is FDA approved for the treatment of renal cell carcinoma. Sunitinib has been reported to exhibit immunomodulatory effects, including reduction of myeloid-derived suppressor cells (Ko et al., 2009; Ko et al., 2010) and reduction of T regulatory suppressor cells (Hipp et al., 2008; Ozao-Choy et al., 2009).

[0138] Sorafenib (Nexavar®) is an orally available, small molecule multikinase inhibitor (cRAF, bRAF, KIT, FLT-3, VEGFR-2, VEGFR-3 and PDGFR-β) approved by the FDA for the treatment of primary renal cancer (advanced renal cell carcinoma), advanced primary liver cancer and radioactive iodine-resistant advanced thyroid cancer. Sorafenib has been reported to inhibit T cell proliferation by inducing apoptosis (Houben et al., 2009; Molhoek et al., 2009), reduce antigen-specific T cell induction and reduce dendritic cell maturation (Hipp et al., 2008).

[0139] The anti-PD-1 mAb, 4H2, is a chimeric rat-mouse anti-mouse PD-1 Ab (U.S. Patent No. 8,008,449) that has been purified and proven to have an endotoxin level of <0.5EU / mg, >95% purity and <5% high molecular weight species.

[0140] Mouse renal carcinoma (Renca) and mouse colon adenocarcinoma (CT-26) tumor cell lines were maintained in vitro for less than 10 passages.

[0141] Efficacy Testing Tumor cells were implanted subcutaneously (SC). The average tumor volume was approximately 100 mm 3 When tumors reached 100 mg / kg, mice were randomized into groups of 8 animals. Vehicle (control), sunitinib (120 mg / kg) or sorafenib (200 mg / kg) were administered orally once daily for 14 days (QD×14, PO). MAb 4H2 was administered intraperitoneally (IP) at 10 mg / kg every 4 days for a total of 4 doses (Q4D×4). Tumors were measured twice weekly and tumor volumes were calculated as length×width. 2 Synergy was defined as the excess of highest single agent (EOHSA), i.e., a combination was considered synergistic if the antitumor effect was significantly greater than that observed with monotherapy (P<0.05, Wilcoxon signed-rank test).

[0142] result The effect of treatment with anti-PD-1 and sunitinib or sorafenib as monotherapy or combination therapy on the median volume of implanted RCC mouse tumors is shown in Figure 1. Sunitinib monotherapy was active in this Renca mouse RCC model, resulting in 85% tumor growth inhibition (TGI) by the last day of treatment (day 33; see Table 1); however, tumors grew progressively after treatment was discontinued (see Figure 1). When administered as monotherapy, anti-PD-1 mAb was inactive in this model, but the combination of anti-PD-1 and sunitinib produced significant antitumor activity (101% TGI) and resulted in complete tumor regression (Table 1) or significant delay in tumor growth (Figure 1) by the end of monitoring 50 days after implantation. These results, observed in two separate studies of the same model, indicate a synergistic interaction of nivolumab murine surrogate anti-PD-1 mAb with sunitinib in a murine RCC (Renca) tumor model, as the effect of this combination in inhibiting tumor growth was significantly greater than the sum of the levels of inhibition exhibited by each therapeutic agent individually.

[0143] In contrast, although sorafenib monotherapy was active in this RCC model (62% TGI; Table 1 ), combination treatment with anti-PD-1 mAb and sorafenib did not significantly increase the antitumor activity of sorafenib in this mouse model and did not result in synergistic efficacy ( Fig. 1 ).

[0144] Figure 2 shows a tumor measuring 500 mm 3 The time (days) taken to reach a volume of 500 mm is shown. Consistent with the data in Figure 1, the growth of tumors treated with the anti-PD-1 / sunitinib combination was significantly inhibited, and these tumors grew to a volume of 500 mm. 3 It took approximately 63 days to reach this milestone, significantly longer than the 19-34 days taken in tumors treated with control, anti-PD-1 Ab, sunitinib, sorafenib, or the anti-PD-1 / sorafenib combination.

[0145] The tumor growth inhibition percentages measured 33 days after implantation are shown in Table 1. These data are consistent with the data shown in Figures 1 and 2, confirming that the combination of anti-PD-1 and sunitinib completely inhibited tumor growth, while the other treatments resulted in low levels of tumor growth inhibition. The superior efficacy shown by the combination of anti-PD-1 and sunitinib over the combination of anti-PD-1 and sorafenib is consistent with the different immunomodulatory effects mediated by sunitinib and sorafenib, respectively, suggesting that sunitinib, but not sorafenib, is suitable for combination with immunotherapeutic approaches for the treatment of cancer patients (see Hipp et al., 2008). However, the efficacy of the combination of anti-PD-1 and sorafenib in human subjects may not fully reflect the efficacy observed in the mouse RCC model, and this combination may be highly effective for the treatment of human patients.

[0146] [Table 1] CR=complete regression;PR=partial regression

[0147] The effect of the various treatments on the overall body weight of the mice is shown in Figure 3. The combination of anti-PD-1 and sunitinib was well tolerated as evidenced by the lack of weight loss in the mice. The combination of anti-PD-1 and sorafenib was also well tolerated, although it was apparently slightly less tolerable than the combination of anti-PD-1 and sunitinib, as the combination with sorafenib caused a small but significant weight loss.

[0148] Example 2 Immune cell infiltration in tumors following treatment with anti-PD-1 and sunitinib material and method Immunohistochemistry and flow cytometry analysis were used to assess immune cell infiltration of tumors.

[0149] immunohistochemistry Frozen sections were fixed in an acetone / methanol mixture and rinsed with phosphate-buffered saline (PBS). Sections were blocked for endogenous peroxidase and primary antibodies (anti-CD8, anti-CD4, anti-FoxP3 and anti-PD-L1) were added and incubated overnight. Chromogenic development was performed using a brown polymer-based detection system (Biocare Medical) and hematoxylin was used as a blue counterstain for visualization of cell nuclei.

[0150] Immunophenotyping by flow cytometry Tumor-infiltrating immune cells were isolated and prepared as single-cell suspensions, then stained and analyzed by flow cytometry for expression of T cell subsets (regulatory and activated) and myeloid cells (i.e., for expression of CD3, CD4, CD8, FoxP3, CD25, and CD69).

[0151] result In control mice, immunohistochemistry (IHC) analysis revealed no CD4 + and CD8 + When animals were treated with sunitinib, we detected predominantly CD4 T cells in the periphery and along the vasculature (Figure 4). + and CD8 +In contrast, when mice were treated with the combination of anti-PD-1 mAb and sunitinib, the entire tumor was enriched in CD4 + and CD8 + A significant infiltration of T cells was observed. The combination of anti-PD-1 mAb and sunitinib significantly increased tumor-infiltrating CD4 T cells compared with either drug alone or vehicle control. + and CD8 + 13 increased the number of T cells (Figure 4). In contrast, increased T cell infiltration into the tumor was not seen with anti-PD-1 mAb or sorafenib alone or with the combination of anti-PD-1 mAb and sorafenib versus the control group (data not shown). Sunitinib and sorafenib have been shown to exhibit immunomodulatory effects, which may be related to the different therapeutic effects seen when these agents are combined with anti-PD-1 Ab immune checkpoint inhibitors.

[0152] Flow cytometry analysis of tumor-infiltrating immune cells showed significant differences in the groups treated with anti-PD-1 mAb in combination with sunitinib or sorafenib (Figures 5-7). Compared with the combination of anti-PD-1 mAb and sorafenib, the combination of anti-PD-1 mAb and sunitinib significantly reduced (i) the proliferation of CD4 + / Ki67 + , CD8 + and CD8 + / Ki67 + (ii) CD11b (P<0.05; Figs. 5 and 6); + / Ly6C hi / Ly6G - monocytic myeloid-derived suppressor cells (P<0.05), CD11b + / Ly6C low / Ly6G - (iii) a decrease in the percentage of monocytic cells expressing CD11b or both (P<0.05; Fig. 7); and (iii) a decrease in the percentage of granulocytic myeloid cells expressing CD11b + / Ly6C - / Ly6G +(P<0.05; Figure 7). Therefore, treatment with the combination of sunitinib and anti-PD-1 mAb induced changes in the composition of the tumor microenvironment that may support an effective antitumor immune response. + / CD4 + / CD25 + / FoxP3 + ) were detected across experimental groups, making it difficult to determine treatment effects (data not shown).

[0153] Example 3 Enhancement of in vivo cytotoxicity of antigen-specific immune responses material and method Cytotoxicity assay The CT26 mouse colon cancer model expressing the AH1 antigen was used in this study to determine whether treatment had any effect on antigen-specific T cells. Following SC implantation of CT26 cells for 5 days, mice were treated with vehicle or IP with anti-PD-1 mAb 10 mg / kg administered twice, 3 days apart (Q3D×2), with or without sunitinib 120 mg / kg once daily for 5 days (QD×5). Cell death was examined 2 and 4 days after the final treatment.

[0154] One day prior to the assay, splenocytes from naive BALB / c donors were incubated with or without CT26 peptide ([H]SPSYVYHQF[OH]; Sigma-Genosys) and labeled with 5(6)-carboxyfluorescein diacetate N-succinimidyl ester (CFSE, Sigma-Aldrich). Antigen-stimulated and unstimulated cells were combined (1:1) and injected into treated mice. The percentage of cell death was determined by flow cytometry 24 hours later. result

[0155] In the CT26 tumor model, the combination of anti-PD-1 mAb and sunitinib resulted in increased in vivo cytotoxicity against antigen-specific immune responses (Figure 8). This enhanced cytotoxicity represented an increase in the immune response component and also correlated with a decrease in tumor volume (Figure 9).

[0156] Example 4 Phase 1 Clinical Trial for Treating Metastatic RCC with Nivolumab in Combination with Sunitinib, Pazopanib, or Ipilimumab Collectively, the data disclosed in Examples 1-3 support the study of combinations of anti-PD-1 Abs, such as nivolumab or pembrolizumab, with anti-angiogenic TKIs, such as sunitinib, pazopanib, sorafenib, axitinib, or tivozanib, to treat subjects with RCC. These data suggest that the immunomodulatory properties of sunitinib may cause changes in the tumor microenvironment that support an effective antitumor immune response, which is expanded and enhanced by anti-PD-1 Abs. Although combination treatment of anti-PD-1 mAb and sorafenib did not significantly enhance the antitumor activity of sorafenib in a mouse RCC model and did not result in synergistic efficacy (Figure 1), the combination was well tolerated and could be effective in human patients. The safety and efficacy of nivolumab in combination with sunitinib, pazopanib or ipilimumab is currently being evaluated in an ongoing Phase 1 study (NCT01472081) in subjects with mRCC. Pazopanib is an orally available multikinase inhibitor (VEGFR-1, VEGFR-2, VEGFR-3, PDGFR-α and -β and c-KIT), and the combination of nivolumab and ipilimumab has shown significantly enhanced efficacy in the treatment of melanoma compared to monotherapy with nivolumab (Wolchok et al., 2013; WO2013 / 173223).

[0157] The primary objective of this Phase 1 study is to evaluate the overall safety and tolerability of nivolumab plus sunitinib, pazopanib or ipilimumab to determine the MTD and recommended Phase 2 dose of each combination in patients with RCC. Secondary objectives are to evaluate the preliminary antitumor activity of the combinations, and an exploratory objective is to determine whether response correlates with baseline PD-L1 expression.

[0158] method Study design A phase 1 open-label study of nivolumab plus sunitinib, pazopanib or ipilimumab in subjects with mRCC is ongoing. The study includes 5 parallel treatment arms: nivolumab plus sunitinib (arm S), nivolumab plus pazopanib (arm P), nivolumab 3 mg / kg plus ipilimumab 1 mg / kg (arm I-1), nivolumab 1 mg / kg plus ipilimumab 3 mg / kg (arm I-3) and nivolumab 3 mg / kg plus ipilimumab 3 mg / kg (arm IN-3). Treatment in arms S and P is conducted in two parts: dose escalation (previously treated subjects) followed by dose expansion (naïve subjects). Treatment in arms I-1 and I-3 is a single initial cohort, with each arm consisting of both previously treated and naïve subjects. The expansion cohorts of arms I-1 and I-3 and arm IN-3 consist of minimally treated (including treatment-naive) subjects. The study designs of arms S and P are summarized in Figure 10A, and the study designs of arms I-1, I-3 and IN-3 are summarized in Figure 10B.

[0159] Study population The study population is subjects ≥18 years of age with histologically confirmed metastatic renal cell carcinoma with clear cell component with at least one measurable disease by Response Evaluation Criteria in Solid Tumors (RECIST) 1.1. Subjects with non-clear cell RCC with measurable disease defined by RECIST 1.1 are admitted to the dose escalation cohorts of arms S and P only. Favorable or intermediate risk Memorial Sloan Kettering Cancer Center (MSKCC) prognostic score and Karnofsky Performance Status (KPS) ≥80% are also required for arms S, P and I-1 and I-3 original cohorts (for arms I-1 and I-3 expansion and IN-3, any MSKCC prognostic score is acceptable).

[0160] Arm S and Arm P Subjects must have received at least one prior systemic treatment regimen in the advanced / metastatic setting to be eligible for the dose-escalation portion of the study, while subjects must be treatment-naïve to be eligible for the dose-expansion portion of the study. Subjects entering the dose-escalation phase who had previously received pazopanib were assigned to receive nivolumab plus sunitinib, while subjects who had previously received sunitinib were assigned to receive nivolumab plus pazopanib. Subjects who had previously received pazopanib and sunitinib were not eligible. Subjects who were chronically treated with systemic steroids or any other immunosuppressive agent or who had previously been treated with anti-PD-1, anti-PD-L1 or anti-PD-L2, anti-CD137, anti-CTLA-4, or similar agents were also excluded from the study.

[0161] Subjects who have not previously received sunitinib or pazopanib can be enrolled in the dose escalation arm and will be assigned to these arms in an alternating fashion when both arms are open. Approximately 36 subjects (18 subjects / arm) are scheduled to be treated in the dose escalation portion of the study. Upon completion of this escalation cohort, the MTD will be evaluated based on a modified toxicity probability interval (mTPI) design. If the MTD of nivolumab is detected to be <5 mg / kg in either arm S or arm P, the arm will not be further expanded to collect safety data in untreated subjects. If the MTD of nivolumab in combination with sunitinib or pazopanib is determined to be 5 mg / kg or greater, another expansion cohort in each arm will be opened for enrollment at the 5 mg / kg dose. In the expansion cohorts of arm S and arm P, only untreated subjects will be enrolled, so that approximately 20 untreated subjects per arm will be treated. Each treatment cycle will be 6 weeks, with nivolumab administered on days 1 and 22, and antiangiogenic therapy administered according to the approved package inserts for sunitinib (50 mg daily; 4 weeks on, 2 weeks off) and pazopanib (800 mg daily).

[0162] Dose escalation in arms S and P Dose levels are shown in Tables 2 and 3. Intrapatient dose escalation of nivolumab is not permitted. [Table 2] a Dose level -1 will be enrolled only if dose reduction from dose level 1 is necessary

[0163] [Table 3] a Dose level -1 will be enrolled only if dose reduction from dose level 1 is necessary

[0164] The starting dose of nivolumab for each dose escalation arm is 2 mg / kg. Initially, 7 eligible subjects per arm will be treated at dose level 1. The decision to treat additional subjects at the same dose level or to escalate to a higher dose level (5 mg / kg) will be guided by the number of dose-limiting toxicity (DLT) observed during the DLT observation period. Subjects who do not complete the DLT period for reasons other than dose-limiting toxicity will be replaced. Dose level-1 (0.3 mg / kg) may be considered if the safety and tolerability profile of 2 mg / kg is not acceptable.

[0165] For the Phase 1 dose escalation study of arms S and P, the sample size for each dose in these arms will depend on observed toxicity and posterior distribution. A total of 18 subjects are expected to be treated in the dose escalation phase of each arm. However, there may be more than 18 subjects per arm due to alternation or less than 18 subjects per arm due to toxicity at low dose levels.

[0166] Dose expansion for Arm S and Arm P Once the safety profile and tolerability of all doses tested have been characterized and the MTD of the combination of nivolumab + sunitinib and nivolumab + pazopanib has been defined for a treatment arm, cohort expansion will begin in that arm at 5 mg / kg only if the nivolumab dose at the MTD is ≥ 5 mg / kg. If the MTD of nivolumab is determined to be < 5 mg / kg in any arm, that arm will not be further expanded. An additional 20 subjects per treatment arm will be treated at the 5 mg / kg dose level for nivolumab in combination with sunitinib or pazopanib to obtain further safety information.

[0167] Arms I-1 and I-3 (First Cohort) Both pretreated and untreated subjects are eligible to participate, with approximately 20 subjects treated in each arm, excluding subjects previously treated with anti-PD-1, anti-PD-L1 or anti-PD-L2, anti-CD137, anti-CTLA-4, or other similar agents. Dosing for the I-1, I-3, and IN-3 arms is detailed in Table 4 below. Intrapatient dose escalation or deescalation will not be permitted for any of the drugs. [Table 4]

[0168] Each treatment cycle is 6 weeks. Nivolumab and ipilimumab are administered every 3 weeks for 4 doses, then nivolumab is administered every 2 weeks starting 3 weeks after the 4th dose of induction therapy or after day 113 if the 4th dose of induction therapy was not administered due to a treatment delay.

[0169] In the initial cohorts of subjects in I-1 and I-3, approximately 20 subjects per arm will be enrolled and treated. These arms will be expanded to a total of 45 subjects in each arm (25 more in each arm in the expansion cohort) for further evaluation of safety and efficacy.

[0170] About Arms I-1, I-3 (expansion cohort) and IN-3 Minimally treated (including untreated) subjects are eligible for enrollment, with approximately 25 subjects treated in each arm. Each treatment cycle is 6 weeks. Nivolumab and ipilimumab are administered every 3 weeks for 4 doses, followed by nivolumab every 2 weeks starting 3 weeks after the 4th dose of induction therapy or after day 113 if the 4th dose of induction therapy was not administered due to a treatment delay.

[0171] Allocation of previously treated subjects to treatment arms Subjects who have received prior treatment in the advanced or metastatic setting are assigned to dose escalation cohorts of arms S and P as described above. Subjects who have received one systemically prior treatment are assigned to at least two ipilimumab combination arms (I-1 and I-3) in an alternating fashion when both of these arms are open, unless they are qualified for escalation arms S and / or P when these arms are open. When only one arm is open, these subjects are assigned to the open arm.

[0172] Allocation of untreated subjects to treatment arms When all 4 arms (expansion arms S and P; I-1 and I-3) are open for enrollment, treatment-naive subjects will be randomly assigned in a 1:1:1:1 ratio to the open arms until enrollment is complete.

[0173] Allocation of Minimally Treated Subjects to Treatment Arms For I-1 and I-3 extensions and IN-3, minimally treated (including treatment-naïve) subjects will be assigned in a 1:1:1 ratio to one of the open arms until enrollment is complete. Adverse events will be staged according to NCI CTCAE v4.0. Disease assessments will be performed every 6 weeks (± 1 week) from the first dose of study drug for the first 4 evaluations, then every 12 weeks (± 1 week) until disease progression. Response assessments will be performed according to RECIST 1.1. For the dose escalation phase in Arm S and Arm P, subjects will be treated until unacceptable toxicity, disease progression, or withdrawal of informed consent.

[0174] Test evaluation Safety outcome measures Safety / tolerability outcome measures and determination of the MTD of the combination are the primary endpoints of the study. Key safety assessments are: (1) occurrence of adverse events (AEs) occurring up to 100 days after the last dose of study drug (or longer for drug-related AEs that do not return to baseline, do not stabilize, do not return, or are considered irreversible until 100 days after the last dose), (2) serious adverse events (SAEs) occurring up to 100 days after the last dose of study drug (or longer for drug-related SAEs that do not return to baseline, do not stabilize, do not return, or are considered irreversible until 100 days after the last dose), and (3) hematologic Frequency of clinical testing by worst toxicity grade, including fluidics, serum chemistry and urinalysis (assessed at screening, baseline, days 1, 8, 22 and 29 of cycles 1-4 and days 1 and 22 of cycle 5 for arms S and P; assessed at screening, baseline, days 1, 8, 22 and 29 of cycles 1-2 and days 1, 15 and 29 of cycle 3+ for arms I-1 and I-3; assessed at end of treatment and follow-up visits 100 days after the last dose for all arms).

[0175] Efficacy outcome measures Antitumor activity is a secondary endpoint of the study and will be measured by objective response rate (ORR) per RECIST 1.1 and duration of response (DOR). Tumor response will be based on tumor assessments at screening for the first 4 assessments, every 6 weeks (± 1 week) from the first dose of study drug, and every 12 weeks (± 1 week) until disease progression.

[0176] Example 5 Antitumor activity demonstrated in the S and P arms Seven subjects were treated in each of arms S N2 and S N5 and received nivolumab (2 mg / kg or 5 mg / kg, respectively) in combination with S (50 mg, 4 weeks on, 2 weeks off) until progression or unacceptable toxicity. No DLTs were observed and the MTD was not reached. Therefore, based on tolerability, the S N5 arm was expanded to 19 additional treatment-naïve patients (total n=33 for S arm). Arm P enrolled 20 subjects in N2 and received nivolumab 2 mg / kg in combination with P (800 mg daily). Four 4 DLTs (ALT / AST elevation [n=3], fatigue [n=1]) were observed, leading to the closure of this arm. The dose escalation performed is illustrated in Figure 11.

[0177] Seven subjects in the S N2 arm, 26 subjects in the S N5 arm, and 20 subjects in the P N2 arm were evaluated for response until database lock in March 2014. Clinical activity (OR: confirmed complete [CR] or partial [PR] response) was observed in all arms as shown in Table 5. The number of evaluable subjects treated with sunitinib and 2 mg / kg nivolumab was small (7 patients), but of these, 1 (14%) had a complete response, 5 (71%) had a partial response, and 1 (14%) had stable disease. Overall, 33 evaluable subjects treated with sunitinib and both concentrations of nivolumab (2 mg / kg and 5 mg / kg) had 18 (55%) confirmed objective responses (OR). Of the 20 evaluable subjects treated with pazopanib and 2 mg / kg nivolumab, 9 (45%) had confirmed ORs. Responses occurred by first evaluation (6 weeks) in 41% (arm S) and 56% (arm P) of responding subjects. ORRs and corresponding two-sided exact 95% CIs were calculated by the method of Clopper and Pearson.

[0178] [Table 5] BOR: best overall response; CR: complete response; PR: partial response; SD: stable disease; PD: progressive disease; UD: indeterminable; CI: confidence interval; (A): CR only; (B): CR + unconfirmed response; unconfirmed response: ongoing subjects with tumor response at last tumor assessment; DOR: duration of response.

[0179] Response durations ranged from 18.1 to 118++ weeks in arm S and 12.1 to 117.1++ weeks in arm P. Stable disease rates were 30% (n=10) in arm S and 35% (n=7) in arm P. Median response durations and corresponding two-sided 95% CIs were determined using the Kaplan-Meier method.

[0180] The effect on tumor burden of evaluable subjects in the S and P arms is illustrated by radar charts (FIGS. 12A-C) and waterfall plots (FIG. 12D).

[0181] The confirmed ORRs shown in Table 5, ranging from 45% in the PAZ+NIV2 arm to 86% in the SUN+NIV2 arm, are significantly higher than those seen with nivolumab or sunitinib or pazopanib monotherapy. For example, WO2013 / 173223 reports an ORR of approximately 28-31% with nivolumab (1 mg / kg or 10 mg / kg) alone for the treatment of RCC in a Phase 1 study. In a more recent analysis, Motzer et al. (2014) report ORRs of 20%, 22% and 20% for nivolumab administered to patients at 0.3 mg / kg, 2 mg / kg and 10 mg / kg, respectively, in a Phase 2 study for the treatment of metastatic RCC. Choueiri et al. (2014) report that nivolumab administered at 0.3 mg / kg, 2 mg / kg and 10 mg / kg produced ORRs of 9%, 23% and 22% in patients with metastatic RCC pretreated with 1-3 prior antiangiogenic therapies, and 10 mg / kg nivolumab produced an ORR of 13% in untreated patients. For TKIs, historical ORRs of approximately 27.5% have been observed with sunitinib alone (see, e.g., Motzer et al., 2007; 2013) and approximately 30% with pazopanib alone (see, e.g., Sternberg et al., 2010; Motzer et al., 2013).

[0182] At the first evaluation (6 weeks), 7 of 17 subjects (41.2%) were responders in the S arm and 5 of 9 subjects (55.6%) were responders in the P arm. Ongoing responders included 10 of 17 subjects (58.8%) in the S arm and 3 of 9 subjects (33.3%) in the P arm.

[0183] Progression-free survival in the S and P arms A plot of progression-free survival (PFS) rate versus time is shown in Figure 13. The PFS rate at 24 weeks was 79% in arm S and 55% in arm P. The median PFS in the S arm (S+N; n=33) was 55.3 weeks (95% CI 47.9-72.4 weeks) and in the P arm (P+N; n=20) was 31.4 weeks (95% CI 12.1-48.1 weeks).

[0184] Objective response rate by baseline PD-L1 expression Tumor tissue collection was retrospective (33 samples were available in the S arm and 20 in the P arm). Surface PD-L1 expression was measured by Dako immunohistochemistry assay using rabbit anti-human PD-L1 mAb, 28-8 (WO2013 / 173223). The cutoff for positive PD-L1 signal was 1% or 5% of tumor membrane staining. The ORR observed in patients in the two arms stratified according to PD-L1 expression status is shown in Table 6.

[0185] [Table 6] a ORR in PD-L1-evaluable patients; ORR includes complete or partial responders as determined by RECIST.

[0186] These data indicate that the response of RCC patients to the combination of nivolumab and sunitinib or pazopanib does not correlate with tumor expression of PD-L1.

[0187] Adverse events in the S and P arms Grade 3–4 treatment-related adverse events (AEs) were observed in 27 / 33 patients (82%) in arm S and 14 / 20 patients (70%) in arm P. Most toxicities were consistent with the known profile of TKIs. The most common treatment-related grade 3–4 AEs were ALT elevation and hypertension (18% each), hyponatremia and lymphocytopenia (15% each) in arm S and ALT and AST elevation and diarrhea (20% each) and fatigue (15%) in arm P. Grade 3 interstitial pneumonitis occurred in 1 patient (arm S, N5). Grade 3–4 treatment-related AEs led to treatment discontinuation in 11 / 33 patients (33%; 3 N2, 8 N5) in arm S and 4 / 20 patients (20%) in arm P. No grade 5 treatment-related AEs were observed. Hepatotoxicity was controllable using the treatment algorithm.

[0188] Overall, the clinical data indicate that the combination of nivolumab and sunitinib shows promising antitumor activity and a manageable safety profile in patients with mRCC. Arm P was closed due to DLT.

[0189] Example 6 Antitumor activity demonstrated in the ipilimumab arm Patients with mRCC (favorable or intermediate MSKCC score; Karnofsky performance status ≥80%; untreated or any number of prior therapies other than prior immuno-oncology treatment) were randomized to various treatment arms. 47 subjects treated with 1 mg / kg ipilimumab and 3 mg / kg nivolumab (I-1 arm), 47 subjects treated with 3 mg / kg ipilimumab and 1 mg / kg nivolumab (I-3 arm), and 6 subjects treated with 3 mg / kg ipilimumab and 3 mg / kg nivolumab (IN-3) were evaluable for response. Most patients (n=54; 54%) had received prior systemic treatment (I-1: 22; I-3: 26; IN-3: 3). ORs were observed in I-1 and I-3 arms as shown in Table 7. ORRs and corresponding two-sided exact 95% CIs were calculated by the Clopper and Pearson method. Overall, 16 (34%) patients in the I-1 arm showed a confirmed OR and 17 (36%) patients in the I-3 arm showed a confirmed OR.

[0190] These ORRs are significantly higher than those seen with nivolumab alone (WO2013 / 173223; Motzer et al., 2014; Choueiri et al., 2014; see Example 5) or ipilimumab alone. Treatment of patients with metastatic RCC with ipilimumab monotherapy in a phase 2 study resulted in an ORR of 5% in cohort A (3 mg / kg loading dose, followed by 1 mg / kg every 3 weeks) and 12.5% ​​in cohort B (3 mg / kg every 3 weeks for all doses), although disparities in characteristics of the two groups prevented direct comparison of efficacy (Yang et al., 2007).

[0191] The median duration of response (DOR) was 53.9 weeks in the I-3 arm and not reached in the I-1 arm. Stable disease (SD) as best overall response was seen in 19 (40%) (I-1) and 18 (38%) (I-3) patients.

[0192] The effect on tumor burden in evaluable subjects is illustrated in radar charts (FIGS. 14A-C) and waterfall plots (FIG. 14D). A comparison of the waterfall plots for subjects treated in the S, P, and I arms is shown in FIG.

[0193] [Table 7] BOR: best overall response; CR: complete response; PR: partial response; SD: stable disease; PD: progressive disease; UD: undeterminable; CI: confidence interval; (A): CR only; (B): CR + unconfirmed response; unconfirmed response: ongoing subjects with tumor response at last tumor assessment; PFS: progression free survival, DOR: duration of response; NR: not reached; a Because of the high percentage of ongoing responses, the median DOR can be misleading. Duration of response is the time between the date of first response and the date of disease progression or death (whichever occurs first).

[0194] Progression-free survival in I-1 and I-3 arms A plot of PFS rate versus time is shown in Figure 16. The median PFS was 30.3 weeks in the I-1 arm and 36.0 weeks in the I-3 arm. PFS rates were estimated by the Kaplan-Meier method, and the corresponding 95% CI was derived based on the Greenwood formula.

[0195] Objective response rate by baseline PD-L1 expression Tumor tissue samples were collected retrospectively and surface PD-L1 expression was measured by Dako immunohistochemistry assay using mAb, 28-8 (WO2013 / 173223). The cutoff for positive PD-L1 signal was 1% or 5% of tumor membrane staining. The ORR observed in different patients stratified according to PD-L1 expression status is shown in Table 8. [Table 8] a ORR in PD-L1-evaluable patients; ORR was determined by RECIST and included complete or partial responders.

[0196] These data indicate that the response of RCC patients to the combination of nivolumab and ipilimumab does not correlate with tumor expression of PD-L1.

[0197] Adverse events in I-1 and I-3 arms Grade 3-4 related adverse events (AEs) were observed in 16 / 47 (34%), 30 / 47 (64%) and 5 / 6 (83%) in the I1, I3 and IN-3 arms, respectively. Five (11%) and 10 (21%) in the I1 and I3 arms, respectively, discontinued treatment due to related AEs. No patients discontinued due to adverse events in the IN-3 arm, however, all 6 patients required steroids for treatment of immune-mediated adverse events, and no further patients were enrolled in the IN3 arm. The most common grade 3-4 related adverse events included lipase elevation (20%, n=20), ALT elevation (11%, n=11), diarrhea (9%, n=9) and AST elevation (6%, n=6). No grade 3-4 interstitial pneumonitis was observed.

[0198] Overall, nivolumab and ipilimumab have shown promising evidence of grade 3-4 events and significant antitumor activity in mRCC, with acceptable safety manageable within established treatment guidelines, with the majority of responses ongoing. ORR suggests greater activity than previously reported with nivolumab or ipilimumab monotherapy in RCC (Topalian et al., 2012a; Motzer et al., 2014; Choueiri et al., 2014; Yang et al., 2007). No median duration of response was achieved in the I-1 arm. Some patients continued to respond after completion of treatment, suggesting an immune memory component to this regimen. Initial and expansion cohorts continue to be evaluated, along with the IN-3 arm. The encouraging antitumor activity reported with the nivolumab + ipilimumab combination is the basis for a planned Phase III combination trial in first-line mRCC.

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Claims

1. 1. A method of treating a subject suffering from renal cancer, comprising administering to the subject a therapeutically effective amount of: (a) an anticancer agent that is an antibody or antigen-binding portion thereof that specifically binds to a programmed cell death (PD-1) receptor and inhibits PD-1 activity; and (b) Other anticancer drugs The method comprises administering a combination of

2. The method of claim 1 , wherein the renal cancer is renal cell carcinoma.

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

4. The method of claim 1, wherein the anti-PD-1 antibody, or antigen-binding portion thereof, is a chimeric, humanized, or human monoclonal antibody, or portion thereof.

5. The method of any of claims 1-4, wherein the anti-PD-1 antibody, or antigen-binding portion thereof, comprises a heavy chain constant region that is of the human IgG1 or IgG4 isotype.

6. 2. The method of claim 1, wherein the anti-PD-1 antibody is nivolumab.

7. 2. The method of claim 1, wherein the anti-PD-1 antibody is pembrolizumab.

8. 2. The method of claim 1, wherein the other anti-cancer agent is an anti-angiogenic tyrosine kinase inhibitor.

9. 9. The method of claim 8, wherein the anti-PD-1 antibody, or antigen-binding portion thereof, is administered at a dose ranging from about 0.1 to about 10.0 mg / kg body weight about once every 2, 3, or 4 weeks.

10. 10. The method of claim 9, wherein the anti-PD-1 antibody, or antigen-binding portion thereof, is administered at a dose of 2 mg / kg body weight or 5 mg / kg body weight once every three weeks.

11. The method of claim 8, wherein the tyrosine kinase inhibitor is sunitinib.

12. 12. The method of claim 11, wherein sunitinib is administered at a dose ranging from 12.5 to 50 mg in cycles comprising 28 days on daily and 14 days off, and the cycles are repeated as long as clinical benefit is observed or until uncontrollable toxicity or disease progression occurs.

13. 12. The method of claim 11, wherein sunitinib is administered at a dose ranging from 12.5 to 50 mg in cycles comprising 28 days on daily and 14 days off, followed by about 37.5 mg of sunitinib administered daily as long as clinical benefit is observed or until uncontrollable toxicity or disease progression occurs.

14. 9. The method of claim 8, wherein the tyrosine kinase inhibitor is pazopanib.

15. 15. The method of claim 14, wherein pazopanib is administered at a dose ranging from 400 to 800 mg daily or every other day as long as clinical benefit is observed or until uncontrollable toxicity or disease progression occurs.

16. The method of claim 1, wherein the other anti-cancer agent is an antibody or an antigen-binding portion thereof that specifically binds to cytotoxic T-lymphocyte antigen-4 (CTLA-4) and inhibits CTLA-4 activity.

17. The method of claim 16, wherein the anti-CTLA-4 antibody, or antigen-binding portion thereof, cross-competes with ipilimumab for binding to human CTLA-4.

18. The method of claim 16, wherein the anti-CTLA-4 antibody or antigen-binding portion thereof is a chimeric, humanized or human monoclonal antibody or portion thereof.

19. The method of any of claims 16 to 18, wherein the anti-CTLA-4 antibody, or antigen-binding portion thereof, comprises a heavy chain constant region that is of the human IgG1 isotype.

20. The method of claim 16, wherein the anti-CTLA-4 antibody is ipilimumab.

21. The method of claim 16, wherein the anti-CTLA-4 antibody is tremelimumab.

22. (a) an induction phase in which 2, 4, 6, 8, or 10 doses of a combination of anti-PD-1 and anti-CTLA-4 antibodies, or antigen-binding portions thereof, are administered, each dose ranging from 0.1 to 10.0 mg / kg body weight administered at least once every 2, 3, or 4 weeks; and (b) followed by a maintenance phase in which the anti-CTLA-4 antibody, or antigen-binding portion thereof, is not administered and the anti-PD-1 antibody, or antigen-binding portion thereof, is administered at a dose of 0.1 to 10.0 mg / kg body weight, repeated once at least every 2, 3, or 4 weeks; 17. The method of claim 16.

23. (a) the induction phase comprises 4 combination doses administered 3 weeks apart, wherein: (i) administering an anti-PD-1 antibody, or antigen-binding portion thereof, at 3 mg / kg body weight and an anti-CTLA-4 antibody, or antigen-binding portion thereof, at 1 mg / kg body weight; (ii) administering an anti-PD-1 antibody, or antigen-binding portion thereof, at 1 mg / kg body weight and an anti-CTLA-4 antibody, or antigen-binding portion thereof, at 3 mg / kg body weight; or (iii) administering an anti-PD-1 antibody, or antigen-binding portion thereof, at 3 mg / kg body weight and an anti-CTLA-4 antibody, or antigen-binding portion thereof, at 3 mg / kg body weight; and 23. The method of claim 22, wherein (b) the maintenance phase comprises repeated administration of the anti-PD-1 antibody, or antigen-binding portion thereof, at a dose of 3 mg / kg every two weeks for as long as clinical benefit is observed or until uncontrollable toxicity or disease progression occurs.

24. The method of claim 22, wherein the anti-PD-1 and anti-CTLA-4 antibodies are formulated for intravenous administration.

25. 23. The method of claim 22, wherein the anti-PD-1 antibody, or antigen-binding portion thereof, and the anti-CTLA-4 antibody, or antigen-binding portion thereof, are administered sequentially to the subject during the induction phase.

26. 26. The method of claim 25, wherein the anti-PD-1 and anti-CTLA-4 antibodies are administered within 30 minutes of each other.

27. (a) administering an anti-PD-1 antibody, or an antigen-binding portion thereof, prior to an anti-CTLA-4 antibody, or an antigen-binding portion thereof; or (b) administering the anti-CTLA-4 antibody, or antigen-binding portion thereof, prior to the anti-PD-1 antibody, or antigen-binding portion thereof; 26. The method of claim 25.

28. 23. The method of claim 22, wherein 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.

29. 23. The method of claim 22, wherein 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.

30. The method of claim 22, wherein the anti-PD-1 antibody, or antigen-binding portion thereof, is administered in a subtherapeutic dose.

31. The method of claim 22, wherein the anti-CTLA-4 antibody, or antigen-binding portion thereof, is administered in a sub-therapeutic dose.

32. 23. The method of claim 22, wherein the anti-PD-1 antibody, or antigen-binding portion thereof, and the anti-CTLA-4 antibody, or antigen-binding portion thereof, are each administered in a subtherapeutic dose.

33. The method of claim 22, wherein administration of the anti-PD-1 antibody in the maintenance phase continues as long as clinical benefit is observed or until uncontrollable toxicity or disease progression occurs.

34. 1. A kit for treating a subject suffering from renal cancer, comprising: (a) a dose in the range of 0.1 to 10 mg / kg body weight of an anticancer agent that is an antibody or antigen-binding portion thereof that specifically binds to the PD-1 receptor and inhibits PD-1 activity; (b) (i) an anti-angiogenic tyrosine kinase inhibitor; or (ii) a dose of another anti-cancer agent that is an antibody or antigen-binding portion thereof that specifically binds and inhibits CTLA-4 in a dose range of 0.1 to 10 mg / kg body weight; and (c) instructions for using the anti-PD-1 antibody and other anti-cancer agent in the method of any of claims 1, 8, or 16. Including the kit.

35. 34. The method of any of claims 1-33, wherein administration of an anti-PD-1 antibody, or antigen-binding portion thereof, and another anti-cancer agent increases T cell infiltration in renal cancer tissue compared to untreated subjects or subjects treated with a monotherapy of the anti-PD-1 antibody, or antigen-binding portion thereof, or another anti-cancer agent.

36. Increased T cell infiltration is seen in CD4 ＋ / Ki67 ＋ T cells, CD8 ＋ T cells, CD8 ＋ / Ki67 ＋ 36. The method of claim 35, characterized by increased infiltration of T cells or any combination thereof.

37. Increased T cell infiltration is seen in CD4 ＋ / Ki67 ＋ T cells, CD8 ＋ / Ki67 ＋ 37. The method of claim 36, characterized by increased infiltration of T cells or both.

38. 38. The method of any of claims 35-37, wherein T cell infiltration is increased 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% or at least about 90%.

39. 39. The method of any of claims 1-33 and 35-38, wherein administration of an anti-PD-1 antibody, or antigen-binding portion thereof, and another anti-cancer agent increases proliferative T cells compared to untreated subjects or subjects treated with a monotherapy of the anti-PD-1 antibody, or antigen-binding portion thereof, or another anti-cancer agent.

40. Proliferative T cells are CD4 ＋ / Ki67 ＋ T cells, CD8 ＋ T cells, CD8 ＋ / Ki67 ＋ 40. The method of claim 39, wherein the T cell is a T cell or any combination thereof.

41. 41. The method of claim 39 or 40, wherein the frequency of proliferating T cells is increased 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% or at least about 90%.

42. 42. The method of any of claims 1-33 and 35-41, wherein administration of an anti-PD-1 antibody, or antigen-binding portion thereof, and another anti-cancer agent reduces the number of monocytic myeloid-derived suppressor cells compared to untreated subjects or subjects treated with a monotherapy of the anti-PD-1 antibody, or antigen-binding portion thereof, or another anti-cancer agent.

43. 43. The method of claim 42, wherein the number of monocytic myeloid-derived suppressor cells is reduced by at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80% or at least about 90%.

44. Monocytic myeloid-derived suppressor cells express CD11b ＋ / Ly6C ｈｉ / Ly6G － Expression or CD11b ＋ / Ly6C ｌｏｗ / Ly6G － 44. The method of claim 42 or 43, characterized by expression.

45. 45. The method of any of claims 1-33 and 35-44, wherein administration of an anti-PD-1 antibody, or antigen-binding portion thereof, and another anti-cancer agent increases the number of granulocytic myeloid cells compared to an untreated subject or a subject treated with a monotherapy of the anti-PD-1 antibody, or antigen-binding portion thereof, or another anti-cancer agent.

46. 45. The method of claim 44, wherein the number of granulocytic myeloid cells is increased by at least about 5%, 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% or at least about 90%.

47. Granulocytic myeloid cells are CD11b ＋ / Ly6C － / Ly6G ＋ 47. The method of claim 45 or 46, characterized by expression.

48. 48. The method of any of claims 1-33 and 35-47, wherein administration of an anti-PD-1 antibody, or antigen-binding portion thereof, and another anti-cancer agent reduces T regulatory cells compared to an untreated subject or a subject treated with a monotherapy of the anti-PD-1 antibody, or antigen-binding portion thereof, or another anti-cancer agent.

49. T regulatory cells are CD45 ＋ / CD4 ＋ / CD25 ＋ / FoxP3 ＋ 49. The method of claim 48, characterized by expression.

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