RU2806210C2 - Antibodies to b7-h1 and ctla-4 for the treatment of non-small cell lung cancer - Google Patents

Abstract

FIELD: pharmaceuticals.

SUBSTANCE: method for treating non-small-cell lung cancer (NSCLC) in a patient, including the administration of MEDI4736 and tremelimumab, where (i) the administration can give an average C_max with a value of 266-300 μg/ml MEDI4736 and C_max with a value of 25-30 μg/ml tremelimumab; (ii) MEDI4736 is administered 1 hour after tremelimumab administration and (iii) MEDI4736 and tremelimumab are administered every 4 weeks for a maximum of 16 weeks.

EFFECT: safety and synergistic effectiveness of the combination.

9 cl, 37 dwg, 9 tbl, 9 ex

Description

CROSS REFERENCE TO RELATED APPLICATION

[0001] This application claims priority and benefit from U.S. Provisional Patent Application Serial No. 61/992658 filed May 13, 2014, U.S. Provisional Patent Application Serial No. 62/105992 filed Jan. 21, 2015, and US Provisional Patent Application Serial No. 62/114336, filed February 10, 2015. The entire contents of each of these applications are hereby incorporated herein by reference.

BACKGROUND OF THE ART

[0002] Cancer is still a major global health problem. Despite progress in cancer treatment, there remains an unmet medical need in the field for more effective and less toxic therapeutic agents, especially for those patients with advanced disease or forms of cancer that are resistant to existing therapies.

[0003] The role of the immune system, in particular T cell-mediated cytotoxicity, in the control of tumors is well known. T cells have been proven to control tumor growth and survival in patients with cancer in both early and late stages of the disease. However, tumor-specific T-cell responses in patients with cancer are difficult to establish and confirm.

[0004] Two T cell pathways currently receiving significant attention signal through cytotoxic T lymphocyte antigen-4 (CTLA-4, CD152) and programmed death ligand 1 (PD-L1, also known as B7-H1 or CD274).

[0005] CTLA4 is expressed on activated T cells and serves as a co-inhibitor to keep T cell responses under control following CD28-mediated T cell activation. CTLA4 is believed to regulate the amplitude of early activation of naïve and memory T cells following TCR engagement and is part of a central inhibitory pathway that impairs both antitumor immunity and autoimmunity. CTLA4 is expressed exclusively on T cells, and expression of its ligands CD80 (B7.1) and CD86 (B7.2) is largely restricted to antigen presenting cells, T cells, and other immune mediating cells. Antagonistic antibodies to CTLA4, which block the CTLA4 signaling pathway, have been reported to enhance T cell activation. One such antibody, ipilimumab, was approved by the FDA in 2011 for the treatment of metastatic melanoma. Another anti-CTLA4 antibody, tremelimumab, has been tested in a phase III trial for the treatment of advanced melanoma, but it does not significantly improve overall patient survival compared with the standard of care (temozolomide or dacarbazine) at the same time.

[0006] PD-L1 is also part of a complex system of receptors and ligands that are involved in controlling T cell activation. In healthy tissue, PD-L1 is expressed on T cells, B cells, dendritic cells, macrophages, mesenchymal stem cells, bone marrow mast cells, as well as on various non-hematopoietic cells. Its normal function is to regulate the balance between T cell activation and tolerance through interaction with its two receptors: programmed death 1 (also known as PD-1 or CD279) and CD80 (also known as B7-1 or B7.1). PD-L1 is also expressed in tumors and acts at multiple sites to help tumors evade detection and elimination by the host immune system. PD-L1 is expressed in a wide range of cancers at high frequency. In some forms of cancer, PD-L1 expression is associated with decreased survival and poor prognosis. Antibodies that block the interaction between B7-H1 and its receptor are capable of attenuating PD-L1-dependent immunosuppressive effects and enhancing the cytotoxic activity of antitumor T cells in vitro . MEDI4736 is a human monoclonal antibody directed against human PD-L1 that is capable of blocking PD-L1 binding to both PD-1 and CD80 receptors.

[0007] Despite significant progress made over the past decade in the development of strategies to combat cancer and other diseases, patients with advanced disease, refractory disease, and metastatic disease have limited clinical options. Chemotherapy, radiation, and high-dose chemotherapy become dose-limiting. There remains a significant unmet need in the field for new, less toxic methods and therapeutic agents that have better therapeutic efficacy, longer-term clinical benefit and improved safety profiles, particularly for those patients with advanced disease or cancers that are resistant to treatment. existing therapy.

SHORT DESCRIPTION

[0008] The present invention provides a method of treating non-small cell lung cancer (NSCLC) in a human patient, comprising administering to the patient 1 mg/kg of MEDI4736 or an antigen-binding fragment thereof and 1 mg/kg of tremelimumab or an antigen-binding fragment thereof.

[0009] The present invention also provides a method of treating NSCLC in a human patient, comprising administering to the patient 3 mg/kg of MEDI4736 or an antigen-binding fragment thereof and 1 mg/kg of tremelimumab or an antigen-binding fragment thereof.

[0010] The present invention also provides a method of treating NSCLC in a human patient, comprising administering to the patient 10 mg/kg of MEDI4736 or an antigen-binding fragment thereof and 1 mg/kg of tremelimumab or an antigen-binding fragment thereof.

[0011] The present invention also provides a method of treating NSCLC in a human patient, comprising administering to the patient 15 mg/kg of MEDI4736 or an antigen-binding fragment thereof and 1 mg/kg of tremelimumab or an antigen-binding fragment thereof.

[0012] The present invention also provides a method of treating NSCLC in a human patient, comprising administering to the patient 10 mg/kg of MEDI4736 or an antigen-binding fragment thereof and 3 mg/kg of tremelimumab or an antigen-binding fragment thereof.

[0013] The present invention also provides a method of treating NSCLC in a human patient, comprising administering to the patient 15 mg/kg of MEDI4736 or an antigen-binding fragment thereof and 3 mg/kg of tremelimumab or an antigen-binding fragment thereof.

[0014] The present invention also provides a method of treating NSCLC in a human patient, comprising administering to the patient 20 mg/kg of MEDI4736 or an antigen-binding fragment thereof and 1 mg/kg of tremelimumab or an antigen-binding fragment thereof.

[0015] The present invention also provides a method of treating NSCLC in a human patient, comprising administering to the patient 15 mg/kg of MEDI4736 or an antigen-binding fragment thereof and 10 mg/kg of tremelimumab or an antigen-binding fragment thereof.

[0016] The present invention also provides a method of treating NSCLC in a human patient, comprising administering to the patient 20 mg/kg of MEDI4736 or an antigen-binding fragment thereof and 3 mg/kg of tremelimumab or an antigen-binding fragment thereof.

[0017] The present invention also provides a method of treating NSCLC in a human patient, comprising administering to the patient 20 mg/kg of MEDI4736 or an antigen-binding fragment thereof and 10 mg/kg of tremelimumab or an antigen-binding fragment thereof.

[0018] The present invention also provides a method of treating NSCLC in a human patient, comprising administering to the patient 10 mg/kg of MEDI4736 or an antigen-binding fragment thereof and 10 mg/kg of tremelimumab or an antigen-binding fragment thereof.

[0019] The present invention also provides a method of treating NSCLC in a human patient, comprising administering to the patient 3 mg/kg of MEDI4736 or an antigen-binding fragment thereof and 3 mg/kg of tremelimumab or an antigen-binding fragment thereof.

[0020] The present invention also provides a method of treating NSCLC in a human patient, comprising administering to the patient 3 mg/kg of MEDI4736 or an antigen-binding fragment thereof and 10 mg/kg of tremelimumab or an antigen-binding fragment thereof.

[0021] The present invention also provides a method of treating NSCLC in a human patient, comprising administering to the patient 1 mg/kg of MEDI4736 or an antigen-binding fragment thereof and 3 mg/kg of tremelimumab or an antigen-binding fragment thereof.

[0022] The present invention also provides a method of treating NSCLC in a human patient, comprising administering to the patient 1 mg/kg of MEDI4736 or an antigen-binding fragment thereof and 10 mg/kg of tremelimumab or an antigen-binding fragment thereof.

[0023] The present invention also provides a method of treatment comprising administration of MEDI4736 or an antigen-binding fragment thereof (eg, at 1 mg/kg, 3 mg/kg, 10 mg/kg, 15 mg/kg, or 20 mg/kg) and tremelimumab or an antigen-binding fragment thereof (eg, at 1 mg/kg, 3 mg/kg, 10 mg/kg) to a patient identified as having PD-L1 ^- or PD-L1 ⁺ NSCLC.

[0024] In specific embodiments of any of the previous methods, MEDI4736 is administered every 4 weeks. In one specific embodiment, MEDI4736 is administered every 4 weeks for 49 weeks. In specific embodiments of any of the previous methods, MEDI4736 is administered every 2 weeks. In another embodiment, a total of 13 doses of MEDI4736 are administered.

[0025] In specific embodiments of the preceding aspects, tremelimumab is administered every 4 weeks for the first 21 weeks. In other embodiments of the previous aspects, tremelimumab is administered every 12 weeks from weeks 25 to 49. In still other embodiments of the previous aspects, a total of 9 doses of tremelimumab are administered.

[0026] The present invention also provides a method of treating NSCLC in a human patient, comprising administering to the patient 10 mg/kg of MEDI4736 or an antigen-binding fragment thereof and 1 mg/kg of tremelimumab or an antigen-binding fragment thereof, with MEDI4736 administered every 2 weeks.

[0027] The present invention also provides a method of treating NSCLC in a human patient, comprising administering to the patient 10 mg/kg of MEDI4736 or an antigen-binding fragment thereof and 3 mg/kg of tremelimumab or an antigen-binding fragment thereof, with MEDI4736 administered every 2 weeks.

[0028] The present invention also provides a method of treating NSCLC in a human patient, comprising administering to the patient 10 mg/kg of MEDI4736 or an antigen-binding fragment thereof and 10 mg/kg of tremelimumab or an antigen-binding fragment thereof, with MEDI4736 administered every 2 weeks.

[0029] In specific embodiments of the preceding aspects, tremelimumab is administered every 4 weeks for the first 25 weeks. In other embodiments of the previous aspects, tremelimumab is administered every 12 weeks from weeks 25 to 49. In still other embodiments of the previous aspects, a total of 9 doses of tremelimumab are administered.

[0030] The present invention also provides a method of treating NSCLC in a human patient, comprising administering to the patient MEDI4736 or an antigen-binding fragment thereof and tremelimumab or an antigen-binding fragment thereof, the administrations resulting in a tumor response.

[0031] In the method of the present invention, administration results in an enhanced tumor response compared to administration of either MEDI4736 or an antigen-binding fragment thereof, or tremelimumab or an antigen-binding fragment thereof alone.

[0032] In some embodiments, tumor response may be detected at 2, 4, 6, 8, or 10 weeks. In other embodiments, tumor response may be detected at 33 weeks. In other embodiments, tumor response may be detected at 50 weeks.

[0033] The present invention also provides a method of treating NSCLC in a human patient, comprising administering to the patient MEDI4736 or an antigen-binding fragment thereof and tremelimumab or an antigen-binding fragment thereof, the administrations improving progression-free survival. In some embodiments, administration results in improved progression-free survival compared to administration of either MEDI4736 or an antigen-binding fragment thereof or tremelimumab or an antigen-binding fragment thereof alone.

[0034] The present invention also provides a method of treating NSCLC in a human patient, comprising administering to the patient MEDI4736 or an antigen-binding fragment thereof and tremelimumab or an antigen-binding fragment thereof, which administrations improve overall survival. In some embodiments, administration results in improved overall survival compared to administration of either MEDI4736 or an antigen-binding fragment thereof or tremelimumab or an antigen-binding fragment thereof alone.

[0035] In some embodiments of the methods of the present invention, administration results in a tumor response. In some embodiments, administration results in an increased tumor response compared to administration of either MEDI4736 or an antigen-binding fragment thereof, or tremelimumab or an antigen-binding fragment thereof alone. In some embodiments, a tumor response may be detected at 8 weeks. In some embodiments, tumor response may be detected at 33 weeks.

[0036] In some embodiments of the methods of the present invention, administration increases progression-free survival. In some embodiments, administration results in improved progression-free survival compared to administration of either MEDI4736 or an antigen-binding fragment thereof or tremelimumab or an antigen-binding fragment thereof alone.

[0037] In some embodiments of the methods of the present invention, administration increases overall survival. In some embodiments, administration results in improved overall survival compared to administration of either MEDI4736 or an antigen-binding fragment thereof or tremelimumab or an antigen-binding fragment thereof alone.

[0038] In specific embodiments of any of the previous methods, MEDI4736 is administered every 4 weeks. In one specific embodiment, MEDI4736 is administered every 4 weeks for 49 weeks. In another embodiment, a total of 13 doses of MEDI4736 are administered.

[0039] In specific embodiments of the preceding aspects, tremelimumab is administered every 4 weeks for the first 21 weeks. In other embodiments of the preceding aspects, tremelimumab is administered every 12 weeks from weeks 25 to 49 or after the first 6 doses. In still other embodiments of the previous aspects, a total of 9 doses of tremelimumab are administered.

[0040] In some embodiments of the method of the present invention, administration of tremelimumab or an antigen-binding fragment thereof occurs approximately every 4 weeks for seven administrations, and then every 12 weeks.

[0041] In some embodiments of the method of the present invention, administration reduces soluble PD-L1. In some embodiments, soluble PD-L1 is reduced by at least about 65%, 80%, 90%, 95%, or 99%.

[0042] In some embodiments of the methods of the present invention, the tumor is resistant to at least one chemotherapeutic agent. Such chemotherapeutic agents may include, without limitation, vemurafenib, erlotinib, afatinib, cetuximab, carboplatin, bevacizumab, erlotinib, gefitinib, or pemetrexed.

[0043] In some embodiments of the methods of the present invention, the patient has an Eastern Cooperative Oncology Group (ECOG) performance status score of 0-1.

[0044] In some embodiments of the methods of the present invention, the patient has not received immunotherapy prior to administration of MEDI4736 or an antigen-binding fragment thereof and tremelimumab or an antigen-binding fragment thereof.

[0045] In some embodiments of the methods of the present invention, the patient received immunotherapy prior to administration of MEDI4736 or an antigen-binding fragment thereof and tremelimumab or an antigen-binding fragment thereof.

[0046] In some embodiments of the methods of the present invention, MEDI4736 or an antigen-binding fragment thereof is administered by intravenous infusion.

[0047] In some embodiments of the methods of the present invention, tremelimumab or an antigen-binding fragment thereof is administered by intravenous infusion.

[0048] In specific embodiments of the methods of the present invention, administrations reduce tumor size by at least about 10%, 25%, 50%, 75%, or 100% relative to baseline.

[0049] In specific embodiments of the methods of the present invention, the human patient has locally advanced, unresectable, or metastatic NSCLC. In specific embodiments of the methods of the present invention, the NSCLC is squamous or non-squamous. In other embodiments of the methods of the present invention, NSCLC involves a KRAS mutation or an EGFR mutation.

[0050] In some embodiments of the methods of the present invention, MEDI4736 or an antigen-binding fragment thereof is MEDI4736, and wherein tremelimumab or an antigen-binding fragment thereof is tremelimumab.

[0051] In another aspect, the present invention provides a method of treating cancer with MEDI4736 or an antigen binding fragment of MEDI4736 in combination with tremelimumab or an antigen binding fragment of tremelimumab. In some embodiments of the present invention, the cancer is selected from prostate cancer, breast cancer, triple negative breast cancer, colon cancer, lung cancer, NSCLC, head and neck cancer, melanoma, gastric cancer, pancreatic cancer, ovarian cancer, hypernephroma and liver cancer.

BRIEF DESCRIPTION OF GRAPHIC MATERIALS/FIGURES

[0052] Figure 1 shows a treatment regimen. MTD=maximum tolerated dose.

[0053] Figure 2 shows dose escalation and dose escalation schemes.

[0054] Figure 3 shows the design of a dose escalation study. DLT=dose limiting toxicity.

[0055] Figure 4 is a graph and table showing the pharmacokinetic (PK) profile of MEDI4736. Q4W=every 4 weeks. LLOQ= lower limit of quantitation. AUC=area under the curve.

[0056] Figure 5 is a graph and table showing the pharmacokinetic profile of tremelimumab.

[0057] Figure 6 is a graph showing the absolute concentration profiles of soluble PD-L1 (sPD-L1).

[0058] Figure 7 is a graph showing percentage inhibition of soluble PD-L1.

[0059] Figure 8 is a graph showing the pharmacokinetic (PK) parameters of MEDI4736 in individual patients.

[0060] Figure 9 is a graph showing the pharmacokinetic (PK) parameters of tremelimumab in individual patients.

[0061] Figures 10A and 10B provide tables showing dose modifications of MEDI4736 and tremelimumab due to grade 1 or grade 2 toxicity (10A) and grade 3 or grade 4 toxicity (10B).

[0062] Figure 11 is a table showing the study criteria.

[0063] Figure 12 shows inclusion in the study.

[0064] Figure 13 shows the relative adverse events (AE) for patients receiving the dose.

[0065] Figure 14 shows the change in tumor size from baseline in the first four cohorts.

[0066] Figure 15 shows study doses and plots for MEDI4736 and tremelimumab.

[0067] Figure 16 shows sPD-L1 suppression in individual profiles given MEDI4736 and tremelimumab in combination.

[0068] Figure 17 shows sPD-L1 suppression in individual profiles given MEDI4736 and tremelimumab in combination combined into a cohort.

[0069] Figure 18 shows the increase in CD4 Ki67+ and CD8 Ki67 proliferating cells from baseline in subjects receiving MEDI4736 and tremelimumab compared to the increases in those receiving MEDI4736 alone. Increases in CD4 Ki67+ and CD8 Ki67 proliferating cells compared with baseline correlated with increasing tremelimumab dose.

[0070] Figure 19 shows the increase in CD4 ICOS+ and CD4 HLADR+ cells from baseline in subjects receiving MEDI4736 and tremelimumab compared to the increases in those receiving MEDI4736 alone. Increases in CD4 ICOS+ and CD4 HLADR+ cells from baseline correlated with increasing tremelimumab dose.

[0071] Figure 20 shows the increase in CD4+ effector T cells from baseline in subjects receiving MEDI4736 and tremelimumab compared to the increase in those receiving MEDI4736 alone. Increases in CD4+ effector T cells from baseline correlated with increasing tremelimumab dose.

[0072] Figure 21 shows the increase in regulatory T cells from baseline in subjects receiving MEDI4736 and tremelimumab compared to the increase in those receiving MEDI4736 alone. Increases in regulatory T cells from baseline correlated with increasing doses of tremelimumab.

[0073] Figure 22 is a table showing the clinical activity of MEDI4736 therapy in combination with tremelimumab compared to MEDI4736 monotherapy.

[0074] Figure 23 presents spider plots showing change in tumor size from baseline in the tremelimumab fixed dose cohorts: 1 mg/kg (top left panel); 10 mg/kg (top right section) and 3 mg/kg (bottom section).

[0075] Figure 24 is a graph showing the time to onset of grade ≥3 relative adverse events (AEs) in the tremelimumab fixed dose cohort combination: 1 mg/kg; 10 mg/kg and 3 mg/kg, compared with MEDI4736 monotherapy.

[0076] Figure 25 presents spider plots showing change in tumor size from baseline in fixed dose cohorts of MEDI4736: 10 mg/kg Q4W (top left panel); 15 mg/kg Q4W (top right section); 20 mg/kg Q4W (lower left section) and 10 mg/kg Q2W (lower right section).

[0077] Figure 26 is a graph showing the time to onset of grade ≥3 relative adverse events (AEs) in the MEDI4736 fixed dose cohort combination: 1 mg/kg; 10 mg/kg and 3 mg/kg, compared with MEDI4736 monotherapy.

[0078] Figure 27 presents spider plots showing change in tumor size from baseline in selected cohorts of NSCLC patients receiving MEDI4736 and tremelimumab (left panel) compared to those receiving MEDI4736 (10 mg/kg Q4W) alone (right panel) .

[0079] Figure 28 presents graphs showing the time to onset of grade ≥3 relative adverse events (AEs) in the selected cohort combination (left and right panels) compared to MEDI4736 alone (left panel).

[0080] Figure 29 presents spider plots showing change in tumor size from baseline in all study cohorts of NSCLC patients receiving MEDI4736 and tremelimumab.

[0081] Figure 30 presents spider plots showing change in tumor size from baseline in selected cohorts of patients with PD-L1 negative NSCLC receiving MEDI4736 and tremelimumab (left panel), compared to those receiving MEDI4736 alone (10 mg/kg ; CP1108) (right section).

[0082] Figure 31 presents spider plots showing change in tumor size from baseline in selected cohorts of patients with PD-L1 positive NSCLC receiving MEDI4736 and tremelimumab (left panel), compared to those receiving MEDI4736 alone (10 mg/kg ; CP1108) (right section).

[0083] Figure 32 shows spider plots with a fixed dose of tremelimumab: 1 mg/kg (left section), 3 mg/kg (center section), and 10 mg/kg (right section), showing the change in tumor size compared to baseline levels in selected cohorts of patients with PD-L1 negative NSCLC receiving MEDI4736 and tremelimumab.

[0084] Figure 33 shows fixed dose spider plots of MEDI4736: 10 mg/kg Q4W (upper left panel), 15 mg/kg (upper right panel), and 20 mg/kg (lower left panel), showing changes in tumor size compared with baseline in selected cohorts of patients with PD-L1 negative NSCLC receiving MEDI4736 and tremelimumab.

[0085] Figures 34A-34D are spider plots showing change in tumor size from baseline in NSCLC patients receiving MEDI4736 and tremelimumab, as shown in Figure 29, grouped according to NSCLC PD-L1 status: all NSCLC patients (34A ); patients identified as having PD-L1 ^- NSCLC (34B); patients identified as having PD-L1 ⁺ NSCLC (34C); and patients with unavailable PD-L1 NSCLC status (34D).

[0086] Figure 35 is a waterfall plot showing the best change in tumor size from baseline in NSCLC patients receiving MEDI4736 and tremelimumab.

[0087] Figure 36 is a waterfall plot showing the best change in tumor size from baseline in NSCLC patients receiving MEDI4736 and tremelimumab, according to Figure 35, identified by NSCLC PD-L1 status.

[0088] Figure 37 is a track graph showing the duration of follow-up of NSCLC patients receiving MEDI4736 and tremelimumab.

DETAILED DESCRIPTION

[0089] It should be noted that the singular form of an object refers to one or more such objects; for example, “antibody” is meant to be one or more antibodies. Accordingly, the singular form of the expressions “one or more” and “at least one” may be used interchangeably herein.

[0090] Presented herein are methods of treating non-small cell lung cancer (NSCLC) using MEDI4736 and tremelimumab. There are three main subtypes of NSCLC: squamous cell carcinoma, adenocarcinoma, and giant cell (undifferentiated) carcinoma. Other subtypes include adenosquamous carcinoma and sarcomatoid carcinoma. NSCLC may involve a mutation in KRAS or the epidermal growth factor receptor. Such mutations are known in the art and are described, for example, in Riely et al., Proc Am Thorac Soc. 2009 Apr 15;6(2):201-5, which is incorporated herein by reference.

[0091] Additionally provided herein are methods of treating cancer or forms of neoplastic growth using MEDI4736 and tremelimumab. As used herein, “cancer” refers to a disease caused by the uncontrolled division of abnormal cells. Examples of cancer include prostate cancer, breast cancer, triple negative breast cancer, colon cancer, lung cancer, NSCLC, head and neck cancer, melanoma, stomach cancer, pancreatic cancer, ovarian cancer, hypernephroma, and liver cancer.

[0092] The present methods involve administering an effective amount of MEDI4736 or an antigen-binding fragment thereof and tremelimumab or an antigen-binding fragment thereof.

[0093] By "MEDI4736" is meant an antibody or an antigen-binding fragment thereof that selectively binds a PD-L1 polypeptide and contains at least a portion of a light chain variable region containing the amino acid sequence of SEQ ID NO: 1, and/or at least part of the heavy chain variable region containing the amino acid sequence of SEQ ID NO:2.

[0094] Information regarding MEDI4736 (or antigen-binding fragments thereof) for use in the methods presented herein can be found in US Pat. No. 8,779,108, the disclosure of which is incorporated herein by reference in its entirety. The crystallizing fragment (Fc) domain from MEDI4736 contains a triple mutation in the IgG1 heavy chain constant domain, which reduces the extent of binding to complement component C1q and Fcγ receptors responsible for mediating antibody-dependent cell-mediated cytotoxicity (ADCC). MEDI4736 is selective for PD-L1 and blocks PD-L1 binding to PD-1 and CD80 receptors. MEDI4736 can alleviate PD-L1-mediated suppression of human T cell activation in vitro, and it inhibits tumor growth in a xenograft model through a T cell-dependent mechanism.

[0095] MEDI4736 for use in the methods provided herein comprises a heavy chain and a light chain or a heavy chain variable region and a light chain variable region. In a specific aspect, MEDI4736 or an antigen binding fragment thereof for use in the methods provided herein comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:1 and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 2. In a specific aspect, MEDI4736 or an antigen binding fragment thereof for use in the methods provided herein comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat defined sequences CDR1, CDR2 and CDR3 of SEQ ID NO :3-5, and wherein the light chain variable region comprises the Kabat-defined sequences CDR1, CDR2 and CDR3 of SEQ ID NO:6-8. Those skilled in the art will readily be able to identify Chothia-defined, Abm-defined, or other CDR definitions known to those skilled in the art. In a specific aspect, MEDI4736 or an antigen binding fragment thereof for use in the methods provided herein comprises the CDR sequences of the heavy chain variable region and the light chain variable region of the antibody 2.14H9OPT disclosed in US Pat. No. 8,779,108, which is incorporated herein by reference. in full.

[0096] By “tremelimumab” is meant an antibody or an antigen-binding fragment thereof that selectively binds a CTLA4 polypeptide and contains at least a portion of a light chain variable region containing the amino acid sequence of SEQ ID NO:9, and/or at least a portion of a light chain variable region a heavy chain region containing the amino acid sequence of SEQ ID NO:10. Exemplary anti-CTLA4 antibodies are described, for example, in US Pat. Nos. 6,682,736, 7,109,003, 7,123,281, 7,411,057, 7,824,679, 8,143,379, 7,807,797, and 8,491,895 (in which tremelimumab is 11.2.1), which are incorporated herein by reference. Tremelimumab is a typical anti-CTLA4 antibody. Tremelimumab sequences are presented in the sequence listing below.

[0097] Information regarding tremelimumab (or antigen-binding fragments thereof) for use in the methods presented herein can be found in US Patent No. 6,682,736 (where it is designated 11.2.1), the disclosure of which is incorporated herein by reference in full. Tremelimumab (also known as CP-675,206, CP-675, CP-675206 and ticilimumab) is a human _IgG2 monoclonal antibody that is highly selective for CTLA4 and blocks CTLA4 binding to CD80 (B7.1) and CD86 (B7.1). 2). It has been shown to lead to immune activation in vitro , and tumor regression has been observed in some patients treated with tremelimumab.

[0098] Tremelimumab and antigen-binding fragments thereof for use in the methods provided herein comprise a heavy chain and a light chain or a heavy chain variable region and a light chain variable region. In a specific aspect, tremelimumab or an antigen binding fragment thereof for use in the methods provided herein comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:9 and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 10. In a specific aspect, tremelimumab or an antigen binding fragment thereof for use in the methods provided herein comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat defined sequences CDR1, CDR2 and CDR3 of SEQ ID NO :11-13, and wherein the light chain variable region comprises the Kabat-defined sequences CDR1, CDR2 and CDR3 of SEQ ID NO:14-16. Those skilled in the art will readily be able to identify Chothia-defined, Abm-defined, or other CDR definitions known to those skilled in the art. In a specific aspect, tremelimumab or an antigen binding fragment thereof for use in the methods provided herein comprises the CDR sequences of the heavy chain variable region and the light chain variable region of antibody 11.2.1 disclosed in US Pat. No. 6,682,736, which is incorporated herein by links in full.

[0099] The term “antigen-binding fragment” refers to a portion of an intact antibody and/or refers to the variable regions of an intact antibody that detect antigens. It is known that the antigen-binding function of an antibody can be represented by fragments of a full-length antibody. Examples of antibody fragments include, but are not limited to, Fab, Fab', F(ab')2 and Fv fragments, linear antibodies, single chain antibodies, diabodies and multispecific antibodies formed from antibody fragments.

[00100] In certain aspects, a patient presenting with NSCLC is administered MEDI4736 or an antigen-binding fragment thereof and tremelimumab or an antigen-binding fragment thereof. MEDI4736 or its antigen-binding fragment and tremelimumab or its antigen-binding fragment can be administered only once or infrequently and still provide benefit to the patient. In additional aspects, additional subsequent doses are administered to the patient. Subsequent doses may be administered at varying intervals depending on the patient's age, weight, clinical tests, tumor burden, and/or other factors, including the judgment of the attending physician.

[00101] Intervals between doses of MEDI4736 or an antigen-binding fragment thereof may be every four weeks. Intervals between doses of tremelimumab or its antigen-binding fragment may be every four weeks. Intervals between doses of tremelimumab or its antigen-binding fragment may be every twelve weeks. Intervals between doses of tremelimumab or its antigen-binding fragment may be every four weeks for six cycles, and then every twelve weeks.

[00102] In certain aspects, MEDI4736 or an antigen-binding fragment thereof is administered approximately as frequently as tremelimumab or an antigen-binding fragment thereof. In certain aspects, MEDI4736 or an antigen-binding fragment thereof is administered approximately three times more frequently than tremelimumab or an antigen-binding fragment thereof.

[00103] In some embodiments, the patient is administered at least two doses of MEDI4736 or an antigen-binding fragment thereof and tremelimumab or an antigen-binding fragment thereof. In some embodiments, the patient may be administered at least three doses, at least four doses, at least five doses, at least six doses, at least seven doses, at least eight doses, at least nine doses, at least ten doses or at least fifteen doses or more. In some embodiments, MEDI4736 or an antigen-binding fragment thereof is administered over a four-week treatment period, over an eight-week treatment period, over a sixteen-week treatment period, over a twenty-week treatment period, over a twenty-four week treatment period, or over a treatment period of one year or more. In some embodiments, tremelimumab or an antigen-binding fragment thereof is administered over a four-week treatment period, over an eight-week treatment period, over a twelve-week treatment period, over a sixteen-week treatment period, over a twenty-week treatment period, over a twenty-four week treatment period, over a thirty-six week treatment period, over a forty-eight week treatment period treatment or over a treatment period of one year or more.

[00104] In some embodiments, MEDI4736 or an antigen-binding fragment thereof and tremelimumab or an antigen-binding fragment thereof are administered on the same day. In some embodiments, MEDI4736 or an antigen-binding fragment thereof is administered concomitantly with tremelimumab or an antigen-binding fragment thereof. In other embodiments, MEDI4736 or an antigen-binding fragment thereof is administered approximately 1 hour after administration of tremelimumab or an antigen-binding fragment thereof.

[00105] The amount of MEDI4736 or an antigen-binding fragment thereof and the amount of tremelimumab or an antigen-binding fragment thereof to be administered to a patient will depend on various parameters, such as the age, weight of the patient, clinical tests, tumor burden and/or other factors, including including the assessment of the attending physician.

[00106] In certain aspects, the patient is administered one or more doses of MEDI4736 or an antigen-binding fragment thereof, where the dose is approximately 1 mg/kg. In certain aspects, the patient is administered one or more doses of MEDI4736 or an antigen-binding fragment thereof, where the dose is approximately 3 mg/kg. In certain aspects, the patient is administered one or more doses of MEDI4736 or an antigen-binding fragment thereof, wherein the dose is approximately 10 mg/kg. In certain aspects, the patient is administered one or more doses of MEDI4736 or an antigen-binding fragment thereof, wherein the dose is approximately 15 mg/kg. In certain aspects, the patient is administered one or more doses of MEDI4736 or an antigen-binding fragment thereof, wherein the dose is approximately 20 mg/kg.

[00107] In certain aspects, the patient is administered at least two doses of MEDI4736 or an antigen-binding fragment thereof, where the dose is approximately 1 mg/kg. In certain aspects, the patient is administered at least two doses of MEDI4736 or an antigen-binding fragment thereof, where the dose is approximately 3 mg/kg. In certain aspects, the patient is administered at least two doses of MEDI4736 or an antigen-binding fragment thereof, where the dose is approximately 10 mg/kg. In certain aspects, the patient is administered at least two doses of MEDI4736 or an antigen-binding fragment thereof, where the dose is approximately 15 mg/kg. In certain aspects, the patient is administered at least two doses of MEDI4736 or an antigen-binding fragment thereof, where the dose is approximately 20 mg/kg. In some embodiments, at least two doses are administered separately, approximately four weeks apart.

[00108] In certain aspects, the patient is administered at least three doses of MEDI4736 or an antigen-binding fragment thereof, where the dose is approximately 1 mg/kg. In certain aspects, the patient is administered at least three doses of MEDI4736 or an antigen-binding fragment thereof, where the dose is approximately 3 mg/kg. In certain aspects, the patient is administered at least three doses of MEDI4736 or an antigen-binding fragment thereof, where the dose is approximately 10 mg/kg. In certain aspects, the patient is administered at least three doses of MEDI4736 or an antigen-binding fragment thereof, where the dose is approximately 15 mg/kg. In certain aspects, the patient is administered at least three doses of MEDI4736 or an antigen-binding fragment thereof, where the dose is approximately 20 mg/kg. In some embodiments, at least three doses are administered separately, approximately four weeks apart.

[00109] In certain aspects, the patient is administered one or more doses of tremelimumab or an antigen-binding fragment thereof, where the dose is approximately 1 mg/kg. In certain aspects, the patient is administered one or more doses of tremelimumab or an antigen-binding fragment thereof, where the dose is approximately 3 mg/kg. In certain aspects, the patient is administered one or more doses of tremelimumab or an antigen-binding fragment thereof, where the dose is approximately 10 mg/kg.

[00110] In certain aspects, the patient is administered at least two doses of tremelimumab or an antigen-binding fragment thereof, where the dose is approximately 1 mg/kg. In certain aspects, the patient is administered at least two doses of tremelimumab or an antigen-binding fragment thereof, where the dose is approximately 3 mg/kg. In certain aspects, the patient is administered at least two doses of tremelimumab or an antigen-binding fragment thereof, where the dose is approximately 10 mg/kg. In some embodiments, at least two doses are administered separately, approximately four weeks apart. In some embodiments, at least two doses are administered separately, approximately twelve weeks apart.

[00111] In certain aspects, the patient is administered at least three doses of tremelimumab or an antigen-binding fragment thereof, where the dose is approximately 1 mg/kg. In certain aspects, the patient is administered at least three doses of tremelimumab or an antigen-binding fragment thereof, where the dose is approximately 3 mg/kg. In certain aspects, the patient is administered at least three doses of tremelimumab or an antigen-binding fragment thereof, where the dose is approximately 10 mg/kg. In some embodiments, at least three doses are administered separately, approximately four weeks apart. In some embodiments, at least three doses are administered separately, approximately twelve weeks apart.

[00112] In certain aspects, administration of MEDI4736 or an antigen-binding fragment thereof and/or tremelimumab or an antigen-binding fragment according to the methods provided herein is accomplished by parenteral administration. For example, MEDI4736 or an antigen-binding fragment thereof and/or tremelimumab or an antigen-binding fragment can be administered by intravenous infusion or by subcutaneous injection. In some embodiments, administration is an intravenous infusion.

[00113] In certain aspects, a patient is administered 1 mg/kg of MEDI4736 or an antigen-binding fragment thereof and 1 mg/kg of tremelimumab or an antigen-binding fragment thereof. In certain aspects, the patient is administered 1 mg/kg of MEDI4736 or an antigen-binding fragment thereof and 3 mg/kg of tremelimumab or an antigen-binding fragment thereof. In certain aspects, the patient is administered 1 mg/kg of MEDI4736 or an antigen-binding fragment thereof and 10 mg/kg of tremelimumab or an antigen-binding fragment thereof.

[00114] In certain aspects, the patient is administered 3 mg/kg of MEDI4736 or an antigen-binding fragment thereof and 1 mg/kg of tremelimumab or an antigen-binding fragment thereof. In certain aspects, the patient is administered 3 mg/kg of MEDI4736 or an antigen-binding fragment thereof and 3 mg/kg of tremelimumab or an antigen-binding fragment thereof. In certain aspects, the patient is administered 3 mg/kg of MEDI4736 or an antigen-binding fragment thereof and 10 mg/kg of tremelimumab or an antigen-binding fragment thereof.

[00115] In certain aspects, a patient is administered 10 mg/kg of MEDI4736 or an antigen-binding fragment thereof and 1 mg/kg of tremelimumab or an antigen-binding fragment thereof. In certain aspects, the patient is administered 10 mg/kg of MEDI4736 or an antigen-binding fragment thereof and 3 mg/kg of tremelimumab or an antigen-binding fragment thereof. In certain aspects, the patient is administered 10 mg/kg of MEDI4736 or an antigen-binding fragment thereof and 10 mg/kg of tremelimumab or an antigen-binding fragment thereof.

[00116] In certain aspects, a patient is administered 15 mg/kg of MEDI4736 or an antigen-binding fragment thereof and 1 mg/kg of tremelimumab or an antigen-binding fragment thereof. In certain aspects, the patient is administered 15 mg/kg of MEDI4736 or an antigen-binding fragment thereof and 3 mg/kg of tremelimumab or an antigen-binding fragment thereof. In certain aspects, the patient is administered 15 mg/kg of MEDI4736 or an antigen-binding fragment thereof and 10 mg/kg of tremelimumab or an antigen-binding fragment thereof.

[00117] In certain aspects, a patient is administered 20 mg/kg of MEDI4736 or an antigen-binding fragment thereof and 1 mg/kg of tremelimumab or an antigen-binding fragment thereof. In certain aspects, the patient is administered 20 mg/kg of MEDI4736 or an antigen-binding fragment thereof and 3 mg/kg of tremelimumab or an antigen-binding fragment thereof. In certain aspects, the patient is administered 20 mg/kg of MEDI4736 or an antigen-binding fragment thereof and 10 mg/kg of tremelimumab or an antigen-binding fragment thereof.

[00118] The methods provided herein can reduce, slow, or stabilize tumor growth. In some aspects, the decrease or slowdown may be statistically significant. Reduction in tumor growth can be measured by comparing a patient's tumor growth at baseline, with expected tumor growth, with expected tumor growth based on a large patient population, or with tumor growth in a control population. In certain aspects, tumor response is measured using Response Evaluation Criteria in Solid Tumors (RECIST).

[00119] In certain aspects, tumor response is detected at 8 weeks. In certain aspects, tumor response is detected at 33 weeks. In certain aspects, tumor response is detected at 50 weeks.

[00120] In certain aspects, a tumor response is detected after administration of two doses of MEDI4736 or an antigen-binding fragment thereof and two doses of tremelimumab or an antigen-binding fragment thereof. In certain aspects, a tumor response is detected after administration of eight doses of MEDI4736 or an antigen-binding fragment thereof and seven doses of tremelimumab or an antigen-binding fragment thereof. In certain aspects, a tumor response is detected after administration of thirteen doses of MEDI4736 or an antigen-binding fragment thereof and nine doses of tremelimumab or an antigen-binding fragment thereof.

[00121] In certain aspects, the patient achieves disease control (DC). Disease control can be complete remission (CR), partial remission (PR), or stable disease (SD).

[00122] “Complete remission” (CR), “partial remission” (PR) and “stable disease” (SD) can be defined as described in Table 1 below.

[00123] In certain aspects, administration of MEDI4736 or an antigen-binding fragment thereof can improve progression-free survival (PFS).

[00124] In certain aspects, administration of MEDI4736 or an antigen-binding fragment thereof may increase overall survival (OS).

[00125] In some embodiments, the patient has been previously treated with at least one chemotherapeutic agent. In some embodiments, the patient has been previously treated with at least two chemotherapeutic agents. The chemotherapy agent may include, without limitation, vemurafenib, erlotinib, afatinib, cetuximab, carboplatin, bevacizumab, erlotinib, gefitinib and/or pemetrexed.

[00126] In some embodiments, the NSCLC is refractory or resistant to at least one chemotherapeutic agent. In some embodiments, the tumor is resistant or resistant to at least two chemotherapeutic agents. The tumor may be resistant or resistant to one or more, for example, but not limited to, vemurafenib, erlotinib, afatinib, cetuximab, carboplatin, bevacizumab, erlotinib, gefitinib and/or pemetrexed. In some embodiments, the NSCLC is PD-L1 negative. In some embodiments, the NSCLC is PD-L1 positive.

[00127] In some embodiments, the patient has an Eastern Cooperative Oncology Group (ECOG) performance status score of 0 or 1 (Oken MM, et al. Am. J. Clin. Oncol. 5 :649-55 (1982)) prior to administration of MEDI4736 or an antigen-binding fragment thereof and tremelimumab or an antigen-binding fragment thereof.

[00128] According to the methods provided herein, administration of MEDI4736 or an antigen-binding fragment thereof and tremelimumab or an antigen-binding fragment thereof can result in desired pharmacokinetic parameters as seen in some early data. Total drug exposure can be assessed using the area under the curve (AUC). “AUC(tau)” refers to the AUC from time 0 to time τ, the dose interval, whereas “AUC(inf)” refers to the AUC to infinity. Administration may provide an AUC (tau) of about 600 to about 3000 μg/mL*day of MEDI4736 or an antigen-binding fragment thereof and from about 250 to about 350 μg/mL*day of tremelimumab or an antigen-binding fragment thereof. Administration may provide a maximum observed concentration (Cmax) of about 60 to about 300 μg/mL of MEDI4736 or an antigen-binding fragment thereof and from about 25 to about 35 μg/mL of tremelimumab or an antigen-binding fragment thereof. Administration may provide a C residual (minimum plasma drug concentration) of about 5 to about 40 μg/mL of MEDI4736 or an antigen-binding fragment thereof and from about 4 to about 6 μg/mL of tremelimumab or an antigen-binding fragment thereof.

[00129] As provided herein, MEDI4736 or an antigen-binding fragment thereof can also reduce levels of free (soluble) PD-L1. Free (soluble) PD-L1 refers to PD-L1 that is not bound (eg, MEDI4736). In some embodiments, PD-L1 levels are reduced by at least 65%. In some embodiments, PD-L1 levels are reduced by at least 80%. In some embodiments, PD-L1 levels are reduced by at least 90%. In some embodiments, PD-L1 levels are reduced by at least 95%. In some embodiments, PD-L1 levels are reduced by at least 99%. In some embodiments, PD-L1 levels are undetectable following administration of MEDI4736 or an antigen-binding fragment thereof and tremelimumab or an antigen-binding fragment thereof.

[00130] In some embodiments, PD-L1 levels are reduced by at least 65% following a single administration of MEDI4736 or an antigen-binding fragment thereof. In some embodiments, PD-L1 levels are reduced by at least 80% following a single administration of MEDI4736 or an antigen-binding fragment thereof. In some embodiments, PD-L1 levels are reduced by at least 90% following a single administration of MEDI4736 or an antigen-binding fragment thereof. In some embodiments, PD-L1 levels are reduced by at least 95% following a single administration of MEDI4736 or an antigen-binding fragment thereof. In some embodiments, PD-L1 levels are reduced by at least 99% following a single administration of MEDI4736 or an antigen-binding fragment thereof. In some embodiments, PD-L1 levels are undetectable following a single administration of MEDI4736 or an antigen-binding fragment thereof.

[00131] In some embodiments, PD-L1 levels are reduced by at least 65% following administration of two doses of MEDI4736 or an antigen-binding fragment thereof. In some embodiments, PD-L1 levels are reduced by at least 80% following administration of two doses of MEDI4736 or an antigen-binding fragment thereof. In some embodiments, PD-L1 levels are reduced by at least 90% following administration of two doses of MEDI4736 or an antigen-binding fragment thereof. In some embodiments, PD-L1 levels are reduced by at least 95% following administration of two doses of MEDI4736 or an antigen-binding fragment thereof. In some embodiments, PD-L1 levels are reduced by at least 99% following administration of two doses of MEDI4736 or an antigen-binding fragment thereof. In some embodiments, PD-L1 levels are undetectable after administration of two doses of MEDI4736 or an antigen-binding fragment thereof.

[00132] Treatment of a patient with a solid tumor using both MEDI4736 or an antigen-binding fragment thereof and tremelimumab or an antigen-binding fragment thereof (ie, co-therapy) provided herein may result in a synergistic effect. As used herein, the term “synergistic” refers to a combination of therapeutics (eg, a combination of MEDI4736 or an antigen-binding fragment thereof and tremelimumab or an antigen-binding fragment thereof) that is more effective than the additive effects of the individual therapeutics.

[00133] The synergistic effect of a combination of therapeutic agents (for example, a combination of MEDI4736 or an antigen-binding fragment thereof and tremelimumab or an antigen-binding fragment thereof) allows the use of lower doses of one or more therapeutic agents and/or less frequent administration of these therapeutic agents to a patient with solid tumor. The ability to use lower doses of therapeutic agents and/or administer these therapeutic agents less frequently reduces the toxicity associated with the administration of these therapeutic agents to a subject without reducing the effectiveness of these therapeutic agents in treating a solid tumor. In addition, the synergistic effect may result in improved effectiveness of the therapeutic agents in controlling, treating or alleviating solid tumors. By using the synergistic effect of a combination of therapeutic agents, the adverse or undesirable side effects associated with the use of any one therapeutic agent can be avoided or reduced.

[00134] In co-therapy, MEDI4736 or an antigen-binding fragment thereof may optionally be included in the same pharmaceutical composition as tremelimumab or an antigen-binding fragment thereof or may be included in a separate pharmaceutical composition. In the latter case, a pharmaceutical composition containing MEDI4736 or an antigen-binding fragment thereof is suitable for administration before, simultaneously with, or after the administration of a pharmaceutical composition containing tremelimumab or an antigen-binding fragment thereof. In certain cases, MEDI4736 or an antigen-binding fragment thereof is administered in parallel with tremelimumab or an antigen-binding fragment thereof in a separate composition.

[00135] Subjects suffering from lung cancer (eg, non-small cell lung cancer) may be tested for expression of a PD-L1 polynucleotide or polypeptide as part of treatment selection. Patients identified as having tumors that are negative for PD-L1 (eg, as determined by Ct score or IHC-M), or having reduced or undetectable PD-L1 levels relative to the reference level, are defined as responders to treatment with the anti-PD antibody combination -L1 and tremelimumab. Such patients are administered MEDI4736 or an antigen-binding fragment thereof in combination with tremelimumab or an antigen-binding fragment thereof.

EXAMPLES

EXAMPLE 1. Patients and methods

(a) SUBJECTS

[00136] Subjects in this study must be 18 years of age or older and have histologically or cytologically confirmed non-small cell lung cancer (NSCLC; squamous and non-squamous) with at least one lesion measured according to Response Evaluation Criteria in Solid Tumors v1.1 guidelines (RECIST), which are incorporated herein by reference in their entirety.

[00137] For both dose escalation and dose expansion phases, the cohort-specific requirements for previously received immunotherapy are as follows: a) immunotherapy-naïve cohort: must not have previously received immunotherapy, such as, but not limited to, other anti-CTLA-4, anti-PD-1 antibodies or to PD-L1, excluding vaccines; and b) immunotherapy-previous cohort: must have previously been exposed to immunotherapy, such as, but not limited to, other anti-CTLA-4, anti-PD-1, or anti-PD-L1 antibodies, excluding vaccines, more than 28 days prior to the first dose of MEDI4736 and tremelimumab.

[00138] Subjects must also fail to respond to standard treatment, relapse after standard treatment, refuse standard treatment, or be ineligible for standard treatment. Subjects will have an Eastern Cooperative Oncology Group (ECOG) performance status score of 0-1.

[00139] Subjects with central nervous system (CNS) metastases should be asymptomatic on day 1 of the study. In addition, by 1 day, at least 28 weeks must have passed without progression of CNS metastases, as confirmed by magnetic resonance imaging (MRI)/computed tomography (CT), after the last day of radiation treatment and at least 14 days after the last dose of corticosteroids.

[00140] Subjects must also have adequate organ (liver and kidney) and bone marrow function. Adequate organ and bone marrow function is defined as follows: hemoglobin≥9 g/dL; absolute number of neutrophils≥1500/ ^mm3 ; platelet count≥100,000/mm ³ ; total bilirubin < 1.5 x upper limit of normal (ULN), except in cases associated with Gilbert's syndrome or liver metastases (for such subjects, total bilirubin at baseline should be ≤3.0 mg/dL); alanine aminotransferase (ALT) and aspartate aminotransferase (AST) should be ≤2.5 × ULN, except in cases associated with liver metastases (for such subjects, ALT and AST should be ≤5 × ULN); and serum creatinine≤2.0 mg/dL.

[00141] Subjects are not eligible to participate if they are receiving any concomitant chemotherapy, immunotherapy, biologic, or hormonal therapy for the treatment of cancer. Subjects are not eligible to participate if they are taking any investigational cancer therapy within 28 days prior to the first dose of MEDI4736 and tremelimumab. Subjects may not participate if they have any pre-existing immune system-related adverse event (irAE) grade ≥3 while receiving immunotherapy, including anti-CTLA-4 antibody treatment, or any unresolved irAE grade >1. Subjects also cannot participate if they underwent a major surgical procedure (as determined by the investigator) within 28 days before the first dose of MEDI4736 and tremelimumab, or if they are still recovering from a previous surgical procedure. Subjects are also ineligible if they exhibit unresolved toxicities from prior anticancer therapy, defined as grade 0 or 1 unresolved according to the National Cancer Institute Common Terminology Criteria for Adverse Events (NCI CTCAE) v4.03, excluding alopecia and laboratory values. listed in the inclusion criteria. Subjects with irreversible toxicities that are not likely to be worsened by MEDI4736 and tremelimumab may be included. Subjects are also ineligible if they are currently using or have used an immunosuppressive drug within 14 days prior to the first dose of MEDI4736 and tremelimumab, with the exception of intranasal and inhaled corticosteroids or systemic corticosteroids at physiological doses not exceeding 10 mg/day of prednisone or equivalent.

[00142] Subjects cannot participate if they have an active or pre-existing autoimmune disease, including inflammatory bowel disease, diverticulitis, irritable bowel disease, celiac disease, Wegener's syndrome and Hashimoto's syndrome, within the last 3 years, excluding vitiligo, alopecia , Graves' disease or psoriasis not requiring systemic treatment (within the last 3 years). Subjects also cannot participate if they have a history of primary immunodeficiency or tuberculosis, or if they have known active or chronic viral hepatitis A, B, or C; if they are infected with human immunodeficiency virus (HIV), have other active severe diseases or uncontrolled intercurrent diseases; received a live, attenuated vaccine within 28 days prior to the first dose of MEDI4736 and tremelimumab; have another invasive malignancy within 5 years or a known allergy or hypersensitivity to the study drug formulations.

[00143] Subjects are also ineligible if they have advanced NSCLC with tumors containing anaplastic lymphoma receptor tyrosine kinase (ALK) gene rearrangements or epidermal growth factor receptor (EGFR)-sensitive mutations, and have not received appropriate tyrosine kinase inhibitor (TKI) therapy ). These subjects may be included after documented progression or intolerance to appropriate TKIs.

(b) STUDY DESIGN

[00144] The study is an open-label, phase 1b study of the combination of MEDI4736 and tremelimumab (Figure 1). Key study criteria for the cohorts are shown in Figure 11. The study will enroll approximately 36 subjects in the dose escalation phase and dose expansion phase, and a minimum of 20 subjects in each of the immunotherapy-naïve and immunotherapy-experienced cohorts. The maximum tolerated dose (MTD) or highest protocol-defined dose for each agent without exceeding the MTD determined for the immunotherapy-naïve cohort during the dose escalation phase will serve as the baseline dose for subjects enrolled in the immunotherapy-naïve cohort as part of the dose escalation phase. . The dose study for the immunotherapy-previous cohort will be conducted in parallel with a dose escalation phase for the immunotherapy-naïve cohort.

DOSE ESCALATION PHASE

Immunotherapy-naïve cohort

[00145] In the dose escalation phase of the study, MEDI4736 and tremelimumab will initially be administered to subsequent cohorts of 3-6 subjects who have not received immunotherapy, with each subject receiving tremelimumab Q4W for 6 doses and every 12 weeks (Q12W) for 3 doses ( i.e., dose 7 of tremelimumab was given at week 4 after dose 6, and dose 8 was given at week 12 after dose 7) and MEDI4736 Q4W 13 doses by IV infusion over a total of one year (Figure 2). Each cohort will include a minimum of 3 subjects according to the standard 3+3 design and the modified zone-based design. Subjects in the first cohort (Cohort 1) will receive a dose of 3 mg/kg MED4736 and 1 mg/kg tremelimumab. If dose limiting toxicities (DLT) ≥2 are observed in the first dose cohort, then the initial dose will be reduced according to Figure 1. If no more than 1 in 6 subjects experience a DLT in a given dose cohort, then dose escalation will continue.

[00146] Subsequent cohorts will receive tremelimumab administered by Q4W for 6 doses followed by Q12W for 3 doses, and MEDI4736 administered by Q4W for 13 doses until the MTD or highest protocol-defined dose for each agent without exceeding is determined MTD.

[00147] The escalation phase for the immunotherapy-naïve cohort (see Figure 1) will begin once the MTD or highest protocol-defined dose for each agent is determined without exceeding the MTD in immunotherapy-naïve subjects during the dose escalation phase. Additional dose(s) from the zone-based bridging cohort(s) that do not exceed the MTD may be assessed in the ramp-up phase based on PK and pharmacodynamics assessments , safety parameters and efficacy in the immunotherapy-naïve cohort from the dose escalation phase.

Previously immunotherapy-treated cohort

[00148] The MTD or highest protocol-defined dose for each agent without exceeding the MTD determined for the immunotherapy-naïve cohort (see Figure 1) during the dose escalation phase will serve as the baseline dose for subjects enrolled in the immunotherapy-naïve cohort as part of the dose escalation phase. A dose study for the immunotherapy-naïve cohort using a 3+3 design will be conducted in parallel with a dose escalation phase for the immunotherapy-naïve cohort. The dose escalation phase for the immunotherapy-previous cohort can begin as soon as the MTD or highest protocol-defined dose for each agent is determined without exceeding the MTD in immunotherapy-previous subjects during the dose escalation phase. Additional dose(s) from the zone-based bridging cohort(s) that do not exceed the MTD may be assessed in the dose escalation phase based on the PK and pharmacodynamics, safety and efficacy parameters in the immunotherapy-previous cohort from the dose escalation phase.

DOSE INCREASE PHASE

[00149] Two dose escalation cohorts will be used: immunotherapy-naïve and immunotherapy-previous (see Figure 1). The dose escalation phase for both cohorts will use the MTD or the highest protocol-defined dose for each agent without exceeding the MTD determined during the dose escalation phase. In addition, dose(s) from the zone-based bridging cohort(s) that do not exceed the MTD may be included as an additional dose(s) for immunotherapy-experienced and immunotherapy-naïve cohorts in the dose escalation phase based on subject-reported data, including safety, PK, pharmacodynamics, biomarker and response data, as well as data from the present trials.

[00150] Initially, approximately 20 subjects with NSCLC will be included in each of the immunotherapy-naïve and immunotherapy-pretreated cohorts. Additional subjects, up to a total of 60 subjects, who may be included in the immunotherapy-naïve cohort in the dose escalation phase depending on data received from the subjects, including safety, PK, pharmacodynamics, biomarker and response data, and data obtained in these tests.

TREATMENT REGIME

[00151] Subjects will be treated in either the dose escalation or dose expansion phase of the study. In the dose escalation and dose expansion phases of the study, tremelimumab will be administered every four weeks (Q4W) for 6 doses and then every twelve weeks (Q12W) for 3 doses, with dose 7 administered 4 weeks after dose 6 and dose 8 administered 12 weeks after dose 7. MEDI4736 will be administered Q4W for 13 doses. Both agents will be administered by IV infusion for a total of one year (Figure 2).

[00152] Tremelimumab will be administered intravenously in 250 ml of 0.9% sodium chloride. MEDI4736 will be administered intravenously in 250 mL of 0.9% sodium chloride. Tremelimumab will be administered first. The MEDI4736 infusion will begin approximately 1 hour after the end of the tremelimumab infusion. In cohort 1, the duration of the first IV infusion of tremelimumab will be approximately 1 hour, and the duration of the first IV infusion of MEDI4736 will be approximately 4 hours; The duration of subsequent infusions for subjects in this cohort and all infusions for the remaining cohorts will be approximately 1 hour.

[00153] Subjects who achieve and maintain disease control (DC) (i.e., complete remission (CR), partial remission (PR), or stable disease (SD)) by the end of the 12-month treatment period with MEDI4736 and tremelimumab , will be enrolled for follow-up. If there is evidence of progressive disease (PD) during follow-up, subjects may be reintroduced to MEDI4736 and tremelimumab for an additional period of up to 12 months with the same guidelines followed during the initial 12-month period, unless the subject is receiving other treatments for their disease and still met the inclusion and exclusion criteria for the study protocol. Only one round of retreatment with MEDI4736 and tremelimumab will be permitted.

[00154] The study uses a 3+3 dose escalation phase and follows the design depicted in Figure 3. A minimum of three subjects will be included in each dose cohort. If 0 of 3 patients in a dose cohort experience dose-limiting toxicity (DLT), then the next dose escalation cohort will be initiated. If 1 of 3 patients is observed to have DLT, then an additional 3 patients will be included in the same cohort. If no more than 1 in 6 subjects experience DLT, then the next dose escalation cohort will begin. If 2 or more patients in a dose cohort experience DLT during the evaluation period, then no additional subjects will be included in that dose cohort.

[00155] The maximum tolerated dose (MTD) is defined as the highest dose of tremelimumab or MEDI4736 in a cohort where no more than 1 in 6 patients experience DLT. It is determined for both the immunotherapy-naïve and treatment-experienced cohorts during the dose escalation phase. Patients in the escalation phase (immunotherapy-naïve and immunotherapy-previous cohorts) will be treated at the MTD determined in the dose escalation phase.

[00156] Administration of MEDI4736 and tremelimumab may be modified or discontinued as a result of toxicities as described in Figures 10A and 10B. Dose modifications will not be required for adverse events (AEs) that are not clearly explained by MEDI4736 or tremelimumab (such as trauma), or for laboratory abnormalities that are not considered clinically significant.

(c) EVALUATION OF PHARMACOKINETIC PARAMETERS, ANTI-TUMOR ACTION, BIOMARKER, SOLUBLE FACTOR AND SAFETY

[00157] Measurements of serum concentrations of MEDI4736 and tremelimumab will be performed using an approved immunoassay.

[00158] Blood samples for pharmacokinetic evaluation will be collected on the following days during the dose escalation phase: day 1 (pre-dose and end of infusion), day 8 ( +1 day), day 15 ( +1 day), day 29 (before dose and end of infusion; +3 days), day 57 (before dose and end of infusion; +3 days), day 85 (before dose and end of infusion; +3 days), day 113 (before dose and end of infusion ;+3 days), day 141 (before dose and end of infusion;+3 days), day 169 (before dose and end of infusion;+3 days), day 197 (before dose and end of infusion;+3 days; MEDI4736 only), day 225 (pre-dose and end of infusion;+3 days; MEDI4736 only), day 253 (pre-dose and end of infusion;+3 days), day 281 (pre-dose and end of infusion;+3 days ; MEDI4736 only), day 309 (pre-dose and end of infusion; +3 days), day 337 (pre-dose and end of infusion; +3 days), at end of treatment, 60 days after end of treatment, and 90 days after end of treatment.

[00159] Blood samples for pharmacokinetic evaluation will be collected on the following days during the dose escalation phase: Day 1 (pre-dose and end of infusion), Day 29 ( +3 days), Day 57 ( +3 days), Day 85 ( + 3 days), day 113 ( + 3 days), day 141 ( + 3 days), day 169 ( + 3 days); day 197 ( + 3 days; MEDI4736 only), day 225 ( + 3 days; MEDI4736 only), day 253 ( + 3 days), day 281 ( + 3 days; MEDI4736 only), day 309 ( + 3 days; MEDI4736 only ), day 337 ( + 3 days), at the end of treatment, 60 days after the end of treatment and 90 days after the end of treatment.

[00160] The presence of anti-drug antibodies (ADA) will be assessed on day 1 and 29 ( +3 days), 85 ( +3 days), 141 ( +3 days), 169 ( +3 days), 253 ( +3 days ) and 337 days ( + 3 days), at the end of treatment, 60 days after the end of treatment and 90 days after the end of treatment. Validated electrochemiluminescence assays using the Meso Scale Discovery platform will be used to detect antibodies to MEDI4736 in human serum and to detect antibodies to tremelimumab in human serum.

[00161] Blood samples will be collected for analysis of circulating soluble factors, including soluble PD-L1 (sPD-L1). During the dose escalation phase, sPD-L1 levels will be assessed on day 1, day 8 ( +1 day), day 15 ( +1 day), day 29 ( +3 days), day 57 ( +3 days), day 85 ( + 3 days), 113 day ( + 3 days), 141 day ( + 3 days), 169 day ( + 3 days); 197 day ( + 3 days), 225 day ( + 3 days), 253 day (+ 3 days), 281 day (+ 3 days), 309 day (+ 3 days), 337 day (+ 3 days), at the end treatment, 60 days after the end of treatment and 90 days after the end of treatment. During the dose escalation phase, sPD-L1 levels will be assessed on day 1, day 29 (+ 3 days), day 57 (+ 3 days), day 85 (+ 3 days), day 113 (+ 3 days), day 141 ( + 3 days), 169 day (+ 3 days); 197 day (+ 3 days), 225 day ( + 3 days), 253 day ( + 3 days), 281 day ( + 3 days), 309 day ( + 3 days), 337 day ( + 3 days), at the end treatment, 60 days after the end of treatment and 90 days after the end of treatment.

[00162] Tumor assessments will be performed at screening (day -28 to -1 day), at week 8 (day 50+3 days) and at week 33 (day 225+3 days) in the dose escalation phase. Tumor assessments will be based on RECIST v1.1 guidelines with modifications and include the following examinations: physical examination (with photographs and measurements of skin lesions, as appropriate), CT or MRI scan of the chest, abdomen, and pelvis; and CT or MRI scans of the brain. Computed tomography or MRI scans of the brain are performed only for screening.

[00163] Estimates of antitumor activity are based on measuring tumor lesions, assessing target lesions, assessing non-target lesions, and the appearance of new lesions.

Assessment of target lesions

[00164] For evaluation of target lesions, complete remission is defined as the disappearance of all target lesions. Any abnormal lymph nodes (either target or non-target) should show a short-axis decline of <10 mm (the sum cannot be “0” if the nodes are target).

[00165] For evaluation of target lesions, partial remission is defined as a reduction of at least 30% in the sum of the diameters of the target lesions, with the initial sum of diameters being the reference.

[00166] For evaluation of target lesions, progressive disease is defined as an increase of at least 20% in the sum of target lesion diameters, with the reference being the smallest sum in the study (this includes the original sum if it is the smallest in the study). In addition to a relative increase of 20% in total, an increase of at least 5 mm must also be visible. The appearance of one or more new lesions is also considered progression.

[00167] For evaluation of target lesions, stable disease (SD) is not sufficient reduction to be PR nor sufficient enhancement to be PD, with the smallest sum of diameters in the study being considered the reference.

Evaluation of non-target lesions

[00168] For evaluation of non-target lesions, complete remission (CR) is defined as the disappearance of all non-target lesions. All lymph nodes should be < 10 mm in short axis.

[00169] For the evaluation of non-target lesions, incomplete remission/non-progressive disease is defined as persistence of one or more non-target lesions and/or maintenance of tumor marker levels above normal limits.

[00170] For the evaluation of non-target lesions, progressive disease is defined as the overall level of significant worsening of the non-target disease such that even in the presence of SD or PR in the target disease, the total tumor burden increases enough to discontinue therapy. In the absence of a measurable disease, the change in the unmeasured disease is compared in magnitude to the increase that would be required to define PD as a measurable disease.

New hearth

[00171] The appearance of new lesions is considered PD according to RECIST v1.1 guidelines. Given the unique response kinetics observed with immunotherapy, new lesions may not reflect actual disease progression. In the absence of rapid clinical deterioration, subjects may continue to receive treatment with MEDI4736 and tremelimumab.

General answer

[00172] Confirmation of CR, PR, as well as PD requires a repeat follow-up assessment no less than 4 weeks from the date of first registration. The overall response assessment based on the assessment of the target lesion, non-target lesion and new lesion is determined as in Table 1 below.

Table 1. Evaluation of the overall response

Target lesions Non-target lesions New outbreaks General answer Complete remission Complete remission No Complete remission Non-target lesion ^a Complete remission No Complete remission Complete remission Inestimable ^b No Partial remission Complete remission Incomplete remission/non-progressive disease No Partial remission Partial remission Non-progressive disease and not assessable ^b No Partial remission Stabilization of the disease Non-progressive disease and not assessable ^b No Stabilization of the disease Not everyone was evaluated Non-progressive disease No Inestimable Non-target lesion ^a Not everyone was evaluated No Inestimable Non-target lesion ^a Incomplete remission/non-progressive disease No Incomplete remission/non-progressive disease Progressive disease Any Yes or no Progressive disease Any Progressive disease Yes or no Progressive disease Any Any Yes Progressive disease Non-target lesion ^a Clearly progressive disease Yes or no Progressive disease Non-target lesion ^a Any Yes Progressive disease

^a Defined as non-target lesions at baseline.

^b Non-assessable is defined as either no measurements or only a subset of lesion measurements taken at assessment.

[00173] Adverse events are monitored following administration of MEDI4736. Other evaluations include a physical examination, monitoring of vital signs, and laboratory measurements.

EXAMPLE 2. Results

(a) ENTRY AND FOLLOW-UP

[00174] Twelve subjects were treated, 3 subjects each in Cohort 1a (1 mg/kg tremelimumab and 3 mg/kg MEDI4736), Cohort 2a (1 mg/kg tremelimumab and 10 mg/kg MEDI4736), Cohort 3a (1 mg /kg tremelimumab and 15 mg/kg MEDI4736) and cohort 3b (3 mg/kg tremelimumab and 10 mg/kg MEDI4736). Two subjects in cohort 1a (one subject withdrew consent after 2 doses of both agents) completed approximately 115 days of follow-up; Cohort 2a subjects completed approximately 56 days of follow-up; and subjects in cohorts 3a and 3b completed 28 days of follow-up (Figure 12).

(b) PHARMACOKINETICS

[00175] Pharmacokinetic data obtained from administration of MEDI4736 at 3 mg/kg and 10 mg/kg (in combination with 1 mg/kg tremelimumab) are summarized in Figure 4. Select PK parameters obtained from administration of MEDI4736 are shown in Figure 8. Among these initial first dose data, the mean Cmax for subjects receiving 3 mg/kg MEDI4736 was 67.36 mcg/mL. The mean AUCτ for subjects receiving 3 mg/kg MEDI4736 was 625.3 μg/mL*day. The mean C _residual for subjects receiving 3 mg/kg MEDI4736 was 8.85 μg/mL. The mean Cmax for subjects receiving 10 mg/kg MEDI4736 was 266.7 μg/mL. The mean AUCτ for subjects receiving 10 mg/kg MEDI4736 was 2860 μg/mL*day. The mean C _residual for subjects receiving 10 mg/kg MEDI4736 was 35.65 μg/mL. MEDI4736 exhibited a more than dose-proportional increase in AUC, indicating a nonlinear PK. See figure 4.

[00176] Pharmacokinetic data obtained from tremelimumab administration at 1 mg/kg (in combination with either 3 mg/kg MEDI4736 or 10 mg/kg MEDI4736) are summarized in Figure 5. Select PK parameters obtained from tremelimumab administration are shown in figure 9.

[00177] The average Cmax for subjects receiving 1 mg/kg tremelimumab in combination with 3 mg/kg MEDI4736 was 27.07 μg/ml. The mean Cmax for subjects receiving 1 mg/kg tremelimumab in combination with 10 mg/kg MEDI4736 was 30.40 mcg/mL.

[00178] The mean AUCτ for subjects receiving 1 mg/kg tremelimumab in combination with 3 mg/kg MEDI4736 was 262.2 μg/mL*day. The mean AUCτ for subjects receiving 1 mg/kg tremelimumab in combination with 10 mg/kg MEDI4736 was 338.8 μg/mL*day.

[00179] The mean _Cmax for subjects receiving 1 mg/kg tremelimumab in combination with 3 mg/kg MEDI4736 was 4.21 μg/ml. The mean _residual C for subjects receiving 1 mg/kg tremelimumab in combination with 10 mg/kg MEDI4736 was 5.33 μg/mL.

(c) ASSESSMENT OF SOLUBLE PD-L1

[00180] sPD-L1 data obtained from MEDI4736 at 3 and 10 mg/kg and tremelimumab at 1 mg/kg are shown in Figures 6 (absolute concentration), 7 and 8 (suppression, % of baseline). Following administration of MEDI4736, complete suppression of sPD-L1 was observed in 5 of 6 subjects. In one subject, sPD-L1 suppression of approximately 65% was observed after the second dose of MEDI4736.

(d) SAFETY

[00181] Ten of the twelve subjects reported a treatment-emergent adverse event (TEAE). The most commonly reported TEAEs were fatigue (41.7%; 5 subjects), elevated amylase (25.0%; 3 subjects), pruritus, and upper respiratory tract infection (16.7%; 2 subjects each). No subjects experienced dose-limiting toxicity. Nine out of ten subjects experiencing TEAE experienced grade 1 or 2 severity. One subject (cohort 2a) had elevated aspartate aminotransferase (AST) and elevated alanine aminotransferase (ALT) grade 3 and fatal (grade 5) myasthenia gravis. Two of the twelve subjects experienced treatment-emergent serious adverse events (SAEs). Two subjects discontinued treatment due to TEAE. Figure 13 shows relevant adverse events.

[00182] Initial results demonstrating change in tumor size from baseline are shown in Figure 14. Reductions in tumor size were observed in patients in the 1 mg/kg (T)+10 mg/kg (M), 1 mg/kg (T) groups +15 mg/kg (M) and 3 mg/kg (T)+10 mg/kg (M).

[00183] This study showed that MEDI4736 and tremelimumab have favorable pharmacokinetic properties, and the combination of agents inhibits sPD-L1. In addition, MEDI4736 is well tolerated by the majority of subjects receiving this treatment.

EXAMPLE 3: Phase 1b open-label study of MEDI4736 in combination

with tremelimumab in patients with advanced NSCLC

[00184] The PD-L1 and CTLA-4 inhibitory pathways play an important role in controlling T cell activation. MEDI4736(M) is a human IgG1 monoclonal antibody that blocks PD-L1 by binding to PD-1 and CD-80. Tremelimumab (T) is a human IgG2 monoclonal antibody that specifically targets CTLA-4. Both MEDI4736 and tremelimumab demonstrated encouraging safety profiles and promising clinical activity as single agents; Because they block various interactions that contribute to immunosuppression, MEDI4736+tremelimumab combination therapy may provide greater antitumor activity in patients (pts) with advanced non-small cell lung cancer compared with either agent alone.

[00185] This phase 1 study evaluates the safety/tolerability, antitumor activity, pharmacokinetics (PK) and immunogenicity of MEDI4736+tremelimumab therapy in patients with advanced NSCLC. The study consists of dose escalation and dose expansion phases.

[00186] Results . As of December 4, 2014, 61 patients were treated ( Table 2 ).

table 2

Cohort Dose 1a 3 mg/kg M q4w+1 mg/kg T 2a 10 mg/kg M q4w+1 mg/kg T 3a 15 mg/kg M q4w+1 mg/kg T 3b 10 mg/kg M q4w+3 mg/kg T 4 20 mg/kg M q4w+1 mg/kg T 4a 15 mg/kg M q4w+3 mg/kg T 5 15 mg/kg M q4w+10 mg/kg T 5a 20 mg/kg M q4w+3 mg/kg T 6 20 mg/kg M q4w+10 mg/kg T 8 10 mg/kg M q2w+1 mg/kg T 9 10 mg/kg M q2w+3 mg/kg T 10 10 mg/kg M q2w+10 mg/kg T

MEDI4736 (M)+tremelimumab (T)

Q4W=every 4 weeks; Q2W=every 2 weeks

[00187] Overall, the most common drug-related AEs of all grades (≥10% pts) were diarrhea, fatigue, pruritus, elevated alanine transaminase (ALT) and amylase levels; which were controlled according to guidelines for standard treatment involving steroids. The most common drug-related grade ≥3 (≥5%) AEs in all patients were colitis, diarrhea, and elevated aspartate aminotransferase (AST) and ALT levels. Drug-related AEs leading to discontinuation were colitis (6.6%), diarrhea (3.3%), pneumonia (3.3%), elevated AST (3.3%), and ALT (1.6 %), cough (1.6%) and shortness of breath (1.6%); There was 1 treatment-related death (polymyositis) in the 10 mg/kg M q4w+1 mg/kg T cohort. Of 31 subjects with at least one scan at 8 weeks, best overall response was recorded in 8 patients (26%); Stabilization of the disease was recorded in 11 pts (36%).

[00188] These promising results indicate that the MEDI4736+tremelimumab combination has a manageable safety profile and early clinical activity.

[00189] Results . As of January 27, 2015, 74 patients were treated in 10 cohorts (Table 3).

Table 3

Demographic and baseline characteristics

Table 3 (continued)

Table 3 (continued)

[00190] Different doses and schedules were examined for MEDI4736 and tremelimumab administered in combination (Figure 15). Doses and schedules for MEDI4736 included 3 mg/kg, 10 mg/kg, 15 mg/kg, and 20 mg/kg every 4 weeks (q4w) and 10 mg/kg every 2 weeks (q2w). Doses and schedules for tremelimumab included 1 mg/kg, 3 mg/kg, and 10 mg/kg every 4 weeks (q4w x 6 doses) and every 12 weeks (q12w x 3 doses).

[00191] Complete suppression of sPD-L1 was observed at doses ≥15 mg/kg Q4W MEDI4736 (Figure 16). One subject in each 10 mg/kg Q4W and 15 mg/kg Q4W groups experienced partial suppression of sPD-L1 at some visits, followed by complete suppression after repeat dosing. In one subject, no suppression was observed at day 29 following 15 mg/kg Q4W. In comparison, complete suppression was observed with single agent MEDI4736 (Study 1108) at doses >3 mg/kg Q2W. When examining individual sPD-L1 profiles, complete suppression of sPD-L1 was observed in >95% of subjects following a dose of 10 mg/kg Q2W (Figure 17).

[00192] Increased T cell proliferation was also observed when MEDI4736 and tremelimumab were administered in combination (Figure 18). Cells were sorted by FACS analysis for CD4 and CD8 markers in combination with Ki67, a cell proliferation marker. A consistent increase in the CD4+Ki67+ cell peak was observed with increasing tremelimumab dose, which was greater than the increase in the CD4+Ki67+ peak observed with MEDI4736 alone. The CD8+ proliferative response was greater with combinations of MEDI4736 and tremelimumab than with MEDI4736 alone.

[00193] Increases in CD4 ICOS+ and CD4 HLADR+ cells were observed in subjects receiving MEDI4736 and tremelimumab compared to MEDI4736 alone (Figure 19). The smallest changes in levels of CD4+ICOS+ cells and CD4+HLADR+ cells were observed only with MEDI4736, and all doses of the combination tested showed a pattern distinct from MEDI4736. Maximum increases in CD4+ICOS+ cells and CD4+HLADR+ cells were observed with a dose of 10 mg/kg tremelimumab. An increase in CD4+ effector T cells and regulatory T cells (the CD25hi CD127lo phenotype may also include some activated T cells) was also observed in subjects receiving MEDI4736 and tremelimumab compared to MEDI4736 alone (Figures 20 and 21).

[00194] Clinical activity in NSCLC was observed with treatment with MEDI4736 and tremelimumab (all doses), showing increased overall response rates compared to treatment with MEDI4736 alone (10 mg/kg Q2W) (Figure 22). Response was assessed in treated patients with measurable disease at baseline+≥1 follow-up scan (including discontinuations due to disease progression or death before first follow-up scan). For MEDI4736 NSCLC (CP1108), only patients with ≥12 weeks of follow-up were included. Overall response rate (ORR) included confirmed and unconfirmed complete remission (CR) or partial remission (PR). For MEDI4736 NSCLC monotherapy (CP1108, 10 mg/kg Q2W), best overall response (BOR) was observed as stable disease (SD) with the shortest duration of 12 weeks. For the combination of MEDI4736 and tremelimumab, BOR was observed as SD with the shortest duration of 7 weeks.

[00195] Reduction or stabilization of tumor size was observed in the majority of NSCLC patients treated with MEDI4736 and tremelimumab at tremelimumab doses of 1 mg/kg, 3 mg/kg, and 10 mg/kg (Figure 23). The occurrence of grade 3 adverse events showed that increasing the dose of tremelimumab also reduced the time to occurrence of the adverse event (Figure 24). Reduction or stabilization of tumor size was observed in the majority of NSCLC patients treated with MEDI4736 and tremelimumab at MEDI4736 doses of 10 mg/kg, 315 mg/kg, and 20 mg/kg Q4W or 10 mg/kg Q2W (Figure 25). The occurrence of grade 3 adverse events showed that increasing the dose of tremelimumab affected the time of occurrence of the adverse event, in contrast to increasing the dosage of MEDI4736 (Figure 26). Therefore, when selecting doses of MEDI4736 and tremelimumab, efficacy and safety should be taken into account.

[00196] Data from tremelimumab as a single agent in cancer therapy support that higher dose exposures (>30 mg/mL) are required for maximum efficacy. Such levels are achieved on average at doses > 3 mg/kg. The exposure achieved after tremelimumab at 1 mg/kg is expected to produce less toxicity than at 3 mg/kg based on an analysis of individual exposure and safety relationships. Without being bound by theory, tremelimumab at 1 mg/kg partially inhibits CTLA4, although immune-mediated side effects are still observed at 1 mg/kg (albeit at a lower incidence). Tremelimumab at 1 mg/kg may be optimal in combination with MEDI4736 if synergistic efficacy is observed and the safety profile improves relative to higher doses of tremelimumab.

[00197] One study observed complete inhibition of sufficient serum dissolution of PD-L1 and sPD-L1 at doses > 3 mg/kg Q2W MEDI4736. The 10 mg/kg dose of Q2W was selected for use as a single agent based on achieving >99% saturation of sufficient dissolution in the majority of subjects with drug levels providing maximum tumor growth inhibition (TGI) in mouse models (100 mg/mL was identified as the lowest target residual concentration). Following 10 mg/kg Q4W in combination with tremelimumab, sPD-L1 suppression was observed in some subjects. Dose density was higher with MEDI4736 Q2W and Cmax was higher with Q4W. A Q4W schedule may provide greater convenience. In other subjects, incomplete suppression was observed, possibly due to upregulation of PD-L1 in combination with suboptimal residual levels achieved at this dose level. MEDI4736 at 10 mg/kg Q4W, evaluated in combination with tremelimumab, provided residual concentrations below the target of 100 mg/mL. However, target levels were achieved with MEDI4736 at 15 and 20 mg/kg Q4W, and higher doses of MEDI4736 maintained sPD-L1 suppression in all subjects evaluated to date. MEDI4736 at 10 mg/kg or higher may be optimal in combination with tremelimumab if synergistic efficacy is observed and the safety profile is improved.

[00198] Overall, the trend for the combination of MEDI4736 and tremelimumab was to reduce or stabilize disease compared with MEDI4736 alone, and both MEDI4736 and tremelimumab at the highest doses tested were associated with an increase in the incidence of adverse events. (Figures 27-29). These results indicate that the combination of MEDI4736 and tremelimumab is well tolerated and has the potential to be an effective therapy for NSCLC.

EXAMPLE 4: Treatment of PD-L1 Negative NSCLC Tumors with MED14736 and Tremelimumab

[00199] Twelve subjects were treated, 3 subjects each in Cohort 1a (1 mg/kg tremelimumab and 3 mg/kg MEDI4736), Cohort 2a (1 mg/kg tremelimumab and 10 mg/kg MEDI4736), Cohort 3a (1 mg /kg tremelimumab and 15 mg/kg MEDI4736) and cohort 3b (3 mg/kg tremelimumab and 10 mg/kg MEDI4736). Two subjects in cohort 1a (one subject withdrew consent after 2 doses of both agents) completed approximately 115 days of follow-up; Cohort 2a subjects completed approximately 56 days of follow-up; and subjects in cohorts 3a and 3b completed 28 days of follow-up.

[00200] Baseline tumor PD-L1 expression data for the 7 subjects in the study are presented in Table 4 (below). Additional information is presented in Table 5.

Table 5

[00201] Test tissue from NSCLC patients was characterized for PD-L1 expression using immunohistochemistry in formalin-embedded, paraffin-embedded tissue samples. A sample was defined as “PD-L1 positive” if the sample contained 25% or more tumor cells with PD-L1 membrane staining. This is expressed as the membrane immunohistochemistry (M) score. All samples were scored “negative” for PD-L1 expression. Tumor evaluations were available for 6 of 7 patients. These patients were treated with a combination of tremelimumab and MEDI4736.

[00202] Two patients were identified as unconfirmed partial remission (PR), two had stable disease (SD), and two had progressive disease (PD). All 6 patients had tumor samples collected >6 months prior to analysis for PD-L1 expression, and 3 of 6 biopsy samples were collected >2 years prior to analysis. These data indicate that the combination of MEDI4736 and tremelimumab is active in PD-L1 negative NSCLC based on stored tissue samples.

[00203] When MEDI4736 and tremelimumab were administered, the majority of patients with PD-L1 negative NSCLC responded to combination therapy and experienced reductions or stabilization of tumor size compared to MEDI4736 alone (CP1108, 10 mg/kg Q2W) (Figure 30 ). Patients with PD-L1 positive NSCLC also responded to the combination of MEDI4736 and tremelimumab as opposed to MEDI4736 alone and experienced reductions or stabilization of tumor size (Figure 31). When grouping the results of patients with PD-L1 negative NSCLC by tremelimumab dose, 1 mg/kg tremelimumab or 3 mg/kg tremelimumab administered in combination with MEDI4736 at 10 mg/kg Q4W or 15 mg/kg Q4W were effective in controlling or attenuating diseases (figure 32). When results were grouped by MEDI4736 dose, the results also showed that tremelimumab at 1 mg/kg - 3 mg/kg, administered in combination with MEDI4736 at 10 mg/kg Q4W - 15 mg/kg Q4W, was effective in controlling or attenuating disease (Figure 33). Analysis of all NSCLC patients receiving MEDI4736 and tremelimumab showed that patients with PD-L1- and PD-L1+ NSCLC responded to treatment (Figures 34A-34D and 35-37; Tables 6-9).

Table 6

Best overall answer - all subjects assessed

Responding population being assessed

Table 7

Best overall answer - PD-L1 positive subjects

Responding population being assessed

Table 8

Best overall answer - PD-L1 negative subjects

Responding population being assessed

Table 9

Best overall answer - NA by PD-L1 subjects

Responding population being assessed

[00204] Numerous equivalents to specific aspects of the disclosure described herein will be apparent to those skilled in the art, or will be able to determine them using no more than routine experimentation. Such equivalents are intended to be covered by the following claims.

[00205] Various publications are cited herein, the disclosures of which are incorporated by reference in their entirety.

[00206] While the foregoing invention has been described in some detail by way of illustration and example to facilitate understanding, it will be apparent that certain changes and modifications may be made within the scope of the appended claims.

LIST OF SEQUENCES

SEQ ID NO:1 MEDI4736 VL

EIVLTQSPGTLLSLSPGERATLSCRASQRVSSSYLAWYQQKPGQAPRLLIYDASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSLPWTFGQGTKVEIK

SEQ ID NO:2 MEDI4736 VH

EVQLVESGGGLVQPGGSLRLSCAASGTFFSRYWMSWVRQAPGKGLEWVANIKQDGSEKYYVDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAREGGWFGELAFDYWGQGTLVTVSS

SEQ ID NO:3 - MEDI4736 VH CDR1

RYWMS

SEQ ID NO:4 - MEDI4736 VH CDR2

NIKQDGSEKYYVDSVKG

SEQ ID NO:5 - MEDI4736 VH CDR3

EGGWFGELAFDY

SEQ ID NO:6 - MEDI4736 VL CDR1

RASQRVSSSYLA

SEQ ID NO:7 - MEDI4736 VL CDR2

DASSRAT

SEQ ID NO:8 - MEDI4736 VL CDR3

QQYGSLPWT

SEQ ID NO:9 VL tremelimumab

PSSLSASVGDRVTITCRASQSINSYLDWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYYSTPFTFGPGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKV

SEQ ID NO:10 VH of tremelimumab

GVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDPRGATLYYYYYGMDVWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVH

SEQ ID NO:11 - CDR1 VH of tremelimumab

GFTFSSYGMH

SEQ ID NO:12 - CDR2 VH of tremelimumab

VIWYDGSNKYYADSV

SEQ ID NO:13 - CDR3 VH of tremelimumab

TAVYYCARDPRGATLYYYYYGMDV

SEQ ID NO:14 - CDR1 VL of tremelimumab

RASQSINSYLD

SEQ ID NO:15 - CDR2 VL of tremelimumab

AASSLQS

SEQ ID NO:16 - CDR3 VL of tremelimumab

QQYYSTPFT

--->

LIST OF SEQUENCES

<110> MEDIMMUNE LIMITED

<120> ANTIBODIES TO B7-H1 AND TO CTLA-4 FOR THE TREATMENT OF NON-SMALL CELL

LUNG CANCER

<130> TRB7-100WO1

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<151> 2015-02-10

<150> 62/105992

<151> 2015-01-21

<150> 61/992658

<151> 2014-05-13

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synthetic

peptide"

<400> 13

Thr Ala Val Tyr Tyr Cys Ala Arg Asp Pro Arg Gly Ala Thr Leu Tyr

1 5 10 15

Tyr Tyr Tyr Tyr Gly Met Asp Val

20

<210> 14

<211> 11

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

synthetic

peptide"

<400> 14

Arg Ala Ser Gln Ser Ile Asn Ser Tyr Leu Asp

1 5 10

<210> 15

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

synthetic

peptide"

<400> 15

Ala Ala Ser Ser Leu Gln Ser

15

<210> 16

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

synthetic

peptide"

<400> 16

Gln Gln Tyr Tyr Ser Thr Pro Phe Thr

15

<---

Claims (12)

1. A method of treating non-small cell lung cancer (NSCLC) in a patient, comprising administering MEDI4736 and tremelimumab, wherein: (i) administration may result in an average _Cmax of 266-300 μg/ml for MEDI4736 and a _Cmax of 25-30 μg/ml for tremelimumab; (ii) MEDI4736 was administered 1 hour after tremelimumab; And (iii) MEDI4736 and tremelimumab are administered every 4 weeks for a maximum of 16 weeks. 2. The method according to claim 1, where the total duration of treatment is 52 weeks. 3. The method of claim 1 or 2, wherein the NSCLC is resistant to at least one chemotherapeutic agent prior to administration of MEDI4736 and tremelimumab. 4. The method of claim 3, wherein the chemotherapeutic agent is Vemurafenib, Erlotinib, Afatinib, Cetuximab, Carboplatin, Bevacizumab, Gefitinib or Pemetrexed. 5. The method according to any one of claims 1 to 4, wherein the patient is immunotherapeutically naïve before administration of MEDI4736 and tremelimumab. 6. The method according to any one of claims 1 to 5, where the administration of MEDI4736 is carried out by intravenous infusion. 7. The method according to any one of claims 1 to 6, where the administration of tremelimumab is carried out by intravenous infusion. 8. The method according to any one of claims 1 to 7, where the patient has locally advanced and metastatic non-small cell lung cancer. 9. Method according to any one of claims 1 to 8, wherein the NSCLC is squamous or non-squamous.