Anti-B7-H1 and anti-CTLA-4 antibodies for the treatment of non-small cell lung cancer

Abstract

The present invention relates to anti-B7-H1 and anti-CTLA-4 antibodies for the treatment of non-small cell lung cancer. Provided herein are methods of treating non-small cell lung cancer comprising administering an effective amount of MEDI4736 or an antigen-binding fragment thereof and tremelimumab or an antigen-binding fragment thereof.

Description

[0001] This application is a divisional application of Chinese patent application 201580024117.6, “Anti-B7-H1 and anti-CTLA-4 antibodies for treating non-small cell lung cancer,” filed on May 12, 2015.

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS

[0003] This application claims priority to and the benefit of U.S. Provisional Patent Application Serial No. 61 / 992,658, filed May 13, 2014, U.S. Provisional Patent Application Serial No. 62 / 105,992, filed January 21, 2015, and U.S. Provisional Patent Application Serial No. 62 / 114,336, filed February 10, 2015. The entire contents of each of these applications are hereby incorporated by reference herein. Background Art

[0004] Cancer continues to be a major global health burden. Despite advances in the treatment of cancer, there continues to be an unmet medical need for more effective and less toxic therapies, especially for those patients with advanced disease or cancer that is resistant to existing therapeutics.

[0005] The role of the immune system, and in particular T cell-mediated cytotoxicity, in tumor control is well established. There is increasing evidence that T cells control tumor growth and survival in cancer patients, both in the early and late stages of the disease. However, tumor-specific T cell responses are difficult to assemble and maintain in cancer patients.

[0006] Two T cell pathways that have received significant attention to date signal through cytotoxic T lymphocyte antigen-4 (CTLA-4, CD152) and programmed death ligand 1 (PD-L1, also known as B7-H1 or CD274).

[0007] CTLA4 is expressed on activated T cells and acts as a co-inhibitor to inhibit T cell responses following CD28-mediated T cell activation. CTLA4 is thought to regulate the amplitude of early activation of naive and memory T cells after TCR engagement and is part of a central inhibitory pathway that affects both anti-tumor immunity and autoimmunity. CTLA4 is expressed exclusively in T cells, and the expression of its ligands CD80 (B7.1) and CD86 (B7.2) is primarily limited to antigen presenting cells, T cells, and other immune-mediated cells. Antagonistic anti-CTLA4 antibodies that 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, was tested in a Phase III clinical trial for the treatment of advanced melanoma, but did not significantly increase overall survival of patients compared to the then-standard treatment (temozolomide or dacarbazine).

[0008] PD-L1 is also part of a complex system of receptors and ligands that control T-cell activation. In normal tissues, PD-L1 is expressed on T cells, B cells, dendritic cells, macrophages, mesenchymal stem cells, bone marrow-derived mast cells, and different 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 by tumors and acts on multiple sites to help tumors avoid detection and elimination by the host immune system. PD-L1 is expressed at high frequencies in a wide range of cancers. In some cancers, the expression of PD-L1 is associated with reduced survival and unfavorable prognosis. Antibodies that block the interaction between B7-H1 and its receptor can relieve PD-L1-dependent immunosuppression in vitro and enhance the cytotoxic activity of anti-tumor T cells. MEDI4736 is a human monoclonal antibody against human PD-L1 that is capable of blocking the binding of PD-L1 to both PD-1 and CD80 receptors.

[0009] Despite significant progress in developing strategies for combating cancer and other diseases over the past decade, patients with advanced, refractory, and metastatic disease have limited clinical options. Chemotherapy, radiation, and high-dose chemotherapy have become dose-limiting. There remains a large unmet need for new less toxic approaches and therapeutic agents with better therapeutic efficacy, longer clinical benefit, and improved safety profiles, particularly for those patients with advanced disease or cancer that are resistant to existing therapeutic agents. Summary of the invention

[0010] The present invention provides a method for treating non-small cell lung cancer (NSCLC) in a human patient, the method 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.

[0011] The present invention also provides a method for treating NSCLC in a human patient, the method 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.

[0012] The present invention also provides a method for treating NSCLC in a human patient, the method 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.

[0013] The present invention also provides a method for treating NSCLC in a human patient, the method 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.

[0014] The present invention also provides a method for treating NSCLC in a human patient, the method 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.

[0015] The present invention also provides a method for treating NSCLC in a human patient, the method 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.

[0016] The present invention also provides a method for treating NSCLC in a human patient, the method 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.

[0017] The present invention also provides a method for treating NSCLC in a human patient, the method 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.

[0018] The present invention also provides a method for treating NSCLC in a human patient, the method 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.

[0019] The present invention also provides a method for treating NSCLC in a human patient, the method 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.

[0020] The present invention also provides a method for treating NSCLC in a human patient, the method 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.

[0021] The present invention also provides a method for treating NSCLC in a human patient, the method 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.

[0022] The present invention also provides a method for treating NSCLC in a human patient, the method 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.

[0023] The present invention also provides a method for treating NSCLC in a human patient, the method 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.

[0024] The present invention also provides a method for treating NSCLC in a human patient, the method 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.

[0025] The present invention also provides a method of treatment, comprising administering MEDI4736 or an antigen-binding fragment thereof (e.g., 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 (e.g., at 1 mg / kg, 3 mg / kg, 10 mg / kg) to a patient identified as having PD-L1 - or PD-L1 + Patients with NSCLC.

[0026] In a specific embodiment of any of the above methods, MEDI4736 is administered once every 4 weeks. In a specific embodiment, MEDI4736 is administered once every 4 weeks for 49 weeks. In a specific embodiment of any of the above methods, MEDI4736 is administered once every 2 weeks. In another embodiment, a total of 13 doses of MEDI4736 are administered.

[0027] In specific embodiments of the above aspects, tremelimumab is administered once every 4 weeks for the first 21 weeks. In other embodiments of the above aspects, tremelimumab is administered once every 12 weeks from weeks 25 to 49. In other embodiments of the above aspects, a total of 9 doses of tremelimumab are administered.

[0028] The present invention also provides a method for treating NSCLC in a human patient, the method comprising administering 10 mg / kg of MEDI4736 or an antigen-binding fragment thereof and 1 mg / kg of tremelimumab or an antigen-binding fragment thereof to the patient, wherein MEDI4736 is administered once every 2 weeks.

[0029] The present invention also provides a method for treating NSCLC in a human patient, the method comprising administering 10 mg / kg of MEDI4736 or an antigen-binding fragment thereof and 3 mg / kg of tremelimumab or an antigen-binding fragment thereof to the patient, wherein MEDI4736 is administered once every 2 weeks.

[0030] The present invention also provides a method for treating NSCLC in a human patient, the method comprising administering 10 mg / kg of MEDI4736 or an antigen-binding fragment thereof and 10 mg / kg of tremelimumab or an antigen-binding fragment thereof to the patient, wherein MEDI4736 is administered once every 2 weeks.

[0031] In specific embodiments of the above aspects, tremelimumab is administered once every 4 weeks for the first 25 weeks. In other embodiments of the above aspects, tremelimumab is administered once every 12 weeks from week 25 to week 49. In other embodiments of the above aspects, a total of 9 doses of tremelimumab are administered.

[0032] The present invention also provides a method of treating NSCLC in a human patient, the method comprising administering MEDI4736 or an antigen-binding fragment thereof and tremelimumab or an antigen-binding fragment thereof to the patient, wherein these administrations result in a tumor response.

[0033] In the methods of the present invention, such administration results in an increased tumor response compared to administration of either MEDI4736 or an antigen-binding fragment thereof alone or tremelimumab or an antigen-binding fragment thereof alone.

[0034] In some embodiments, a tumor response can be detected at week 2, 4, 6, 8, or 10. In other embodiments, a tumor response can be detected at week 33. In other embodiments, a tumor response can be detected at week 50.

[0035] The present invention also provides a method of treating NSCLC in a human patient, the method comprising administering MEDI4736 or an antigen-binding fragment thereof and tremelimumab or an antigen-binding fragment thereof to the patient, wherein these administrations increase progression-free survival. In some embodiments, these administrations result in an increase in progression-free survival compared to administration of MEDI4736 or an antigen-binding fragment thereof alone or tremelimumab or an antigen-binding fragment thereof alone.

[0036] The present invention also provides a method of treating NSCLC in a human patient, the method comprising administering MEDI4736 or an antigen-binding fragment thereof and tremelimumab or an antigen-binding fragment thereof to the patient, wherein these administrations increase overall survival. In some embodiments, these administrations result in an increase in overall survival compared to administration of MEDI4736 or an antigen-binding fragment thereof alone or tremelimumab or an antigen-binding fragment thereof alone.

[0037] In some embodiments of the methods of the invention, these administrations result in a tumor response. In some embodiments, these administrations result in an increase in tumor response compared to administration of MEDI4736 or an antigen-binding fragment thereof alone or tremelimumab or an antigen-binding fragment thereof alone. In some embodiments, a tumor response can be detected at week 8. In some embodiments, a tumor response can be detected at week 33.

[0038] In some embodiments of the methods of the invention, these administrations increase progression-free survival. In some embodiments, these administrations result in an increase in progression-free survival compared to administration of MEDI4736 or an antigen-binding fragment thereof alone or tremelimumab or an antigen-binding fragment thereof alone.

[0039] In some embodiments of the methods of the invention, these administrations increase overall survival. In some embodiments, these administrations result in an increase in overall survival compared to administration of MEDI4736 or an antigen-binding fragment thereof alone or tremelimumab or an antigen-binding fragment thereof alone.

[0040] In a specific embodiment of any of the above methods, MEDI4736 is administered once every 4 weeks. In a specific embodiment, MEDI4736 is administered once every 4 weeks for 49 weeks. In another embodiment, a total of 13 doses of MEDI4736 are administered.

[0041] In specific embodiments of the above aspects, tremelimumab is administered once every 4 weeks for the first 21 weeks. In other embodiments of the above aspects, tremelimumab is administered once every 12 weeks from week 25 to 49 or after the first 6 doses. In other embodiments of the above aspects, a total of 9 doses of tremelimumab are administered.

[0042] In some embodiments of the methods of the invention, tremelimumab or an antigen-binding fragment thereof is administered once every 4 weeks for 7 times and then once every 12 weeks.

[0043] In some embodiments of the methods of the invention, these administrations reduce soluble PD-L1. In some embodiments, soluble PD-L1 is reduced by at least about 65%, 80%, 90%, 95%, or 99%.

[0044] In some embodiments of the methods of the invention, the tumor is refractory to at least one chemotherapeutic agent. Such chemotherapeutic agents may include, but are not limited to, vemurafenib, erlotinib, afatinib, cetuximab, carboplatin, bevacizumab, erlotinib, gefitinib, or pemetrexed.

[0045] In some embodiments of the methods of the invention, the patient has an Eastern Cooperative Oncology Group (ECOG) performance status of 0-1.

[0046] In some embodiments of the methods of the invention, the patient is immunotherapy naive prior to administration of MEDI4736 or an antigen-binding fragment thereof and tremelimumab or an antigen-binding fragment thereof.

[0047] In some embodiments of the methods of the invention, the patient has received tumor therapy prior to administration of MEDI4736 or an antigen-binding fragment thereof and tremelimumab or an antigen-binding fragment thereof.

[0048] In some embodiments of the methods of the invention, administration of MEDI4736 or an antigen-binding fragment thereof is by intravenous infusion.

[0049] In some embodiments of the methods of the invention, administration of tremelimumab or an antigen-binding fragment thereof is by intravenous infusion.

[0050] In specific embodiments of the methods of the invention, such administrations reduce tumor size by at least about 10%, 25%, 50%, 75%, or 100% relative to baseline.

[0051] In a specific embodiment of the method of the present invention, the human patient has locally advanced unresectable or metastatic NSCLC. In a specific embodiment of the method of the present invention, the NSCLC is squamous or non-squamous. In other embodiments of the method of the present invention, the NSCLC comprises a KRAS mutation or an EGFR mutation.

[0052] In some embodiments of the methods of the invention, the MEDI4736 or an antigen-binding fragment thereof is MEDI4736, and wherein the tremelimumab or an antigen-binding fragment thereof is tremelimumab.

[0053] In another aspect, the invention provides a method of treating cancer in which MEDI4736 or an antigen-binding fragment of MEDI4736 is combined with tremelimumab or an antigen-binding fragment of tremelimumab. In some embodiments of the invention, the cancer is selected from prostate cancer, breast cancer, triple-negative breast cancer, colon cancer, lung cancer, non-small cell lung cancer, head and neck cancer, melanoma, gastric cancer, pancreatic cancer, ovarian cancer, renal cell carcinoma, and liver cancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0054] Figure 1 Treatment schedule is shown. MTD = maximum tolerated dose.

[0055] Figure 2 The dose escalation and dose expansion plans are shown.

[0056] Figure 3 The dose escalation study design is shown. DLT = dose limiting toxicity.

[0057] Figure 4 Figures and tables showing the pharmacokinetic (PK) profile of MEDI4736 are provided. Q4W = once every 4 weeks. LLOQ = lower limit of quantitation. AUC = area under the curve.

[0058] Figure 5 Graphs and tables showing the pharmacokinetic (PK) profile of tremelimumab are provided.

[0059] Figure 6 It is a graph showing the absolute concentration curve of soluble PD-L1 (sPD-L1).

[0060] Figure 7 is a graph showing the inhibition percentage of soluble PD-L1.

[0061] Figure 8 is a graph showing the pharmacokinetic (PK) parameters of MEDI4736 in individual patients.

[0062] Fig. 9 is a graph showing the pharmacokinetic (PK) parameters of tremelimumab in individual patients.

[0063] Fig. 10A and 10B is a table showing dose modifications of MEDI4736 and tremelimumab due to grade 1 or 2 toxicity (10A) and grade 3 or 4 toxicity (10B).

[0064] Fig.11 is a table showing the research criteria.

[0065] Fig.12 Study enrollment is shown.

[0066] Fig.13 Relevant adverse events (AEs) for dosed patients are shown.

[0067] Fig.14 Shown are the changes in tumor size from baseline in the first four cohorts.

[0068] Fig.15 The MEDI4736 and tremelimumab doses and schedules studied are shown.

[0069] Fig.16 Inhibition of sPD-L1 is shown in individual patients receiving the combination of MEDI4736 and tremelimumab.

[0070] Fig.17 Inhibition of sPD-L1 is shown in individual patients grouped by cohort who received the combination of MEDI4736 and tremelimumab.

[0071] Fig.18 Shown are increases from baseline in CD4 Ki67+ and CD8 Ki67 proliferative cells in subjects receiving MEDI4736 and tremelimumab compared to increases in those subjects receiving MEDI4736 alone. Increases from baseline in CD4 Ki67+ and CD8 Ki67 proliferative cells were associated with increasing doses of tremelimumab.

[0072] Fig.19 Shown are increases from baseline in CD4 ICOS+ and CD4 HLADR+ cells in subjects receiving MEDI4736 and tremelimumab compared to increases in those subjects receiving MEDI4736 alone. Increases from baseline in CD4 ICOS+ and CD4 HLADR+ cells were associated with increasing doses of tremelimumab.

[0073] Fig. 20 Shown are increases from baseline in CD4+ T effector cells in subjects receiving MEDI4736 and tremelimumab compared to increases in those subjects receiving MEDI4736 alone. Increases from baseline in CD4+ T effector cells were associated with increasing doses of tremelimumab.

[0074] Fig.21 Shown is an increase in Treg cells from baseline in subjects receiving MEDI4736 and tremelimumab compared to increases in those subjects receiving MEDI4736 alone. The increase in Treg cells from baseline was associated with increasing doses of tremelimumab.

[0075] Fig. 22 is a table showing the clinical activity of MEDI4736 combined with tremelimumab therapy compared to MEDI4736 monotherapy.

[0076] Fig.23 is a spider plot showing the change in tumor size from baseline in cohorts anchored by tremelimumab dose: 1 mg / kg (upper left panel); 10 mg / kg (upper right panel); and 3 mg / kg (lower panel).

[0077] Fig.24 is a graph showing the time to onset of relevant grade ≥3 adverse events (AEs) in combination cohorts anchored by tremelimumab dose (1 mg / kg; 10 mg / kg; and 3 mg / kg) compared to MEDI4736 monotherapy.

[0078] Fig.25 is a spider plot showing the change in tumor size from baseline in cohorts anchored by MEDI4736 dose: 10 mg / kg Q4W (upper left panel); 15 mg / kg Q4W (upper right panel); 20 mg / kg Q4W (lower left panel); and 10 mg / kg Q2W (lower right panel).

[0079] Fig.26 is a graph showing the time to onset of relevant grade ≥3 adverse events (AEs) in combination cohorts anchored by MEDI4736 dose (1 mg / kg; 10 mg / kg; and 3 mg / kg) compared to MEDI4736 monotherapy.

[0080] Fig. 27 is a spider plot showing the change in tumor size from baseline in a selective cohort of NSCLC patients receiving MEDI4736 and tremelimumab (left panel) compared to those receiving MEDI4736 alone (10 mg / kg Q4W) (right panel).

[0081] Fig.28 is a graph showing the time to onset of relevant grade ≥3 adverse events (AEs) in select combination cohorts (left and right panels) compared to MEDI4736 monotherapy (left panel).

[0082] Fig.29 is a spider plot showing the change in tumor size from baseline in all study cohorts of patients with NSCLC who received MEDI4736 and tremelimumab.

[0083] Fig.30 is a spider plot showing the change in tumor size from baseline in a selective cohort of PD-L1 -negative NSCLC patients receiving MEDI4736 and tremelimumab (left panel) compared to those receiving MEDI4736 (10 mg / kg; CP1108) alone (right panel).

[0084] Fig.31 is a spider plot showing the change in tumor size from baseline in a selective cohort of PD-L1 -positive NSCLC patients receiving MEDI4736 and tremelimumab (left panel) compared to those receiving MEDI4736 (10 mg / kg; CP1108) alone (right panel).

[0085] Fig.32 Figure 2 is a spider plot anchored by tremelimumab dose (1 mg / kg (left panel), 3 mg / kg (middle panel), and 10 mg / kg (right panel)) showing the change in tumor size from baseline in a selective cohort of PD-L1-negative NSCLC patients receiving MEDI4736 and tremelimumab.

[0086] Fig.33 Figure 2 is a spider plot anchored by MEDI4736 dose (10 mg / kg Q4W (upper left panel), 15 mg / kg (upper right panel), and 20 mg / kg (lower left panel)) showing the change in tumor size from baseline in a selective cohort of PD-L1-negative NSCLC patients receiving MEDI4736 and tremelimumab.

[0087] Figures 34A-34D The results were shown in the NSCLC grouped by PD-L1 status. Fig.29 Spider plot of change in tumor size from baseline in NSCLC patients receiving MEDI4736 and tremelimumab: All NSCLC patients (34A); patients identified as having PD-L1 - Patients with NSCLC (34B); identified as having PD-L1 + patients with NSCLC (34C); and patients with NSCLC whose PD-L1 status is unknown (34D).

[0088] Fig.35 is a waterfall plot showing the best change in tumor size from baseline in NSCLC patients receiving MEDI4736 and tremelimumab.

[0089] Fig.36 Is shown in the identification of NSCLC according to PD-L1 status Fig.35 Waterfall plot of the best change in tumor size from baseline in NSCLC patients receiving MEDI4736 and tremelimumab.

[0090] Fig.37 is a lane plot showing the duration of follow-up in NSCLC patients receiving MEDI4736 and tremelimumab.

[0091] Detailed description

[0092] It should be noted that the term "a or an" entity refers to one or more of the entity; for example, "an antibody" should be understood to represent one or more antibodies. Therefore, the terms "a" (or "a kind"), "one or more", and "at least one" can be used interchangeably herein.

[0093] Provided herein are methods for treating non-small cell lung cancer (NSCLC) using MEDI4736 and tremelimumab. There are three major subtypes of NSCLC: squamous cell carcinoma, adenocarcinoma, and large cell (undifferentiated) carcinoma. Other subtypes include adenosquamous carcinoma and sarcomatoid carcinoma. NSCLC may include mutations in KRAS or epidermal growth factor receptor. Such mutations are known in the art and described in, for example, Riley et al., Proc Am Thorac Soc. 2009 Apr 15; 6(2): 201-5, which is incorporated herein by reference.

[0094] In addition, provided herein are methods of using MEDI4736 and tremelimumab to treat cancer or tumor proliferation. "Cancer" as used herein 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, non-small cell lung cancer, head and neck cancer, melanoma, gastric cancer, pancreatic cancer, ovarian cancer, renal cell carcinoma, and liver cancer.

[0095] The methods provided include administering effective amounts of MEDI4736 or an antigen-binding fragment thereof and tremelimumab or an antigen-binding fragment thereof.

[0096] “MEDI4736” means an antibody or antigen-binding fragment thereof that selectively binds to a PD-L1 polypeptide and comprises at least a portion of a light chain variable region and / or at least a portion of a heavy chain variable region, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 1 and the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 2.

[0097] Information about MEDI4736 (or its antigen-binding fragment) used in the methods provided herein can be found in U.S. Pat. No. 8,779,108, the disclosure of which is incorporated herein by reference in its entirety. The fragment crystallizable (Fc) domain of MEDI4736 contains a triple mutation in the constant domain of the IgG1 heavy chain, which reduces 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 the binding of PD-L1 to PD-1 and CD80 receptors. MEDI4736 can relieve PD-L1-mediated inhibition of human T-cell activation in vitro and inhibit tumor growth in xenograft models through a T-cell-dependent mechanism.

[0098] MEDI4736 used 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 its antigen-binding fragment used in the methods provided herein comprises a light chain variable region and a heavy chain variable region, the light chain variable region comprises an amino acid sequence of SEQ ID NO: 1, and the heavy chain variable region comprises an amino acid sequence of SEQ ID NO: 2. In a specific aspect, MEDI4736 or its antigen-binding fragment used in the methods provided herein comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises a CDR1, CDR2 and CDR3 sequence defined by Kabat of SEQ ID NO: 3-5, and wherein the light chain variable region comprises a CDR1, CDR2 and CDR3 sequence defined by Kabat of SEQ ID NO: 6-8. Those skilled in the art will be able to easily identify Chothia's definition, Abm's definition 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 variable heavy chain and variable light chain CDR sequences of the 2.14H90PT antibody as disclosed in US Pat. No. 8,779,108, which is incorporated herein by reference in its entirety.

[0099] "Tremelimumab" means an antibody or antigen-binding fragment thereof that selectively binds to a CTLA4 polypeptide and comprises at least a portion of a light chain variable region comprising the amino acid sequence of SEQ ID NO: 9 and / or at least a portion of a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 10. Exemplary anti-CTLA4 antibodies are described, for example, in U.S. 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 (wherein Tremelimumab is 11.2.1), which are incorporated herein by reference. Tremelimumab is an exemplary anti-CTLA4 antibody. The Tremelimumab sequence is provided in the sequence listing below.

[0100] Information about tremelimumab (or an antigen-binding fragment thereof) used in the methods provided herein can be found in US6,682,736 (where it is referred to as 11.2.1), the disclosure of which is incorporated herein by reference in its entirety. 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 the binding of CTLA4 to CD80 (B7.1) and CD86 (B7.2). It has been shown to cause immune activation in vitro, and some patients treated with tremelimumab have shown tumor regression.

[0101] Tremelimumab and its antigen-binding fragment used in these methods provided herein include heavy chain and light chain or heavy chain variable region and light chain variable region. In a specific aspect, tremelimumab or its antigen-binding fragment used in these methods provided herein include light chain variable region and heavy chain variable region, the light chain variable region includes SEQ ID NO:9 amino acid sequence, the heavy chain variable region includes SEQ ID NO:10 amino acid sequence. In a specific aspect, tremelimumab or its antigen-binding fragment used in these methods provided herein include heavy chain variable region and light chain variable region, wherein the heavy chain variable region includes SEQ ID NO:11-13 Kabat (Kabat) defined CDR1, CDR2 and CDR3 sequence, and wherein the light chain variable region includes SEQ ID NO:14-16 Kabat defined CDR1, CDR2 and CDR3 sequence. Those skilled in the art will be able to easily identify Chothia definition, Abm definition 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 variable heavy chain and variable light chain CDR sequences of the 11.2.1 antibody as disclosed in US Pat. No. 6,682,736, which is incorporated herein by reference in its entirety.

[0102] The term "antigen-binding fragment" refers to a portion of an intact antibody and / or refers to the antigen-determining variable region of an intact antibody. It is known that the antigen-binding function of an antibody can be performed by a fragment 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.

[0103] In certain aspects, patients presenting with NSCLC are given MEDI4736 or its antigen binding fragment and tremelimumab or its antigen binding fragment. MEDI4736 or its antigen binding fragment and tremelimumab or its antigen binding fragment can be given only once or occasionally, and still provide benefits to the patient. In other aspects, the patient is given additional subsequent doses. Subsequent doses may be given at different time intervals depending on the patient's age, weight, clinical evaluation, tumor load and / or other factors (including the judgment of the attending physician).

[0104] The interval between doses of MEDI4736 or an antigen-binding fragment thereof may be once every 4 weeks. The interval between doses of tremelimumab or an antigen-binding fragment thereof may be once every 4 weeks. The interval between doses of tremelimumab or an antigen-binding fragment thereof may be once every 12 weeks. The interval between doses of tremelimumab or an antigen-binding fragment thereof may be once every 4 weeks for 6 cycles, then once every 12 weeks.

[0105] In certain aspects, MEDI4736 or an antigen-binding fragment thereof is administered about as frequently as tremelimumab or an antigen-binding fragment thereof. In certain aspects, MEDI4736 or an antigen-binding fragment thereof is administered about 3 times as frequently as tremelimumab or an antigen-binding fragment thereof.

[0106] In some embodiments, at least two doses of MEDI4736 or an antigen-binding fragment thereof and tremelimumab or an antigen-binding fragment thereof are administered to the patient. In some embodiments, 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 may be administered to the patient. In some embodiments, MEDI4736 or an antigen-binding fragment thereof is administered within a four-week treatment cycle, within an eight-week treatment cycle, within a sixteen-week treatment cycle, within a twenty-week treatment cycle, within a twenty-four-week treatment cycle, or within a year or longer treatment cycle. In some embodiments, tremelimumab or an antigen-binding fragment thereof is administered within a four-week treatment cycle, within an eight-week treatment cycle, within a twelve-week treatment cycle, within a sixteen-week treatment cycle, within a twelve-week treatment cycle, within a twenty-four-week treatment cycle, within a thirty-six-week treatment cycle, within a forty-eight-week treatment cycle, or within a year or longer treatment cycle.

[0107] 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 and tremelimumab or an antigen-binding fragment thereof are administered simultaneously. In other embodiments, MEDI4736 or an antigen-binding fragment thereof is administered about 1 hour after tremelimumab or an antigen-binding fragment thereof is administered.

[0108] 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 the patient will depend on various parameters, such as the patient's age, weight, clinical evaluation, tumor burden and / or other factors, including the judgment of the attending physician.

[0109] In certain aspects, the patient is given one or more doses of MEDI4736 or its antigen binding fragment, wherein the dose is about 1 mg / kg. In certain aspects, the patient is given one or more doses of MEDI4736 or its antigen binding fragment, wherein the dose is about 3 mg / kg. In certain aspects, the patient is given one or more doses of MEDI4736 or its antigen binding fragment, wherein the dose is about 10 mg / kg. In certain aspects, the patient is given one or more doses of MEDI4736 or its antigen binding fragment, wherein the dose is about 15 mg / kg. In certain aspects, the patient is given one or more doses of MEDI4736 or its antigen binding fragment, wherein the dose is about 20 mg / kg.

[0110] In certain aspects, the patient is given at least two doses of MEDI4736 or its antigen binding fragment, wherein the dose is about 1 mg / kg. In certain aspects, the patient is given at least two doses of MEDI4736 or its antigen binding fragment, wherein the dose is about 3 mg / kg. In certain aspects, the patient is given at least two doses of MEDI4736 or its antigen binding fragment, wherein the dose is about 10 mg / kg. In certain aspects, the patient is given at least two doses of MEDI4736 or its antigen binding fragment, wherein the dose is about 15 mg / kg. In certain aspects, the patient is given at least two doses of MEDI4736 or its antigen binding fragment, wherein the dose is about 20 mg / kg. In some embodiments, the at least two doses are given about four weeks apart.

[0111] In certain aspects, the patient is given at least three doses of MEDI4736 or its antigen binding fragment, wherein the dose is about 1 mg / kg. In certain aspects, the patient is given at least three doses of MEDI4736 or its antigen binding fragment, wherein the dose is about 3 mg / kg. In certain aspects, the patient is given at least three doses of MEDI4736 or its antigen binding fragment, wherein the dose is about 10 mg / kg. In certain aspects, the patient is given at least three doses of MEDI4736 or its antigen binding fragment, wherein the dose is about 15 mg / kg. In certain aspects, the patient is given at least three doses of MEDI4736 or its antigen binding fragment, wherein the dose is about 20 mg / kg. In some embodiments, the at least three doses are given about four weeks apart.

[0112] In certain aspects, the patient is administered one or more doses of tremelimumab or an antigen-binding fragment thereof, wherein the dose is about 1 mg / kg. In certain aspects, the patient is administered one or more doses of tremelimumab or an antigen-binding fragment thereof, wherein the dose is about 3 mg / kg. In certain aspects, the patient is administered one or more doses of tremelimumab or an antigen-binding fragment thereof, wherein the dose is about 10 mg / kg.

[0113] In certain aspects, the patient is given at least two doses of tremelimumab or an antigen-binding fragment thereof, wherein the dose is about 1 mg / kg. In certain aspects, the patient is given at least two doses of tremelimumab or an antigen-binding fragment thereof, wherein the dose is about 3 mg / kg. In certain aspects, the patient is given at least two doses of tremelimumab or an antigen-binding fragment thereof, wherein the dose is about 10 mg / kg. In some embodiments, the at least two doses are given about four weeks apart. In some embodiments, the at least two doses are given about twelve weeks apart.

[0114] In certain aspects, the patient is given at least three doses of tremelimumab or its antigen-binding fragment, wherein the dose is about 1 mg / kg. In certain aspects, the patient is given at least three doses of tremelimumab or its antigen-binding fragment, wherein the dose is about 3 mg / kg. In certain aspects, the patient is given at least three doses of tremelimumab or its antigen-binding fragment, wherein the dose is about 10 mg / kg. In some embodiments, the at least three doses are given about four weeks apart. In some embodiments, the at least three doses are given about twelve weeks apart.

[0115] In certain aspects, administration of MEDI4736 or an antigen-binding fragment thereof and / or tremelimumab or an antigen-binding fragment thereof according to the methods provided herein is by parenteral administration. For example, MEDI4736 or an antigen-binding fragment thereof and / or tremelimumab or an antigen-binding fragment thereof can be administered by intravenous infusion or by subcutaneous injection. In some embodiments, the administration is by intravenous infusion.

[0116] In certain aspects, the 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.

[0117] 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.

[0118] In certain aspects, the 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.

[0119] In certain aspects, the 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.

[0120] In certain aspects, the 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.

[0121] These methods provided herein can weaken, delay or stabilize tumor growth.In some aspects, this reduction or delay can be statistically significant.The minimization of tumor growth can be measured by comparing with the growth of the patient's tumor at baseline, for the tumor growth expected, for the tumor growth expected based on a large patient population or for the tumor growth of a control population.In some aspects, tumor response is measured using solid tumor response evaluation criteria (RECIST).

[0122] In some aspects, a tumor response is detectable at week 8. In some aspects, a tumor response is detectable at week 33. In some aspects, a tumor response is detectable at week 50.

[0123] In certain aspects, a tumor response is detectable after administration of MEDI4736, or an antigen-binding fragment thereof, and tremelimumab, or an antigen-binding fragment thereof. In certain aspects, a tumor response is detectable after administration of 8 doses of MEDI4736, or an antigen-binding fragment thereof, and 7 doses of tremelimumab, or an antigen-binding fragment thereof. In certain aspects, a tumor response is detectable after administration of 13 doses of MEDI4736, or an antigen-binding fragment thereof, and 9 doses of tremelimumab, or an antigen-binding fragment thereof.

[0124] In certain aspects, the patient achieves disease control (DC).Disease control can be a complete response (CR), a partial response (PR), or stable disease (SD).

[0125] "Complete response" (CR), "partial response" (PR), and "stable disease" (SD) may be determined as defined in Table 1 below.

[0126] In certain aspects, administration of MEDI4736 or an antigen-binding fragment thereof can increase progression-free survival (PFS).

[0127] In certain aspects, administration of MEDI4736 or an antigen-binding fragment thereof can increase overall survival (OS).

[0128] In some embodiments, the patient has previously received treatment with at least one chemotherapeutic agent. In some embodiments, the patient has previously received treatment with at least two chemotherapeutic agents. The chemotherapeutic agent can be, for example and without limitation, vemurafenib, erlotinib, afatinib, cetuximab, carboplatin, bevacizumab, erlotinib, gefitinib, and / or pemetrexed.

[0129] In some embodiments, the NSCLC is refractory to at least one chemotherapeutic agent or the NSCLC is resistant to at least one chemotherapeutic agent. In some embodiments, the tumor is refractory to at least two chemotherapeutic agents or the tumor is resistant to at least two chemotherapeutic agents. The tumor may be refractory to one or more of the following or the tumor is resistant to one or more of the following: for example and not limited to vemurafenib, erlotinib, afatinib, cetuximab, carboplatin, bevacizumab, erlotinib, gefitinib, and / or pemetrexed. In some embodiments, the NSCLC is negative for PD-L1. In some embodiments, the NSCLC is positive for PD-L1.

[0130] In some embodiments, the patient has an Eastern Cooperative Oncology Group (ECOG) (Oken MM et al., Am. J. Clin. Oncol., 5:649-55 (1982)) performance status of 0 or 1 prior to administration of MEDI4736 or an antigen-binding fragment thereof and tremelimumab or an antigen-binding fragment thereof.

[0131] According to these methods provided herein, as shown in some early data, administration of MEDI4736 or its antigen-binding fragment and tremelimumab or its antigen-binding fragment can result in ideal pharmacokinetic parameters. The "area under the curve" (AUC) can be used to assess total drug exposure. "AUC (τ)" refers to the AUC of the dosing interval from time 0 to time τ, while "AUC (∞)" refers to the AUC until infinite time. Administration can produce an AUC (τ) of about 600 to about 3,000 μg / mL*day for MEDI4736 or its antigen-binding fragment and an AUC (τ) of about 250 to about 350 μg / mL*day for tremelimumab or its antigen-binding fragment. Administration can produce a maximum observed concentration (Cmax) of about 60 to about 300 μg / mL MEDI4736 or its antigen-binding fragment and about 25 to about 35 μg / mL tremelimumab or its antigen-binding fragment. Dosing can produce a Cnadir (minimum blood drug concentration) of about 5 to about 40 μg / mL MEDI4736 or an antigen-binding fragment thereof and about 4 to about 6 μg / mL tremelimumab or an antigen-binding fragment thereof.

[0132] As provided herein, MEDI4736 or its antigen-binding fragment can also reduce the level of free (soluble) PD-L1. Free (soluble) PD-L1 refers to PD-L1 that is not bound (e.g., by 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, after administration of MEDI4736 or its antigen-binding fragment and tremelimumab or its antigen-binding fragment, PD-L1 levels are undetectable.

[0133] In some embodiments, after administration of MEDI4736 or an antigen-binding fragment thereof, PD-L1 levels are reduced by at least 65%. In some embodiments, after a single administration of MEDI4736 or an antigen-binding fragment thereof, PD-L1 levels are reduced by at least 80%. In some embodiments, after a single administration of MEDI4736 or an antigen-binding fragment thereof, PD-L1 levels are reduced by at least 90%. In some embodiments, after a single administration of MEDI4736 or an antigen-binding fragment thereof, PD-L1 levels are reduced by at least 95%. In some embodiments, after a single administration of MEDI4736 or an antigen-binding fragment thereof, PD-L1 levels are reduced by at least 99%. In some embodiments, after a single administration of MEDI4736 or an antigen-binding fragment thereof, PD-L1 levels are undetectable.

[0134] In some embodiments, after two doses of MEDI4736 or an antigen-binding fragment thereof are administered, PD-L1 levels are reduced by at least 65%. In some embodiments, after two doses of MEDI4736 or an antigen-binding fragment thereof are administered, PD-L1 levels are reduced by at least 80%. In some embodiments, after two doses of MEDI4736 or an antigen-binding fragment thereof are administered, PD-L1 levels are reduced by at least 90%. In some embodiments, after two doses of MEDI4736 or an antigen-binding fragment thereof are administered, PD-L1 levels are reduced by at least 95%. In some embodiments, after two doses of MEDI4736 or an antigen-binding fragment thereof are administered, PD-L1 levels are reduced by at least 99%. In some embodiments, after two doses of MEDI4736 or an antigen-binding fragment thereof are administered, PD-L1 levels are undetectable.

[0135] Treatment of patients with solid tumors using both MEDI4736 or an antigen-binding fragment thereof and tremelimumab or an antigen-binding fragment thereof (i.e., combination therapy) can result in a synergistic effect. The term "synergistic" as used herein means that the combination therapy (e.g., MEDI4736 or an antigen-binding fragment thereof combined with tremelimumab or an antigen-binding fragment thereof) is more effective than the additive effects of monotherapy.

[0136] The synergistic effect of the combination therapy (e.g., MEDI4736 or an antigen-binding fragment thereof in combination with tremelimumab or an antigen-binding fragment thereof) allows the use of lower doses of one or more of these therapeutic agents to patients with solid tumors and / or the administration of the therapeutic agent to patients with solid tumors less frequently. The ability to utilize lower doses of the therapeutic agent and / or to administer the therapy less frequently reduces the toxicity associated with administering the therapy to the subject without reducing the efficacy of the therapy in the treatment of solid tumors. In addition, the synergistic effect can result in an increase in the efficacy of the therapeutic agent in the management, treatment, or improvement of solid tumors. The synergistic effect of the combination of therapeutic agents can avoid or reduce adverse or unwanted side effects associated with the use of any single therapy.

[0137] In combination therapy, MEDI4736 or an antigen binding fragment thereof may optionally be included in the same pharmaceutical composition as tremelimumab or its antigen binding fragment, or may be included in a separate pharmaceutical composition. In the latter case, the pharmaceutical composition comprising MEDI4736 or its antigen binding fragment is suitable for administration before, simultaneously with, or after administration of a pharmaceutical composition comprising tremelimumab or its antigen binding fragment. In some cases, MEDI4736 or its antigen binding fragment is administered during a period overlapping with tremelimumab or its antigen binding fragment in a separate composition.

[0138] Subjects with lung cancer (e.g., non-small cell lung cancer) can be tested for PD-L1 polynucleotide or polypeptide expression during the selection of a treatment method. Patients identified as having tumors that are negative for PD-L1 (e.g., as defined by a Ct or IHC-M score) or patients identified by having a reduced or undetectable PD-L1 level relative to a reference level are identified as responding to a combination treatment of an anti-PD-L1 antibody and tremelimumab. Such patients are given a combination of MEDI4736 or an antigen-binding fragment thereof and tremelimumab or an antigen-binding fragment thereof. DETAILED DESCRIPTION

[0139] Examples

[0140] Example 1: Patients and Methods

[0141] (a) Subjects

[0142] Subjects in this study were required to 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 measurable lesion according to the Response Evaluation Criteria in Solid Tumors (RECIST) guidelines v1.1, which are incorporated herein by reference in its entirety.

[0143] For both the dose escalation and dose expansion phases, the cohort-specific pre-immunotherapy requirements are as follows: a) Immunotherapy-naive cohort: must have no prior exposure to immunotherapy, such as, but not limited to, other anti-CTLA-4, anti-PD-1, or anti-PD-L1 antibodies excluding vaccines; and b) Immunotherapy-pretreated cohort: must have prior exposure to immunotherapy for greater than 28 days prior to the first dose of MEDI4736 and tremelimumab, which immunotherapies are, such as, but not limited to, other anti-CTLA-4, anti-PD-1, or anti-PD-L1 antibodies excluding vaccines.

[0144] Subjects were also required to have failed to respond to standard therapy, had relapsed after standard therapy, had refused standard therapy, or were not eligible for standard therapy.Subjects had an Eastern Cooperative Oncology Group (ECOG) performance status of 0-1.

[0145] Subjects with central nervous system (CNS) metastases must be asymptomatic on study day 1. In addition, on day 1, there must be at least 28 weeks without progression of CNS metastases, as demonstrated by magnetic resonance imaging (MRI) / computed tomography (CT) after the last day of radiation therapy and at least 14 days from the last dose of corticosteroids.

[0146] Subjects were also required to have adequate organ (liver and kidney) and hematopoietic function. Adequate organ and hematopoietic function were defined as: hemoglobin ≥ 9 g / dL; absolute neutrophil count ≥ 1500 cells / mm 3 ; Platelet count ≥100,000 / mm 3 ; total bilirubin ≤1.5× upper limit of normal (ULN), unless associated with Gilbert's syndrome or liver metastases (for these subjects, baseline total bilirubin must be ≤3.0 mg / dL); alanine aminotransferase (ALT) and aspartate aminotransferase (AST) must be ≤2.5×ULN, unless associated with liver metastases (for these subjects, ALT and AST must be ≤5×ULN); and serum creatinine ≤2.0 mg / dL.

[0147] If the subject is undergoing any concurrent chemotherapy, immunotherapy, biological or hormonal therapy for cancer treatment, they cannot participate. If the subject has been subjected to any experimental anticancer treatment within 28 days before the first dose of MEDI4736 and tremelimumab, they cannot participate. If the subject has any prior ≥ grade 3 immune-related adverse events (irAE) or any unresolved irAE> grade 1 while receiving immunotherapy (including anti-CTLA-4 therapy), they cannot participate. If the subject has undergone major surgery (as defined by the investigator) within 28 days before the first dose of MEDI4736 and tremelimumab or if they are still recovering from a previous surgery, they cannot participate. In addition to the alopecia and laboratory values ​​listed in the inclusion criteria, if the subject has unresolved toxicity from previous anticancer treatment, the toxicity is defined as unresolved to grade 0 or 1 of the National Cancer Institute Common Terminology Criteria for Adverse Events (NCI CTCAE) V4.03. Subjects with irreversible toxicity that is reasonably expected not to be exacerbated by MEDI4736 and tremelimumab may be included. Subjects were also excluded if they were currently using immunosuppressive medications or had used immunosuppressive medications within 14 days prior to the first dose of MEDI4736 and tremelimumab, with the exception of intranasal and inhaled glucocorticoids or systemic corticosteroids at physiological doses not exceeding 10 mg / day of prednisone or equivalent.

[0148] Subjects were not eligible to participate if they had active or prior autoimmune disease within the past 3 years, including inflammatory bowel disease, diverticulitis, irritable bowel disease, celiac disease, Wegener's syndrome, and Hashimoto's syndrome, except for vitiligo, alopecia areata, Graves' disease, or psoriasis (within the past 3 years) not requiring systemic treatment. Subjects were also not eligible to participate if they had a history of primary immunodeficiency or tuberculosis; if subjects had known active or chronic viral hepatitis A, B, or C; if subjects had human immunodeficiency virus (HIV); other active serious illness or uncontrolled intercurrent illness; had received a live attenuated vaccine within 28 days prior to the first dose of MEDI4736 and tremelimumab; had other invasive malignancies within 5 years; or known hypersensitivity or allergy to the study drug formulations.

[0149] Subjects are also not eligible if they have advanced NSCLC with tumors harboring anaplastic lymphoma receptor tyrosine kinase (ALK) gene rearrangements or epidermal growth factor receptor (EGFR) sensitizing mutations and have not received appropriate tyrosine kinase inhibitor (TKI) therapy. These subjects may be enrolled after documented progression or intolerance to an appropriate TKI.

[0150] (b) Research design

[0151] This study is an open-label Phase 1b study of the combination of MEDI4736 and tremelimumab ( Figure 1 ). The key research criteria for the cohort are shown in Fig.11 The study will enroll approximately 36 subjects in the dose-escalation phase and the dose-expansion phase, with at least 20 subjects in each of the immunotherapy-naive and immunotherapy-pretreated cohorts. The maximum tolerated dose (MTD) determined for the immunotherapy-naive cohort in the dose-escalation phase or the highest protocol-defined dose that does not exceed the MTD for each agent will serve as the starting dose for subjects enrolled in the immunotherapy-pretreated cohort as part of the dose-escalation phase. Dose-finding for the immunotherapy-pretreated cohort will be conducted in parallel with the dose-expansion phase for the immunotherapy-naive cohort.

[0152] Dose escalation phase

[0153] Immunotherapy-naive group

[0154] In the dose-escalation phase of the study, MEDI4736 and tremelimumab will initially be administered in consecutive cohorts of 3-6 immunotherapy-naive subjects, with each subject receiving tremelimumab Q4W for 6 doses and once every 12 weeks (Q12W) for 3 doses (i.e., tremelimumab dose 7 is given 4 weeks after dose 6 and dose 8 is given 12 weeks after dose 7) and MEDI4736 Q4W for 13 doses via intravenous infusion for a total of 1 year ( Figure 2 ). According to the standard 3+3 design and the modified area-based design, each cohort will include at least 3 subjects. Subjects in the first cohort (Cohort 1) will receive a dose of 3 mg / kg MED4736 and 1 mg / kg tremelimumab. If ≥2 dose-limiting toxicities (DLTs) are observed in the first dose cohort, the starting dose will be based on Figure 1 Dose escalation will continue if no more than 1 of 6 subjects in a given dose cohort experiences a DLT.

[0155] Subsequent cohorts will receive tremelimumab (6 doses given Q4W followed by 3 doses given Q12W) and MEDI4736 (13 doses given Q4W) until the MTD or the highest protocol-defined dose that does not exceed the MTD for each agent is identified.

[0156] Once the MTD or the highest protocol-defined dose that did not exceed the MTD for each agent was established in immunotherapy-naive subjects during the dose-escalation phase, the immunotherapy-naive cohort (see Figure 1Based on the evaluation of PK and pharmacodynamics, safety and efficacy parameters in the immunotherapy-naive cohort from the dose expansion phase, one or more additional doses that do not exceed the MTD from one or more intermediate regional-based cohorts may be evaluated in the expansion phase.

[0157] Immunotherapy pretreatment group

[0158] In the dose-escalation phase, the immunotherapy-naive group (see Figure 1 ) The MTD determined by the immunotherapy pretreatment group or the highest protocol defined dose that does not exceed the MTD for each agent will be used as the starting dose for the subjects included in the immunotherapy pretreatment group as part of the dose escalation phase. The dose exploration for the immunotherapy pretreatment group using the 3+3 design will be carried out in parallel with the dose expansion phase of the immunotherapy naive group. Once the MTD is determined in the immunotherapy pretreatment subjects in the dose escalation phase or the highest protocol defined dose that does not exceed the MTD for each agent, the dose expansion phase of the immunotherapy pretreatment group can begin. Based on the evaluation of PK and pharmacodynamics, safety and efficacy parameters in the immunotherapy pretreatment group from the dose expansion phase, one or more additional doses that do not exceed the MTD from one or more intermediate regional-based groups can be evaluated in the dose expansion phase.

[0159] Dose expansion phase

[0160] Two dose expansion cohorts will be used: immune-naive and immune-pretreated (see Figure 1 ). The MTD identified in the dose escalation phase or the highest protocol-defined dose that does not exceed the MTD for each agent will be used for both cohorts in the dose expansion phase. In addition, based on the formed subject data, including safety, PK, pharmacodynamics, biomarkers, and responses from ongoing trials together with data, one or more doses from one or more intermediate region-based cohorts that do not exceed the MTD may be included as one or more additional doses for immunotherapy pretreatment and immunotherapy naive cohorts in the dose expansion phase.

[0161] Initially, approximately 20 NSCLC subjects will be enrolled in each of the immunotherapy-naive and immunotherapy-pretreated cohorts. Depending on the evolving subject data, including safety, PK, pharmacodynamics, biomarkers, and responses and data from ongoing trials, additional subjects may be enrolled in each of the immunotherapy-naive cohorts in the dose expansion phase, up to a total of 60 subjects.

[0162] Treatment options

[0163] Subjects will be treated in either the dose escalation or dose expansion phase of the study. In the dose escalation and dose expansion phase of the study, tremelimumab will be administered every four weeks (Q4W) for 6 doses, followed by 3 doses every twelve weeks (Q12W), with dose 7 being administered 4 weeks after dose 6 and dose 8 being 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 1 year ( Figure 2 ).

[0164] 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 first IV infusion of Tremelimumab will last approximately 1 hour and the first IV infusion of MEDI4736 will last approximately 4 hours; subsequent infusions for subjects in this cohort and all infusions for the remaining cohorts will last approximately 1 hour.

[0165] Subjects who achieve and maintain disease control (DC) (i.e., complete remission (CR), partial remission (PR), or stable disease (SD)) will enter follow-up until the end of the 12-month MEDI4736 and tremelimumab treatment period. Subjects may be re-administered with MEDI4736 and tremelimumab for up to an additional 12 months with the same treatment guidelines during the initial 12-month period, based on evidence of disease progression (PD) during the follow-up period, if the subject has not received other treatment for their disease and still meets the inclusion and exclusion criteria of the study protocol. Only one round of retreatment with MEDI4736 and tremelimumab is allowed.

[0166] The study utilizes a 3+3 dose escalation phase and will follow Figure 3 The regimen shown in . At least 3 subjects will be included in each dose cohort. If no patient has dose-limiting toxicity (DLT) among the 3 patients in a dose cohort, the next dose escalation cohort will be initiated. If 1 patient has DLT among the 3 patients in a dose cohort, an additional 3 patients will be included in the same cohort. If no more than 1 of the 6 subjects experiences DLT, the next dose escalation cohort will be initiated. If 2 or more patients experience DLT in a dose cohort during the evaluation period, no additional subjects will be included in that dose cohort.

[0167] The maximum tolerated dose (MTD) is defined as the highest dose of tremelimumab or MEDI4736 in the cohort in which no more than 1 patient out of 6 patients experiences a DLT. This is determined for both the immunotherapy-naive and pre-treated cohorts in the dose-escalation phase. Patients in the expansion phase (immunotherapy-naive and immunotherapy-pre-treated cohorts) will be treated at the MTD level determined in the dose-escalation phase.

[0168] Because as in Fig. 10A and 10B MEDI4736 and tremelimumab dosing may be modified or discontinued for toxicities described in Table 1. Dose modifications are not necessary for adverse events (AEs) not clearly attributable to MEDI4736 or tremelimumab (e.g., unexpected events) or for laboratory abnormalities that are not considered clinically significant.

[0169] (c) Pharmacokinetics, anti-tumor, biomarkers, soluble factors, and safety assessment

[0170] Determination of MEDI4736 and tremelimumab concentrations in serum will be performed using validated immunoassays.

[0171] Blood samples for pharmacokinetic assessments will be collected on the following days during the dose escalation phase: Day 1 (before dosing and end of infusion), Day 8 (±1 day), Day 15 (±1 day), Day 29 (before dosing and end of infusion; ±3 days), Day 57 (before dosing and end of infusion; ±3 days), Day 85 (before dosing and end of infusion; ±3 days), Day 113 (before dosing and end of infusion; ±3 days), Day 141 (before dosing and end of infusion; ±3 days), Day 169 (before dosing and end of infusion; ±3 days) ), Day 197 (prior to dosing and end of infusion; ±3 days; MEDI4736 only), Day 225 (prior to dosing and end of infusion; ±3 days; MEDI4736 only), Day 253 (prior to dosing and end of infusion; ±3 days), Day 281 (prior to dosing and end of infusion; ±3 days; MEDI4736 only), Day 309 (prior to dosing and end of infusion; ±3 days), Day 337 (prior to dosing and end of infusion; ±3 days), at the end of treatment, 60 days after the end of treatment, and 90 days after the end of treatment.

[0172] Blood samples for pharmacokinetic assessments will be collected on the following days during the dose expansion 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.

[0173] The presence of anti-drug antibodies (ADA) will be assessed on the following days: Day 1 and Day 29 (±3 days), Day 85 (±3 days), Day 141 (±3 days), Day 169 (±3 days), Day 253 (±3 days), and Day 337 (±3 days), at the end of treatment, 60 days after the end of treatment, and 90 days after the end of treatment. Validated electrochemiluminescent assays using the mesoscale discovery platform will be used for the determination of anti-MEDI4736 antibodies in human serum and for the determination of anti-tremelimumab antibodies in human serum.

[0174] 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 the following days: Day 1, Day 8 (±1 day), Day 15 (±1 day), 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), Day 225 (±3 days), Day 253 (±3 days), Day 281 (±3 days), Day 309 (±3 days), Day 337 (±3 days), at the end of treatment, 60 days after the end of treatment, and 90 days after the end of treatment. During the dose-expansion phase, sPD-L1 levels will be assessed on the following days: Day 1, 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), Day 225 (±3 days), Day 253 (±3 days), Day 281 (±3 days), Day 309 (±3 days), Day 337 (±3 days), at the end of treatment, 60 days after the end of treatment, and 90 days after the end of treatment.

[0175] Tumor assessments will be performed during screening (Day -28 to Day -1), at Week 8 (Day 50 ± 3 days), at Week 33 (Day 225 ± 3 days), and during the dose escalation phase. Tumor evaluation will be based on modified RECIST criteria v1.1 and include the following assessments: physical examination (with photographs and measurements of skin lesions if applicable), CT or MRI scans of the chest, abdomen, and pelvis, and CT or MRI scans of the brain. Computed tomography or MRI scans of the brain will be performed only at screening.

[0176] Evaluation of antitumor activity was based on measurement of tumor lesions, assessment of target lesions, assessment of non-target lesions, and the appearance of new lesions.

[0177] Target lesion assessment

[0178] For target lesion assessment, complete response was defined as disappearance of all target lesions. Any pathological lymph node (whether target or non-target) must be reduced to <10 mm in the short axis (the sum may not be "0" if there are target nodes).

[0179] For target lesion assessment, partial response was defined as a reduction of at least 30% in the sum of diameters of target lesions, with the baseline sum diameter as reference.

[0180] For the assessment of target lesions, disease progression was defined as an increase of at least 20% in the sum of diameters of target lesions, taking the smallest sum on study as reference (this included the baseline sum if it was the smallest sum on study). In addition to the 20% relative increase, the sum must also show an absolute increase of at least 5 mm. The appearance of one or more new lesions was also considered progression.

[0181] For target lesion assessment, stable disease (SD) was neither sufficient shrinkage to qualify as PR nor sufficient increase to qualify as PD, with the smallest sum of diameters used as reference in this study.

[0182] Non-target lesion assessment

[0183] For the evaluation of non-target lesions, complete response (CR) was defined as the disappearance of all non-target lesions. All lymph nodes must be <10 mm in the short axis.

[0184] For the assessment of non-target lesions, non-complete response / non-progressive disease was defined as the persistence of one or more non-target lesions and / or tumor marker levels remaining above the normal range.

[0185] For the assessment of nontarget lesions, progressive disease was defined as an overall level of substantial worsening of nontarget disease such that the overall tumor burden had increased sufficiently to warrant discontinuation of treatment, even in the presence of SD or PR in target disease. In the absence of measurable disease, the magnitude of the change in nonmeasurable disease was approximately comparable to the increase in measurable disease that would require a declaration of PD.

[0186] New lesions

[0187] The appearance of new lesions is considered PD according to RECIST guidelines v1.1. Given the unique response kinetics observed for immunotherapy, new lesions may not represent true disease progression. In the absence of rapid clinical worsening, subjects can continue to receive treatment with MEDI4736 and tremelimumab.

[0188] Overall Response

[0189] Confirmation of CR, PR, together with PD, may be required by repeated serial assessments no less than four weeks from the date of first documentation.As in Table 1 below, the assessment of overall response based on the assessment of target lesions, non-target lesions, and new lesions is determined.

[0190] Table 1: Assessment of overall response

[0191]

[0192]

[0193] a Defined as non-target lesions at baseline.

[0194] b Non-evaluable was defined as no lesion measurements or only a small portion of the lesion measurements were performed during the evaluation.

[0195] Adverse events were monitored following administration of MEDI4736. Other evaluations included physical examinations, vital sign monitoring, and laboratory measurements.

[0196] Example 2: Results

[0197] (a) Registration and follow-up

[0198] Twelve subjects were treated in groups of 3 subjects: Group 1a (1 mg / kg tremelimumab and 3 mg / kg MEDI4736), Group 2a (1 mg / kg tremelimumab and 10 mg / kg MEDI4736), Group 3a (1 mg / kg tremelimumab and 15 mg / kg MEDI4736), and Group 3b (3 mg / kg tremelimumab and 10 mg / kg MEDI4736). Two subjects in Group 1a (1 subject withdrew consent after 2 doses of both agents) completed approximately 115 days of follow-up; subjects in Group 2a completed approximately 56 days of follow-up; and subjects in Groups 3a and 3b completed 28 days of follow-up ( Fig.12 ).

[0199] (b) Pharmacokinetics

[0200] The pharmacokinetic data obtained from dosing MEDI4736 at 3 mg / kg and 10 mg / kg (in combination with 1 mg / kg tremelimumab) are summarized in Figure 4 The individual PK parameters resulting from administration of MEDI4736 are shown in Figure 8 In this early data from the first dose, the mean Cmax for subjects receiving 3 mg / kg MEDI4736 was 67.36 μg / ml. The mean AUCτ for subjects receiving 3 mg / kg MEDI4736 was 625.3 μg / ml*day. The mean C 最低值 (C trough ) 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 最低值 (C trough ) was 35.65 μg / ml. MEDI4736 showed an increase in AUC that exceeded dose proportionality, indicating nonlinear PK. Figure 4 .

[0201] The pharmacokinetic data obtained from dosing of tremelimumab at 1 mg / kg (in combination with 3 mg / kg MEDI4736 or 10 mg / kg MEDI4736) are summarized in Figure 5 The individual PK parameters resulting from administration of tremelimumab are shown in Fig. 9 middle.

[0202] The mean 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 μg / ml.

[0203] 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.

[0204] The mean C for subjects receiving 1 mg / kg tremelimumab in combination with 3 mg / kg MEDI4736 最低值 (C trough The mean C for subjects receiving 1 mg / kg tremelimumab in combination with 10 mg / kg MEDI4736 was 4.21 μg / ml. 最低值 It is 5.33μg / ml.

[0205] (c) Evaluation of soluble PD-L1

[0206] The sPD-L1 data obtained from administration of 3 mg / kg and 10 mg / kg MEDI4736 and 1 mg / kg tremelimumab are shown in Figure 6 (absolute concentration), Figure 7 and Figure 8 (Inhibition, % of baseline). Complete sPD-L1 inhibition was observed in 5 of 6 subjects following MEDI4736 administration. In 1 subject, approximately 65% ​​sPD-L1 inhibition was observed at the second dose of MEDI4736.

[0207] (d) Security

[0208] Treatment-emergent adverse events (TEAEs) were reported in 10 of 12 subjects. The most frequently reported TEAEs were fatigue (41.7%; 5 subjects), increased amylase (25.0%; 3 subjects), pruritus, and upper respiratory tract infection (16.7%; 2 subjects each). No subject experienced dose-limiting toxicities. Nine of the 10 subjects who experienced TEAEs had events of Grade 1 or 2 severity. One subject (Cohort 2a) experienced Grade 3 increased aspartate transferase (AST) and increased alanine aminotransferase (ALT) and fatal (Grade 5) myasthenia gravis. Two of the 12 subjects experienced treatment-emergent serious adverse events (SAEs). Two subjects discontinued treatment due to TEAEs. Fig.13 Relevant adverse events are shown.

[0209] Early results showing changes in tumor size from baseline were shown in Fig.14 Reductions in tumor size were seen in patients in the 1 mg / kg(T)+10 mg / kg(M), 1 mg / kg(T)+15 mg / kg(M) and 3 mg / kg(T)+10 mg / kg(M) groups.

[0210] This study demonstrates that MEDI4736 and tremelimumab have favorable pharmacokinetic properties and that the combination of agents inhibits sPD-L1. In addition, MEDI4736 is well tolerated in the majority of subjects currently being treated.

[0211] Example 3: Phase 1b open-label study of MEDI4736 in combination with tremelimumab in patients with advanced NSCLC

[0212] The inhibitory PD-L1 and CTLA-4 pathways play a major role in controlling T-cell activation. MEDI4736 (M) is a human IgG1 monoclonal antibody that blocks the binding of PD-L1 to PD-1 and CD-80. Tremelimumab (T) is a human IgG2 monoclonal antibody that targets CTLA-4. As single agents, both MEDI4736 and tremelimumab showed encouraging safety profiles and promising clinical activity; because they prevent different interactions that promote immunosuppression, the MEDI4736 + tremelimumab combination therapy may provide higher anti-tumor activity in patients (pts) with advanced non-small cell lung cancer compared to either agent alone.

[0213] This Phase 1 study is evaluating the safety / tolerability, antitumor activity, pharmacokinetics (PK), and immunogenicity of the MEDI4736 + tremelimumab combination therapy in patients with advanced NSCLC. The study has dose escalation and expansion phases.

[0214] Results: As of December 4, 2014, 61 patients had been treated (Table 2).

[0215] Table 2

[0216] group dose 1a 3mg / kg M q4w+1mg / kg T 2a 10mg / kg M q4w+1mg / kg T 3a 15mg / kg M q4w + 1mg / kg T 3b 10mg / kg M q4w+3mg / kg T 4 20 mg / kg M q4w + 1 mg / kg T 4a 15mg / kg M q4w + 3mg / 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 10mg / kg M q2w+1mg / kg T 9 10mg / kg M q2w+3mg / kg T 10 10mg / kg M q2w+10mg / kg T

[0217] MEDI4736(M)+Tremelimumab(T)

[0218] Q4W = once every 4 weeks; Q2W = once every 2 weeks

[0219] Overall, the most common drug-related adverse events of all grades (≥10% of patients) were diarrhea, fatigue, pruritus, increased alanine aminotransferase (ALT), and amylase; they were manageable using standard treatment guidelines including steroids. The most common grade ≥3 drug-related AEs (≥5%) in all patients were colitis, diarrhea, increased aspartate aminotransferase (AST), and ALT. Drug-related AEs leading to discontinuation were colitis (6.6%), diarrhea (3.3%), pneumonia (3.3%), increased AST (3.3%) and ALT (1.6%), cough (1.6%), and dyspnea (1.6%); there was 1 treatment-related death (polymyositis) in the 10 mg / kg M q4w + 1 mg / kg T group. Of the 31 subjects who had at least one 8-week scan, best overall response was reported in 8 patients (26%); stable disease was reported in 11 patients (36%).

[0220] These promising results suggest that the MEDI4736+tremelimumab combination has a manageable safety profile and early clinical activity.

[0221] Results: As of January 27, 2015, 74 patients in 10 cohorts had been treated (Table 3).

[0222]

[0223]

[0224]

[0225] Various doses and schedules of MEDI4736 and tremelimumab given in combination were examined ( Fig.15 ). MEDI4736 doses and schedules include 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). Tremelimumab doses and schedules include 1 mg / kg, 3 mg / kg, and 10 mg / kg every 4 weeks (q4wx6 doses) and every 12 weeks (q12wx3 doses).

[0226] Complete sPD-L1 inhibition was observed at doses ≥15 mg / kg Q4W MEDI4736 ( Fig.16). One subject each at 10 mg / kg Q4W and 15 mg / kg Q4W showed partial sPD-L1 inhibition in some settings, followed by complete inhibition after repeat dosing. One subject was not inhibited at Day 29 after 15 mg / kg Q4W. In contrast, complete inhibition was observed with single agent MEDI4736 at doses >3 mg / kg Q2W (Study 1108). When individual sPD-L1 profiles were examined by cohort, complete sPD-L1 inhibition was observed in >95% of subjects after dosing at 10 mg / kg Q2W ( Fig.17 ).

[0227] Increased proliferation of T cells was also observed when MEDI4736 and tremelimumab were given in combination ( Fig.18 ). Cells were sorted by FACS analysis of a combination of markers CD4 and CD8 with Ki67 (a marker of cell proliferation). When the dose of tremelimumab was increased, a monotonic increase in the CD4+Ki67+ cell peak was observed, which was greater than the increase in the CD4+Ki67+ cell peak when MEDI4736 alone was used. The CD8+ proliferation response using the combination of MEDI4736 and tremelimumab was greater than that using MEDI4736 alone.

[0228] Increases in CD4 ICOS+ and CD4 HLADR+ cells were observed in subjects receiving MEDI4736 and tremelimumab compared to MEDI4736 alone ( Fig.19 ). Minimal changes in CD4+ICOS+ cells and CD4+HLADR+ cells were observed with MEDI4736 alone, and all combination doses tested had patterns that were distinguishable from MEDI4736. The greatest increases in CD4+ICOS+ cells and CD4+HLADR+ cells were observed with the 10 mg / kg tremelimumab dose. Increases in CD4+ T effector cells and regulatory T cells (CD25hi Cd127lo phenotype can also include some activated T cells) were observed in subjects receiving MEDI4736 and tremelimumab compared to MEDI4736 alone ( Fig. 20 and 21 ).

[0229] Clinical activity in NSCLC was observed with treatment with MEDI4736 and tremelimumab (all doses), which showed an increased overall response rate (ORR) compared to treatment with MEDI4736 monotherapy (10 mg / kg Q2W). Fig. 22). Responses were evaluable at baseline + ≥1 follow-up scan (including discontinuation due to disease progression or death before the first follow-up scan) in treated patients with measurable disease. For MEDI4736 NSCLC (CP1108), only patients with ≥12 weeks of follow-up were included. The overall response rate (ORR) includes confirmed and unconfirmed complete remission (CR) or partial remission (PR). For MEDI4736 NSCLC monotherapy (CP1108, 10 mg / kg Q2W), the best overall response (BOR) of disease stability (SD) with a minimum duration of 12 weeks was presented. For the combination of MEDI4736 and tremelimumab, a BOR of SD with a minimum duration of 7 weeks was presented.

[0230] Reduction and stabilization of tumor size were seen in the majority of NSCLC patients treated with MEDI4736 and tremelimumab (tremelimumab doses were 1 mg / kg; 3 mg / kg; and 10 mg / kg). Fig.23 The occurrence of grade 3 adverse events suggests that increasing the dose of tremelimumab also reduces the time to adverse events ( Fig.24 Reduction and stabilization of tumor size were seen in the majority of NSCLC patients given MEDI4736 and tremelimumab (MEDI4736 at doses of 10 mg / kg; 315 mg / kg; and 20 mg / kg (Q4W) or 10 mg / kg (Q2W)). Fig.25 The occurrence of grade 3 adverse events suggests that increasing the dose of tremelimumab affected the timing of adverse events compared with increasing the dose of MEDI4736 ( Fig.26 ). Therefore, the selection of MEDI4736 and tremelimumab doses will take into account efficacy and safety.

[0231] In cancer treatment Single agent The exposure data of tremelimumab have supported the need for higher exposure (>30mg / mL) to maximize efficacy. On average, these levels are obtained at doses>3mg / kg. Based on the analysis of single-agent exposure-safety relationships, the exposure achieved after administration of 1mg / kg of tremelimumab is expected to produce less toxicity than 3mg / kg. Without being limited to a specific theory, 1mg / kg of tremelimumab partially inhibits CTLA4 because immune-mediated adverse reactions are still observed at 1mg / kg (although at a lower incidence). If a synergistic effect is observed and the safety profile is improved relative to a higher dose of tremelimumab, 1mg / kg of tremelimumab in combination with MEDI4736 may be optimal.

[0232] In one study, a decrease in PD-L1 and complete inhibition of sPD-L1 in serum were observed at doses >3mg / kg Q2W MEDI4736. Based on the achievement of >99% decrease saturation in most subjects, a dose of 10mg / kgQ2W was selected for use as a single agent using the drug level that produced maximum tumor growth inhibition (TGI) in a mouse model (100mg / mL was identified as the minimum target minimum concentration). Inhibition of sPD-L1 was observed in some subjects after combining 10mg / kg Q4W with tremelimumab. The dose density with MEDI4736 Q2W was higher and the Cmax with Q4W was higher. The Q4W plan may provide greater convenience. Incomplete inhibition was observed in other subjects, possibly due to upregulation of PD-L1 and suboptimal minimum levels achieved at this dose level. MEDI4736 at 10mg / kg Q4W, evaluated in combination with tremelimumab, produced a minimum concentration below the 100mg / mL target. However, target levels were achieved with MEDI4736 at 15 mg / kg and 20 mg / kg Q4W, and higher doses of MEDI4736 maintained sPD-L1 inhibition in all subjects evaluated to date. If a synergistic effect is observed and the safety profile is improved, MEDI4736 at 10 mg / kg or higher in combination with tremelimumab may be optimal.

[0233] Overall, the combination of MEDI4736 and tremelimumab trended toward reduced or stable disease compared with MEDI4736 monotherapy, and was associated with an increased incidence of adverse events at the highest doses of MEDI4736 and tremelimumab tested ( Figure 27-29 ). These results suggest that the combination of MEDI4736 and tremelimumab is well tolerated and has the potential to be an effective therapy for NSCLC.

[0234] Example 4: Treatment of PD-L1 negative tumors in NSCLC using MED14736 and tremelimumab.

[0235] Twelve subjects were treated in groups of 3 subjects each: Group 1a (1 mg / kg tremelimumab and 3 mg / kg MEDI4736), Group 2a (1 mg / kg tremelimumab and 10 mg / kg MEDI4736), Group 3a (1 mg / kg tremelimumab and 15 mg / kg MEDI4736), and Group 3b (3 mg / kg tremelimumab and 10 mg / kg MEDI4736). Two subjects in Group 1a (1 subject withdrew consent after 2 doses of both agents) completed approximately 115 days of follow-up; subjects in Group 2a completed approximately 56 days of follow-up; and subjects in Groups 3a and 3b completed 28 days of follow-up.

[0236] Baseline levels of PD-L1 tumor expression data for the 7 subjects in the study are provided in Table 4 (below). Additional information is provided in Table 5.

[0237]

[0238]

[0239] The PD-L1 expression of subject tissues of NSCLC patients was characterized by immunohistochemistry in formalin-fixed and paraffin-embedded tissue samples. A sample was determined to be "PD-L1 positive" if it contained 25% or more tumor cells with PD-L1 membrane staining. This is expressed as the immunohistochemistry membrane (M)-score. All samples were scored as "negative" for PD-L1 expression. Tumor evaluation was available in 6 of 7 patients. Three patients were treated with a combination of tremelimumab and MEDI4736.

[0240] Two patients were identified as having an unconfirmed partial response (PR), 2 with stable disease (SD), and 2 with progressive disease (PD). In all 6 patients, tumor samples were collected >6 months before analysis of PD-L1 expression, and 3 of 6 biopsy samples were collected more than 2 years before analysis. These data suggest that the combination of MEDI4736 and tremelimumab is active in PD-L1-negative NSCLC based on stored tissue samples.

[0241] When given MEDI4736 and tremelimumab, the majority of patients with PD-L1-negative NSCLC responded to the combination therapy and showed a decrease or stabilization of tumor size compared to MEDI4736 monotherapy (CP1108, 10 mg / kg Q2W). Fig.30Patients with PD-L1-positive NSCLC also responded to the combination of MEDI4736 and tremelimumab and showed a decrease or stabilization of tumor size compared to MEDI4736 monotherapy ( Fig.31 When the results for patients with PD-L1-negative NSCLC were grouped by dose of tremelimumab, 1 mg / kg tremelimumab or 3 mg / kg tremelimumab given in combination with MEDI4736 at 10 mg / kg Q4W or 15 mg / kg Q4W were effective in controlling or reducing disease ( Fig.32 When the results were grouped by dose of MEDI4736, the results also showed that 1 mg / kg to 3 mg / kg of tremelimumab given in combination with MEDI4736 at 10 mg / kg Q4W or 15 mg / kg Q4W was effective in controlling or reducing disease ( Fig.33 An analysis of all NSCLC patients who received MEDI4736 and tremelimumab showed that both PD-L1- and PD-L1+ NSCLC patients responded to treatment ( Figures 34A-34D and Figure 35-37 ; Table 6-9).

[0242]

[0243]

[0244]

[0245]

[0246] Those skilled in the art will recognize or be able to ascertain using no more than routine experimentation numerous equivalents to the specific aspects of the disclosure described herein. Such equivalents are intended to be encompassed by the following claims.

[0247] Various publications are cited herein, the disclosures of which are incorporated by reference in their entireties.

[0248] Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, it will be apparent that certain changes and modifications may be practiced within the scope of the appended claims.

[0249] Sequence Listing

[0250] SEQ ID NO:1 MEDI4736 VL

[0251]

[0252] SEQ ID NO:2 MEDI4736 VH

[0253]

[0254] SEQ ID NO:3-MEDI4736 VH CDR1

[0255] RYWMS

[0256] SEQ ID NO:4-MEDI4736 VH CDR2

[0257] NIKQDGSEKYYVDSVKG

[0258] SEQ ID NO:5-MEDI4736 VH CDR3

[0259] EGGWFGELAFDY

[0260] SEQ ID NO:6-MEDI4736 VL CDR1

[0261] RASQRVSSSYLA

[0262] SEQ ID NO:7-MEDI4736 VL CDR2

[0263] DASSRAT

[0264] SEQ ID NO:8-MEDI4736 V LCDR3

[0265] QYGSLPWT

[0266] SEQ ID NO:9 Tremelimumab VL

[0267]

[0268] SEQ ID NO: 10 Tremelimumab VH

[0269]

[0270] SEQ ID NO: 11 - Tremelimumab VH CDR1

[0271] GFTFSSYGMH

[0272] SEQ ID NO: 12-Tremelimumab VH CDR2

[0273] VIWYDGSNKYYADSV

[0274] SEQ ID NO: 13 - Tremelimumab VH CDR3

[0275] TAVYYCARDPRGATLYYYYYGMDV

[0276] SEQ ID NO: 14 - Tremelimumab VL CDR1

[0277] RASQSINSYLD

[0278] SEQ ID NO: 15 - Tremelimumab VL CDR2

[0279] AASSLQS

[0280] SEQ ID NO: 16 - Tremelimumab VL CDR3

[0281] QQYYSTPFT Sequence Listing <110> MEDIMMUNE LIMITED <120> Anti-B7-H1 and anti-CTLA-4 antibodies for the treatment of non-small cell lung cancer <130> TRB7-100WO1 <140> <141> <150> 62 / 114,336 <151> 2015-02-10 <150> 62 / 105,992 <151> 2015-01-21 <150> 61 / 992,658 <151> 2014-05-13 <160> 16 <170> PatentIn Version 3.5 <210> 1 <211> 108 <212> PRT <213> Artificial sequence <220> <221> source <223> / Annotation="Artificial sequence description: synthetic peptide" <400> 1 Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Arg Val Ser Ser Ser 20 25 30 Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45 Ile Tyr Asp Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60 Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu 65 70 75 80 Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Leu Pro 85 90 95 Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 2 <211> 121 <212> PRT <213> Artificial sequence <220> <221> source <223> / Annotation="Artificial sequence description: synthetic peptide" <400> 2 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Arg Tyr 20 25 30 Trp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Asn Ile Lys Gln Asp Gly Ser Glu Lys Tyr Tyr Val Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Gly Gly Trp Phe Gly Glu Leu Ala Phe Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 3 <211> 5 <212> PRT <213> Artificial sequence <220> <221> source <223> / Annotation="Artificial sequence description: synthetic peptide" <400> 3 Arg Tyr Trp Met Ser 1 5 <210> 4 <211> 17 <212> PRT <213> Artificial sequence <220> <221> source <223> / Annotation="Artificial sequence description: synthetic peptide" <400> 4 Asn Ile Lys Gln Asp Gly Ser Glu Lys Tyr Tyr Val Asp Ser Val Lys 1 5 10 15 Gly <210> 5 <211> 12 <212> PRT <213> Artificial sequence <220> <221> source <223> / Annotation="Artificial sequence description: synthetic peptide" <400> 5 Glu Gly Gly Trp Phe Gly Glu Leu Ala Phe Asp Tyr 1 5 10 <210> 6 <211> 12 <212> PRT <213> Artificial sequence <220> <221> source <223> / Annotation="Artificial sequence description: synthetic peptide" <400> 6 Arg Ala Ser Gln Arg Val Ser Ser Ser Tyr Leu Ala 1 5 10 <210> 7 <211> 7 <212> PRT <213> Artificial sequence <220> <221> source <223> / Annotation="Artificial sequence description: synthetic peptide" <400> 7 Asp Ala Ser Ser Arg Ala Thr 1 5 <210> 8 <211> 9 <212> PRT <213> Artificial sequence <220> <221> source <223> / Annotation="Artificial sequence description: synthetic peptide" <400> 8 Gln Gln Tyr Gly Ser Leu Pro Trp Thr 1 5 <210> 9 <211> 139 <212> PRT <213> Artificial sequence <220> <221> source <223> / Annotation="Artificial sequence description: synthetic peptide" <400> 9 Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys 1 5 10 15 Arg Ala Ser Gln Ser Ile Asn Ser Tyr Leu Asp Trp Tyr Gln Gln Lys 20 25 30 Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr Ala Ala Ser Ser Leu Gln 35 40 45 Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe 50 55 60 Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr 65 70 75 80 Cys Gln Gln Tyr Tyr Ser Thr Pro Phe Thr Phe Gly Pro Gly Thr Lys 85 90 95 Val Glu Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro 100 105 110 Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu 115 120 125 Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val 130 135 <210> 10 <211> 167 <212> PRT <213> Artificial sequence <220> <221> source <223> / Annotation="Artificial sequence description: synthetic peptide" <400> 10 Gly Val Val Gln Pro Gly Arg Ser Leu Arg Leu Ser Cys Ala Ala Ser 1 5 10 15 Gly Phe Thr Phe Ser Ser Tyr Gly Met His Trp Val Arg Gln Ala Pro 20 25 30 Gly Lys Gly Leu Glu Trp Val Ala Val Ile Trp Tyr Asp Gly Ser Asn 35 40 45 Lys Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp 50 55 60 Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu 65 70 75 80 Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Pro Arg Gly Ala Thr Leu 85 90 95 Tyr Tyr Tyr Tyr Tyr Gly Met Asp Val Trp Gly Gln Gly Thr Thr Val 100 105 110 Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala 115 120 125 Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu 130 135 140 Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly 145 150 155 160 Ala Leu Thr Ser Gly Val His 165 <210> 11 <211> 10 <212> PRT <213> Artificial sequence <220> <221> source <223> / Annotation="Artificial sequence description: synthetic peptide" <400> 11 Gly Phe Thr Phe Ser Ser Tyr Gly Met His 1 5 10 <210> 12 <211> 15 <212> PRT <213> Artificial sequence <220> <221> source <223> / Annotation="Artificial sequence description: synthetic peptide" <400> 12 Val Ile Trp Tyr Asp Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val 1 5 10 15 <210> 13 <211> twenty four <212> PRT <213> Artificial sequence <220> <221> source <223> / Annotation="Artificial sequence description: 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> PRT <213> Artificial sequence <220> <221> source <223> / Annotation="Artificial sequence description: synthetic peptide" <400> 14 Arg Ala Ser Gln Ser Ile Asn Ser Tyr Leu Asp 1 5 10 <210> 15 <211> 7 <212> PRT <213> Artificial sequence <220> <221> source <223> / Annotation="Artificial sequence description: synthetic peptide" <400> 15 Ala Ala Ser Ser Leu Gln Ser 1 5 <210> 16 <211> 9 <212> PRT <213> Artificial sequence <220> <221> source <223> / Annotation="Artificial sequence description: synthetic peptide" <400> 16 Gln Gln Tyr Tyr Ser Thr Pro Phe Thr 1 5

Claims

1. Use of a combination comprising MEDI4736 or an antigen-binding fragment thereof and tremelimumab or an antigen-binding fragment thereof in the preparation of a medicament for treating non-small cell lung cancer (NSCLC) in a patient, wherein the medicament is suitable for administering MEDI4736 or an antigen-binding fragment thereof at a dose of 1500 mg and tremelimumab or an antigen-binding fragment thereof at a dose of 75 mg, wherein the patient has metastatic NSCLC.

2. The use according to claim 1, wherein 1500 mg of MEDI4736 or an antigen-binding fragment thereof and 75 mg of tremelimumab or an antigen-binding fragment thereof are co-administered every 2 to 4 weeks.

3. The use according to claim 1 or claim 2, wherein at least 4 doses of MEDI4736 or an antigen-binding fragment thereof and tremelimumab or an antigen-binding fragment thereof are administered.

4. The use according to claim 1 or claim 2, further comprising administering 1500 mg of MEDI4736 or an antigen-binding fragment thereof every 4 weeks after administering at least 4 doses of 1500 mg of MEDI4736 or an antigen-binding fragment thereof and 75 mg of tremelimumab or an antigen-binding fragment thereof.

5. The use according to claim 4, further comprising administering a 75 mg dose of tremelimumab or an antigen-binding fragment thereof at least 7 weeks after administering 4 doses of 1500 mg MEDI4736 or an antigen-binding fragment thereof and 75 mg tremelimumab or an antigen-binding fragment thereof.

6. The use according to any one of claims 1-2 and 5, wherein MEDI4736 or an antigen-binding fragment thereof is administered every 2 to 4 weeks for up to 49 weeks or for 49 weeks.

7. The use according to any one of claims 1-2 and 5, wherein the NSCLC is refractory to at least one chemotherapeutic agent prior to administration of MEDI4736 or an antigen-binding fragment thereof and tremelimumab or an antigen-binding fragment thereof.

8. The use according to claim 7, wherein the chemotherapeutic agent is vemurafenib, erlotinib, afatinib, cetuximab, carboplatin, bevacizumab, erlotinib, gefitinib or pemetrexed.

9. The use according to any one of claims 1-2, 5 and 8, wherein the patient is immunotherapy naive prior to the administration of MEDI4736 or an antigen-binding fragment thereof and tremelimumab or an antigen-binding fragment thereof.

10. The use according to any one of claims 1-2, 5 and 8, wherein the administration of MEDI4736 or an antigen-binding fragment thereof is by intravenous infusion.

11. The use according to any one of claims 1-2, 5 and 8, wherein the administration of tremelimumab or an antigen-binding fragment thereof is by intravenous infusion.

12. The use according to any one of claims 1-2, 5 and 8, wherein the administration reduces the tumor size by at least 10% relative to the tumor size measured before administration of MEDI4736 or an antigen-binding fragment thereof and tremelimumab or an antigen-binding fragment thereof.

13. The use according to any one of claims 1-2, 5 and 8, wherein the administration reduces the tumor size by at least 25% or 50% relative to the tumor size measured before administration of MEDI4736 or an antigen-binding fragment thereof and tremelimumab or an antigen-binding fragment thereof.

14. The use according to any one of claims 1-2, 5 and 8, wherein the NSCLC is squamous or non-squamous.

Images