COMBINED ANTI-PLD-1 AND ANTI-CTLA-4 ANTIBODIES FOR TREATING NON-SMALL LUNG CANCER

Abstract

A method for treating lung cancer (e.g., non-small cell lung cancer) with an anti-PD-L1 antibody (MEDI4736), alone or in combination with an anti-CTLA4 antibody (Tremelimumab), in a patient identified using a polynucleotide marker of IFNgamma.

Claims

1. A method of treatment comprising administering an anti-PD-L1 antibody, or an antigen binding fragment thereof, to a patient identified as having a squamous cell carcinoma or non-squamous cell carcinoma non-small cell lung cancer tumor that expresses an IFNgamma gene.

2. The method of claim 1, wherein the anti-PD-L1 antibody is MEDI4736.

3. The method of any of claims 1-2, wherein the tumor expresses IFNgamma mRNA.

4. A method of treatment comprising administering MEDI4736 or an antigen binding fragment thereof to a patient identified as having a squamous cell carcinoma or non-squamous cell carcinoma non-small cell lung cancer tumor that expresses an IFNgamma mRNA.

5. The method of any of claims 1-4, wherein the patient is identified as responsive to MEDI4736.

6. The method of any of claims 1-5, wherein interferon gamma gene expression is detected in a Real-Time PCR assay.

7. The method of any of claims 1-6, wherein the IFNgamma mRNA is detected in a blood sample.

8. The method of any of claims 1-7, wherein at least about 0.1, about 0.3, about 1, about 3, about 10, about 15 mg/kg, or about 20 mg/kg MEDI4736, or an antigen-binding fragment thereof, is administered.

9. The method of claim 8, wherein about 1 mg/kg MEDI4736, or an antigen-binding fragment thereof, is administered.

10. The method of claim 8, wherein about 3 mg/kg MEDI4736, or an antigen-binding fragment thereof, is administered.

11. The method of claim 8, wherein about 10 mg/kg MEDI4736 or an antigen-binding fragment thereof is administered.

12. The method of claim 8, wherein about 15 mg/kg MEDI4736, or an antigen-binding fragment, thereof is administered.

13. The method of claim 8, wherein about 20 mg/kg MEDI4736, or an antigen-binding fragment, thereof is administered.

14. The method of any one of claims 1-8, wherein the administration is repeated about every 14, 21, or 28 days.

15. The method of any one of claims 1-8, wherein at least two doses is administered.

16. The method of any one of claims 1-8, wherein at least three doses is administered.

17. The method of any one of claims 1-8, wherein at least five doses is administered.

18. A method of identifying a subject having a squamous cell carcinoma or non-squamous cell carcinoma non-small cell lung cancer responsive to anti-PD-L1 therapy, the method comprising detecting an increase in the level of IFNgamma gene expression in a non-small cell lung cancer tumor of the subject, relative to a reference, thereby identifying said non-small cell lung cancer as responsive to anti-PD-L1 therapy.

19. The method of claim 18, wherein the IFNgamma gene expression is detected by real-time PCR.

20. A method of treatment comprising administering a combination of an anti-PD-L1 antibody and an anti-CTLA4 antibody, or antigen binding fragments thereof, to a patient identified as having a squamous cell carcinoma or non-squamous cell carcinoma non-small cell lung cancer tumor that expresses IFNgamma.

21. The method of claim 20, wherein the anti-PD-L1 antibody is MEDI4736 and the anti-CTLA4 antibody is Tremelimumab.

22. A method of treatment comprising administering a combination of MEDI4736 and Tremelimumab, or antigen binding fragments thereof, to a patient identified as having a squamous cell carcinoma or non-squamous cell carcinoma non-small cell lung cancer tumor that expresses IFNgamma.

23. The method of any one of claims 20-22, wherein the patient is identified as responsive to MEDI4736 and Tremelimumab.

24. The method of any one of claims 20-22, wherein interferon gamma gene expression is detected in a Real-Time PCR assay.

25. The method of any one of claims 20-22, wherein interferon gamma mRNA is present in a tumor or blood sample.

26. The method of any one of claims 20-22, wherein at least about 0.1, about 0.3, about 1, about 3, about 10, about 15 mg/kg, or about 20 mg/kg MEDI4736, or an antigen-binding fragment thereof, is administered.

27. The method of claim 26, wherein about 1 mg/kg MEDI4736, or an antigen-binding fragment thereof, is administered.

28. The method of claim 26, wherein about 3 mg/kg MEDI4736, or an antigen-binding fragment thereof, is administered.

29. The method of claim 26, wherein about 10 mg/kg MEDI4736 or an antigen-binding fragment thereof is administered.

30. The method of claim 26, wherein about 15 mg/kg MEDI4736, or an antigen-binding fragment, thereof is administered.

31. The method of claim 26, wherein about 20 mg/kg MEDI4736, or an antigen-binding fragment, thereof is administered.

32. The method of any of claims 20-31, wherein at least 1 mg/kg, at least about 3 mg/kg, or about 10 mg/kg Tremelimumab, or an antigen-binding fragment thereof, is administered.

33. The method of claim 32, wherein about 1 mg/kg Tremelimumab, or an antigen-binding fragment, thereof is administered.

34. The method of claim 32, wherein about 3 mg/kg Tremelimumab, or an antigen-binding fragment, thereof is administered.

35. The method of claim 32, wherein about 10 mg/kg Tremelimumab, or an antigen-binding fragment, thereof is administered.

36. The method of claim 32, wherein about 1 mg/kg Tremelimumab and about 20 mg/kg MEDI4736 or an antigen-binding fragment thereof is administered.

37. The method of claim 32, wherein about 3 mg/kg Tremelimumab and about 15 mg/kg MEDI4736 or an antigen-binding fragment thereof is administered.

38. The method of claim 32, wherein about 10 mg/kg Tremelimumab and about 15 mg/kg MEDI4736 or an antigen-binding fragment thereof is administered.

39. The method of claim 32, wherein about 3 mg/kg Tremelimumab and about 10 mg/kg MEDI4736 or an antigen-binding fragment thereof is administered.

40. The method of any one of claims 20-39, wherein the administration is repeated about every 14, 21, or 28 days.

41. The method of any of claims 32-40, wherein at least two doses is administered.

42. The method of any of claims 32-40, wherein at least three doses is administered.

43. The method of any of claims 32-40, wherein at least five doses is administered.

44. A method of identifying a subject having a squamous cell carcinoma or non-squamous cell carcinoma non-small cell lung cancer responsive to a combination of an anti-PD-L1 therapy and an anti-CTLA4 therapy, the method comprising detecting an increase in the level of IFNgamma gene expression in a non-small cell lung cancer tumor of the subject, relative to a reference, thereby identifying said non-small cell lung cancer as responsive to an anti-PD-L1 therapy and an anti-CTLA4 therapy.

45. The method of any one of claims 32-44, wherein the markers are detected by real-time PCR.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0051] FIG. 1 is a bar graph that shows interferon gamma gene expression at baseline in tumor and blood samples obtained from patients with squamous cell carcinoma (SCC) non-small cell lung cancer (NSCLC). The patients were subsequently treated with an anti-PD-L1 antibody. Subjects were characterized as having SCC NSCLC that was confirmed or unconfirmed as having a partial response (PR) to anti-PD-L1 treatment, as having stable disease (SD), as having progressive disease (PD) or as not evaluable (NE).

[0052] FIG. 2 provides two scatter plots showing baseline tumor and blood interferon gamma (IFGN mRNA) expression in patients subsequently treated with an anti-PD-L1 antibody. dCT indicates the difference in interferon gamma expression prior to and following anti-PD-L1 antibody therapy. Sens=sensitivity; Spec=specificity; PPV=positive predictive value; NPV=negative predictive value; PR=partial responder; PD=progressive disease; NE=not evaluable.

[0053] FIG. 3 provides a scatter plot and two bar graphs showing a correlation between baseline tumor and blood interferon gamma mRNA expression in patients subsequently treated with an anti-PD-L1 antibody.

[0054] FIG. 4 provides two scatter plots showing tumor size change and dCT IFNgamma mRNA in partial responder (PR) patients having non-squamous cell carcinoma non-small cell lung cancer (SCC NSCLC) treated with a combination of anti-CTLA4 and anti-PD-L1 antibodies. Dosages for the MEDI4736 and Tremelimumab (Treme) are shown in the bottom panel: Treme 1 mg/kg, MEDI4736 20 mg/kg Q4 W, Treme 3 mg/kg, MEDI4736 15 mg/kg Q4w; Treme 10 mg/kg, MEDI4736 15 mg/kg Q4 W; Treme 3 mg/kg, MEDI4736 10 mg/kg Q2W.

[0055] FIG. 5 includes two pie charts showing clinical response status of interferon gamma mRNA positive and negative patients treated with a combination of anti-CTLA4 and anti-PD-L1 antibodies.

LIST OF SEQUENCES

[0056] MEDI4736 light chain variable region amino acid sequence: SEQ ID NO:1

[0057] MEDI4736 heavy chain variable region amino acid sequence: SEQ ID NO:2.

[0058] MEDI4736 heavy chain variable region amino acid sequence of CDR1, CDR2, and CDR3: SEQ ID NOs:3-5.

[0059] MEDI4736 light chain variable region amino acid sequence of CDR1, CDR2, and CDR3: SEQ ID NOs:6-8.

DETAILED DESCRIPTION OF THE INVENTION

[0060] The present invention provides methods for treating lung cancer (e.g., non-squamous cell or squamous cell non-small cell lung cancer) with an anti-PD-L1 antibody or a combination of an anti-PD-L1 antibody and an anti-CTLA4 antibody in a patient identified by detecting IFNgamma polynucleotide expression in a tumor or blood sample of the patient.

[0061] The invention is based, at least in part, on the discovery that patients having lung cancer (e.g., non-squamous cell or squamous cell non-small cell lung cancer) that is responsive to treatment with an anti-PD-L1 antibody or a combination of an anti-PD-L1 antibody and an anti-CTLA4 antibody may be identified by detecting increased levels of interferon gamma mRNA in a tumor or blood sample. Accordingly, the invention provides methods for identifying subjects that have lung cancer that is likely to respond to anti-PD-L1 antibody treatment, alone or in combination with an anti-CTLA4 antibody, based on the presence or level of IFNgamma mRNA in a subject tumor or blood sample.

B7-H1/PD-L1

[0062] B7-H1, also known as PD-L1, is a type I transmembrane protein of approximately 53 kDa in size. In humans B7-H1 is expressed on a number of immune cell types including activated and anergic/exhausted T cells, on nave and activated B cells, as well as on myeloid dendritic cells (DC), monocytes and mast cells. It is also expressed on non-immune cells including islets of the pancreas, Kupffer cells of the liver, vascular endothelium and selected epithelia, for example airway epithelia and renal tubule epithelia, where its expression is enhanced during inflammatory episodes. B7-H1 expression is also found at increased levels on a number of tumours including, but not limited to breast, colon, colorectal, lung, renal, including renal cell carcinoma, gastric, bladder, non-small cell lung cancer (NSCLC), hepatocellular cancer (HCC), and pancreatic cancer, as well as melanoma.

[0063] B7-H1 is known to bind two alternative ligands, the first of these, PD-1, is a 50-55 kDa type I transmembrane receptor that was originally identified in a T cell line undergoing activation-induced apoptosis. PD-1 is expressed on activated T cells, B cells, and monocytes, as well as other cells of the immune system and binds both B7-H1 (PD-L1) and the related B7-DC (PD-L2). The second is the B7 family member B7-1, which is expressed on activated T cells, B cells, monocytes and antigen presenting cells.

[0064] Signaling via the PD-1/B7-H1 axis is believed to serve important, non-redundant functions within the immune system, by negatively regulating T cell responses. B7-H1 expression on tumor cells is believed to aid tumors in evading detection and elimination by the immune system. B7-H1 functions in this respect via several alternative mechanisms including driving exhaustion and anergy of tumour infiltrating T lymphocytes, stimulating secretion of immune repressive cytokines into the tumour micro-environment, stimulating repressive regulatory T cell function and protecting B7-H1 expressing tumor cells from lysis by tumor cell specific cytotoxic T cells.

Anti-PD-L1 Antibodies

[0065] Antibodies that specifically bind and inhibit PD-L1 activity (e.g., binding to PD-1 and/or CD80) are useful for the treatment of lung cancer (e.g., non-small cell lung cancer

[0066] MEDI4736 is an exemplary anti-PD-L1 antibody that is selective for B7-H1 and blocks the binding of B7-H1 to the PD-1 and CD80 receptors. MEDI4736 can relieve B7-H1-mediated suppression of human T-cell activation in vitro and inhibits tumor growth in a xenograft model via a T-cell dependent mechanism. Other agents that could be used include agents that inhibit PD-L1 and/or PD-1 (AB or other).

[0067] Information regarding MEDI4736 (or fragments thereof) for use in the methods provided herein can be found in International Application Publication No. WO 2011/066389 A1, 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 that reduces binding to the complement component C1q and the Fc receptors responsible for mediating antibody-dependent cell-mediated cytotoxicity (ADCC).

[0068] MEDI4736 and antigen-binding fragments thereof for use in the methods provided herein comprises a heavy chain and a light chain or a heavy chain variable region and a light chain variable region. In a specific aspect, MEDI4736 or an antigen-binding fragment thereof for use in the methods provided herein comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:1 and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:2. In a specific aspect, MEDI4736 or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs:3-5, and wherein the light chain variable region comprises the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs:6-8. Those of ordinary skill in the art would easily be able to identify Chothia-defined, Abm-defined or other CDR definitions known to those of ordinary skill 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 2.14H9OPT antibody as disclosed in WO 2011/066389 A1, which is herein incorporated by reference in its entirety.

CTLA4 Blocking Antibody: Tremelimumab

[0069] Accordingly, in one embodiment therapeutic combinations of the invention comprise a CTLA4 blocking antibody (e.g., Tremelimumab) and/or antibodies that reduce PD1/PD-L1 interactions. Two T cell modulatory pathways receiving significant attention to date signal through cytotoxic T lymphocyte antigen-4 (CTLA4, CD152) and programmed death ligand 1 (PD-L1, also known as B7H-1 or CD274).

[0070] CTLA4 is expressed on activated T cells and serves as a co-inhibitor to keep T cell responses in check following CD28-mediated T cell activation. CTLA4 is believed to regulate the amplitude of the early activation of nave and memory T cells following TCR engagement and to be part of a central inhibitory pathway that affects both antitumor immunity and autoimmunity. CTLA4 is expressed primarily on T cells, and the expression of its ligands CD80 (B71) and CD86 (B7.2), is largely restricted to antigen-presenting cells, T cells, and other immune mediating 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 phase III trials for the treatment of advanced melanoma but did not significantly increase the overall survival of patients compared to the standard of care (temozolomide or dacarbazine) at that time.

[0071] Information regarding tremelimumab (or antigen-binding fragments thereof) for use in the methods provided herein can be found in U.S. Pat. No. 6,682,736 (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 binding of CTLA4 to CD80 (B7.1) and CD86 (B7.2). It has been shown to result in immune activation in vitro and some patients treated with tremelimumab have shown tumor regression.

[0072] Tremelimumab for use in the methods provided herein comprises a heavy chain and a light chain or a heavy chain variable region and a light chain variable region. In a specific aspect, tremelimumab or an antigen-binding fragment thereof for use in the methods provided herein comprises a light chain variable region and a heavy chain variable region. In a specific aspect, tremelimumab or an antigen-binding fragment thereof for use in the methods provided herein comprises a heavy chain variable region and a light chain variable region identified herein. Those of ordinary skill in the art would easily be able to identify Chothia-defined, Abm-defined or other CDR definitions known to those of ordinary skill 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 U.S. Pat. No. 6,682,736, which is herein incorporated by reference in its entirety.

Characterizing Responsiveness to Anti-PD-L1 Antibody and/or Anti-CTLA4 Therapy

[0073] In characterizing the responsiveness of lung cancer in a subject to an anti-PD-L1 antibody treatment alone or in combination with an anti-CTLA4 antibody treatment, the level of IFNgamma expression is measured in different types of biologic samples (e.g., tumor or blood samples).

[0074] IFNgamma polynucleotide expression is higher in a tumor or blood sample obtained from a subject that is responsive to anti-PD-L1 antibody treatment or treatment with a combination of the invention comprising an anti-PD-L1 antibody and an anti-CTLA4 antibody than the level of expression in a non-responsive subject (e.g., a subject with progressive disease). In one embodiment, an alteration in expression is calculated using Ct, where Ct represents cycle threshold values. For example, the Ctvalue of an IFNgamma gene is obtained and from that value the Ct value of a reference gene (e.g., B2M, ACTB, GAPDH) is subtracted from the mean Ct value for IFNgamma gene to obtain a Delta-Ct value. Then this value is multiplied by 1 to retain directionality. In other embodiments, expression of a marker of the invention is increased by at least about 2, 3, 4, 5 or 10-fold in a responsive patient relative to the level in a non-responsive subject (e.g., a subject with progressive disease). IFNgamma polynucleotide fold change values are determined using any method known in the art, including but not limited to quantitative PCR, RT-PCR, Northern blotting, in situ hybridization, fluorescence in situ hybridization (FISH) and/or RNA sequencing.

[0075] In particular embodiments, the responsiveness of lung cancer in a subject to anti-PD-L1 antibody treatment or treatment with a combination of the invention comprising an anti-PD-L1 antibody and an anti-CTLA4 antibody, is assayed by detecting IFNgamma expression (e.g., mRNA expression).

Selection of a Treatment Method

[0076] Subjects suffering from lung cancer (e.g., squamous or non-squamous cell carcinoma non-small cell lung cancer) may be tested for IFNgamma polynucleotide expression in the course of selecting a treatment method. Patients characterized as having high expression (e.g., as defined by Ct score) or increased expression relative to a reference level are identified as responsive to anti-PD-L1 treatment or treatment with a combination of the invention comprising an anti-PD-L1 antibody and an anti-CTLA4 antibody.

Treatment with an Anti-PD-L1 Antibody

[0077] Patients identified as having tumors or blood samples that express IFNgamma, particularly at high levels, are likely to be responsive to anti-PD-L1 antibody therapy or treatment with a combination of the invention comprising an anti-PD-L1 antibody and an anti-CTLA4 antibody. Such patients are administered an anti-PD-L1 antibody, such as MEDI4736, or an antigen-binding fragment thereof, alone or in combination with Tremelimumab. MEDI4736, alone or in combination with Tremelimumab, or an antigen-binding fragment thereof can be administered only once or infrequently while still providing benefit to the patient. In further aspects the patient is administered additional follow-on doses. Follow-on doses can be administered at various time intervals depending on the patient's age, weight, clinical assessment, tumor burden, and/or other factors, including the judgment of the attending physician.

[0078] In some embodiments, at least two doses of MEDI4736, alone or in combination with 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 can be administered to the patient. In some embodiments, MEDI4736, alone or in combination with Tremelimumab, or an antigen-binding fragment thereof is administered over a two-week treatment period, over a four-week treatment period, over a six-week treatment period, over an eight-week treatment period, over a twelve-week treatment period, over a twenty-four-week treatment period, or over a one-year or more treatment period. In some embodiments, MEDI4736, alone or in combination with Tremelimumab, or an antigen-binding fragment thereof is administered over a three-week treatment period, a six-week treatment period, over a nine-week treatment period, over a twelve-week treatment period, over a twenty-four-week treatment period, or over a one-year or more treatment period. In some embodiments, MEDI4736, alone or in combination with Tremelimumab, or an antigen-binding fragment thereof is administered over a two-month treatment period, over a four-month treatment period, or over a six-month or more treatment period (e.g., during a maintenance phase).

[0079] The amount of MEDI4736, alone or in combination with 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 assessment, tumor burden and/or other factors, including the judgment of the attending physician.

[0080] In certain aspects the patient is administered one or more doses of MEDI4736 or an antigen-binding fragment thereof wherein the dose is about 0.1 mg/kg. In certain aspects the patient is administered one or more doses of MEDI4736 or an antigen-binding fragment thereof wherein the dose is about 0.3 mg/kg. In certain aspects the patient is administered one or more doses of MEDI4736 or an antigen-binding fragment thereof wherein the dose is about 1 mg/kg. In certain aspects the patient is administered one or more doses of MEDI4736 or an antigen-binding fragment thereof wherein the dose is about 3 mg/kg. In certain aspects the patient is administered one or more doses of MEDI4736 or an antigen-binding fragment thereof wherein the dose is about 10 mg/kg. In certain aspects the patient is administered one or more doses of MEDI4736 or an antigen-binding fragment thereof wherein the dose is about 15 mg/kg. In certain aspects the patient is administered one or more doses of MEDI4736 or an antigen-binding fragment thereof wherein the dose is about 20 mg/kg MEDI4736, or an antigen-binding fragment thereof.

[0081] In certain aspects the patient is administered at least two doses of MEDI4736 or an antigen-binding fragment thereof wherein the dose is about 0.1 mg/kg. In certain aspects the patient is administered at least two doses of MEDI4736 or an antigen-binding fragment thereof wherein the dose is about 0.3 mg/kg. In certain aspects the patient is administered at least two doses of MEDI4736 or an antigen-binding fragment thereof wherein the dose is about 1 mg/kg. In certain aspects the patient is administered at least two doses of MEDI4736 or an antigen-binding fragment thereof wherein the dose is about 3 mg/kg. In certain aspects the patient is administered at least two doses of MEDI4736 or an antigen-binding fragment thereof wherein the dose is about 10 mg/kg. In certain aspects the patient is administered at least two doses of MEDI4736 or an antigen-binding fragment thereof wherein the dose is about 15 mg/kg. In certain aspects the patient is administered at least two doses of MEDI4736 or an antigen-binding fragment thereof wherein the dose is about 20 mg/kg. In some embodiments, the at least two doses are administered about two weeks apart. In some embodiments, the at least two doses are administered about three weeks apart. In some embodiments, the at least two doses are administered about four weeks apart.

[0082] In certain aspects the patient is administered at least three doses of MEDI4736 or an antigen-binding fragment thereof wherein the dose is about 0.1 mg/kg. In certain aspects the patient is administered at least three doses of MEDI4736 or an antigen-binding fragment thereof wherein the dose is about 0.3 mg/kg. In certain aspects the patient is administered at least three doses of MEDI4736 or an antigen-binding fragment thereof wherein the dose is about 1 mg/kg. In certain aspects the patient is administered at least three doses of MEDI4736 or an antigen-binding fragment thereof wherein the dose is about 3 mg/kg. In certain aspects the patient is administered at least three doses of MEDI4736 or an antigen-binding fragment thereof wherein the dose is about 10 mg/kg. In certain aspects the patient is administered at least three doses of MEDI4736 or an antigen-binding fragment thereof wherein the dose is about 15 mg/kg. In certain aspects the patient is administered at least two doses of MEDI4736 or an antigen-binding fragment thereof wherein the dose is about 20 mg/kg. In some embodiments, the at least three doses are administered about two weeks apart. In some embodiments, the at least three doses are administered about three weeks apart. In some embodiments, the at least three doses are administered about four

[0083] In certain aspects, administration of MEDI4736, alone or in combination with Tremelimumab, or an antigen-binding fragment thereof according to the methods provided herein is through parenteral administration. For example, MEDI4736, alone or in combination with 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. 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.

[0084] In certain aspects, MEDI4736, alone or in combination with Tremelimumab, or an antigen-binding fragment thereof is administered according to the methods provided herein in combination or in conjunction with additional cancer therapies. Such therapies include, without limitation, chemotherapeutic agents such as Vemurafenib, Erlotinib, Afatinib, Cetuximab, Carboplatin, Bevacizumab, Erlotinib, or Pemetrexed, or other chemotherapeutic agents, as well radiation or any other anti-cancer treatments.

[0085] The methods provided herein can decrease tumor size, retard tumor growth or maintain a steady state. In certain aspects the reduction in tumor size can be significant based on appropriate statistical analyses. A reduction in tumor size can be measured by comparison to the size of patient's tumor at baseline, against an expected tumor size, against an expected tumor size based on a large patient population, or against the tumor size of a control population. In certain aspects provided herein, the administration of MEDI4736, alone or in combination with Tremelimumab, can reduce a tumor size by at least 25%. In certain aspects provided herein, the administration of MEDI4736 alone or in combination with Tremelimumab, can reduce a tumor size by at least 25% within about 6 weeks of the first treatment. In certain aspects provided herein, the administration of MEDI4736, alone or in combination with Tremelimumab, can reduce a tumor size by at least 50%. In certain aspects provided herein, the administration of MEDI4736, alone or in combination with Tremelimumab, can reduce a tumor size by at least 50% within about 10 weeks of the first treatment. In certain aspects provided herein, the administration of MEDI4736, alone or in combination with Tremelimumab, can reduce a tumor size by at least 75%. In certain aspects provided herein, the administration of MEDI4736, alone or in combination with Tremelimumab, can reduce a tumor size by at least 75% within about 10 weeks of the first treatment.

[0086] In certain aspects, use of the methods provided herein, i.e., administration of MEDI4736, alone or in combination with Tremelimumab, or an antigen-binding fragment thereof can decrease tumor size within 6 weeks, within 7 weeks, within 8 weeks, within 9 weeks, within 10 weeks, within 12 weeks, within 16 weeks, within 20 weeks, within 24 weeks, within 28 weeks, within 32 weeks, within 36 weeks, within 40 weeks, within 44 weeks, within 48 weeks, or within 52 weeks of the first treatment.

[0087] In some embodiments, administration of 1 mg/kg of MEDI4736, alone or in combination with Tremelimumab, or an antigen-binding fragment thereof (e.g., at least one dose, at least two doses, 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 more every two weeks or every three weeks) can be sufficient to reduce tumor size. However, as provided herein, larger doses can also be administered, for example, to optimize efficacy, number of doses necessary, or certain pharmacokinetic parameters.

[0088] The methods provided herein can decrease or retard tumor growth. In some aspects the reduction or retardation can be statistically significant. A reduction in tumor growth can be measured by comparison to the growth of patient's tumor at baseline, against an expected tumor growth, against an expected tumor growth based on a large patient population, or against the tumor growth of a control population.

[0089] In certain aspects, a patient achieves disease control (DC). Disease control can be a complete response (CR), partial response (PR), or stable disease (SD).

[0090] A complete response (CR) refers to the disappearance of all lesions, whether measurable or not, and no new lesions. Confirmation can be obtained using a repeat, consecutive assessment no less than four weeks from the date of first documentation. New, non-measurable lesions preclude CR.

[0091] A partial response (PR) refers to a decrease in tumor burden 50% relative to baseline. Confirmation can be obtained using a consecutive repeat assessment at least 4 weeks from the date of first documentation

[0092] Progressive disease (PD) refers to an increase in tumor burden 25% relative to the minimum recorded (nadir). Confirmation can be obtained by a consecutive repeat assessment at least 4 weeks from the date of first documentation. New, non-measurable lesions do not define PD.

[0093] Stable disease (SD) refers to not meeting the criteria for CR, PR, or PD.

[0094] In certain aspects, administration of MEDI4736, alone or in combination with Tremelimumab, or an antigen-binding fragment thereof can increase progression-free survival (PFS).

[0095] In certain aspects, administration of MEDI4736, alone or in combination with Tremelimumab, or an antigen-binding fragment thereof can increase overall survival (OS).

[0096] According to the methods provided herein, administration of MEDI4736, alone or in combination with Tremelimumab, or an antigen-binding fragment thereof can result in desirable pharmacokinetic parameters. Total drug exposure can be estimated using the area under the curve (AUC). AUC (tau) refers to AUC until the end of the dosing period, whereas AUC (inf) refers to the AUC until infinite time. The administration can produce AUC (tau) of about 100 to about 2,500 d.Math.g/mL. The administration can produce a maximum observed concentration (Cmax) of about 15 to about 350 g/mL. The half-life of the MEDI4736 or the antigen-binding fragment thereof can be about 5 to about 25 days. In addition, the clearance of the MEDI4736 or the antigen-binding fragment thereof can be about 1-10 ml/day/kg.

[0097] As provided herein, MEDI4736 or an antigen-binding fragment thereof can also decrease free B7-H1 levels. Free B7-H1 refers to B7-H1 that is not bound (e.g., by MEDI4736). In some embodiments, B7-H1 levels are reduced by at least 80%. In some embodiments, B7-H1 levels are reduced by at least 90%. In some embodiments, B7-H1 levels are reduced by at least 95%. In some embodiments, B7-H1 levels are reduced by at least 99%. In some embodiments, B7-H1 levels are eliminated following administration of MEDI4736 or an antigen-binding fragment thereof. In some embodiments, administration of MEDI4736 or an antigen-binding fragment thereof reduces the rate of increase of B7-H1 levels as compared, e.g., to the rate of increase of B7-H1 levels prior to the administration of MEDI4736 or an antigen-binding fragment thereof.

Kits

[0098] The invention provides kits for characterizing the responsiveness of a subject to anti-PD-L1 antibody treatment or treatment with a combination of the invention comprising an anti-PD-L1 antibody and an anti-CTLA4 antibody. In one embodiment, the kit includes a therapeutic composition containing an effective amount of an antibody that specifically binds a PD-L1 polypeptide in unit dosage form alone or in combination with an anti-CTLA4 antibody.

[0099] A diagnostic kit of the invention provides a reagent (e.g., TaqMan primers/probes for an IFNgamma polynucleotide and housekeeping reference genes) for measuring relative expression of an IFNgamma polynucleotide.

[0100] In some embodiments, the kit comprises a sterile container which contains a therapeutic and/or diagnostic composition; such containers can be boxes, ampoules, bottles, vials, tubes, bags, pouches, blister-packs, or other suitable container forms known in the art. Such containers can be made of plastic, glass, laminated paper, metal foil, or other materials suitable for holding medicaments.

[0101] In one embodiment, a kit of the invention comprises reagents for measuring an IFNgamma polynucleotide expression and a therapeutic anti-PD-L1 antibody or a combination of an anti-PD-L1 antibody and an anti-CTLA4 antibody. If desired, the kit further comprises instructions for measuring an IFNgamma polynucleotide expression and/or instructions for administering the anti-PD-L1 antibody or a combination of an anti-PD-L1 antibody and an anti-CTLA4 antibody to a subject having a lung cancer (e.g., squamous cell or non-squamous cell carcinoma non-small cell lung cancer) selected as responsive to anti-PD-L1 antibody treatment or treatment with a combination of the invention comprising an anti-PD-L1 antibody and an anti-CTLA4 antibody. In particular embodiments, the instructions include at least one of the following: description of the therapeutic agent; dosage schedule and administration for treatment or prevention of lung cancer (e.g., non-small cell lung cancer, small cell lung cancer) or symptoms thereof; precautions; warnings; indications; counter-indications; over dosage information; adverse reactions; animal pharmacology; clinical studies; and/or references. The instructions may be printed directly on the container (when present), or as a label applied to the container, or as a separate sheet, pamphlet, card, or folder supplied in or with the container.

[0102] The practice of the present invention employs, unless otherwise indicated, conventional techniques of molecular biology (including recombinant techniques), microbiology, cell biology, biochemistry and immunology, which are well within the purview of the skilled artisan. Such techniques are explained fully in the literature, such as, Molecular Cloning: A Laboratory Manual, second edition (Sambrook, 1989); Oligonucleotide Synthesis (Gait, 1984); Animal Cell Culture (Freshney, 1987); Methods in Enzymology Handbook of Experimental Immunology (Weir, 1996); Gene Transfer Vectors for Mammalian Cells (Miller and Calos, 1987); Current Protocols in Molecular Biology (Ausubel, 1987); PCR: The Polymerase Chain Reaction, (Mullis, 1994); Current Protocols in Immunology (Coligan, 1991). These techniques are applicable to the production of the polynucleotides and polypeptides of the invention, and, as such, may be considered in making and practicing the invention. Particularly useful techniques for particular embodiments will be discussed in the sections that follow.

[0103] The following examples are put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the assay, screening, and therapeutic methods of the invention, and are not intended to limit the scope of what the inventors regard as their invention.

EXAMPLES

Example 1: Interferon Gamma mRNA in Blood Predicts Clinical Response to Anti-PD-L1 Antibody Treatment in Squamous Cell Carcinomaa Non-Small Cell Lung Cancer (SCC NSCLC)

[0104] Interferon gamma (IFN) mRNA in blood showed a positive trend that indicated that it likely predicts clinical response to anti-PD-L1 antibody treatment in squamous cell carcinoma non-small cell lung cancer (SCC NSCLC). Baseline levels of interferon gamma expression in blood and tumor are shown in FIGS. 1 and 2. Levels of interferon gamma mRNA correlate in blood and tumor indicating that either can be used to evaluate clinical responsiveness to anti-PD-L1 antibody treatment. Patients that respond to anti-PD-L1 therapy have higher interferon gamma expression in blood and tumor than SCC NSCLC patients who do not respond to anti-PD-L1 therapy. The trend with tumor shrinkage is also prevalent among those patients with higher interferon gamma expression in both blood and tumor. FIG. 3 shows that levels of interferon gamma mRNA in tumor and in blood had a modest correlation. In sum, IFN mRNA in blood shows positive predictive value for anti-PD-L1 antibody (MEDI4736) clinical response in SCC NSCLC.

Example 2: IFN mRNA is Predictive of Clinical Response for Anti-PD-L1 and Anti-CTLA4 Combination Therapy in Non-SCC NSCLC

[0105] Nineteen patients have baseline high quality IFN mRNA to evaluate for predictive value. Eighteen patients have non squamous cell carcinoma non-small cell lung cancer (NSCC NSCLC, FIG. 4). The remaining patient does not have histology information available. Similar bimodal distribution of interferon gamma mRNA is observed in non-SCC NSCLC, similar to that observed in patients treated with anti-PD-L1 for NSCLC of the same histology. Of the nine non-squamous cell non-small cell lung cancer patients that are interferon gamma mRNA high, five have evaluable clinical outcome and three are partially responsive (PRs). One, three or ten mg/kg (mpk) of an anti-CTLA4 antibody (tremelimumab treme) was administered in combination with ten, fifteen, or twenty mg/kg of an anti-PD-L1 antibody (MEDI4736) (bottom panel). Doses were administered at two or four week intervals. Interestingly, three out of five partially responsive patients were interferon gamma positive at baseline while only one in nine partially responsive patients was interferon gamma negative (FIG. 5). In sum, IFN mRNA is predictive of clinical response for anti-PD-L1 and anti-CTLA4 combination therapy in non-SCC NSCLC.

Other Embodiments

[0106] From the foregoing description, it will be apparent that variations and modifications may be made to the invention described herein to adopt it to various usages and conditions. Such embodiments are also within the scope of the following claims.

[0107] The recitation of a listing of elements in any definition of a variable herein includes definitions of that variable as any single element or combination (or subcombination) of listed elements. The recitation of an embodiment herein includes that embodiment as any single embodiment or in combination with any other embodiments or portions thereof.

[0108] All patents and publications mentioned in this specification are herein incorporated by reference to the same extent as if each independent patent and publication was specifically and individually indicated to be incorporated by reference.

TABLE-US-00014 Sequencesforanillustrativeexampleananti- PDL1antibody,MEDI4736. >PCT/US2010/058007_77Sequence77fromPCT/US2010/ 058007Organism:Homosapiens EIVLTQSPGTLSLSPGERATLSCRASQRVSSSYLAWYQQKPGQAPRLLIY DASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSLPWTFG QGTKVEIK >PCT/US2010/058007_72Sequence72fromPCT/US2010/ 058007Organism:Homosapiens EVQLVESGGGLVQPGGSLRLSCAASGFTFSRYWMSWVRQAPGKGLEWVAN IKQDGSEKYYVDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAREG GWFGELAFDYWGQGTLVTVSS MEDI4736VHCDR1 GFTFSRYWMS MEDI4736VHCDR2 NIKQDGSEKYYVDSVKG MEDI4736VHCDR3 EGGWFGELAFDY MEDI4736VLCDR1 RASQRVSSSYLA MEDI4736VLCDR2 DASSRAT MEDI4736VLCDR3 QQYGSLPWT