Antibody molecule against PD-L1 and its use

Abstract

To consider the importance of an immune checkpoint route in regulating an immune response and provide a novel agent that modulates the activity of immunoinhibitory proteins, such as PD-L1, thus leading to activation of the immune system.SOLUTION: The present disclosure provides use of an isolated antibody molecule that can bind to human Programmed Death Ligand 1 (PD-L1) in a combination with an AKT inhibitor, in the production of a pharmaceutical to treat cancer or infectious disease in a subject.SELECTED DRAWING: Figure 3

Description

Technical Field

[0001] Cross - Reference to Related Applications This application claims the benefit of U.S. Provisional Patent Application No. 62 / 063,852, filed October 14, 2014; U.S. Provisional Patent Application No. 62 / 094,847, filed December 19, 2014; U.S. Provisional Patent Application No. 62 / 198,545, filed July 29, 2015; and U.S. Provisional Patent Application No. 62 / 213,076, filed September 1, 2015, the entire contents of which are hereby incorporated by reference herein.

[0002] Sequence Listing This application contains a Sequence Listing that has been submitted electronically in ASCII format and is hereby incorporated by reference herein in its entirety. The ASCII copy created on October 13, 2015, is named C2160 - 7003WO_SL.txt and is 310,216 bytes in size.

Background Art

[0003] The ability of T cells to mediate an immune response against an antigen requires two distinct signaling interactions [Viglietta, V. et al. (2007) Neurotherapeutics 4:666 - 675; Korman, A. J. et al. (2007) Adv. Immunol. 90:297 - 339]. First, an antigen arrayed on the surface of an antigen - presenting cell (APC) is presented to antigen - specific naive CD4 + T cells. Such presentation transmits a signal via the T - cell receptor (TCR), thereby directing the T cell to mount an immune response specific for the presented antigen. Second, various co - stimulatory and inhibitory signals mediated by the interaction between the APC and distinct T - cell surface molecules induce T - cell activation and proliferation and ultimately its inhibition.

[0004] The immune system is tightly regulated by a network of co-stimulatory and co-inhibitory ligands and receptors. These molecules provide a second signal for T cell activation, resulting in a network with a balanced positive and negative signal that limits autoimmunity while maximizing the immune response to infection [Wang, L. et al. (Epub Mar. 7, 2011) J. Exp. Med. 208(3):577-92; Lepenies, B. et al. (2008) Endocrine, Metabolic & Immune Disorders--Drug Targets 8:279-288]. Examples of co-stimulatory signals include the binding between the B7.1 (CD80) and B7.2 (CD86) ligands of APCs and the CD28 and CTLA-4 receptors of CD4 + T lymphocytes [Sharpe, A. H. et al. (2002) Nature Rev. Immunol. 2:116-126; Lindley, P. S. et al. (2009) Immunol. Rev. 229:307-321]. Binding of B7.1 or B7.2 to CD28 stimulates T cell activation, while binding of B7.1 or B7.2 to CTLA-4 inhibits such activation [Dong, C. et al. (2003) Immunolog. Res. 28(1):39-48; Greenwald, R. J. et al. (2005) Ann. Rev. Immunol. 23:515-548]. CD28 is constitutively expressed on the surface of T cells [Gross, J., et al. (1992) J. Immunol. 149:380-388], while CTLA-4 expression is rapidly upregulated after T cell activation [Linsley, P. et al. (1996) Immunity 4:535-543].

[0005] Other ligands of the CD28 receptor include a group of related B7 molecules, also known as the "B7 superfamily" [Coyle, A. J. et al. (2001) Nature Immunol. 2(3):203-209; Sharpe, A. H. et al. (2002) Nature Rev. Immunol. 2:116-126; Collins, M. et al. (2005) Genome Biol. 6:223.1-223.7; Korman, A. J. et al. (2007) Adv. Immunol. 90:297-339]. Several members of the B7 superfamily are known, including B7.1 (CD80), B7.2 (CD86), inducible costimulatory ligand (ICOS-L), programmed death-1 ligand (PD-L1; B7-H1), programmed death-2 ligand (PD-L2; B7-DC), B7-H3, B7-H4 and B7-H6 [Collins, M. et al. (2005) Genome Biol. 6:223.1-223.7].

[0006] Programmed death 1 (PD-1) protein is an inhibitory member of the extended CD28 / CTLA-4 family of T cell regulators [Okazaki et al. (2002) Curr Opin Immunol 14: 391779-82; Bennett et al. (2003) J. Immunol. 170:711-8]. Other members of the CD28 family include CD28, CTLA-4, ICOS and BTLA. Two cell surface glycoprotein ligands for PD-1, Programmed Death ligand 1 (PD-L1) and Programmed Death ligand 2 (PD-L2), have been identified. PD-L1 and PD-L2 have been shown to downregulate T cell activation and cytokine secretion after binding to PD-1 [Freeman et al. (2000) J Exp Med 192:1027-34; Latchman et al. (2001) Nat Immunol 2:261-8; Carter et al. (2002) Eur J Immunol 32:634-43; Ohigashi et al. (2005) Clin Cancer Res 11:2947-53].

[0007] PD-L1 (also known as surface antigen classification 274 (CD274) or B7 homolog 1 (B7-H1)) is a 40 kDa type I transmembrane protein. PD-L1 binds to its receptor, PD-1, which is found on activated T cells, B cells, and myeloid cells, and modulates activation or inhibition. Both PD-L1 and PD-L2 are B7 homologs that bind to PD-1 but not to CD28 or CTLA-4 [Blank et al. (2005) Cancer Immunol Immunother. 54:307-14]. The binding of PD-L1 to its receptor PD-1 on T cells transmits signals that inhibit TCR-mediated activation of IL-2 production and T cell proliferation. This mechanism involves inhibition of ZAP70 phosphorylation and its association with CD3ζ [Sheppard et al. (2004) FEBS Lett. 574:37-41]. PD-1 signaling attenuates the PKC-θ activation loop phosphorylation resulting from TCR signaling, which is required for activation of the transcription factors NF-κB and AP-1 and production of IL-2. PD-L1 also binds to the costimulatory molecule CD80 (B7-1) but not to CD86 (B7-2) [Butte et al. (2008) Mol Immunol. 45:3567-72].

[0008] The expression of PD-L1 on the cell surface has been shown to be upregulated by IFN-γ stimulation. PD-L1 expression has been found in many cancers, including human lung, ovarian and colon cancers, as well as various myelomas, and is often associated with poor prognosis [Iwai et al. (2002) PNAS 99:12293-7; Ohigashi et al. (2005) Clin Cancer Res 11:2947-53; Okazaki et al. (2007) Intern. Immun. 19:813-24; Thompson et al. (2006) Cancer Res. 66:3381-5]. PD-L1 has been suggested to play a role in tumor immunity by increasing apoptosis of antigen-specific T cell clones [Dong et al. (2002) Nat Med 8:793-800]. It has also been suggested that PD-L1 may be involved in intestinal mucosal inflammation and that inhibition of PD-L1 suppresses wasting diseases associated with colitis [Kanai et al. (2003) J Immunol 171:4156-63]. SUMMARY OF THE INVENTION PROBLEMS TO BE SOLVED BY THE INVENTION

[0009] Considering the importance of immune checkpoint pathways in the regulation of immune responses, there is a need to develop novel agents that modulate the activity of immune inhibitory proteins, such as PD-L1, and thereby bring about activation of the immune system. Such agents can be used, for example, in cancer immunotherapy and the treatment of other conditions such as chronic infections. MEANS FOR SOLVING THE PROBLEMS

[0010] Antibody molecules (e.g., humanized antibody molecules) that bind to programmed death-ligand 1 (PD-L1) with high affinity and specificity are disclosed herein. In certain embodiments, the anti-PD-L1 antibody molecules comprise novel combinations of framework regions (e.g., FW1, FW2, FW3, and / or FW4), e.g., novel combinations of heavy chain framework regions and / or light chain framework regions. Nucleic acid molecules, expression vectors, host cells, and methods for making the antibody molecules are also provided. Immunoconjugates, multispecific or bispecific antibody molecules, and pharmaceutical compositions comprising the antibody molecules are also provided. The anti-PD-L1 antibody molecules disclosed herein can be used (alone or in combination with other agents or therapeutic modalities) to treat, prevent, and / or diagnose disorders such as cancerous disorders (e.g., solid and soft tissue tumors) and infectious diseases (e.g., chronic infectious disorders or sepsis). Moreover, methods and compositions comprising combinations of two or more therapeutic agents selected from one, two, or all of the following categories (i)-(iii) are disclosed herein: (i) agents that enhance antigen presentation (e.g., tumor antigen presentation); (ii) agents that enhance effector cell responses (e.g., B cell and / or T cell activation and / or recruitment); or (iii) agents that reduce tumor immunosuppression. In certain embodiments, the combination comprises an inhibitor of PD-L1 (e.g., an anti-PD-L1 antibody molecule described herein). Thus, compositions and methods for detecting PD-L1 and methods for treating various disorders including cancer and / or infectious diseases using anti-PD-L1 antibody molecules, as well as combinations thereof, are disclosed herein.

[0011] Accordingly, in one aspect, the invention features antibody molecules (e.g., isolated or recombinant antibody molecules) having one or more of the following characteristics: (i) High affinity, e.g., at least about 10 7 M -1 , typically about 10 8 M -1 , more typically about 10 9 M -1 ~1010 M -1 binds to PD-L1, e.g., human PD-L1, with an equal or stronger affinity constant; (ii) does not substantially bind to CD28, CTLA-4, ICOS or BTLA; (iii) inhibits or reduces the binding of PD-L1 to a receptor, e.g., PD-1 or CD80 (B7-1), or both; (iv) specifically binds to an epitope in PD-L1 that is the same as or similar to the epitope recognized by an epitope, e.g., mouse monoclonal antibody BAP058 or chimeric antibody BAP058, e.g., BAP058-chi; (v) exhibits the same or similar binding affinity and / or specificity as any of BAP058-hum01, BAP058-hum02, BAP058-hum03, BAP058-hum04, BAP058-hum05, BAP058-hum06, BAP058-hum07, BAP058-hum08, BAP058-hum09, BAP058-hum10, BAP058-hum11, BAP058-hum12, BAP058-hum13, BAP058-hum14, BAP058-hum15, BAP058-hum16, BAP058-hum17, BAP058-clone-K, BAP058-clone-L, BAP058-clone-M, BAP058-clone-N, or BAP058-clone-O; (vi) exhibits the same or similar binding affinity and / or specificity as the antibody molecules (e.g., heavy chain variable region and light chain variable region) described in Table 1; (vii) exhibits the same or similar binding affinity and / or specificity as the antibody molecules (e.g., heavy chain variable region and light chain variable region) having the amino acid sequences shown in Table 1; (viii) exhibits the same or similar binding affinity and / or specificity as the antibody molecules (e.g., heavy chain variable region and light chain variable region) encoded by the nucleotide sequences shown in Table 1; (ix) inhibiting the binding of a second antibody molecule to PD-L1, for example, competitively inhibiting, where this second antibody molecule is an antibody molecule selected, for example, from any of the antibody molecules described herein, such as BAP058-hum01, BAP058-hum02, BAP058-hum03, BAP058-hum04, BAP058-hum05, BAP058-hum06, BAP058-hum07, BAP058-hum08, BAP058-hum09, BAP058-hum10, BAP058-hum11, BAP058-hum12, BAP058-hum13, BAP058-hum14, BAP058-hum15, BAP058-hum16, BAP058-hum17, BAP058-clone-K, BAP058-clone-L, BAP058-clone-M, BAP058-clone-N, or BAP058-clone-O;

[0012] (x) binding to the same or overlapping epitopes as a second antibody molecule to PD-L1, where this second antibody molecule is an antibody molecule selected, for example, from any of the antibody molecules described herein, such as BAP058-hum01, BAP058-hum02, BAP058-hum03, BAP058-hum04, BAP058-hum05, BAP058-hum06, BAP058-hum07, BAP058-hum08, BAP058-hum09, BAP058-hum10, BAP058-hum11, BAP058-hum12, BAP058-hum13, BAP058-hum14, BAP058-hum15, BAP058-hum16, BAP058-hum17, BAP058-clone-K, BAP058-clone-L, BAP058-clone-M, BAP058-clone-N, or BAP058-clone-O; (xi) Competing with respect to binding to a second antibody molecule against PD-L1 and / or binding to the same epitope as it, and this second antibody molecule is an antibody molecule selected, for example, from any of the antibody molecules described herein, such as BAP058-hum01, BAP058-hum02, BAP058-hum03, BAP058-hum04, BAP058-hum05, BAP058-hum06, BAP058-hum07, BAP058-hum08, BAP058-hum09, BAP058-hum10, BAP058-hum11, BAP058-hum12, BAP058-hum13, BAP058-hum14, BAP058-hum15, BAP058-hum16, BAP058-hum17, BAP058-clone-K, BAP058-clone-L, BAP058-clone-M, BAP058-clone-N, or BAP058-clone-O; (xii) Having one or more biological properties of an antibody molecule selected, for example, from any of the antibody molecules described herein, such as BAP058-hum01, BAP058-hum02, BAP058-hum03, BAP058-hum04, BAP058-hum05, BAP058-hum06, BAP058-hum07, BAP058-hum08, BAP058-hum09, BAP058-hum10, BAP058-hum11, BAP058-hum12, BAP058-hum13, BAP058-hum14, BAP058-hum15, BAP058-hum16, BAP058-hum17, BAP058-clone-K, BAP058-clone-L, BAP058-clone-M, BAP058-clone-N, or BAP058-clone-O; (xiii) having one or more pharmacokinetic properties of the antibody molecules described herein, for example, selected from any of BAP058-hum01, BAP058-hum02, BAP058-hum03, BAP058-hum04, BAP058-hum05, BAP058-hum06, BAP058-hum07, BAP058-hum08, BAP058-hum09, BAP058-hum10, BAP058-hum11, BAP058-hum12, BAP058-hum13, BAP058-hum14, BAP058-hum15, BAP058-hum16, BAP058-hum17, BAP058-clone-K, BAP058-clone-L, BAP058-clone-M, BAP058-clone-N, or BAP058-clone-O; (xiv) inhibiting one or more activities of PD-L1, for example, resulting in one or more of the following: an increase in tumor-infiltrating lymphocytes, an increase in T cell receptor-mediated proliferation, or a decrease in immune evasion by cancerous cells; or (xv) binding to human PD-L1 and being cross-reactive with cynomolgus PD-L1.

[0013] In certain embodiments, the antibody molecule binds to PD-L1 with high affinity, for example, with a K D that is approximately the same as or at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% higher or lower than the K D of a murine or chimeric anti-PD-L1 antibody molecule, for example, a murine or chimeric anti-PD-L1 antibody molecule described herein. In certain embodiments, the K D of the murine or chimeric anti-PD-L1 antibody molecule is less than about 0.4, 0.3, 0.2, 0.1, or 0.05 nM, as measured by, for example, the Biacore method. In certain embodiments, the K D of the murine or chimeric anti-PD-L1 antibody molecule is less than about 0.2 nM, for example, about 0.171 nM. In other embodiments, the K Dis less than about 10, 5, 3, 2 or 1 nM, as measured by binding in cells expressing PD-L1 (e.g., 300.19 cells). In certain embodiments, the K D of a murine or chimeric anti-PD-L1 antibody molecule is less than about 1 nM, e.g., about 0.285 nM.

[0014] In certain embodiments, the anti-PD-L1 antibody molecule binds to PD-L1 with a K -4 that is slower than 1×10 -5 , 5×10 -5 s -1 , e.g., about 6.33×10 -5 s -1 . In certain embodiments, the anti-PD-L1 antibody molecule binds to PD-L1 with a K d that is faster than 1×10 4 , 5×10 4 , 1×10 5 or 5×10 5 M -1 s -1 , e.g., about 3.07×10 5 M -1 s -1 . a

[0015] In certain embodiments, the expression level of the antibody molecule is higher than that of a murine or chimeric antibody molecule, e.g., a murine or chimeric anti-PD-L1 antibody molecule described herein, e.g., at least about 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10-fold higher. In certain embodiments, the antibody molecule is expressed in CHO cells.

[0016] In certain embodiments, the anti-PD-L1 antibody molecule has an IC 50 that is approximately the same as or lower than that of a murine or chimeric anti-PD-L1 antibody molecule, e.g., a murine or chimeric anti-PD-L1 antibody molecule described herein, e.g., at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90% lower IC 50 ​(At the concentration for 50% inhibition), it reduces one or more PD-L1-related activities. In certain embodiments, the IC 50 of a murine or chimeric anti-PD-L1 antibody molecule is less than about 6, 5, 4, 3, 2, or 1 nM, as measured by binding in cells expressing PD-L1 (e.g., 300.19 cells). In certain embodiments, the IC 50 of a murine or chimeric anti-PD-L1 antibody molecule is less than about 4 nM, e.g., about 3.40 nM (or about 0.51 μg / mL). In certain embodiments, the PD-L1-related activity that is reduced is the binding of PD-L1 and / or PD-L2 to PD-1. In certain embodiments, the anti-PD-L1 antibody molecule binds to peripheral blood mononuclear cells (PBMCs) activated by Staphylococcus enterotoxin B (SEB). In other embodiments, the anti-PD-L1 antibody molecule increases the expression of IL-2 in whole blood activated by SEB. For example, the anti-PD-L1 antibody increases the expression of IL-2 by at least about 2, 3, 4, or 5-fold compared to the expression of IL-2 when an isotype control (e.g., IgG4) is used.

[0017] In certain embodiments, the anti-PD-L1 antibody molecule has improved stability, e.g., is at least about 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10-fold more stable in vivo or in vitro than a murine or chimeric anti-PD-L1 antibody molecule, e.g., a murine or chimeric anti-PD-L1 antibody molecule described herein.

[0018] In certain embodiments, the anti-PD-L1 antibody molecule is a humanized antibody molecule and has a risk score based on T cell epitope analysis of 300 - 700, 400 - 650, 450 - 600 or a risk score described herein.

[0019] In another aspect, the anti-PD-L1 antibody molecule is an antibody selected from the antibodies described herein, such as any of BAP058-hum01, BAP058-hum02, BAP058-hum03, BAP058-hum04, BAP058-hum05, BAP058-hum06, BAP058-hum07, BAP058-hum08, BAP058-hum09, BAP058-hum10, BAP058-hum11, BAP058-hum12, BAP058-hum13, BAP058-hum14, BAP058-hum15, BAP058-hum16, BAP058-hum17, BAP058-clone-K, BAP058-clone-L, BAP058-clone-M, BAP058-clone-N, or BAP058-clone-O; or an antibody described in Table 1 or an antibody encoded by the nucleotide sequence in Table 1; or at least one antigen-binding region derived from a sequence that is substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or more identical) to any of the foregoing sequences, such as its variable region or an antigen-binding fragment.

[0020] In another aspect, the anti-PD-L1 antibody molecule is an antibody selected from the antibodies described herein, such as BAP058-hum01, BAP058-hum02, BAP058-hum03, BAP058-hum04, BAP058-hum05, BAP058-hum06, BAP058-hum07, BAP058-hum08, BAP058-hum09, BAP058-hum10, BAP058-hum11, BAP058-hum12, BAP058-hum13, BAP058-hum14, BAP058-hum15, BAP058-hum16, BAP058-hum17, BAP058-clone-K, BAP058-clone-L, BAP058-clone-M, BAP058-clone-N or BAP058-clone-O; or an antibody described in Table 1 or an antibody encoded by the nucleotide sequence in Table 1; or at least 1, 2, 3 or 4 variable regions derived from a sequence that is substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or more identical) to any of the foregoing sequences.

[0021] In another aspect, the anti-PD-L1 antibody molecule is an antibody selected from the antibodies described herein, such as BAP058-hum01, BAP058-hum02, BAP058-hum03, BAP058-hum04, BAP058-hum05, BAP058-hum06, BAP058-hum07, BAP058-hum08, BAP058-hum09, BAP058-hum10, BAP058-hum11, BAP058-hum12, BAP058-hum13, BAP058-hum14, BAP058-hum15, BAP058-hum16, BAP058-hum17, BAP058-clone-K, BAP058-clone-L, BAP058-clone-M, BAP058-clone-N or BAP058-clone-O; or an antibody described in Table 1 or an antibody encoded by the nucleotide sequence in Table 1; or at least one or two heavy chain variable regions derived from a sequence that is substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or more identical) to any of the foregoing sequences.

[0022] In another aspect, the anti-PD-L1 antibody molecule is an antibody selected from the antibodies described herein, such as BAP058-hum01, BAP058-hum02, BAP058-hum03, BAP058-hum04, BAP058-hum05, BAP058-hum06, BAP058-hum07, BAP058-hum08, BAP058-hum09, BAP058-hum10, BAP058-hum11, BAP058-hum12, BAP058-hum13, BAP058-hum14, BAP058-hum15, BAP058-hum16, BAP058-hum17, BAP058-clone-K, BAP058-clone-L, BAP058-clone-M, BAP058-clone-N or BAP058-clone-O; or an antibody described in Table 1 or an antibody encoded by the nucleotide sequence in Table 1; or at least one or two light chain variable regions derived from a sequence that is substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or more identical) to any of the foregoing sequences.

[0023] In yet another aspect, the anti-PD-L1 antibody molecule comprises an IgG4, for example, the heavy chain constant region of human IgG4. In one aspect, the human IgG4 comprises a substitution at position 228 (e.g., a substitution from Ser to Pro). In yet another aspect, the anti-PD-L1 antibody molecule comprises an IgG1, for example, the heavy chain constant region of human IgG1. In one aspect, the human IgG1 comprises a substitution at position 297 (e.g., a substitution from Asn to Ala). In one aspect, the human IgG1 comprises a substitution at position 265, a substitution at position 329, or both (e.g., a substitution from Asp to Ala at position 265 and / or a substitution from Pro to Ala at position 329). In one aspect, the human IgG1 comprises a substitution at position 234, a substitution at position 235, or both (e.g., a substitution from Leu to Ala at position 234 and / or a substitution from Leu to Ala at position 235). In one aspect, the heavy chain constant region comprises the amino acid sequence set forth in Table 3, or a sequence that is substantially identical thereto (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or more identical).

[0024] In yet another aspect, the anti-PD-L1 antibody molecule comprises a kappa light chain constant region, for example, the human kappa light chain constant region. In one aspect, the light chain constant region comprises the amino acid sequence set forth in Table 3, or a sequence that is substantially identical thereto (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or more identical).

[0025] In another aspect, the anti-PD-L1 antibody molecule comprises heavy and light chain constant regions comprising IgG4, e.g., the heavy chain constant region of human IgG4 and the kappa light chain constant region, e.g., the human kappa light chain constant region, e.g., the amino acid sequence set forth in Table 3, or a sequence that is substantially identical thereto (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or more identical). In yet another aspect, the anti-PD-L1 antibody molecule comprises heavy and light chain constant regions comprising IgG1, e.g., the heavy chain constant region of human IgG1 and the kappa light chain constant region, e.g., the human kappa light chain constant region, e.g., the amino acid sequence set forth in Table 3, or a sequence that is substantially identical thereto (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or more identical). In one aspect, human IgG1 comprises a substitution at position 297 (e.g., substitution from Asn to Ala). In one aspect, human IgG1 comprises a substitution at position 265, a substitution at position 329 or both (e.g., substitution from Asp to Ala at position 265 and / or substitution from Pro to Ala at position 329). In one aspect, human IgG1 comprises a substitution at position 234, a substitution at position 235 or both (e.g., substitution from Leu to Ala at position 234 and / or substitution from Leu to Ala at position 235).

[0026] In another aspect, the anti-PD-L1 antibody molecule comprises a heavy chain variable domain and a constant region, a light chain variable domain and a constant region or both, comprising the amino acid sequence of BAP058-clone-K, BAP058-clone-L, BAP058-clone-M, BAP058-clone-N or BAP058-clone-O; or the amino acid sequence described in Table 1 or the amino acid sequence encoded by the nucleotide sequence in Table 1; or a sequence that is substantially identical to any of the foregoing sequences (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or more identical). The anti-PD-1 antibody molecule may comprise a leader sequence derived from the heavy chain, the light chain or both.

[0027] In another aspect, the anti-PD-L1 antibody molecule is an antibody selected from the antibodies described herein, such as BAP058-hum01, BAP058-hum02, BAP058-hum03, BAP058-hum04, BAP058-hum05, BAP058-hum06, BAP058-hum07, BAP058-hum08, BAP058-hum09, BAP058-hum10, BAP058-hum11, BAP058-hum12, BAP058-hum13, BAP058-hum14, BAP058-hum15, BAP058-hum16, BAP058-hum17, BAP058-clone-K, BAP058-clone-L, BAP058-clone-M, BAP058-clone-N, or BAP058-clone-O; or an antibody described in Table 1 or an antibody encoded by the nucleotide sequence in Table 1; or at least 1, 2, or 3 complementarity-determining regions (CDRs) derived from the heavy-chain variable region of a sequence that is substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or more identical) to any of the foregoing sequences.

[0028] In another aspect, the anti-PD-L1 antibody molecule comprises at least 1, 2, or 3 CDRs (or all CDRs together) derived from the heavy-chain variable region comprising the amino acid sequence shown in Table 1 or the amino acid sequence encoded by the nucleotide sequence shown in Table 1. In one aspect, one or more (or all CDRs together) of the CDRs have 1, 2, 3, 4, 5, 6 or more changes, e.g., amino acid substitutions or deletions, compared to the amino acid sequence shown in Table 1 or the amino acid sequence encoded by the nucleotide sequence shown in Table 1.

[0029] In another aspect, the anti-PD-L1 antibody molecule is an antibody selected from the antibodies described herein, such as BAP058-hum01, BAP058-hum02, BAP058-hum03, BAP058-hum04, BAP058-hum05, BAP058-hum06, BAP058-hum07, BAP058-hum08, BAP058-hum09, BAP058-hum10, BAP058-hum11, BAP058-hum12, BAP058-hum13, BAP058-hum14, BAP058-hum15, BAP058-hum16, BAP058-hum17, BAP058-clone-K, BAP058-clone-L, BAP058-clone-M, BAP058-clone-N, or BAP058-clone-O; or an antibody described in Table 1 or an antibody encoded by the nucleotide sequence in Table 1; or at least 1, 2, or 3 CDRs derived from the light chain variable region of a sequence that is substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or more identical) to any of the foregoing sequences.

[0030] In another aspect, the anti-PD-L1 antibody molecule comprises at least 1, 2, or 3 CDRs (or all CDRs together) derived from the light chain variable region comprising the amino acid sequence shown in Table 1 or the amino acid sequence encoded by the nucleotide sequence shown in Table 1. In one aspect, one or more (or all CDRs together) of the CDRs have 1, 2, 3, 4, 5, 6 or more changes, such as amino acid substitutions or deletions, compared to the amino acid sequence shown in Table 1 or the amino acid sequence encoded by the nucleotide sequence shown in Table 1. In a particular aspect, the anti-PD-L1 antibody molecule comprises substitutions in the light chain CDRs, e.g., one or more substitutions in CDR1, CDR2, and / or CDR3 of the light chain.

[0031] In another aspect, the anti-PD-L1 antibody molecule comprises at least 1, 2, 3, 4, 5, or 6 CDRs (or all CDRs together) derived from the heavy and light chain variable regions comprising the amino acid sequence shown in Table 1 or the amino acid sequence encoded by the nucleotide sequence shown in Table 1. In certain aspects, one or more (or all together) of the CDRs have 1, 2, 3, 4, 5, 6 or more changes, such as amino acid substitutions or deletions, compared to the amino acid sequence shown in Table 1 or the amino acid sequence encoded by the nucleotide sequence shown in Table 1.

[0032] In one aspect, the anti-PD-L1 antibody molecule is an antibody selected from the antibodies described herein, such as BAP058-hum01, BAP058-hum02, BAP058-hum03, BAP058-hum04, BAP058-hum05, BAP058-hum06, BAP058-hum07, BAP058-hum08, BAP058-hum09, BAP058-hum10, BAP058-hum11, BAP058-hum12, BAP058-hum13, BAP058-hum14, BAP058-hum15, BAP058-hum16, BAP058-hum17, BAP058-clone-K, BAP058-clone-L, BAP058-clone-M, BAP058-clone-N, or BAP058-clone-O; or all six CDRs derived from the antibodies described in Table 1 or antibodies encoded by the nucleotide sequences in Table 1, or related CDRs, such as CDRs that are identical or have at least one amino acid change, provided 2, 3, or 4 or fewer changes (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions). In one aspect, the anti-PD-L1 antibody molecule can comprise any of the CDRs described herein. In a particular aspect, the anti-PD-L1 antibody molecule comprises substitutions in the light chain CDRs, e.g., one or more substitutions in light chain CDR1, CDR2, and / or CDR3.In another aspect, the anti-PD-L1 antibody molecule is an antibody selected from the antibodies described herein, such as BAP058-hum01, BAP058-hum02, BAP058-hum03, BAP058-hum04, BAP058-hum05, BAP058-hum06, BAP058-hum07, BAP058-hum08, BAP058-hum09, BAP058-hum10, BAP058-hum11, BAP058-hum12, BAP058-hum13, BAP058-hum14, BAP058-hum15, BAP058-hum16, BAP058-hum17, BAP058-clone-K, BAP058-clone-L, BAP058-clone-M, BAP058-clone-N, or BAP058-clone-O; or an antibody described in Table 1 or an antibody encoded by the nucleotide sequence in Table 1; or a sequence that is substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or more identical) to any of the foregoing sequences; or a sequence having at least one amino acid change compared to 1, 2 or 3 CDRs according to Kabat et al. shown in Table 1, provided that there are 2, 3 or 4 or fewer changes (e.g., substitutions, deletions or insertions, e.g., conservative substitutions), and comprises at least 1, 2 or 3 CDRs (e.g., at least 1, 2 or 3 CDRs according to the Kabat definition presented in Table 1) derived from the heavy chain variable region of the sequence.

[0033] In another aspect, the anti-PD-L1 antibody molecule is an antibody selected from the antibodies described herein, such as BAP058-hum01, BAP058-hum02, BAP058-hum03, BAP058-hum04, BAP058-hum05, BAP058-hum06, BAP058-hum07, BAP058-hum08, BAP058-hum09, BAP058-hum10, BAP058-hum11, BAP058-hum12, BAP058-hum13, BAP058-hum14, BAP058-hum15, BAP058-hum16, BAP058-hum17, BAP058-clone-K, BAP058-clone-L, BAP058-clone-M, BAP058-clone-N, or BAP058-clone-O; or an antibody described in Table 1 or an antibody encoded by the nucleotide sequence in Table 1; or a sequence that is substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or more identical) to any of the foregoing sequences; or a sequence having at least one amino acid change compared to one, two or three CDRs according to Kabat et al. shown in Table 1, provided that there are two, three or four or fewer changes (e.g., substitutions, deletions or insertions, e.g., conservative substitutions), and comprises at least one, two or three CDRs (e.g., at least one, two or three CDRs according to the Kabat definition presented in Table 1) derived from the light chain variable region of the sequence.

[0034] In another aspect, the anti-PD-L1 antibody molecule is an antibody selected from the antibodies described herein, such as BAP058-hum01, BAP058-hum02, BAP058-hum03, BAP058-hum04, BAP058-hum05, BAP058-hum06, BAP058-hum07, BAP058-hum08, BAP058-hum09, BAP058-hum10, BAP058-hum11, BAP058-hum12, BAP058-hum13, BAP058-hum14, BAP058-hum15, BAP058-hum16, BAP058-hum17, BAP058-clone-K, BAP058-clone-L, BAP058-clone-M, BAP058-clone-N, or BAP058-clone-O; or an antibody described in Table 1 or an antibody encoded by the nucleotide sequence in Table 1; or a sequence that is substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or more identical) to any of the foregoing sequences; or at least one amino acid change compared to 1, 2, 3, 4, 5 or 6 CDRs according to Kabat et al. shown in Table 1, provided that there are 2, 3 or 4 or fewer changes (e.g., substitutions, deletions or insertions, e.g., conservative substitutions), and includes at least 1, 2, 3, 4, 5 or 6 CDRs (e.g., at least 1, 2, 3, 4, 5 or 6 CDRs according to the Kabat definition presented in Table 1) derived from the heavy and light chain variable regions of a sequence having such changes.

[0035] In another aspect, the anti-PD-L1 antibody molecule is an antibody selected from the antibodies described herein, such as BAP058-hum01, BAP058-hum02, BAP058-hum03, BAP058-hum04, BAP058-hum05, BAP058-hum06, BAP058-hum07, BAP058-hum08, BAP058-hum09, BAP058-hum10, BAP058-hum11, BAP058-hum12, BAP058-hum13, BAP058-hum14, BAP058-hum15, BAP058-hum16, BAP058-hum17, BAP058-clone-K, BAP058-clone-L, BAP058-clone-M, BAP058-clone-N, or BAP058-clone-O; or an antibody described in Table 1 or an antibody encoded by the nucleotide sequence in Table 1; or a sequence that is substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or more identical) to any of the foregoing sequences; or a sequence having at least one amino acid change compared to all 6 CDRs according to Kabat et al. shown in Table 1, provided that there are 2, 3 or 4 or fewer changes (e.g., substitutions, deletions or insertions, e.g., conservative substitutions), and includes all 6 CDRs according to Kabat et al. (e.g., all 6 CDRs according to the Kabat definition presented in Table 1) derived from the heavy and light chain variable regions of the sequence. In one aspect, the anti-PD-L1 antibody molecule can comprise any of the CDRs described herein.

[0036] In another aspect, the anti-PD-L1 antibody molecule is an antibody selected from the antibodies described herein, such as BAP058-hum01, BAP058-hum02, BAP058-hum03, BAP058-hum04, BAP058-hum05, BAP058-hum06, BAP058-hum07, BAP058-hum08, BAP058-hum09, BAP058-hum10, BAP058-hum11, BAP058-hum12, BAP058-hum13, BAP058-hum14, BAP058-hum15, BAP058-hum16, BAP058-hum17, BAP058-clone-K, BAP058-clone-L, BAP058-clone-M, BAP058-clone-N, or BAP058-clone-O; or an antibody described in Table 1 or an antibody encoded by the nucleotide sequence in Table 1; or at least the amino acids derived from the high-frequency variable loops that contact PD-L1; or at least 1, 2, or 3 Chothia high-frequency variable loops (e.g., at least 1, 2, or 3 high-frequency variable loops according to the Chothia definition presented in Table 1) derived from the heavy chain variable region of a sequence having at least 1 amino acid change compared to 1, 2, or 3 high-frequency variable loops according to Chothia et al. shown in Table 1, provided that there are 2, 3, or 4 or fewer changes (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions).

[0037] In another aspect, the anti-PD-L1 antibody molecule is an antibody selected from the antibodies described herein, such as BAP058-hum01, BAP058-hum02, BAP058-hum03, BAP058-hum04, BAP058-hum05, BAP058-hum06, BAP058-hum07, BAP058-hum08, BAP058-hum09, BAP058-hum10, BAP058-hum11, BAP058-hum12, BAP058-hum13, BAP058-hum14, BAP058-hum15, BAP058-hum16, BAP058-hum17, BAP058-clone-K, BAP058-clone-L, BAP058-clone-M, BAP058-clone-N, or BAP058-clone-O; or an antibody described in Table 1 or an antibody encoded by the nucleotide sequence in Table 1; or at least an amino acid derived from a highly variable loop that contacts PD-L1; or at least one amino acid change compared to 1, 2, or 3 highly variable loops according to Chothia et al. shown in Table 1, provided that there are 2, 3, or 4 or fewer changes (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions), and includes at least 1, 2, or 3 Chothia highly variable loops (e.g., at least 1, 2, or 3 highly variable loops according to the Chothia definition presented in Table 1) of the light chain variable region of a sequence having such changes.

[0038] In another aspect, the anti-PD-L1 antibody molecule is an antibody selected from the antibodies described herein, such as BAP058-hum01, BAP058-hum02, BAP058-hum03, BAP058-hum04, BAP058-hum05, BAP058-hum06, BAP058-hum07, BAP058-hum08, BAP058-hum09, BAP058-hum10, BAP058-hum11, BAP058-hum12, BAP058-hum13, BAP058-hum14, BAP058-hum15, BAP058-hum16, BAP058-hum17, BAP058-clone-K, BAP058-clone-L, BAP058-clone-M, BAP058-clone-N, or BAP058-clone-O; or an antibody described in Table 1 or an antibody encoded by the nucleotide sequence in Table 1; or at least the amino acids derived from the high-frequency variable loops that contact PD-L1; or at least one amino acid change compared to 1, 2, 3, 4, 5, or 6 high-frequency variable loops according to Chothia et al. shown in Table 1, provided that there are 2, 3, or 4 or fewer changes (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions), and includes at least 1, 2, 3, 4, 5, or 6 high-frequency variable loops (e.g., at least 1, 2, 3, 4, 5, or 6 high-frequency variable loops according to the Chothia definition presented in Table 1) derived from the heavy and light chain variable regions of a sequence having such changes.

[0039] In one aspect, the anti-PD-L1 antibody molecule is an antibody selected from any of the antibodies described herein, such as BAP058-hum01, BAP058-hum02, BAP058-hum03, BAP058-hum04, BAP058-hum05, BAP058-hum06, BAP058-hum07, BAP058-hum08, BAP058-hum09, BAP058-hum10, BAP058-hum11, BAP058-hum12, BAP058-hum13, BAP058-hum14, BAP058-hum15, BAP058-hum16, BAP058-hum17, BAP058-clone-K, BAP058-clone-L, BAP058-clone-M, BAP058-clone-N, or BAP058-clone-O, and includes all six high-frequency variable loops (e.g., all six high-frequency variable loops according to the Chothia definition presented in Table 1) of the antibody, or closely related high-frequency variable loops, e.g., loops that are identical or have at least one amino acid change, provided that the change is 2, 3, or 4 or fewer (e.g., substitution, deletion, or insertion, e.g., conservative substitution); or includes high-frequency variable loops having at least one amino acid change compared to all six high-frequency variable loops according to Chothia et al. shown in Table 1, provided that the change is 2, 3, or 4 or fewer (e.g., substitution, deletion, or insertion, e.g., conservative substitution). In one aspect, the anti-PD-L1 antibody molecule can include any of the high-frequency variable loops described herein.

[0040] In yet another aspect, the anti-PD-L1 antibody molecule comprises at least 1, 2, or 3 hypervariable loops having the same canonical structure as the corresponding hypervariable loops of an antibody selected from the antibodies described herein, such as BAP058-hum01, BAP058-hum02, BAP058-hum03, BAP058-hum04, BAP058-hum05, BAP058-hum06, BAP058-hum07, BAP058-hum08, BAP058-hum09, BAP058-hum10, BAP058-hum11, BAP058-hum12, BAP058-hum13, BAP058-hum14, BAP058-hum15, BAP058-hum16, BAP058-hum17, BAP058-clone-K, BAP058-clone-L, BAP058-clone-M, BAP058-clone-N, or BAP058-clone-O; for example, the same canonical structure as at least loop 1 and / or loop 2 of the heavy and / or light chain variable domains of the antibodies described herein. For a description of hypervariable loop canonical structures, see, for example, Chothia et al., (1992) J. Mol. Biol. 227:799-817; Tomlinson et al., (1992) J. Mol. Biol. 227:776-798. These structures can be determined by examining the tables described in these references.

[0041] In certain aspects, the anti-PD-L1 antibody molecule comprises a combination of CDRs or hypervariable loops defined according to Kabat et al. and Chothia et al.

[0042] In one aspect, the anti-PD-L1 antibody molecule comprises at least one, two or three CDRs or hypervariable loops (e.g., at least one, two or three CDRs or hypervariable loops according to the Kabat and Chothia definitions presented in Table 1) derived from the heavy chain variable region of an antibody selected from any of the antibodies described herein according to the Kabat and Chothia definitions, such as BAP058-hum01, BAP058-hum02, BAP058-hum03, BAP058-hum04, BAP058-hum05, BAP058-hum06, BAP058-hum07, BAP058-hum08, BAP058-hum09, BAP058-hum10, BAP058-hum11, BAP058-hum12, BAP058-hum13, BAP058-hum14, BAP058-hum15, BAP058-hum16, BAP058-hum17, BAP058-clone-K, BAP058-clone-L, BAP058-clone-M, BAP058-clone-N, or BAP058-clone-O; or a sequence encoded by the nucleotide sequence in Table 1; or a sequence that is substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or more identical) to any of the foregoing sequences; or a sequence having at least one amino acid change compared to one, two or three CDRs or hypervariable loops according to Kabat and / or Chothia shown in Table 1, provided that there are 2, 3 or 4 or fewer changes (e.g., substitutions, deletions or insertions, e.g., conservative substitutions).

[0043] For example, an anti-PD-L1 antibody molecule can include, for example, a VH CDR1 according to Kabat et al., a VH hypervariable loop 1 according to Chothia et al., or a combination thereof as shown in Table 1. In certain embodiments, a combination of the Kabat and Chothia CDRs of VH CDR1 comprises an amino acid sequence GYTFTSYWMY (SEQ ID NO: 195) or an amino acid sequence that is substantially identical thereto [e.g., having at least one amino acid change, but no more than 2, 3, or 4 changes (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions)]. The anti-PD-L1 antibody molecule can further include, for example, VH CDR2-3 according to Kabat et al. and VL CDR1-3 according to Kabat et al. as shown in Table 1. Thus, in certain embodiments, the framework regions are defined based on a combination of CDRs defined according to Kabat et al. and hypervariable loops defined according to Chothia et al. For example, an anti-PD-L1 antibody molecule can include a VH FR1 defined based on VH hypervariable loop 1 according to Chothia et al. and a VH FR2 defined based on VH CDR1-2 according to Kabat et al. as shown in Table 1. The anti-PD-L1 antibody molecule can further include VH FR3-4 defined based on VH CDR2-3 according to Kabat et al. and VL FR1-4 defined based on VL CDR1-3 according to Kabat et al.

[0044] An anti-PD-L1 antibody molecule can contain any combination of CDRs or hypervariable loops according to the Kabat and Chothia definitions. In certain embodiments, the anti-PD-L1 antibody molecule comprises at least 1, 2, or 3 CDRs (e.g., at least 1, 2, or 3 CDRs according to the Kabat and Chothia definitions presented in Table 1) derived from the light chain variable region of an antibody selected from any of the antibodies described herein according to the Kabat and Chothia definitions, such as BAP058-hum01, BAP058-hum02, BAP058-hum03, BAP058-hum04, BAP058-hum05, BAP058-hum06, BAP058-hum07, BAP058-hum08, BAP058-hum09, BAP058-hum10, BAP058-hum11, BAP058-hum12, BAP058-hum13, BAP058-hum14, BAP058-hum15, BAP058-hum16, BAP058-hum17, BAP058-clone-K, BAP058-clone-L, BAP058-clone-M, BAP058-clone-N, or BAP058-clone-O.

[0045] Aspects, for example, in the aspects including variable regions, CDRs (e.g., Chothia CDRs or Kabat CDRs) or other sequences as referred to herein, for example, in Table 1, the antibody molecule is a monospecific antibody molecule, a bispecific antibody molecule, or an antigen-binding fragment of an antibody, for example, a half antibody or an antigen-binding fragment of a half antibody. In an aspect, the antibody molecule is a bispecific antibody molecule having a first binding specificity for PD-L1 and a second binding specificity for TIM-3, LAG-3, CEACAM (e.g., CEACAM-1 and / or CEACAM-5), PD-1 or PD-L2. In an aspect, the second binding specificity for TIM-3, LAG-3 and / or PD-1 includes the amino acid sequences described herein or is encoded by the nucleotide sequences described herein [e.g., as disclosed in the section entitled "Inhibitors of Immune Checkpoint Molecules" starting on page 218 hereinafter (including any publications mentioned therein)].

[0046] In one aspect, the anti-PD-L1 antibody molecule is (i) a heavy chain variable region (VH) comprising a VHCDR1 amino acid sequence selected from SEQ ID NO: 1, SEQ ID NO: 4 or SEQ ID NO: 195, a VHCDR2 amino acid sequence of SEQ ID NO: 2, and a VHCDR3 amino acid sequence of SEQ ID NO: 3, and (ii) a light chain variable region (VL) comprising a VLCDR1 amino acid sequence of SEQ ID NO: 9, a VLCDR2 amino acid sequence of SEQ ID NO: 10, and a VLCDR3 amino acid sequence of SEQ ID NO: 11 comprising.

[0047] In another aspect, the anti-PD-L1 antibody molecule is (i) a heavy chain variable region (VH) comprising a VHCDR1 amino acid sequence selected from SEQ ID NO: 1, SEQ ID NO: 4 or SEQ ID NO: 195, a VHCDR2 amino acid sequence of SEQ ID NO: 5, and a VHCDR3 amino acid sequence of SEQ ID NO: 3, and (ii) A light chain variable region (VL) comprising the VLCDR1 amino acid sequence of SEQ ID NO: 12, the VLCDR2 amino acid sequence of SEQ ID NO: 13, and the VLCDR3 amino acid sequence of SEQ ID NO: 14 comprising.

[0048] In certain embodiments, the anti-PD-L1 antibody molecule comprises the VHCDR1 amino acid sequence of SEQ ID NO: 1. In another embodiment, the anti-PD-L1 antibody molecule comprises the VHCDR1 amino acid sequence of SEQ ID NO: 4. In yet another embodiment, the anti-PD-L1 antibody molecule comprises the VHCDR1 amino acid sequence of SEQ ID NO: 195.

[0049] In certain embodiments, the light or heavy chain variable framework of the anti-PD-L1 antibody molecule (e.g., the region encompassing at least FR1, FR2, FR3 and optionally FR4) is (a) an amino acid residue derived from a human light or heavy chain variable framework, e.g., a light or heavy chain variable framework residue from a human mature antibody, human germline sequence or human consensus sequence, comprising at least 80%, 85%, 87%, 90%, 92%, 93%, 95%, 97%, 98% or preferably 100% of the light or heavy chain variable framework; (b) an amino acid residue derived from a human light or heavy chain variable framework, e.g., a light or heavy chain variable framework residue from a human mature antibody, human germline sequence or human consensus sequence, comprising 20% - 80%, 40% - 60%, 60% - 90% or 70% - 95% of the light or heavy chain variable framework; (c) a non-human framework (e.g., a rodent framework); or (d) can be selected from, for example, a non-human framework modified to remove, e.g., antigenic or cytotoxic determinants, e.g., a deimmunized or partially humanized non-human framework. In certain embodiments, the light or heavy chain variable framework region (particularly FR1, FR2 and / or FR3) comprises a light or heavy chain variable framework sequence that is at least 70%, 75%, 80%, 85%, 87%, 88%, 90%, 92%, 94%, 95%, 96%, 97%, 98%, 99% identical to the framework of the VL or VH segment of a human germline gene.

[0050] In certain embodiments, the anti-PD-L1 antibody molecule comprises a heavy chain variable domain having, for example, at least 1, 2, 3, 4, 5, 6, 7, 10, 15, 20 or more changes, such as amino acid substitutions or deletions, from the amino acid sequence of BAP058-chi-HC shown in FIGS. 8A-8B or SEQ ID NO: 16, for example, from the amino acid sequences of the FR regions throughout the variable region. In some embodiments, the anti-PD-L1 antibody molecule comprises a heavy chain variable domain having one or more of the following: for example, Q at position 1, I at position 2, T at position 3, V or K at position 5, P at position 9, T at position 10, V at position 11, K at position 12, T at position 15, E or Q at position 16, T at position 17, L at position 18, R or T at position 19, I or V at position 20, T at position 21, T at position 23, G, V or F at position 24, I at position 37, R at position 38, A or P or S at position 40, T or R at position 41, S at position 42, Q or K at position 43, M or L or V at position 48, R at position 67, F or V or L at position 68, I at position 70, S at position 71, A, K or R at position 72, D or T or N at position 74, T or K at position 76, N at position 77, Q at position 78, F or V or L at position 79, S or V at position 80, L at position 81, E or K or T at position 82, M at position 83, T or N at position 84, N at position 85, V or M at position 86, K or R or D at position 87, T or A or P at position 88, A or V at position 89, T at position 91 or T at position 93, from the amino acid sequence of BAP058-chi-HC shown in FIGS. 8A-8B or SEQ ID NO: 16, for example, from the amino acid sequences of the FR regions throughout the variable region.

[0051] Alternatively, or in combination with the heavy chain replacement of BAP058-chi-HC described herein, the anti-PD-L1 antibody molecule comprises a light chain variable domain having at least 1, 2, 3, 4, 5, 6, 7, 10, 15, 20 or more amino acid changes, such as amino acid substitutions or deletions, from the amino acid sequence of BAP058-chi-LC, e.g., the amino acid sequence shown in FIGS. 9A-9B or SEQ ID NO: 17. In certain embodiments, the anti-PD-L1 antibody molecule comprises a heavy chain variable domain having one or more of the following: E or A at position 1, V at position 2, V or Q at position 3, L at position 4, T at position 7, P at position 8, D or L or S or A at position 9, S or T at position 10, Q or L at position 11, P at position 12, V or L or A at position 13, T at position 14, P or L at position 15, K at position 16, Q or E at position 17, K or P at position 18, A at position 19, T at position 20, L at position 21, S at position 22, L at position 37, A at position 43, R or Q at position 45, I at position 58, A or S or P at position 60, S at position 63, Y at position 67, E at position 70, F at position 73, K at position 74, N at position 76, S or R at position 77, I or L at position 78, E at position 79, P or A at position 80, D at position 81, F or I or V or A at position 83, G at position 84, T or V or Y at position 85 or Y at position 87 of the amino acid sequence of BAP058-chi-LC, e.g., the amino acid sequence shown in FIGS. 10A-10B or SEQ ID NO: 24 or 26.

[0052] In other embodiments, the anti-PD-L1 antibody molecule comprises one, two, three or four heavy chain framework regions (e.g., the VHFW amino acid sequence shown in Table 2 or the VHFW amino acid sequence encoded by the nucleotide sequence shown in Table 2) or a sequence that is substantially identical thereto.

[0053] In other embodiments, the anti-PD-L1 antibody molecule comprises one, two, three or four light chain framework regions (e.g., the VLFW amino acid sequence shown in Table 2 or the VLFW amino acid sequence encoded by the nucleotide sequence shown in Table 2) or a sequence that is substantially identical thereto.

[0054] In other embodiments, the anti-PD-L1 antibody molecule comprises one, two, three or four heavy chain framework regions (e.g., the VHFW amino acid sequence shown in Table 2 or the VHFW amino acid sequence encoded by the nucleotide sequence shown in Table 2) or a sequence that is substantially identical thereto; and one, two, three or four light chain framework regions (e.g., the VLFW amino acid sequence shown in Table 2 or the VLFW amino acid sequence encoded by the nucleotide sequence shown in Table 2) or a sequence that is substantially identical thereto.

[0055] In one embodiment, the anti-PD-L1 antibody molecule comprises the heavy chain framework region 1 (VHFW1) of BAP058-hum01, BAP058-hum02, BAP058-hum07, BAP058-hum14 or BAP058-hum16 (e.g., SEQ ID NO: 124). In one embodiment, the antibody molecule comprises the heavy chain framework region 1 (VHFW1) of BAP058-hum04, BAP058-hum06, BAP058-hum08, BAP058-hum09, BAP058-hum12, BAP058-hum15, BAP058-hum17, BAP058-clone-L or BAP058-clone-M (e.g., SEQ ID NO: 126). In one embodiment, the antibody molecule comprises the heavy chain framework region 1 (VHFW1) of BAP058-hum03, BAP058-hum05, BAP058-hum11, BAP058-hum13, BAP058-clone-K, BAP058-clone-N or BAP058-clone-O (e.g., SEQ ID NO: 128). In one embodiment, the antibody molecule comprises the heavy chain framework region 1 (VHFW1) of BAP058-hum10 (e.g., SEQ ID NO: 130).

[0056] In certain embodiments, the anti-PD-L1 antibody molecule comprises the heavy chain framework region 2 (VHFW2) (e.g., SEQ ID NO: 132) of BAP058-hum01, BAP058-hum02, BAP058-hum03, BAP058-hum11, BAP058-hum14, BAP058-clone-K or BAP058-clone-N. In certain embodiments, the antibody molecule comprises the heavy chain framework region 2 (VHFW2) (e.g., SEQ ID NO: 134) of BAP058-hum04, BAP058-hum12 or BAP058-clone-L. In certain embodiments, the antibody molecule comprises the heavy chain framework region 2 (VHFW2) (e.g., SEQ ID NO: 136) of BAP058-hum06, BAP058-hum09, BAP058-hum15 or BAP058-clone-M. In certain embodiments, the antibody molecule comprises the heavy chain framework region 2 (VHFW2) (e.g., SEQ ID NO: 138) of BAP058-hum05, BAP058-hum07 or BAP058-hum16. In certain embodiments, the antibody molecule comprises the heavy chain framework region 2 (VHFW2) (e.g., SEQ ID NO: 140) of BAP058-hum08, BAP058-hum13, BAP058-hum17 or BAP058-clone-O. In certain embodiments, the antibody molecule comprises the heavy chain framework region 2 (e.g., SEQ ID NO: 142) of BAP058-hum10.

[0057] In certain embodiments, the anti-PD-L1 antibody molecule comprises the variable heavy chain framework region 3 (VHFW3) (e.g., SEQ ID NO: 144) of BAP058-hum01, BAP058-hum02, BAP058-hum07, BAP058-hum14 or BAP058-hum16. In certain embodiments, the antibody molecule comprises the variable heavy chain framework region 3 (VHFW3) (e.g., SEQ ID NO: 146) of BAP058-hum03, BAP058-hum06, BAP058-hum09, BAP058-hum10, BAP058-hum11, BAP058-hum15, BAP058-clone-K, BAP058-clone-M or BAP058-clone-N. In certain embodiments, the antibody molecule comprises the variable heavy chain framework region 3 (VHFW3) (e.g., SEQ ID NO: 148) of BAP058-hum04, BAP058-hum12 or BAP058-clone-L. In certain embodiments, the antibody molecule comprises the variable heavy chain framework region 3 (VHFW3) (e.g., SEQ ID NO: 150) of BAP058-hum05, BAP058-hum08 or BAP058-hum17. In certain embodiments, the antibody molecule comprises the variable heavy chain framework region 3 (VHFW3) (e.g., SEQ ID NO: 152) of BAP058-hum13 or BAP058-clone-O. In certain embodiments, the anti-PD-L1 antibody molecule comprises the variable heavy chain framework region 4 (VHFW4) (e.g., SEQ ID NO: 154) of BAP058-hum01, BAP058-hum02, BAP058-hum03, BAP058-hum04, BAP058-hum05, BAP058-hum06, BAP058-hum07, BAP058-hum08, BAP058-hum09, BAP058-hum10, BAP058-hum11, BAP058-hum12, BAP058-hum13, BAP058-hum14, BAP058-hum15, BAP058-hum16, BAP058-hum17, BAP058-clone-K, BAP058-clone-L, BAP058-clone-M, BAP058-clone-N, or BAP058-clone-O.

[0058] In certain embodiments, the anti-PD-L1 antibody molecule comprises the light chain framework region 1 (VLFW1) (e.g., SEQ ID NO: 156) of BAP058-hum03, BAP058-hum04, BAP058-hum05, BAP058-hum06, BAP058-hum14, BAP058-hum15, BAP058-hum16, BAP058-hum17, BAP058-clone-K, BAP058-clone-L or BAP058-clone-M. In certain embodiments, the antibody molecule comprises the light chain framework region 1 (VLFW1) (e.g., SEQ ID NO: 158) of BAP058-BAPhum08, BAP058-hum10, BAP058-hum11 or BAP058-clone-N. In certain embodiments, the antibody molecule comprises the light chain framework region 1 (VLFW1) (e.g., SEQ ID NO: 160) of BAP058-hum01 or BAP058-hum09. In certain embodiments, the antibody molecule comprises the light chain framework region 1 (VLFW1) (e.g., SEQ ID NO: 162) of BAP058-hum02 or BAP058-hum12. In certain embodiments, the antibody molecule comprises the light chain framework region 1 (VLFW1) (e.g., SEQ ID NO: 164) of BAP058-hum07. In certain embodiments, the antibody molecule comprises the light chain framework region 1 (VLFW1) (e.g., SEQ ID NO: 166) of BAP058-hum13 or BAP058-clone-O.

[0059] In certain embodiments, the anti-PD-L1 antibody molecule comprises the light chain framework region 2 (VLFW2) (e.g., SEQ ID NO: 168) of BAP058-hum08, BAP058-hum10, BAP058-hum11, BAP058-hum12, BAP058-hum14, BAP058-hum15, BAP058-hum16, BAP058-hum17 or BAP058-clone-N. In certain embodiments, the antibody molecule comprises the light chain framework region 2 (VLFW2) (e.g., SEQ ID NO: 170) of BAP058-hum01, BAP058-hum02, BAP058-hum03, BAP058-hum04, BAP058-hum05, BAP058-hum06, BAP058-hum07, BAP058-hum09, BAP058-hum13, BAP058-clone-K, BAP058-clone-L, BAP058-clone-M or BAP058-clone-O.

[0060] In certain embodiments, the anti-PD-L1 antibody molecule comprises the light chain framework region 3 (VLFW3) (e.g., SEQ ID NO: 172) of BAP058-hum08, BAP058-hum10, BAP058-hum11, BAP058-hum14, BAP058-hum15, BAP058-hum16, BAP058-hum17 or BAP058-clone-N. In certain embodiments, the antibody molecule comprises the light chain framework region 3 (VLFW3) (e.g., SEQ ID NO: 174) of BAP058-hum04, BAP058-hum05, BAP058-hum06, BAP058-hum12, BAP058-clone-L or BAP058-clone-M. In certain embodiments, the antibody molecule comprises the light chain framework region 3 (VLFW3) (e.g., SEQ ID NO: 176) of BAP058-hum01 or BAP058-hum09. In certain embodiments, the antibody molecule comprises the light chain framework region 3 (VLFW3) (e.g., SEQ ID NO: 178) of BAP058-hum02. In certain embodiments, the antibody molecule comprises the light chain framework region 3 (VLFW3) (e.g., SEQ ID NO: 180) of BAP058-hum03 or BAP058-clone-K. In certain embodiments, the antibody molecule comprises the light chain framework region 3 (VLFW3) (e.g., SEQ ID NO: 182) of BAP058-hum07. In certain embodiments, the antibody molecule comprises the light chain framework region 3 (VLFW3) (e.g., SEQ ID NO: 184) of BAP058-hum13 or BAP058-clone-O.

[0061] In one aspect, the anti-PD-L1 antibody molecule comprises the light chain framework region 4 (VLFW4) of BAP058-hum01, BAP058-hum02, BAP058-hum03, BAP058-hum04, BAP058-hum05, BAP058-hum06, BAP058-hum07, BAP058-hum08, BAP058-hum09, BAP058-hum10, BAP058-hum11, BAP058-hum12, BAP058-hum13, BAP058-hum14, BAP058-hum15, BAP058-hum16, BAP058-hum17, BAP058-clone-K, BAP058-clone-L, BAP058-clone-M, BAP058-clone-N, or BAP058-clone-O (e.g., SEQ ID NO: 186).

[0062] In certain embodiments, the anti-PD-L1 antibody molecule comprises heavy chain framework regions 1-3 of BAP058-hum01, BAP058-hum02, or BAP058-hum14 [e.g., SEQ ID NO: 124 (VHFW1), SEQ ID NO: 132 (VHFW2), and SEQ ID NO: 144 (VHFW3)]. In certain embodiments, the antibody molecule comprises heavy chain framework regions 1-3 of BAP058-hum07 or BAP058-hum16 [e.g., SEQ ID NO: 124 (VHFW1), SEQ ID NO: 138 (VHFW2), and SEQ ID NO: 144 (VHFW3)]. In certain embodiments, the antibody molecule comprises heavy chain framework regions 1-3 of BAP058-hum04, BAP058-hum12, or BAP058-clone-L [e.g., SEQ ID NO: 126 (VHFW1), SEQ ID NO: 134 (VHFW2), and SEQ ID NO: 148 (VHFW3)]. In certain embodiments, the antibody molecule comprises heavy chain framework regions 1-3 of BAP058-hum06, BAP058-hum09, BAP058-hum15, or BAP058-clone-M [e.g., SEQ ID NO: 126 (VHFW1), SEQ ID NO: 136 (VHFW2), and SEQ ID NO: 146 (VHFW3)]. In certain embodiments, the antibody molecule comprises heavy chain framework regions 1-3 of BAP058-hum08 or BAP058-hum17 [e.g., SEQ ID NO: 126 (VHFW1), SEQ ID NO: 140 (VHFW2), and SEQ ID NO: 150 (VHFW3)]. In certain embodiments, the antibody molecule comprises heavy chain framework regions 1-3 of BAP058-hum03, BAP058-hum11, BAP058-clone-K, or BAP058-clone-N [e.g., SEQ ID NO: 128 (VHFW1), SEQ ID NO: 132 (VHFW2), and SEQ ID NO: 146 (VHFW3)]. In certain embodiments, the antibody molecule comprises heavy chain framework regions 1-3 of BAP058-hum13 or BAP058-clone-O [e.g., SEQ ID NO: 128 (VHFW1), SEQ ID NO: 140 (VHFW2), and SEQ ID NO: 152 (VHFW3)]. In certain embodiments, the antibody molecule comprises heavy chain framework regions 1-3 of BAP058-hum05 [e.g., SEQ ID NO: 128 (VHFW1), SEQ ID NO: 138 (VHFW2), and SEQ ID NO: 150 (VHFW3)].In one aspect, the antibody molecule comprises heavy chain framework regions 1-3 of BAP058-hum10 [e.g., SEQ ID NO: 130 (VHFW1), SEQ ID NO: 142 (VHFW2), and SEQ ID NO: 146 (VHFW3)]. In one aspect, the antibody molecule further comprises heavy chain framework region 4 (VHFW4) of BAP058-hum01, BAP058-hum02, BAP058-hum03, BAP058-hum04, BAP058-hum05, BAP058-hum06, BAP058-hum07, BAP058-hum08, BAP058-hum09, BAP058-hum10, BAP058-hum11, BAP058-hum12, BAP058-hum13, BAP058-hum14, BAP058-hum15, BAP058-hum16, BAP058-hum17, BAP058-clone-K, BAP058-clone-L, BAP058-clone-M, BAP058-clone-N, or BAP058-clone-O (e.g., SEQ ID NO: 154).

[0063] In one aspect, the anti-PD-L1 antibody molecule comprises light chain framework regions 1 to 3 of BAP058-hum01 or BAP058-hum09 [e.g., SEQ ID NO: 160 (VLFW1), SEQ ID NO: 170 (VLFW2), and SEQ ID NO: 176 (VLFW3)]. In one aspect, the antibody molecule comprises light chain framework regions 1 to 3 of BAP058-hum14, BAP058-hum15, BAP058-hum16, or BAP058-hum17 [e.g., SEQ ID NO: 156 (VLFW1), SEQ ID NO: 168 (VLFW2), and SEQ ID NO: 172 (VLFW3)]. In one aspect, the antibody molecule comprises light chain framework regions 1 to 3 of BAP058-hum04, BAP058-hum05, BAP058-hum06, BAP058-clone-L, or BAP058-clone-M [e.g., SEQ ID NO: 156 (VLFW1), SEQ ID NO: 170 (VLFW2), and SEQ ID NO: 174 (VLFW3)]. In one aspect, the antibody molecule comprises light chain framework regions 1 to 3 of BAP058-hum08, BAP058-hum10, or BAP058-hum11 [e.g., SEQ ID NO: 158 (VLFW1), SEQ ID NO: 168 (VLFW2), and SEQ ID NO: 172 (VLFW3)]. In one aspect, the antibody molecule comprises light chain framework regions 1 to 3 of BAP058-hum13 or BAP058-clone-O [e.g., SEQ ID NO: 166 (VLFW1), SEQ ID NO: 170 (VLFW2), and SEQ ID NO: 184 (VLFW3)]. In one aspect, the antibody molecule comprises light chain framework regions 1 to 3 of BAP058-hum02 [e.g., SEQ ID NO: 162 (VLFW1), SEQ ID NO: 170 (VLFW2), and SEQ ID NO: 178 (VLFW3)]. In one aspect, the antibody molecule comprises light chain framework regions 1 to 3 of BAP058-hum03 [e.g., SEQ ID NO: 156 (VLFW1), SEQ ID NO: 170 (VLFW2), and SEQ ID NO: 180 (VLFW3)]. In one aspect, the antibody molecule comprises light chain framework regions 1 to 3 of BAP058-hum07 [e.g., SEQ ID NO: 164 (VLFW1), SEQ ID NO: 170 (VLFW2), and SEQ ID NO: 182 (VLFW3)].In certain embodiments, the antibody molecule comprises light chain framework regions 1-3 of BAP058-hum12 [e.g., SEQ ID NO: 162 (VLFW1), SEQ ID NO: 168 (VLFW2), and SEQ ID NO: 174 (VLFW3)]. In certain embodiments, the antibody molecule comprises light chain framework regions 1-3 of BAP058-clone-K [e.g., SEQ ID NO: 156 (VLFW1), SEQ ID NO: 168 (VLFW2), and SEQ ID NO: 180 (VLFW3)]. In certain embodiments, the antibody molecule comprises light chain framework regions 1-3 of BAP058-clone-N [e.g., SEQ ID NO: 158 (VLFW1), SEQ ID NO: 170 (VLFW2), and SEQ ID NO: 172 (VLFW3)]. In certain embodiments, the antibody molecule further comprises light chain framework region 4 (VLFW4) of BAP058-hum01, BAP058-hum02, BAP058-hum03, BAP058-hum04, BAP058-hum05, BAP058-hum06, BAP058-hum07, BAP058-hum08, BAP058-hum09, BAP058-hum10, BAP058-hum11, BAP058-hum12, BAP058-hum13, BAP058-hum14, BAP058-hum15, BAP058-hum16, BAP058-hum17, BAP058-clone-K, BAP058-clone-L, BAP058-clone-M, BAP058-clone-N, or BAP058-clone-O (e.g., SEQ ID NO: 186).

[0064] In certain embodiments, the anti-PD-L1 antibody molecule comprises heavy chain framework regions 1-3 of BAP058-hum01 [e.g., SEQ ID NO: 124 (VHFW1), SEQ ID NO: 132 (VHFW2), and SEQ ID NO: 144 (VHFW3)] and light chain framework regions 1-3 of BAP058-hum01 [e.g., SEQ ID NO: 160 (VLFW1), SEQ ID NO: 170 (VLFW2), and SEQ ID NO: 176 (VLFW3)].

[0065] In one aspect, the anti-PD-L1 antibody molecule comprises the heavy chain framework regions 1-3 of BAP058-hum02 [e.g., SEQ ID NO: 124 (VHFW1), SEQ ID NO: 132 (VHFW2), and SEQ ID NO: 144 (VHFW3)] and the light chain framework regions 1-3 of BAP058-hum02 [e.g., SEQ ID NO: 162 (VLFW1), SEQ ID NO: 170 (VLFW2), and SEQ ID NO: 178 (VLFW3)].

[0066] In one aspect, the anti-PD-L1 antibody molecule comprises the heavy chain framework regions 1-3 of BAP058-hum03 [e.g., SEQ ID NO: 128 (VHFW1), SEQ ID NO: 132 (VHFW2), and SEQ ID NO: 146 (VHFW3)] and the light chain framework regions 1-3 of BAP058-hum03 [e.g., SEQ ID NO: 156 (VLFW1), SEQ ID NO: 170 (VLFW2), and SEQ ID NO: 180 (VLFW3)].

[0067] In one aspect, the anti-PD-L1 antibody molecule comprises the heavy chain framework regions 1-3 of BAP058-hum04 or BAP058-clone-L [e.g., SEQ ID NO: 126 (VHFW1), SEQ ID NO: 134 (VHFW2), and SEQ ID NO: 148 (VHFW3)] and the light chain framework regions 1-3 of BAP058-hum04 or BAP058-clone-L [e.g., SEQ ID NO: 156 (VLFW1), SEQ ID NO: 170 (VLFW2), and SEQ ID NO: 174 (VLFW3)].

[0068] In one aspect, the anti-PD-L1 antibody molecule comprises the heavy chain framework regions 1-3 of BAP058-hum05 [e.g., SEQ ID NO: 128 (VHFW1), SEQ ID NO: 138 (VHFW2), and SEQ ID NO: 150 (VHFW3)] and the light chain framework regions 1-3 of BAP058-hum05 [e.g., SEQ ID NO: 156 (VLFW1), SEQ ID NO: 170 (VLFW2), and SEQ ID NO: 174 (VLFW3)].

[0069] In one aspect, the anti-PD-L1 antibody molecule comprises the heavy chain framework regions 1-3 of BAP058-hum06 or BAP058-clone-M [e.g., SEQ ID NO: 126 (VHFW1), SEQ ID NO: 136 (VHFW2), and SEQ ID NO: 146 (VHFW3)] and the light chain framework regions 1-3 of BAP058-hum06 or BAP058-clone-M [e.g., SEQ ID NO: 156 (VLFW1), SEQ ID NO: 170 (VLFW2), and SEQ ID NO: 174 (VLFW3)].

[0070] In one aspect, the anti-PD-L1 antibody molecule comprises the heavy chain framework regions 1-3 of BAP058-hum07 [e.g., SEQ ID NO: 124 (VHFW1), SEQ ID NO: 138 (VHFW2), and SEQ ID NO: 144 (VHFW3)] and the light chain framework regions 1-3 of BAP058-hum07 [e.g., SEQ ID NO: 164 (VLFW1), SEQ ID NO: 170 (VLFW2), and SEQ ID NO: 182 (VLFW3)].

[0071] In one aspect, the anti-PD-L1 antibody molecule comprises the heavy chain framework regions 1-3 of BAP058-hum08 [e.g., SEQ ID NO: 126 (VHFW1), SEQ ID NO: 140 (VHFW2), and SEQ ID NO: 150 (VHFW3)] and the light chain framework regions 1-3 of BAP058-hum08 [e.g., SEQ ID NO: 158 (VLFW1), SEQ ID NO: 168 (VLFW2), and SEQ ID NO: 172 (VLFW3)].

[0072] In one aspect, the anti-PD-L1 antibody molecule comprises the heavy chain framework regions 1-3 of BAP058-hum09 [e.g., SEQ ID NO: 126 (VHFW1), SEQ ID NO: 136 (VHFW2), and SEQ ID NO: 146 (VHFW3)] and the light chain framework regions 1-3 of BAP058-hum09 [e.g., SEQ ID NO: 160 (VLFW1), SEQ ID NO: 170 (VLFW2), and SEQ ID NO: 176 (VLFW3)].

[0073] In one aspect, the anti-PD-L1 antibody molecule comprises framework regions 1-3 of the heavy chain of BAP058-hum10 [e.g., SEQ ID NO: 130 (VHFW1), SEQ ID NO: 142 (VHFW2), and SEQ ID NO: 146 (VHFW3)] and framework regions 1-3 of the light chain of BAP058-hum10 [e.g., SEQ ID NO: 158 (VLFW1), SEQ ID NO: 168 (VLFW2), and SEQ ID NO: 172 (VLFW3)].

[0074] In one aspect, the anti-PD-L1 antibody molecule comprises framework regions 1-3 of the heavy chain of BAP058-hum11 [e.g., SEQ ID NO: 128 (VHFW1), SEQ ID NO: 132 (VHFW2), and SEQ ID NO: 146 (VHFW3)] and framework regions 1-3 of the light chain of BAP058-hum11 [e.g., SEQ ID NO: 158 (VLFW1), SEQ ID NO: 168 (VLFW2), and SEQ ID NO: 172 (VLFW3)].

[0075] In one aspect, the anti-PD-L1 antibody molecule comprises framework regions 1-3 of the heavy chain of BAP058-hum12 [e.g., SEQ ID NO: 126 (VHFW1), SEQ ID NO: 134 (VHFW2), and SEQ ID NO: 148 (VHFW3)] and framework regions 1-3 of the light chain of BAP058-hum12 [e.g., SEQ ID NO: 162 (VLFW1), SEQ ID NO: 168 (VLFW2), and SEQ ID NO: 174 (VLFW3)].

[0076] In one aspect, the anti-PD-L1 antibody molecule comprises framework regions 1-3 of the heavy chain of BAP058-hum13 or BAP058-clone-O [e.g., SEQ ID NO: 128 (VHFW1), SEQ ID NO: 140 (VHFW2), and SEQ ID NO: 152 (VHFW3)] and framework regions 1-3 of the light chain of BAP058-hum13 or BAP058-clone-O [e.g., SEQ ID NO: 166 (VLFW1), SEQ ID NO: 170 (VLFW2), and SEQ ID NO: 184 (VLFW3)].

[0077] In one aspect, the anti-PD-L1 antibody molecule comprises framework regions 1-3 of the heavy chain of BAP058-hum14 [e.g., SEQ ID NO: 124 (VHFW1), SEQ ID NO: 132 (VHFW2), and SEQ ID NO: 144 (VHFW3)] and framework regions 1-3 of the light chain of BAP058-hum14 [e.g., SEQ ID NO: 156 (VLFW1), SEQ ID NO: 168 (VLFW2), and SEQ ID NO: 172 (VLFW3)].

[0078] In one aspect, the anti-PD-L1 antibody molecule comprises framework regions 1-3 of the heavy chain of BAP058-hum15 [e.g., SEQ ID NO: 126 (VHFW1), SEQ ID NO: 136 (VHFW2), and SEQ ID NO: 146 (VHFW3)] and framework regions 1-3 of the light chain of BAP058-hum15 [e.g., SEQ ID NO: 156 (VLFW1), SEQ ID NO: 168 (VLFW2), and SEQ ID NO: 172 (VLFW3)].

[0079] In one aspect, the anti-PD-L1 antibody molecule comprises framework regions 1-3 of the heavy chain of BAP058-hum16 [e.g., SEQ ID NO: 124 (VHFW1), SEQ ID NO: 138 (VHFW2), and SEQ ID NO: 144 (VHFW3)] and framework regions 1-3 of the light chain of BAP058-hum16 [e.g., SEQ ID NO: 156 (VLFW1), SEQ ID NO: 168 (VLFW2), and SEQ ID NO: 172 (VLFW3)].

[0080] In one aspect, the anti-PD-L1 antibody molecule comprises framework regions 1-3 of the heavy chain of BAP058-hum17 [e.g., SEQ ID NO: 126 (VHFW1), SEQ ID NO: 140 (VHFW2), and SEQ ID NO: 150 (VHFW3)] and framework regions 1-3 of the light chain of BAP058-hum17 [e.g., SEQ ID NO: 156 (VLFW1), SEQ ID NO: 168 (VLFW2), and SEQ ID NO: 172 (VLFW3)].

[0081] In certain embodiments, the anti-PD-L1 antibody molecule comprises the heavy chain framework regions 1-3 of BAP058-clone-N [e.g., SEQ ID NO: 128 (VHFW1), SEQ ID NO: 132 (VHFW2), and SEQ ID NO: 146 (VHFW3)] and the light chain framework regions 1-3 of BAP058-clone-N [e.g., SEQ ID NO: 158 (VLFW1), SEQ ID NO: 170 (VLFW2), and SEQ ID NO: 172 (VLFW3)].

[0082] In certain embodiments, the anti-PD-L1 antibody molecule further comprises the heavy chain framework region 4 (VHFW4) of BAP058-hum01, BAP058-hum02, BAP058-hum03, BAP058-hum04, BAP058-hum05, BAP058-hum06, BAP058-hum07, BAP058-hum08, BAP058-hum09, BAP058-hum10, BAP058-hum11, BAP058-hum12, BAP058-hum13, BAP058-hum14, BAP058-hum15, BAP058-hum16, BAP058-hum17, BAP058-clone-K, BAP058-clone-L, BAP058-clone-M, BAP058-clone-N, or BAP058-clone-O (e.g., SEQ ID NO: 154) and the light chain framework region 4 (VLFW4) of BAP058-hum01, BAP058-hum02, BAP058-hum03, BAP058-hum04, BAP058-hum05, BAP058-hum06, BAP058-hum07, BAP058-hum08, BAP058-hum09, BAP058-hum10, BAP058-hum11, BAP058-hum12, BAP058-hum13, BAP058-hum14, BAP058-hum15, BAP058-hum16, BAP058-hum17, BAP058-clone-K, BAP058-clone-L, BAP058-clone-M, BAP058-clone-N, or BAP058-clone-O (e.g., SEQ ID NO: 186).

[0083] In one aspect, the anti-PD-L1 antibody molecule comprises a heavy chain framework region having a combination of framework regions FW1, FW2, and FW3 shown in FIG. 4 or FIG. 6. In another aspect, the antibody molecule comprises a light chain framework region having a combination of framework regions FW1, FW2, and FW3 shown in FIG. 4 or FIG. 6. In yet another aspect, the antibody molecule comprises a heavy chain framework region having a combination of framework regions FW1, FW2, and FW3 shown in FIG. 4 or FIG. 6, and a light chain framework region having a combination of framework regions FW1, FW2, and FW3 shown in FIG. 4 or FIG. 6.

[0084] In one aspect, the heavy and / or light chain variable domain(s) of the anti-PD-L1 antibody molecule is / are substantially identical to the amino acids disclosed herein, e.g., an antibody selected from the antibodies described herein, e.g., BAP058-hum01, BAP058-hum02, BAP058-hum03, BAP058-hum04, BAP058-hum05, BAP058-hum06, BAP058-hum07, BAP058-hum08, BAP058-hum09, BAP058-hum10, BAP058-hum11, BAP058-hum12, BAP058-hum13, BAP058-hum14, BAP058-hum15, BAP058-hum16, BAP058-hum17, BAP058-clone-K, BAP058-clone-L, BAP058-clone-M, BAP058-clone-N, or BAP058-clone-O; or at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or more identical to the variable region of an antibody described in Table 1 or an antibody encoded by the nucleotide sequence in Table 1, or an amino acid sequence that contains at least 1 or 5 residues different from the variable region of an antibody described herein, provided that less than 40, 30, 20 or 10 residues are different.

[0085] In one aspect, the heavy and / or light chain variable regions of the anti-PD-L1 antibody molecule, or both, are the amino acid sequences encoded by a nucleic acid sequence described herein, or, for example, under low stringency, medium stringency or high stringency, or other hybridization conditions described herein, a nucleic acid sequence described herein (e.g., the nucleic acid sequences shown in Tables 1 and 2) or its complement that hybridizes to the nucleic acid.

[0086] In another aspect, the anti-PD-L1 antibody molecule comprises at least 1, 2, 3 or 4 antigen-binding regions, e.g., variable regions, having an amino acid sequence set forth in Table 1 or a sequence that is substantially identical thereto (e.g., a sequence that is at least about 85%, 90%, 95%, 99% or more identical thereto, or a sequence that differs from the sequence shown in Table 1 by 1, 2, 5, 10 or 15 or fewer amino acid residues). In another aspect, the anti-PD-L1 antibody molecule comprises VH and / or VL domains encoded by a nucleic acid having a nucleotide sequence set forth in Table 1 or a sequence that is substantially identical thereto (e.g., a sequence that is at least about 85%, 90%, 95%, 99% or more identical thereto, or a sequence that differs from the sequence shown in Table 1 by 3, 6, 15, 30 or 45 or fewer nucleotides).

[0087] In yet another aspect, the anti-PD-L1 antibody molecule comprises at least one, two, or three CDRs derived from a heavy chain variable region having the amino acid sequence set forth in Table 1 or a sequence that is substantially homologous thereto (e.g., a sequence that is at least about 85%, 90%, 95%, 99% or more identical thereto and / or has one, two, three or more substitutions, insertions or deletions, e.g., conservative substitutions). In yet another aspect, the anti-PD-L1 antibody molecule comprises at least one, two, or three CDRs derived from a light chain variable region having the amino acid sequence set forth in Table 1 or a sequence that is substantially homologous thereto (e.g., a sequence that is at least about 85%, 90%, 95%, 99% or more identical thereto and / or has one, two, three or more substitutions, insertions or deletions, e.g., conservative substitutions). In yet another aspect, the anti-PD-L1 antibody molecule comprises at least one, two, three, four, five or six CDRs derived from heavy and light chain variable regions having the amino acid sequence set forth in Table 1 or a sequence that is substantially homologous thereto (e.g., a sequence that is at least about 85%, 90%, 95%, 99% or more identical thereto and / or has one, two, three or more substitutions, insertions or deletions, e.g., conservative substitutions).

[0088] In one aspect, the anti-PD-L1 antibody molecule comprises at least one, two or three CDRs and / or hypervariable loops derived from a heavy chain variable region having the amino acid sequence of an antibody selected from the antibodies described herein, such as BAP058-hum01, BAP058-hum02, BAP058-hum03, BAP058-hum04, BAP058-hum05, BAP058-hum06, BAP058-hum07, BAP058-hum08, BAP058-hum09, BAP058-hum10, BAP058-hum11, BAP058-hum12, BAP058-hum13, BAP058-hum14, BAP058-hum15, BAP058-hum16, BAP058-hum17, BAP058-clone-K, BAP058-clone-L, BAP058-clone-M, BAP058-clone-N, or BAP058-clone-O summarized in Table 1, or a sequence substantially identical thereto (e.g., a sequence that is at least about 85%, 90%, 95%, 99% or more identical thereto and / or has one, two, three or more substitutions, insertions or deletions, such as conservative substitutions).In another aspect, the anti-PD-L1 antibody molecule comprises at least one, two, or three CDRs and / or hypervariable loops derived from the light chain variable region having the amino acid sequence of an antibody selected from the antibodies described herein, such as BAP058-hum01, BAP058-hum02, BAP058-hum03, BAP058-hum04, BAP058-hum05, BAP058-hum06, BAP058-hum07, BAP058-hum08, BAP058-hum09, BAP058-hum10, BAP058-hum11, BAP058-hum12, BAP058-hum13, BAP058-hum14, BAP058-hum15, BAP058-hum16, BAP058-hum17, BAP058-clone-K, BAP058-clone-L, BAP058-clone-M, BAP058-clone-N, or BAP058-clone-O summarized in Table 1, or a sequence substantially identical thereto (e.g., a sequence that is at least about 85%, 90%, 95%, 99% or more identical thereto and / or has one, two, three or more substitutions, insertions or deletions, such as conservative substitutions). In one aspect, the anti-PD-L1 antibody molecule comprises all six CDRs and / or hypervariable loops described herein, such as those described in Table 1.

[0089] In one aspect, the anti-PD-L1 antibody molecule has a variable region that is identical in sequence to the variable regions described herein (e.g., the FR regions disclosed herein) or that differs from it by one, two, three, or four amino acids.

[0090] In one aspect, the anti-PD-L1 antibody molecule is a full antibody or a fragment thereof [e.g., Fab, F(ab’)2, Fv or single-chain Fv fragment (scFv)]. In certain aspects, the anti-PD-L1 antibody molecule is a monoclonal antibody or an antibody having a single specificity. The anti-PD-L1 antibody molecule can also be a humanized, chimeric, camelid, shark or in vitro-produced antibody molecule. In one aspect, such an anti-PD-L1 antibody molecule is a humanized antibody molecule. The heavy and light chains of the anti-PD-L1 antibody molecule can be full-length (e.g., the antibody can comprise at least one, preferably two full heavy chains and at least one, preferably two full light chains), or antigen-binding fragments [e.g., Fab, F(ab’)2, Fv, single-chain Fv fragment, single-domain antibody, diabody (dAb), bivalent or bispecific antibody, or fragments thereof, single-domain variants thereof, or camelid antibodies].

[0091] In certain embodiments, the anti-PD-L1 antibody molecule is in the form of a bispecific or multispecific antibody molecule. In some embodiments, the bispecific antibody molecule has a first binding specificity for PD-L1 and a second binding specificity for TIM-3, LAG-3, CEACAM (e.g., CEACAM-1, CEACAM-3, and / or CEACAM-5), PD-1, or PD-L2. In some embodiments, the bispecific antibody molecule binds to PD-L1 and TIM-3. In another embodiment, the bispecific antibody molecule binds to PD-L1 and LAG-3. In another embodiment, the bispecific antibody molecule binds to PD-L1 and CEACAM (e.g., CEACAM-1, CEACAM-3, and / or CEACAM-5). In another embodiment, the bispecific antibody molecule binds to PD-L1 and CEACAM-1. In yet another embodiment, the bispecific antibody molecule binds to PD-L1 and CEACAM-5. In another embodiment, the bispecific antibody molecule binds to PD-L1 and PD-1. In yet another embodiment, the bispecific antibody molecule binds to PD-L1 and PD-L2. In a multispecific antibody molecule, any combination of the aforementioned molecules may be made, for example, a trispecific antibody comprising a first binding specificity for PD-L1 and second and third binding specificities for one or more of TIM-3, LAG-3, CEACAM (e.g., CEACAM-1, CEACAM-3, and / or CEACAM-5), PD-1, or PD-L2. In some embodiments, the second and / or third binding specificities for TIM-3, LAG-3, and / or PD-1 include the amino acid sequences disclosed herein or are encoded by the nucleotide sequences disclosed herein [e.g., as disclosed in the section entitled "Inhibitors of Immune Checkpoint Molecules" beginning on page 218 hereinafter (including any publications mentioned therein)].

[0092] In other embodiments, the anti-PD-L1 antibody molecule is used in combination with a bispecific molecule comprising one or more of TIM-3, LAG-3, CEACAM (e.g., CEACAM-1, CEACAM-3, and / or CEACAM-5), PD-1, or PD-L2. In certain embodiments, the bispecific antibody molecule used in combination binds to CEACAM (e.g., CEACAM-1, CEACAM-3, and / or CEACAM-5) and LAG-3. In another embodiment, the bispecific antibody molecule used in combination binds to CEACAM (e.g., CEACAM-1, CEACAM-3, and / or CEACAM-5) and TIM-3. In another embodiment, the bispecific antibody molecule used in combination binds to LAG-3 and TIM-3.

[0093] In still other embodiments, the anti-PD-L1 antibody molecule is selected from, for example, the heavy chain constant regions (Fc) of IgG1, IgG2, IgG3, IgG4, IgM, IgA1, IgA2, IgD, and IgE, and in particular, for example, the heavy chain constant regions of IgG1, IgG2, IgG3, and IgG4, and more specifically, has a heavy chain constant region selected from the heavy chain constant regions of IgG1 or IgG2 (e.g., human IgG1, IgG2, or IgG4). In one embodiment, the heavy chain constant region is human IgG1. In another embodiment, the anti-PD-L1 antibody molecule has a light chain constant region selected from, for example, kappa or lambda, preferably kappa (e.g., human kappa). In one embodiment, the constant region is altered, for example, mutated, so as to modify the properties of the anti-PD-L1 antibody molecule (e.g., to increase or decrease one or more of Fc receptor binding, antibody glycosylation, the number of cysteine residues, effector cell function, or complement function). For example, the constant region is mutated at positions 296 (M to Y), 298 (S to T), 300 (T to E), 477 (H to K), and 478 (N to F) to alter Fc receptor binding [e.g., the mutated positions correspond to positions 132 (M to Y), 134 (S to T), 136 (T to E), 313 (H to K), and 314 (N to F) of SEQ ID NO: 212 or 214; or positions 135 (M to Y), 137 (S to T), 139 (T to E), 316 (H to K), and 317 (N to F) of SEQ ID NO: 215, 216, 217, or 218]. In another embodiment, the heavy chain constant region of IgG4, e.g., human IgG4, is mutated at position 228 (e.g., S to P), as shown in Table 3, for example. In a particular embodiment, the anti-PD-L1 antibody molecule comprises, for example, human IgG4 mutated at position 228 (e.g., S to P) as shown in Table 3; and, for example, a kappa light chain constant region as shown in Table 3. In yet another embodiment, the heavy chain constant region of IgG1, e.g., human IgG1, is mutated at one or more of positions 297 (e.g., N to A), 265 (e.g., D to A), 329 (e.g., P to A), 234 (e.g., L to A), or 235 (e.g., L to A), as shown in Table 3, for example.In certain embodiments, the anti-PD-L1 antibody molecule is, for example, a human IgG1 in which one or more of the aforementioned positions are mutated, as shown in Table 3; and includes, for example, a kappa light chain constant region, as shown in Table 3.

[0094] In one embodiment, the anti-PD-L1 antibody molecule is isolated or recombinant. In one embodiment, the anti-PD-L1 antibody molecule is a humanized antibody molecule.

[0095] The present invention also features nucleic acid molecules encoding one or both of the heavy and light chain variable regions, CDRs, hypervariable loops, and framework regions of the anti-PD-L1 antibody molecules described herein. In certain embodiments, the nucleotide sequences encoding the anti-PD-L1 antibody molecules are codon-optimized. For example, the present invention features first and second nucleic acids encoding the heavy and light chain variable regions, respectively, of an anti-PD-L1 antibody molecule selected from one or more of, or substantially identical to, BAP058-hum01, BAP058-hum02, BAP058-hum03, BAP058-hum04, BAP058-hum05, BAP058-hum06, BAP058-hum07, BAP058-hum08, BAP058-hum09, BAP058-hum10, BAP058-hum11, BAP058-hum12, BAP058-hum13, BAP058-hum14, BAP058-hum15, BAP058-hum16, BAP058-hum17, BAP058-clone-K, BAP058-clone-L, BAP058-clone-M, BAP058-clone-N, or BAP058-clone-O, as summarized in Table 1. For example, the nucleic acid can include a nucleotide sequence as set forth in Tables 1 and 2 or a sequence substantially identical thereto (e.g., a sequence that is at least about 85%, 90%, 95%, 99% or more identical thereto, or a sequence that differs from the sequences shown in Tables 1 and 2 by 3, 6, 15, 30 or 45 or fewer nucleotides).

[0096] In other embodiments, the nucleic acid molecule comprises a nucleotide sequence encoding a heavy chain variable domain and / or a heavy chain constant region that has the amino acid sequence of BAP058-clone-K, BAP058-clone-L, BAP058-clone-M, BAP058-clone-N or BAP058-clone-O; or the amino acid sequence described in Table 1; or the nucleotide sequence in Table 1; or a sequence that is substantially identical to any of the foregoing sequences (e.g., a sequence that is at least about 85%, 90%, 95%, 99% or more identical).

[0097] In other embodiments, the nucleic acid molecule comprises a nucleotide sequence encoding a light chain variable domain and / or a light chain constant region that has the amino acid sequence of BAP058-clone-K, BAP058-clone-L, BAP058-clone-M, BAP058-clone-N or BAP058-clone-O; or the amino acid sequence described in Table 1; or the nucleotide sequence in Table 1; or a sequence that is substantially identical to any of the foregoing sequences (e.g., a sequence that is at least about 85%, 90%, 95%, 99% or more identical).

[0098] The foregoing nucleotide sequences encoding the anti-PD-L1 heavy chain and light chain variable domains and constant regions can be present in separate nucleic acid molecules or the same nucleic acid molecule. In certain embodiments, the nucleic acid molecule comprises a nucleotide sequence encoding a leader sequence.

[0099] In certain embodiments, the nucleic acid molecule comprises a nucleotide sequence encoding at least one, two or three CDRs or hypervariable loops derived from the heavy chain variable region that has the amino acid sequence set forth in Table 1 or a sequence that is substantially homologous thereto (e.g., a sequence that is at least about 85%, 90%, 95%, 99% or more identical thereto and / or has one, two, three or more substitutions, insertions or deletions, e.g., conservative substitutions).

[0100] In another aspect, the nucleic acid molecule comprises a nucleotide sequence encoding at least one, two or three CDRs or hypervariable loops derived from a variable light chain region having an amino acid sequence set forth in Table 1 or a sequence substantially homologous thereto (e.g., a sequence that is at least about 85%, 90%, 95%, 99% or more identical thereto and / or has one, two, three or more substitutions, insertions or deletions, e.g., conservative substitutions).

[0101] In yet another aspect, the nucleic acid molecule comprises a nucleotide sequence encoding at least one, two, three, four, five or six CDRs or hypervariable loops derived from variable heavy and light chain regions having an amino acid sequence set forth in Table 1 or a sequence substantially homologous thereto (e.g., a sequence that is at least about 85%, 90%, 95%, 99% or more identical thereto and / or has one, two, three or more substitutions, insertions or deletions, e.g., conservative substitutions).

[0102] In another aspect, the nucleic acid molecule comprises one or more heavy chain framework regions of a sequence that is any one of or substantially identical to BAP058-hum01, BAP058-hum02, BAP058-hum03, BAP058-hum04, BAP058-hum05, BAP058-hum06, BAP058-hum07, BAP058-hum08, BAP058-hum09, BAP058-hum10, BAP058-hum11, BAP058-hum12, BAP058-hum13, BAP058-hum14, BAP058-hum15, BAP058-hum16, BAP058-hum17, BAP058-clone-K, BAP058-clone-L, BAP058-clone-M, BAP058-clone-N, or BAP058-clone-O summarized in Tables 1 and 2 [e.g., any one of VHFW1 (type a), VHFW1 (type b), VHFW1 (type c), VHFW1 (type d), VHFW2 (type a), VHFW2 (type a'), VHFW2 (type b), VHFW2 (type c), VHFW2 (type d), VHFW2 (type e), VHFW3 (type a), VHFW3 (type b), VHFW3 (type c), VHFW3 (type d), VHFW3 (type e), or VHFW4, or any combination thereof, e.g., the framework combinations described herein]. For example, the nucleic acid molecule can comprise a nucleotide sequence set forth in Tables 1 and 2 or a sequence substantially identical thereto (e.g., a sequence that is at least about 85%, 90%, 95%, 99% or more identical thereto, or a sequence that differs from the sequences shown in Tables 1 and 2 by 3, 6, 15, 30 or 45 or fewer nucleotides).

[0103] In another aspect, the nucleic acid molecule comprises one or more light chain framework regions of a sequence that is any one of or substantially identical to BAP058-hum01, BAP058-hum02, BAP058-hum03, BAP058-hum04, BAP058-hum05, BAP058-hum06, BAP058-hum07, BAP058-hum08, BAP058-hum09, BAP058-hum10, BAP058-hum11, BAP058-hum12, BAP058-hum13, BAP058-hum14, BAP058-hum15, BAP058-hum16, BAP058-hum17, BAP058-clone-K, BAP058-clone-L, BAP058-clone-M, BAP058-clone-N, or BAP058-clone-O, as summarized in Tables 1 and 2 [e.g., VLFW1 (type a), VLFW1 (type b), VLFW1 (type c), VLFW1 (type d), VLFW1 (type e), VLFW1 (type f), VLFW2 (type a), VLFW2 (type c), VLFW3 (type a), VLFW3 (type b), VLFW3 (type c), VLFW3 (type d), VLFW3 (type e), VLFW3 (type f), VLFW3 (type g), or VLFW4, or any combination thereof, e.g., the framework combinations described herein]. For example, the nucleic acid molecule can comprise a nucleotide sequence as set forth in Tables 1 and 2 or a sequence that is substantially identical thereto (e.g., a sequence that is at least about 85%, 90%, 95%, 99% or more identical thereto, or a sequence that differs from the sequences shown in Tables 1 and 2 by 3, 6, 15, 30, or 45 or fewer nucleotides).

[0104] In another aspect, the nucleic acid molecule comprises one or more heavy chain framework regions and one or more light chain framework regions as described herein. The heavy chain and light chain framework regions can be present in the same vector or in separate vectors.

[0105] In another aspect, the present application features host cells and vectors containing the nucleic acids described herein. The nucleic acids can be present in a single vector, or in separate vectors present in the same host cell or separate host cells. The host cell can be a eukaryotic cell, such as a mammalian cell, an insect cell, a yeast cell, or a prokaryotic cell, such as E. coli. For example, the mammalian cell can be a cultured cell or a cell line. Exemplary mammalian cells include lymphocyte cell lines (e.g., NSO), Chinese hamster ovary cells (CHO), COS cells, oocytes, and cells derived from transgenic animals, such as mammary epithelial cells.

[0106] In one aspect, the present invention features a method of preparing an antibody molecule described herein. The method includes providing a PD-L1 antigen (e.g., an antigen comprising at least a portion of a PD-L1 epitope); obtaining an antibody molecule that specifically binds to the PD-L1 polypeptide; and evaluating whether the antibody molecule specifically binds to the PD-L1 polypeptide, or evaluating the effectiveness of the antibody molecule in modulating, e.g., inhibiting, the activity of PD-L1. The method can further include administering the antibody molecule to a subject, such as a human or non-human animal.

[0107] In another aspect, the present invention provides a composition, such as a pharmaceutical composition, comprising a pharmaceutically acceptable carrier, excipient or stabilizer, and at least one of the anti-PD-L1 antibody molecules described herein. In one embodiment, the composition, such as a pharmaceutical composition, comprises the antibody molecule and one or more agents, such as a combination of a therapeutic agent or another antibody molecule described herein. In one embodiment, the antibody molecule is conjugated to a label or a therapeutic agent.

[0108] The anti-PD-L1 antibody molecules disclosed herein can inhibit, reduce or neutralize one or more activities of PD-L1, resulting in blockade or reduction of immune checkpoints. In certain embodiments, the antibody molecules result in one or more of the following: an increase in tumor-infiltrating lymphocytes, an increase in T cell receptor-mediated proliferation, a decrease in immune evasion by cancerous cells, restoration of effector cell function (e.g., one or more of T cell proliferation, IFN-γ secretion or cytolytic function), inhibition of regulatory T cell function, or effects on the activities of multiple cell types such as regulatory T cells, effector T cells and NK cells. Thus, such antibody molecules can be used to treat or prevent disorders in which enhancement of the immune response in a subject is desired.

[0109] Use of anti-PD-L1 antibody molecules Accordingly, in another aspect, a method of modulating an immune response in a subject is provided. The method comprises administering to the subject an anti-PD-L1 antibody molecule (e.g., a therapeutically effective amount of an anti-PD-L1 antibody molecule) disclosed herein, alone or in combination with one or more agents or procedures, such that the immune response in the subject is modulated. In certain embodiments, the antibody molecule enhances, stimulates or increases the immune response in the subject. The subject can be a mammal, e.g., a primate, preferably a higher primate, e.g., a human (e.g., a patient having or at risk of having a disorder described herein). In certain embodiments, the subject is in need of enhancement of the immune response. In certain embodiments, the subject has or is at risk of having a disorder described herein, e.g., a cancer or an infectious disorder described herein. In certain particular embodiments, the subject is immunosuppressed or at risk of becoming immunosuppressed. For example, the subject has received or is receiving chemotherapy treatment and / or radiation therapy. Alternatively or in combination therewith, the subject is immunosuppressed or at risk of becoming immunosuppressed as a result of an infection.

[0110] In one aspect, a method of treating cancer or a tumor in a subject (e.g., reducing, inhibiting, or delaying progression of one or more thereof) is provided. The method includes administering to the subject an anti-PD-L1 antibody molecule described herein, e.g., a therapeutically effective amount of the anti-PD-L1 antibody molecule alone or in combination with one or more agents or procedures. In certain embodiments, the anti-PD-L1 antibody molecule is administered in combination with, e.g., a modulator of a costimulatory molecule (e.g., an agonist of a costimulatory molecule) or a modulator of an inhibitory molecule (e.g., an inhibitor of an immune checkpoint inhibitor) described herein.

[0111] In certain embodiments, cancers treatable by the anti-PD-L1 antibody molecule include, but are not limited to, solid tumors, hematological cancers (e.g., leukemia, lymphoma, myeloma, e.g., multiple myeloma), and metastatic lesions. In one embodiment, the cancer is a solid tumor. Examples of solid tumors include malignant lesions, e.g., sarcomas and carcinomas, e.g., lung, breast, ovarian, lymphatic, gastrointestinal tract (e.g., colon), anal, genital and urogenital (e.g., kidney, urothelial, bladder cell, prostate), pharyngeal, CNS (e.g., brain, nerve or glial cells), head and neck, skin (e.g., melanoma), nasopharyngeal cancer, e.g., differentiated or undifferentiated metastatic or locally recurrent nasopharyngeal carcinoma), and carcinomas and adenocarcinomas of various organ systems affected by pancreas, etc., and adenocarcinomas including malignant lesions such as colon cancer, rectal cancer, renal cell carcinoma, liver cancer, non-small cell lung cancer, small intestine cancer, and esophageal cancer. The cancer can be of early, intermediate, advanced stage or metastatic cancer.

[0112] In one aspect, the cancer is selected from lung cancer [e.g., non-small cell lung cancer (NSCLC) (e.g., NSCLC by squamous and / or non-squamous histological examination, or NSCLC adenocarcinoma)], melanoma (e.g., advanced melanoma), kidney cancer (e.g., renal cell carcinoma), liver cancer, myeloma (e.g., multiple myeloma), prostate cancer, breast cancer (e.g., breast cancer that does not express one, two, or all of estrogen receptor, progesterone receptor, or Her2 / neu, e.g., triple negative breast cancer), colorectal cancer, pancreatic cancer, head and neck cancer (e.g., head and neck squamous cell carcinoma (HNSCC), anal cancer, gastroesophageal cancer, thyroid cancer, cervical cancer, lymphoproliferative disorders (e.g., post-transplant lymphoproliferative disorder) or hematological cancer, T cell lymphoma, B cell lymphoma, non-Hodgkin lymphoma or leukemia (e.g., myeloid leukemia or lymphocytic leukemia)).

[0113] In another aspect, the cancer is selected from carcinoma (e.g., advanced or metastatic carcinoma), melanoma or lung cancer, e.g., non-small cell lung cancer.

[0114] In one aspect, the cancer is lymphoma, e.g., diffuse large B cell lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma.

[0115] In one aspect, the cancer is breast cancer, e.g., metastatic breast cancer. In one aspect, the cancer is leukemia, e.g., chronic myeloid leukemia.

[0116] In one aspect, the cancer is head and neck cancer, e.g., head and neck squamous cell carcinoma (HNSCC). In one aspect, the cancer is myelodysplastic syndrome.

[0117] In one aspect, the cancer is bladder cancer (e.g., transitional cell carcinoma). In one aspect, the cancer is colon cancer.

[0118] In one aspect, the cancer is lung cancer, such as non-small cell lung cancer (NSCLC), such as stage IV or recurrent NSCLC, NSCLC adenocarcinoma or NSCLC squamous cell carcinoma or small cell lung cancer.

[0119] In one aspect, the cancer is skin cancer, such as melanoma (e.g., stage III or IV melanoma) or Merkel cell carcinoma. In one aspect, the cancer is melanoma, such as advanced melanoma. In one aspect, the cancer is advanced or inoperable melanoma that does not respond to other therapies. In other aspects, the cancer is melanoma having a BRAF mutation (e.g., BRAF V600 mutation). In still other aspects, the anti-PD-L1 antibody molecule is administered after treatment with a BRAF inhibitor (e.g., vemurafenib or dabrafenib) with or without an anti-CTLA-4 antibody (e.g., ipilimumab).

[0120] In another aspect, the cancer is hepatocellular carcinoma, such as advanced hepatocellular carcinoma with or without viral infection, such as chronic viral hepatitis.

[0121] In another aspect, the cancer is prostate cancer, such as advanced prostate cancer. In yet another aspect, the cancer is myeloma, such as multiple myeloma.

[0122] In yet another aspect, the cancer is kidney cancer, such as renal cell carcinoma (RCC) [e.g., metastatic RCC or clear cell renal cell carcinoma (ccRCC), such as advanced or metastatic clear cell renal cell carcinoma].

[0123] In one aspect, the cancer microenvironment has elevated levels of PD-L1 expression. Alternatively or in combination therewith, the cancer microenvironment can have increased IFNγ and / or CD8 expression. In one aspect, alternatively or in combination therewith, the subject has elevated levels of Bim expression (e.g., in PD-1+CD8+ T cells, compared to PD-1-CD8+ T cells).

[0124] In one aspect, the subject is identified as having or being having a tumor having high PD-L1 level or expression or tumor infiltrating lymphocytes (TIL)+ (e.g., having an increased number of TIL), or one or more of both. In certain aspects, the subject is identified as having or being having a tumor having high PD-L1 level or expression and being TIL+. In one aspect, the methods described herein further comprise identifying a subject based on having a tumor having high PD-L1 level or expression or TIL+, or one or more of both. In certain aspects, the methods described herein further comprise identifying a subject based on having a tumor having high PD-L1 level or expression and being TIL+. In one aspect, the tumor that is TIL+ is positive for CD8 and IFNγ. In one aspect, the subject is identified as having or being having a high percentage of cells that are positive for one, two or more of PD-L1, CD8 and / or IFNγ. In certain aspects, the subject is identified as having or being having a high percentage of cells that are positive for all of PD-L1, CD8 and IFNγ. The subject can be identified before, during or after receiving a treatment, e.g., an anti-PD-L1 antibody molecule treatment and / or another treatment described herein. In one aspect, the subject is identified before receiving a treatment, e.g., a treatment described herein (e.g., before the start of treatment or during an inter-treatment interval).

[0125] In one aspect, the methods described herein further include identifying a subject based on having a high percentage of cells that are positive for one, two or more of PD-L1, CD8 and / or IFNγ. In certain aspects, the methods described herein further include identifying a subject based on having a high percentage of cells that are positive for all of PD-L1, CD8 and IFNγ. In one aspect, the subject has or is identified as having one or more of one, two or more of PD-L1, CD8 and / or IFNγ, and lung cancer, such as squamous cell lung cancer or adenocarcinoma of the lung; head and neck cancer; squamous cell cervical cancer; gastric cancer; esophageal cancer; thyroid cancer; melanoma and / or nasopharyngeal cancer (NPC). In certain aspects, the methods described herein are further described for identifying a subject based on having one or more of one, two or more of PD-L1, CD8 and / or IFNγ, and lung cancer, such as squamous cell lung cancer or adenocarcinoma of the lung; head and neck cancer; squamous cell cervical cancer; gastric cancer; thyroid cancer; melanoma and / or nasopharyngeal cancer. The subject can be identified before, during or after receiving a treatment, such as the anti-PD-L1 antibody molecule treatment described herein and / or another treatment. In one aspect, the subject is identified before receiving a treatment, such as the treatment described herein (e.g., before the start of treatment or during a treatment interval).

[0126] The methods and compositions disclosed herein are useful for treating metastatic lesions associated with the cancers described above.

[0127] In a further aspect, the invention provides a method of treating an infectious disease in a subject, the method comprising administering to the subject a therapeutically effective amount of an anti-PD-L1 antibody molecule described herein alone or in combination with one or more agents or procedures. In one aspect, the infectious disease is selected from hepatitis (e.g., hepatitis C infection) or sepsis.

[0128] Furthermore, the present invention provides a method for enhancing an immune response against an antigen in a subject, the method comprising administering to the subject (i) an antigen; and (ii) an anti-PD-L1 antibody molecule such that the immune response against the antigen in the subject is enhanced. The antigen can be, for example, a tumor antigen, a viral antigen, a bacterial antigen, or an antigen derived from a pathogen.

[0129] The anti-PD-L1 antibody molecule can be administered to the subject systemically (e.g., orally, parenterally, subcutaneously, intravenously, rectally, intramuscularly, intraperitoneally, intranasally, transdermally, or by inhalation or intracavitary instillation), locally, or by application to mucous membranes such as the nose, pharynx, and bronchus.

[0130] The dosage and treatment regimen of the anti-PD-L1 antibody molecule can be determined by those skilled in the art. In certain embodiments, the anti-PD-L1 antibody molecule is administered by injection (e.g., subcutaneously or intravenously) at a dose of about 1 to 30 mg / kg, such as about 5 to 25 mg / kg, about 10 to 20 mg / kg, about 1 to 5 mg / kg, or about 3 mg / kg. The dosing schedule can vary, for example, from once a week to once every 2, 3, or 4 weeks. In one embodiment, the anti-PD-L1 antibody molecule is administered at a dose of about 10 to 20 mg / kg once a week. In one embodiment, the anti-PD-L1 antibody molecule is administered alone or in combination (e.g., in combination with an anti-LAG-3 antibody molecule) at a dose of about 5 mg / kg or less; about 4 mg / kg or less; about 3 mg / kg or less; about 2 mg / kg or less; about 1 mg / kg or less once a week. In one embodiment, the anti-PD-L1 antibody molecule is administered at a dose of 1 to 5 mg / kg once a week; 1 to 4 mg / kg once a week, 1 to 3 mg / kg once a week, or 1 to 2 mg / kg once a week. In one embodiment, the anti-LAG-3 antibody molecule is administered alone or in combination (e.g., in combination with an anti-PD-L1 antibody molecule) at a dose of 1 to 5 mg / kg once a week; 1 to 4 mg / kg once a week, 1 to 3 mg / kg once a week, or 1 to 2 mg / kg once a week.

[0131] The antibody molecules described herein can be used in the methods described herein, but other anti-PD-L1 antibodies can be used instead of or in combination with the anti-PD-L1 antibody molecules of the invention.

[0132] Combination therapy The methods and compositions described herein can be used in combination with other agents or treatment modalities. In certain embodiments, the methods described herein comprise administering to a subject an anti-PD-L1 antibody molecule described herein, in an amount effective to treat or prevent a disorder, in combination with an agent or treatment procedure or modality. The anti-PD-L1 antibody molecule and the agent or treatment procedure or modality can be administered simultaneously or sequentially in either order. Any combination and order of the anti-PD-L1 antibody molecule and other therapeutic agents, procedures or modalities (e.g., as described herein) can be used. The antibody molecule and / or other therapeutic agents, procedures or modalities can be administered during the period of an active disorder or during a period of remission or low activity of the disease. The antibody molecule can be administered before, concurrently with, after the other treatment, or during remission of the disorder.

[0133] In certain embodiments, the methods and compositions described herein are administered in combination with one or more of other antibody molecules, chemotherapy, other anti-cancer therapies [e.g., targeted anti-cancer therapies, gene therapy, viral therapy, RNA therapy, bone marrow transplantation, nano-therapy or oncolytic agents], cytotoxic agents, immune-based therapies (e.g., cytokine or cell-based immunotherapies), surgical procedures (e.g., breast tumor excision or mastectomy) or radiation procedures, or combinations of any of the foregoing. The additional therapy can be in the form of adjuvant or neoadjuvant therapy. In one embodiment, the additional therapy is an enzyme inhibitor (e.g., a small molecule enzyme inhibitor) or a metastasis inhibitor. Exemplary cytotoxic agents that can be administered in combination include microtubule inhibitory agents, topoisomerase inhibitors, antimetabolites, mitotic inhibitors, alkylating agents, anthracyclines, vinca alkaloids, intercalating agents, agents that can interfere with signal transduction pathways, agents that promote apoptosis, proteasome inhibitors and radiation (e.g., local or total body irradiation (e.g., gamma irradiation)). In other embodiments, the additional therapy is surgery or radiation or a combination thereof. In other embodiments, the additional therapy is a therapy that targets one or more of the PI3K / AKT / mTOR pathway, HSP90 inhibitors or tubulin inhibitors.

[0134] Alternatively, or in combination with the foregoing combinations, the methods and compositions described herein can be administered in combination with one or more of: immunomodulatory agents (e.g., activators or inhibitory molecules of costimulatory molecules, e.g., inhibitors of immune checkpoint molecules); vaccines, e.g., therapeutic cancer vaccines; or other forms of cellular immunotherapy.

[0135] Exemplary non-limiting combinations and uses of anti-PD-L1 antibody molecules include the following.

[0136] In certain embodiments, the anti-PD-L1 antibody molecule is administered in combination with a costimulatory or inhibitory molecule, e.g., a modulator of a coinhibitory ligand or receptor.

[0137] In certain embodiments, the anti-PD-L1 antibody molecule is administered in combination with a modulator of a costimulatory molecule, such as an agonist. In certain embodiments, the agonist of the costimulatory molecule is selected from agonists of OX40, CD2, CD27, CD8, ICAM-1, LFA-1 (CD11a / CD18), ICOS (CD278), 4-1BB (CD137), GITR, CD30, CD40, BAFFR, HVEM, CD7, LIGHT, NKG2C, SLAMF7, NKp80, CD160, B7-H3 or CD83 ligand (e.g., an agonist antibody or an antigen-binding fragment thereof or a soluble fusion).

[0138] In certain embodiments, the anti-PD-L1 antibody molecule is administered in combination with an inhibitor of an inhibitory (or immune checkpoint) molecule selected from PD-1, PD-L2, CTLA-4, TIM-3, LAG-3, CEACAM (e.g., CEACAM-1, CEACAM-3 and / or CEACAM-5), VISTA, BTLA, TIGIT, LAIR1, CD160, 2B4 and / or TGFR beta. Inhibition of the inhibitory molecule can be effected by inhibition at the DNA, RNA or protein level. In embodiments, an inhibitory nucleic acid (e.g., dsRNA, siRNA or shRNA) can be used to inhibit the expression of the inhibitory molecule. In other embodiments, the inhibitor of the inhibitory signal is a polypeptide that binds to the inhibitory molecule, such as a soluble ligand or an antibody or an antigen-binding fragment thereof. In certain embodiments, the inhibitor is a soluble ligand (e.g., CTLA-4-Ig) or an antibody or antibody fragment that binds to PD-1, PD-L2 or CTLA-4. For example, the anti-PD-L1 antibody molecule can be administered in combination with an anti-CTLA-4 antibody, such as ipilimumab, to treat, for example, a cancer (e.g., melanoma, such as metastatic melanoma; lung cancer, such as non-small cell lung cancer; or prostate cancer). In certain embodiments, the anti-PD-1 antibody molecule is administered after treatment with a BRAF inhibitor (e.g., vemurafenib or dabrafenib) with or without an anti-CTLA-4 antibody (e.g., ipilimumab).

[0139] In another aspect, the anti-PD-L1 antibody molecule is administered in combination with an anti-LAG-3 antibody or an antigen-binding fragment thereof.

[0140] In another aspect, the anti-PD-L1 antibody molecule is administered in combination with an anti-TIM-3 antibody or an antigen-binding fragment thereof.

[0141] In yet another aspect, the anti-PD-L1 antibody molecule is administered in combination with an anti-LAG-3 antibody and an anti-TIM-3 antibody (or antigen-binding fragments thereof).

[0142] In another aspect, the anti-PD-L1 antibody is administered in combination with a CEACAM inhibitor (e.g., CEACAM-1, CEACAM-3, and / or CEACAM-5), e.g., an anti-CEACAM antibody molecule. In another aspect, the anti-PD-L1 antibody molecule is administered in combination with a CEACAM-1 inhibitor, e.g., an anti-CEACAM-1 antibody molecule. In another aspect, the anti-PD-L1 antibody molecule is administered in combination with a CEACAM-5 inhibitor, e.g., an anti-CEACAM-5 antibody molecule.

[0143] The combinations of antibodies recited herein can be administered separately, e.g., as separate antibodies or antigen-binding fragments thereof, or linked, e.g., as bispecific or trispecific antibody molecules. In one aspect, a bispecific antibody comprising an anti-PD-L1 antibody molecule and an anti-TIM-3, anti-CEACAM (e.g., CEACAM-1, CEACAM-3, and / or CEACAM-5), or anti-LAG-3 antibody or antigen-binding fragment thereof is administered. In certain aspects, the combinations of antibodies recited herein are used for the treatment of cancer, e.g., cancer as described herein (e.g., solid tumors or hematological malignancies).

[0144] In one aspect, antibody molecules (e.g., single, dual or triple specific antibodies) against TIM-3, LAG-3 and / or PD-1 used in any of the methods and compositions disclosed herein comprise the amino acid sequences described herein or are encoded by the nucleotide sequences described herein [e.g., as disclosed in the section entitled "Inhibitors of Immune Checkpoint Molecules" starting on page 218 hereinafter (including any publications mentioned therein)].

[0145] In other aspects, anti-PD-L1 antibody molecules are administered in combination with a cytokine. The cytokine can be administered as a fusion to the anti-PD-L1 antibody molecule or as a separate composition. In one aspect, the anti-PD-L1 antibody is administered in combination with 1, 2 or more than 3 cytokines, for example, as a fusion molecule or as a separate composition. In one aspect, the cytokine is an interleukin (IL) selected from 1, 2 or more than 3 of IL-1, IL-2, IL-15 or IL-21. In a particular aspect, the combination of the anti-PD-L1 antibody molecule and the cytokine described herein is used for the treatment of cancer, for example, cancer (e.g., solid tumors) described herein.

[0146] In a particular aspect, the anti-PD-L1 antibody molecule is administered in combination with an antibody specific for HLA C, for example, an antibody specific for a killer cell immunoglobulin-like receptor (also referred to herein as an "anti-KIR antibody"). In a particular aspect, the combination of the anti-PD-L1 antibody molecule and the anti-KIR antibody is used for the treatment of cancer, for example, cancer (e.g., solid tumors, e.g., advanced solid tumors) described herein.

[0147] In one aspect, the anti-PD-L1 antibody molecule is administered in combination with cellular immunotherapy (e.g., Provenge® (e.g., Sipuleucel-T)) and optionally in combination with cyclophosphamide. In certain aspects, the combination of the anti-PD-L1 antibody molecule, Provenge® and / or cyclophosphamide is used for the treatment of cancer, such as the cancers described herein (e.g., prostate cancer, e.g., advanced prostate cancer).

[0148] In another aspect, the anti-PD-L1 antibody molecule is administered in combination with a vaccine, such as a cancer vaccine (e.g., dendritic cell renal cell carcinoma (DC-RCC) vaccine). In one aspect, the vaccine is peptide-based, DNA-based, RNA-based or antigen-based or combinations thereof. In an aspect, the vaccine comprises one or more peptides, nucleic acids (e.g., DNA or RNA), antigens or combinations thereof. In certain aspects, the combination of the anti-PD-L1 antibody molecule and the DC-RCC vaccine is used for the treatment of cancer, such as the cancers described herein [e.g., renal cancer, e.g., metastatic renal cell carcinoma (RCC) or clear cell renal cell carcinoma (CCRCC)].

[0149] In another aspect, the anti-PD-1 antibody molecule is administered in combination with an adjuvant.

[0150] In yet another aspect, the anti-PD-L1 antibody molecule is administered in combination with chemotherapy and / or immunotherapy. For example, the anti-PD-L1 antibody molecule can be used alone or in combination with one or more of the following to treat multiple myeloma: chemotherapy or other anti-cancer agents (e.g., thalidomide analogs, e.g., lenalidomide), anti-TIM-3 antibodies, dendritic cells pulsed with tumor antigens, fusions of tumor cells and dendritic cells (e.g., electrofusions), or vaccination with immunoglobulins idiotypes produced by malignant plasma cells. In one aspect, the anti-PD-L1 antibody molecule is used in combination with an anti-TIM-3 antibody to treat multiple myeloma, such as multiple myeloma.

[0151] In one aspect, an anti-PD-L1 antibody molecule is used in combination with chemotherapy to treat lung cancer, such as non-small cell lung cancer. In one aspect, an anti-PD-L1 antibody molecule is used in combination with standard lung, e.g., NSCLC chemotherapy, e.g., platinum doublet therapy, to treat lung cancer. In yet another aspect, in a subject having progressive or metastatic cancer [e.g., a patient having metastatic and recurrent NSCL cancer], the anti-PD-L1 antibody molecule is used in combination with an indoleamine-pyrrole 2,3-dioxygenase (IDO) inhibitor {e.g., (4E)-4-[(3-chloro-4-fluoroanilino)-nitrosomethylidene]-1,2,5-oxadiazol-3-amine (also known as INCB24360), indoximod (1-methyl-D-tryptophan), α-cyclohexyl-5H-imidazo[5,1-a]isoindole-5-ethanol (also known as NLG919), etc.}.

[0152] In yet another aspect, the anti-PD-L1 antibody molecule is used in combination with one or more of the following: immune-based strategies (e.g., interleukin-2 or interferon-α), targeted agents (e.g., monoclonal antibodies against VEGF, etc., VEGF inhibitors); VEGF tyrosine kinase inhibitors such as sunitinib, sorafenib, axitinib, and pazopanib; RNAi inhibitors; or inhibitors of downstream mediators of VEGF signaling, e.g., inhibitors of the mammalian target of rapamycin (mTOR) of rapamycin, e.g., everolimus and temsirolimus. Any such combination can be used to treat kidney cancer, such as renal cell carcinoma (RCC) [e.g., clear cell renal cell carcinoma (CCRCC)] or metastatic RCC.

[0153] In one aspect, an anti-PD-L1 antibody molecule, such as the anti-PD-L1 antibody molecules described herein, is used in combination with a MEK inhibitor (e.g., a MEK inhibitor described herein). In one aspect, a combination of an anti-PD-L1 antibody and a MEK inhibitor is used for the treatment of cancer (e.g., a cancer described herein). In one aspect, the cancer treated by this combination is selected from melanoma, colorectal cancer, non-small cell lung cancer, ovarian cancer, breast cancer, prostate cancer, pancreatic cancer, hematological malignancies or renal cell carcinoma. In certain aspects, the cancer comprises a BRAF mutation (e.g., a BRAF V600E mutation), BRAF wild-type, KRAS wild-type or an activated KRAS mutation. The cancer can be of an early, intermediate or late stage.

[0154] In another aspect, the anti-PD-L1 antibody molecule is used in combination with one, two or all of oxaliplatin, leucovorin or 5-FU (e.g., FOLFOX co-treatment). Alternatively or in combination therewith, the combination further comprises a VEGF inhibitor (e.g., a VEGF inhibitor disclosed herein). In one aspect, a combination of an anti-PD-L1 antibody, FOLFOX co-treatment and a VEGF inhibitor is used for the treatment of cancer (e.g., a cancer described herein). In one aspect, the cancer treated by this combination is selected from melanoma, colorectal cancer, non-small cell lung cancer, ovarian cancer, breast cancer, prostate cancer, pancreatic cancer, hematological malignancies or renal cell carcinoma. The cancer can be of an early, intermediate or late stage.

[0155] In other aspects, the anti-PD-L1 antibody molecule is administered with a tyrosine kinase inhibitor (e.g., axitinib) to treat renal cell carcinoma and other solid tumors.

[0156] In other embodiments, the anti-PD-L1 antibody molecule is administered with a 4-1BB receptor targeting agent [e.g., an antibody that stimulates signaling by 4-1BB (CD-137), e.g., PF-2566]. In certain embodiments, the anti-PD-L1 antibody molecule is administered in combination with a tyrosine kinase inhibitor (e.g., axitinib) and a 4-1BB receptor targeting agent.

[0157] The anti-PD-L1 antibody molecule can be conjugated to a substance, such as a cytotoxic agent or moiety (e.g., a therapeutic agent; a compound that emits radiation; a molecule of plant, fungal or bacterial origin; or a biological protein (e.g., a protein toxin) or particle (e.g., a recombinant viral particle, e.g., via a viral coat protein). For example, the antibody can be conjugated to a radioisotope, such as an α-, β- or γ-emitter or a β- and γ-emitter.

[0158] Any combination and order of the anti-PD-L1 antibody molecule and other therapeutic agents, procedures or modalities (e.g., as described herein) may be used. The antibody molecule and / or other therapeutic agents, procedures or modalities can be administered during a period of active disease or during a period of remission or low activity of the disease. The antibody molecule can be administered before, concurrently with, after, or during remission of the disorder, other treatments.

[0159] Additional combination therapies The methods and compositions described herein (e.g., anti-PD-L1 antibodies and methods of using the same) can be used in combination with other agents or treatment modalities, e.g., a second therapeutic agent selected from one or more of the agents listed in Table 6. In certain embodiments, the methods described herein comprise administering to a subject an anti-PD-L1 antibody molecule [an inhibitor of one or more of PD-1, LAG-3, TIM-3, CEACAM (e.g., CEACAM-1, CEACAM-3, and / or CEACAM-5), or CTLA-4 as described herein], and further comprise administering, in an amount effective to treat or prevent a disorder, e.g., a disorder described herein, e.g., cancer, a second therapeutic agent selected from one or more of the agents listed in Table 6. When administered in combination, the anti-PD-L1 antibody molecule, additional agent (e.g., a second or third agent), or all of them can be administered individually at an amount or dosage that is more, less, or the same as the amount or dosage of each agent used as monotherapy, e.g., individually. In certain embodiments, the amount or dosage of the anti-PD-L1 antibody, additional agent (e.g., a second or third agent), or all of them administered is individually less (e.g., at least 20%, at least 30%, at least 40%, or at least 50%) than the amount or dosage of each agent used as monotherapy, e.g., individually. In other embodiments, the amount or dosage of the anti-PD-L1 antibody, additional agent (e.g., a second or third agent), or all of them that produces the desired effect (e.g., treatment of cancer) is less (e.g., at least 20%, at least 30%, at least 40%, or at least 50% less).

[0160] In other embodiments, the second therapeutic agent is selected from one or more of the agents listed in Table 6. In certain embodiments, the cancer is selected from lung cancer (e.g., non-small cell lung cancer (NSCLC) (e.g., NSCLC by squamous and / or non-squamous histological examination, or NSCLC adenocarcinoma), or is disclosed in the publications listed in Table 6.In one aspect, the second therapeutic agent is selected from one or more of: 1) a protein kinase C (PKC) inhibitor; 2) a heat shock protein 90 (HSP90) inhibitor; 3) an inhibitor of phosphoinositide 3-kinase (PI3K) and / or mammalian target of rapamycin (mTOR); 4) an inhibitor of cytochrome P450 (e.g., a CYP17 inhibitor or a 17alpha-hydroxylase / C17-20 lyase inhibitor); 5) an iron chelating agent; 6) an aromatase inhibitor; 7) an inhibitor of p53, e.g., an inhibitor of the p53 / Mdm2 interaction; 8) an apoptosis inducer; 9) an angiogenesis inhibitor; 10) an aldosterone synthase inhibitor; 11) a smoothened (SMO) receptor inhibitor; 12) a prolactin receptor (PRLR) inhibitor; 13) a Wnt signaling inhibitor; 14) a CDK4 / 6 inhibitor; 15) a fibroblast growth factor receptor 2 (FGFR2) / fibroblast growth factor receptor 4 (FGFR4) inhibitor; 16) an inhibitor of macrophage colony-stimulating factor (M-CSF); 17) an inhibitor of one or more of c-KIT, histamine release, Flt3 (e.g., FLK2 / STK1) or PKC; 18) an inhibitor of one or more of VEGFR-2 (e.g., FLK-1 / KDR), PDGFR beta, c-KIT or Raf kinase C; 19) a somatostatin agonist and / or a growth hormone release inhibitor; 20) an anaplastic lymphoma kinase (ALK) inhibitor; 21) an insulin-like growth factor 1 receptor (IGF-1R) inhibitor; 22) a P-glycoprotein 1 inhibitor; 23) a vascular endothelial growth factor receptor (VEGFR) inhibitor; 24) a BCR-ABL kinase inhibitor; 25) an FGFR inhibitor; 26) an inhibitor of CYP11B2; 27) an HDM2 inhibitor, e.g., an inhibitor of the HDM2-p53 interaction; 28) an inhibitor of tyrosine kinase; 29) an inhibitor of c-MET; 30) an inhibitor of JAK; 31) an inhibitor of DAC; 32) an inhibitor of 11beta-hydroxylase; 33) an inhibitor of IAP; 34) an inhibitor of PIM kinase; 35) an inhibitor of Porcupine; 36) an inhibitor of BRAF, e.g., BRAF V600E or wild-type BRAF; 37) an inhibitor of HER3; 38) an inhibitor of MEK; or 39) an inhibitor of lipid kinase, e.g., selected from one or more of those described herein and in Table 6.

[0161] In one embodiment, the second therapeutic agent is selected from one or more of compound A8, compound A17, compound A23, compound A24, compound A27, compound A29, compound A33, and compound A13.

[0162] In another embodiment, the second therapeutic agent is selected from one or more of compound A5, compound A8, compound A17, compound A23, compound A24, compound A29, and compound A40.

[0163] In another embodiment, the second therapeutic agent is selected from one or more of compound A9, compound A16, compound A17, compound A21, compound A22, compound A25, compound A28, compound A48, and compound 49.

[0164] In one embodiment, the second therapeutic agent is administered at a therapeutic dose or a dose lower than the therapeutic dose. In certain embodiments, the concentration of the second therapeutic agent required to achieve inhibition, e.g., growth inhibition, is lower when the second therapeutic agent is administered in combination with an anti-PD-L1 antibody molecule than when the second therapeutic agent is administered individually. In certain embodiments, the concentration of the anti-PD-L1 antibody molecule required to achieve inhibition, e.g., growth inhibition, is lower when the anti-PD-L1 antibody molecule is administered in combination with the second therapeutic agent than when the anti-PD-L1 antibody molecule is administered individually. In certain embodiments, in the combination therapy, the concentration of the second therapeutic agent required to achieve inhibition, e.g., growth inhibition, is lower than the therapeutic dose of the second therapeutic agent as a monotherapy, e.g., 10 - 20%, 20 - 30%, 30 - 40%, 40 - 50%, 50 - 60%, 60 - 70%, 70 - 80%, or 80 - 90% lower. In certain embodiments, in the combination therapy, the concentration of the anti-PD-L1 antibody molecule required to achieve inhibition, e.g., growth inhibition, is lower than the therapeutic dose of the anti-PD-L1 antibody molecule as a monotherapy, e.g., 10 - 20%, 20 - 30%, 30 - 40%, 40 - 50%, 50 - 60%, 60 - 70%, 70 - 80%, or 80 - 90% lower.

[0165] Additional features and embodiments Alternatively, or in combination with the methods disclosed herein, the present invention features methods of treating (e.g., inhibiting, reducing, alleviating or preventing) a disorder in a subject, such as a proliferative condition or disorder (e.g., cancer). The methods include administering to the subject a combination of two or more therapeutic agents selected from one, two or all of the following categories (i)-(iii): (i) an agent that enhances antigen (e.g., tumor antigen) presentation; (ii) an agent that enhances effector cell responses (e.g., B cell and / or T cell activation and / or recruitment); or (iii) an agent that reduces tumor immunosuppression, thereby treating a disorder, such as a proliferative condition or disorder (e.g., cancer). In one aspect, the combination includes a PD-L1 inhibitor (e.g., an anti-PD-L1 antibody molecule described herein). The cancer to be treated can be, for example, lung cancer (squamous), lung cancer (adenocarcinoma), head and neck cancer, cervical cancer (squamous), gastric cancer, thyroid cancer, melanoma, nasopharyngeal cancer or breast cancer, etc., as described herein.

[0166] In another aspect, the present invention features methods of reducing the activity (e.g., growth, survival or viability or all thereof) of proliferating (e.g., cancerous) cells. The methods include contacting the cells with a combination of two or more therapeutic agents selected from one, two or all of the following categories (i)-(iii): (i) an agent that enhances antigen (e.g., tumor antigen) presentation; (ii) an agent that enhances effector cell responses (e.g., B cell and / or T cell activation and / or recruitment); or (iii) an agent that reduces tumor immunosuppression, thereby reducing the activity in proliferating cells. In one aspect, the combination includes a PD-L1 inhibitor (e.g., an anti-PD-L1 antibody molecule described herein). The methods can be performed in a subject, for example, as part of a treatment protocol. The cancer cells can be, for example, cells derived from cancers such as lung cancer (squamous), lung cancer (adenocarcinoma), head and neck cancer, cervical cancer (squamous), gastric cancer, thyroid cancer, melanoma, nasopharyngeal cancer or breast cancer, etc., as described herein.

[0167] In certain aspects of the methods disclosed herein, the method further includes determining the level and / or distribution of immune cell (e.g., T cell) infiltration in a subject [e.g., the level and / or distribution of tumor infiltrating lymphocytes (TIL)]. In one aspect, the level and / or distribution of immune cell infiltration is determined in vivo, e.g., non-invasively [e.g., by detecting an antibody to a T cell marker detectably labeled using a suitable imaging technique, e.g., a positron emission tomography (PET) scan]. In other aspects, the level of immune cell infiltration is determined in a sample obtained from the subject (e.g., a tumor biopsy) (e.g., using immunohistochemical techniques). In one aspect, an elevated level and / or a more extensive distribution of TIL in a cancer, e.g., a tumor (e.g., as compared to a reference or control) indicates a better prognosis, e.g., a more favorable treatment outcome, for the subject. In one aspect, a decreased level and / or a more restricted distribution of TIL in a cancer, e.g., a tumor (e.g., as compared to a reference or control) indicates a worse prognosis, e.g., a more unfavorable treatment outcome, for the subject. In one aspect, the reference is the subject at different time intervals, e.g., prior to or at an earlier stage of the treatment regimen. In one aspect, in response to low or undetectable tumor infiltration in the subject, one or more agents of category (i) or (ii) or both (i) and (ii) are administered. In other aspects, in response to detectable or elevated levels of tumor infiltration in the subject, one or more agents of category (iii) are administered. The detection step can also be used to monitor, e.g., the efficacy of a therapeutic agent described herein. For example, the detection step can be used to monitor the efficacy of therapeutic agents of categories (i), (ii), and / or (iii).

[0168] In another aspect, the present invention features a composition (e.g., one or more compositions or dosage forms) comprising a combination of two or more therapeutic agents selected from one, two or all of the following categories (i)-(iii): (i) an agent that enhances antigen (e.g., tumor antigen) presentation; (ii) an agent that enhances effector cell responses (e.g., activation and / or recruitment of B cells and / or T cells); or (iii) an agent that reduces tumor immunosuppression. In certain embodiments, this combination comprises a PD-L1 inhibitor (e.g., an anti-PD-L1 antibody molecule described herein).

[0169] In yet another aspect, the present invention features a composition [e.g., one or more compositions or dosage forms described herein (hereom)] for use in the treatment of a disorder, such as cancer. In certain embodiments, the composition for use comprises a combination of two or more therapeutic agents selected from one, two or all of the following categories (i)-(iii): (i) an agent that enhances antigen (e.g., tumor antigen) presentation; (ii) an agent that enhances effector cell responses (e.g., activation and / or recruitment of B cells and / or T cells); or (iii) an agent that reduces tumor immunosuppression. In certain embodiments, the combination used comprises a PD-L1 inhibitor (e.g., an anti-PD-L1 antibody molecule described herein). The cancer can be, for example, lung cancer (squamous cell), lung cancer (adenocarcinoma), head and neck cancer, cervical cancer (squamous cell), gastric cancer, thyroid cancer, melanoma, nasopharyngeal cancer or breast cancer, etc., as described herein.

[0170] Formulations, such as dosage formulations and kits, such as therapeutic kits, comprising a combination of two or more therapeutic agents selected from one, two or all of the following categories (i)-(iii): (i) agents that enhance antigen (e.g., tumor antigen) presentation; (ii) agents that enhance effector cell responses (e.g., activation and / or recruitment of B cells and / or T cells); or (iii) agents that reduce tumor immunosuppression and thereby reduce activity in cells, and (optionally) instructions for use are also disclosed. In one aspect, this combination comprises a PD-L1 inhibitor (e.g., an anti-PD-L1 antibody molecule as described herein).

[0171] The combinations of therapeutic agents disclosed herein comprise two or more therapeutic agents as described herein. As described below, the therapeutic agents in the combination can belong to the same category, e.g., can be two or more therapeutic agents of category (i), or can comprise at least one agent of two or more categories [e.g., a therapeutic agent of category (i) combined with a therapeutic agent of category (ii)]. A particular therapeutic agent can belong to two or more of categories (i)-(iii). For example, therapeutic agents (e.g., especially GITR agonists, IDO antagonists, TGF-β inhibitors) can act as therapeutic agents in multiple categories.

[0172] In certain embodiments, the combinations disclosed herein include one, two, three, four or more therapeutic agents (referred to herein as "antigen presentation combinations") that enhance antigen (e.g., tumor antigen) presentation. In certain embodiments, the antigen presentation combinations include one or more of the following: agents that enhance antigen presentation (e.g., vaccines, e.g., cell- or antigen-based vaccines); agents that enhance lysis of tumor cells (e.g., oncolytic viruses); agents that stimulate (e.g., derepress) phagocytic cells, e.g., type I interferon (IFN) activators [e.g., TLR agonists, RIG-I-like receptor agonists (RLRs)], and / or agents that activate and / or recruit dendritic cells or macrophages (e.g., macrophage I), e.g., bispecific or trispecific cell engaging factors.

[0173] In one aspect, the antigen-presenting combination comprises 1, 2, 3, 4, 5 or more therapeutic agents selected from the following: (i) an agonist of the stimulator of interferon genes (STING agonist), (ii) an agonist of a Toll-like receptor (TLR) (e.g., an agonist of TLR-3, -4, -5, -7, -8 or -9), (iii) a TIM-3 modulator (e.g., an anti-TIM-3 antibody molecule), (iv) a vascular endothelial growth factor receptor (VEGFR) inhibitor, (v) a c-Met inhibitor, (vi) a TGFβ inhibitor (e.g., an anti-TGFβ antibody), (vii) an IDO / TDO inhibitor, (viii) an A2AR antagonist, (ix) an oncolytic virus, (x) a vaccine (e.g., a scaffold vaccine) or (xi) a bispecific or trispecific cell-engaging factor. Any combination of the aforementioned agents (i)-(xi) can be used in the antigen-presenting combination. In one exemplary aspect, the antigen-presenting combination comprises a STING agonist. In another exemplary aspect, the antigen-presenting combination comprises a TLR agonist (e.g., a TLR7 agonist). In another exemplary aspect, the antigen-presenting combination comprises a STING agonist and a TLR agonist (e.g., a TLR7 agonist). In one aspect, the antigen-presenting combination comprises a STING agonist, a TLR agonist, an A2AR antagonist or an oncolytic virus or a combination thereof, and optionally one or more selected from (iii)-(vii) or (x)-(xi). In one aspect, the antigen-presenting combination comprises a STING agonist or a TLR agonist or a combination of both, and optionally one or more selected from (iii)-(xi). In another aspect, the antigen-presenting combination comprises a STING agonist, a TLR agonist (e.g., a TLR7 agonist) and a TIM-3 modulator (e.g., an anti-TIM-3 inhibitor). In another aspect, the antigen-presenting combination comprises a STING agonist, a TLR agonist (e.g., a TLR7 agonist) and a VEGFR inhibitor. In another aspect, the antigen-presenting combination comprises a STING agonist, a TLR agonist (e.g., a TLR7 agonist) and a c-MET inhibitor. In still another aspect, the antigen-presenting combination comprises an oncolytic virus.In other aspects, the antigen presenting combination comprises a tumor-lytic virus and one or more of cytokines such as GM-CSF or CSF (e.g., CSF1 or CSF2), wherein the tumor-lytic virus expresses the cytokine(s). In certain aspects, the antigen presenting combination comprises a dual or triple specific cell engaging factor, e.g., a dual or triple specific antibody molecule against CD47 and CD19, with or without an Fc domain. In certain aspects, the antigen presenting combination comprises a TGFb inhibitor (e.g., an anti-TGFb antibody). In other aspects, the antigen presenting combination comprises an IDO / TDO inhibitor. In still other aspects, the antigen presenting combination comprises an A2AR antagonist. In still other aspects, the antigen presenting combination comprises a vaccine {e.g., IL-2 in combination with MUC1, or a dendritic cell-based vaccine [e.g., Provenge®]}. In still other aspects, the antigen presenting combination comprises a vaccine and a TLR agonist (e.g., a TLR agonist described herein). In certain specific aspects, the antigen presenting combination comprises a vaccine and a STING agonist. In certain specific aspects, the antigen presenting combination comprises a vaccine, a STING agonist and a TLR agonist.

[0174] In certain embodiments, the combination comprises 1, 2, 3, 4, 5 or more therapeutic agents (referred to herein as "effector cell combinations") that enhance an effector cell response. In some embodiments, the effector cell combination comprises a lymphocyte activating factor, such as an NK cell activating factor and / or a T cell activating factor. In some embodiments, the effector cell combination activates (e.g., unleashes) tumor infiltrating lymphocytes (TILs), such as NK cells or T cells. In some embodiments, the effector cell combination comprises a modulator of an NK receptor (e.g., an antibody molecule) (e.g., one or more modulators of NKG2A, KIR3DL, NKp46, MICA or CEACAM1); an interleukin or interleukin variant (e.g., IL-2, IL-15, IL-21, IL-13R or IL-12 cytokine or variants thereof, or combinations thereof); a bispecific or trispecific cell engaging factor (e.g., a bispecific antibody molecule of NKG2A and CD138, or a bispecific antibody molecule of CD3 and TCR); an NK cell therapy; or an NK cell modulator selected from a vaccine comprising NK cells and an antigen / immunostimulatory substance. In some embodiments, the effector cell combination comprises an immunomodulatory agent (e.g., one or more of the activators of co-stimulatory molecules or inhibitors of immune checkpoint molecules described herein). In some embodiments, the effector cell combination comprises a T cell modulator selected from inhibitors of checkpoint inhibitors (e.g., inhibitors of one or more of PD-1, PD-L1, TIM-3, LAG-3, VISTA, DKG-α, B7-H3, B7-H4, TIGIT, CTLA-4, BTLA, CD160, TIM1, IDO, LAIR1, IL-12 or combinations thereof, e.g., an inhibitor of PD-1 and TIM-3, or an inhibitor of PD-1 and LAG-3). In some embodiments, the inhibitor of the checkpoint inhibitor is an antibody molecule (e.g., a single or bispecific antibody or fragments thereof described herein).For example, the inhibitor of the checkpoint inhibitor is an antibody molecule against PD-1, PD-L1, TIM-3, LAG-3, VISTA, B7-H4, CTLA-4 or TIGIT or any combination thereof (e.g., the combinations described herein). In certain embodiments, the effector cell combination comprises a T cell modulator selected from agonists or activators of costimulatory molecules. In certain embodiments, the agonist of the costimulatory molecule is an agonist of GITR, OX40, ICOS, SLAM (e.g., SLAMF7), HVEM, LIGHT, CD2, CD27, CD28, CDS, ICAM-1, LFA-1 (CD11a / CD18), ICOS (CD278), 4-1BB (CD137), CD30, CD40, BAFFR, CD7, NKG2C, NKp80, CD160, B7-H3 or CD83 ligand (e.g., an agonist antibody or an antigen-binding fragment thereof, or a soluble fusion). In other embodiments, the effector cell combination comprises a bispecific T cell engaging factor [e.g., a bispecific antibody molecule that binds CD3 and a tumor antigen (e.g., especially EGFR, PSCA, PSMA, EpCAM, HER2)].

[0175] In one aspect, the effector cell combination comprises 1, 2, 3, 4, 5 or more therapeutic agents selected from the following: (i) a GITR modulator (e.g., a GITR agonist), (ii) a PD-L1 inhibitor (e.g., an anti-PD-L1 antibody molecule as described herein), (iii) a PD-1 inhibitor, (iv) an inhibitor of IAP (inhibitor of apoptosis protein), (v) an inhibitor of EGFR (epidermal growth factor receptor), (vi) an inhibitor of mammalian target of rapamycin (mTOR), (vii) IL-15 or a variant thereof, (viii) a CTLA-4 inhibitor, (ix) a bispecific T cell engaging factor [e.g., a bispecific antibody molecule that binds CD3 and a tumor antigen (e.g., especially EGFR, PSCA, PSMA, EpCAM, HER2)], (x) a CD40 agonist (e.g., an anti-CD40 antibody molecule), (xi) an OX40 agonist (e.g., an anti-OX40 antibody molecule) or (xii) a CD27 agonist (e.g., an anti-CD27 antibody molecule). Any combination of the aforementioned agents can be used in the effector cell combination. In one exemplary aspect, the effector cell combination comprises a GITR agonist. In another aspect, the effector cell combination comprises a PD-L1 inhibitor (e.g., an anti-PD-L1 antibody molecule as described herein). In another aspect, the effector cell combination comprises a PD-1 inhibitor. In other aspects, the effector cell combination comprises a GITR agonist and a PD-L1 inhibitor (e.g., an anti-PD-L1 antibody molecule as described herein). In other aspects, the effector cell combination comprises a GITR agonist and a PD-1 inhibitor. In other aspects, the effector cell combination comprises a GITR agonist, a PD-L1 inhibitor (e.g., an anti-PD-L1 antibody molecule as described herein) and a PD-1 inhibitor. In other aspects, the effector cell combination comprises a PD-L1 inhibitor (e.g., an anti-PD-L1 antibody molecule as described herein) and a PD-1 inhibitor. In one aspect, the effector cell combination comprises a GITR agonist and an inhibitor of IAP. In another aspect, the effector cell combination comprises a GITR agonist and an inhibitor of an EGFR inhibitor. In yet another aspect, the effector cell combination comprises a GITR agonist and an inhibitor of an mTOR inhibitor.In one aspect, the effector cell combination comprises IL-15 or a variant thereof. In one aspect, the effector cell combination comprises a CTLA-4 inhibitor. In one aspect, the effector cell combination comprises a bispecific T cell engaging factor [e.g., a bispecific antibody molecule that binds to CD3 and a tumor antigen (e.g., EGFR, PSCA, PSMA, EpCAM, HER2, among others)]. In one aspect, the effector cell combination comprises a CD40 agonist (e.g., an anti-CD40 antibody molecule). In one aspect, the effector cell combination comprises an OX40 agonist (e.g., an anti-OX40 antibody molecule). In one aspect, the effector cell combination comprises a CD27 agonist (e.g., an anti-CD27 antibody molecule).

[0176] In certain aspects, the combination comprises 1, 2, 3, 4, 5 or more therapeutic agents that reduce tumor immunosuppression (referred to herein as an "anti-tumor immunosuppression combination"). In one aspect, this combination reg modulates the activity or level of one or more of T regIncrease one or more of dehydration or T cell recruitment. In certain embodiments, the anti-tumor immunosuppressive combination comprises 1, 2, 3, 4, 5 or more therapeutic agents selected from the following: (i) immunomodulatory agents [e.g., activators of co-stimulatory molecules described herein (e.g., GITR agonists) or inhibitors of immune checkpoint molecules (e.g., one or more of PD-L1, PD-1, LAG-3, TIM-3 or CTLA-4)], (ii) CSF-1 / 1R inhibitors [e.g., inhibitors of macrophage colony-stimulating factor (M-CSF)], (iii) IL-17 inhibitors, (iv) IL-1 beta inhibitors, (v) CXCR2 inhibitors, (vi) inhibitors of phosphoinositide 3-kinase (PI3K, e.g., PI3K delta or PI3K gamma), (vii) BAFF-R inhibitors, (viii) MALT-1 / BTK inhibitors, (ix) JAK inhibitors, (x) CRTH2 inhibitors, (xi) VEGFR inhibitors, (xiii) IL-15 or variants thereof, (xiv) CTLA-4 inhibitors, (xv) IDO / TDO inhibitors, (xvi) A2AR antagonists, (xvii) TGFb inhibitors, or (xviii) PFKFB3 inhibitors. In certain embodiments, the immunomodulatory agent is an inhibitor of an immune checkpoint molecule [e.g., an inhibitor of PD-L1, PD-1, LAG-3, TIM-3, CEACAM (e.g., CEACAM-1, -3 and / or -5) or CTLA-4 or any combination thereof]. Any combination of the foregoing agents can be used in the tumor immunosuppressive combination. In an exemplary embodiment, the anti-tumor immunosuppressive combination comprises 1, 2, 3, 4, 5 or more therapeutic agents selected from a PD-L1 inhibitor (e.g., an anti-PD-L1 antibody molecule described herein), a PD-1 inhibitor, a LAG-3 inhibitor, a TIM-3 modulator (e.g., an anti-TIM-3 inhibitor), a GITR agonist, a CSF-1 / 1R inhibitor (e.g., an M-CSF inhibitor), an IL-17 inhibitor, an IL-1 beta inhibitor or a CXCR2 inhibitor. In certain embodiments, the anti-tumor immunosuppressive combination comprises one, two or all of a CSF-1 / 1R inhibitor (e.g., an M-CSF inhibitor), an IL-17 inhibitor, an IL-1 beta inhibitor.In one aspect, the anti-tumor immunosuppressive combination comprises an IL-17 inhibitor, a CXCR2 inhibitor, a CRTH2 inhibitor, an A2AR antagonist or a PFKFB3 inhibitor, or a combination thereof.

[0177] In one aspect, the combination comprises one or more therapeutic agents of an antigen presenting combination. In other aspects, the combination comprises one or more therapeutic agents of an effector cell combination. In still other aspects, the combination comprises one or more therapeutic agents of an anti-tumor immunosuppressive combination. In other aspects, the combination comprises one or more therapeutic agents of an antigen presenting combination and one or more therapeutic agents of an effector cell combination. In other aspects, the combination comprises one or more therapeutic agents of an antigen presenting combination and one or more therapeutic agents of an anti-tumor immunosuppressive combination. In other aspects, the combination comprises one or more therapeutic agents of an antigen presenting combination, one or more therapeutic agents of an effector cell combination and one or more therapeutic agents of an anti-tumor immunosuppressive combination. In other aspects, the combination comprises one or more therapeutic agents of an antigen presenting combination, one or more therapeutic agents of an effector cell combination and one or more therapeutic agents of an anti-tumor immunosuppressive combination.

[0178] In certain aspects, the combination comprises: (i) one or more therapeutic agents of an antigen presenting combination selected from one, two or all of a STING agonist, a TLR agonist (e.g., a TLR7 agonist) or a TIM-3 modulator (e.g., a TIM-3 inhibitor); (ii) one or more therapeutic agents of an effector cell combination selected from one, two or all of a GITR modulator (e.g., a GITR agonist), a PD-L1 inhibitor (e.g., an anti-PD-L1 antibody molecule described herein) or a PD-1 inhibitor; (iii) one or more therapeutic agents of an anti-tumor immunosuppressive combination selected from one, two or all of a CSF-1 / 1R inhibitor (e.g., an M-CSF inhibitor), an IL-17 inhibitor or an IL-1 beta inhibitor: (iv) a combination of (i) and (ii); (v) (i) and (iii) combination; (vi) (ii) and (iii) combination; or (vii) (i), (ii) and (iii) combination.

[0179] Using the combination, cancers described herein such as lung cancer (squamous cell), lung cancer (adenocarcinoma), head and neck cancer, cervical cancer (squamous cell), gastric cancer, thyroid cancer, melanoma (e.g., advanced melanoma), nasopharyngeal cancer or breast cancer can be treated.

[0180] In other embodiments, the combination comprises a therapeutic agent derived from an antigen-presenting combination (e.g., one or more of a STING agonist, TLR agonist, vaccine or oncolytic virus) combined with a therapeutic agent derived from an effector cell and / or anti-tumor immunosuppressive combination [e.g., an inhibitor of a checkpoint inhibitor, e.g., PD-L1, PD-1, LAG-3, TIM-3, CEACAM (e.g., CEACAM-1, -3 and / or -5) or CTLA-4 or an inhibitor of any combination thereof]. In one embodiment, one or more of a STING agonist, TLR agonist, vaccine or oncolytic virus is administered in combination with an anti-PD-L1 antibody molecule described herein. In one embodiment, a STING agonist and / or vaccine is administered in combination with an anti-PD-L1 antibody molecule described herein. In one embodiment, an oncolytic virus is administered in combination with an anti-PD-L1 antibody molecule described herein. Using the combination, cancers described herein such as lung cancer (squamous cell), lung cancer (adenocarcinoma), head and neck cancer, cervical cancer (squamous cell), gastric cancer, thyroid cancer, melanoma (e.g., advanced melanoma), nasopharyngeal cancer or breast cancer can be treated.

[0181] In certain embodiments, the combination comprises a combination of therapeutic agents presented in the section entitled "Exemplary Combinations of Antigen-Presenting Combinations, Effector Cell Combinations and Anti-Tumor Immunosuppressive Combinations" presented in the detailed description.

[0182] The combinations disclosed herein can be administered together in a single composition or separately in two or more different compositions, such as the compositions or dosage forms described herein. The administration of the therapeutic agents can be done in either order. The first agent and additional agents (e.g., second, third agents) can be administered by the same or different routes of administration. For example, the first therapeutic agent can be administered concurrently with, before, or after the additional agent. In certain embodiments, the first agent is administered locally, e.g., such that the administration of the first agent is localized to a desired site, e.g., a tumor site (e.g., a dendritic cell-enriched site), and any of the therapeutic agents of categories (i)-(iii) can be conjugated to a tumor targeting agent, e.g., a tumor targeting antibody (e.g., to form an antibody-drug conjugate) or any other delivery agent (e.g., a targeting formulation, etc., a formulation). In one embodiment, the therapeutic agent is an antigen (e.g., a vaccine, e.g., an intraepithelial cancer vaccine) that is targeted to the tumor environment and thereby results in the activation of dendritic cells. The therapeutic agent can also be administered locally, e.g., by injection into the tumor site (e.g., intratumoral or peritumoral administration). Localized delivery or administration of the therapeutic agent can reduce one or more side effects or toxicities that might otherwise accompany systemic administration of the therapeutic agent. In an exemplary embodiment, a therapeutic agent (e.g., STING or TLR) is conjugated to a tumor-binding antibody (e.g., an antibody that binds to HER2) such that the therapeutic agent can be delivered to HER-2 expressing cells.

[0183] Detection / Ceranostics In another aspect, the present invention features a method for detecting the presence of PD-L1 in a sample, e.g., in vitro or in vivo (e.g., a biological sample such as serum, semen or urine, or a tissue biopsy derived from, e.g., hyperplasia or a neoplastic lesion). The method can be used to assess (e.g., monitor the treatment or progression of a disorder described herein in a subject, e.g., a hyperplasia or neoplastic disorder, diagnose and / or stage it). The method includes (i) contacting the antibody molecules described herein with the sample (and optionally, a reference, e.g., a control sample) or administering it to a subject under conditions in which an interaction occurs, and (ii) detecting the formation of a complex between the antibody molecules and the sample (and optionally, a reference, e.g., a control sample). The formation of the complex indicates the presence of PD-L1 and can indicate the suitability or need for a treatment described herein. In one embodiment, PD-L1 is detected prior to treatment, e.g., prior to initial treatment, or prior to treatment after a treatment interval. Detection can involve immunohistochemical examination, immunocytochemical examination, FACS, magnetic beads complexed with antibody molecules, ELISA assay, PCR techniques (e.g., RT-PCR) or in vivo imaging techniques. Typically, the antibody molecules used in in vivo and in vitro detection methods are labeled directly or indirectly with a detectable substance to facilitate detection of bound or unbound binding agents. Suitable detectable substances include various biologically active enzymes, avidin family molecules, fluorescent materials, luminescent materials, paramagnetic (e.g., nuclear magnetic resonance active) materials and radioactive materials. In another embodiment, the antibody molecules are detected in vivo, e.g., using in vivo imaging techniques (e.g., PET imaging) described herein.

[0184] Additional aspects provide a method of treating cancer that includes identifying the presence of one, two, or all of PD-L1, CD8, or IFN-γ in a subject, such as a sample (e.g., a sample of a subject that includes cancer cells and optionally immune cells such as TILs), thereby providing a value for one, two, or all of PD-L1, CD8, and IFN-γ. The method can further include comparing the PD-L1, CD8, and / or IFN-γ value to a reference value, such as a control value. When the PD-L1, CD8, and / or IFN-γ value is greater than the reference value, such as the control value, a therapeutically effective amount of an anti-PD-L1 antibody (e.g., an anti-PD-L1 antibody described herein) is administered to the subject, optionally in combination with one or more other agents, thereby treating the cancer. In one aspect, the subject is identified prior to treatment, such as prior to initial treatment or prior to treatment after a treatment interval. The cancer can be, for example, lung cancer (squamous), lung cancer (adenocarcinoma), head and neck cancer, cervical cancer (squamous), gastric cancer, thyroid cancer, melanoma, nasopharyngeal cancer, or breast cancer, such as TN breast cancer, such as IM-TN breast cancer, etc., as described herein. In one aspect, the cancer is ER+ breast cancer or pancreatic cancer.

[0185] Also provided is a method of treating cancer that includes examining a subject, such as a sample (e.g., a sample of a subject that includes cancer cells), for the presence of PD-L1, thereby identifying a PD-L1 value; comparing the PD-L1 value to a control value; and when the PD-L1 value is greater than the control value, administering to the subject a therapeutically effective amount of an anti-PD-L1 antibody (e.g., an anti-PD-L1 antibody described herein), optionally in combination with one or more other agents, thereby treating the cancer. The cancer can be, for example, non-small cell lung (NSCLC) adenocarcinoma (ACA), NSCLC squamous cell carcinoma (SCC), or hepatocellular carcinoma (HCC), etc., as described herein.

[0186] Without being limited by theory, subjects that exhibit an existing immune response to cancer, such as an immune response prior to immunomodulatory pharmacotherapy (e.g., checkpoint molecule inhibitor therapy), are thought to be able to have an extended and / or more robust response to the therapy compared to subjects that do not have the same immune response. Thus, in one aspect, the assessment of the state of a subject with activated immune cells (e.g., T cells) prior to immunomodulatory pharmacotherapy can function as a means for assessing and / or monitoring the responsiveness of the subject to immunomodulatory pharmacotherapy. In one aspect, such an assessment can be used to identify, select, and / or stratify a subject (e.g., a patient or patient population) as being likely or unlikely to respond to immunomodulatory pharmacotherapy.

[0187] Accordingly, provided are methods for assessing the status of a subject for immune cell (e.g., T cell) activation, alternatively or in combination with the methods described herein, e.g., for assessing the likelihood of response of a subject to immunomodulatory therapy. The methods include determining the level and / or distribution of T cell activation in a subject. In certain embodiments, the level and / or distribution of T activation includes measurement of one or more of the following levels and / or distributions: CD8, PD-L1, or other checkpoint inhibitors [e.g., one or more of PD-1, LAG-3, TIM-3, CEACAM (e.g., CEACAM-1, -3, and / or -5), or CTLA-4] or any combination thereof. For example, the level and / or distribution of CD8-expressing cells can be evaluated as a marker of activated T cells. In other embodiments, the level and / or distribution of cells expressing PD-L1 or other checkpoint inhibitors can be evaluated. The subject can be evaluated before, during, or after administration of immunomodulatory therapy. In certain embodiments, the subject is evaluated before immunomodulatory therapy (e.g., checkpoint molecule inhibitor therapy), e.g., before initial treatment or before treatment after a treatment interval. In certain embodiments, an increase in the level of one or more of CD8, PD-L1, or other checkpoint inhibitors in a subject (e.g., relative to a reference, e.g., compared to a control) indicates an increased responsiveness of the subject to the therapy (also referred to herein as a “positive immune activation state”). In another embodiment, a decrease in the level of one or more of CD8, PD-L1, or other checkpoint inhibitors in a subject (e.g., relative to a reference, e.g., compared to a control) indicates a decreased responsiveness of the subject to the therapy (also referred to herein as a “negative immune activation state”). If the subject is determined to have a positive immune activation state, the method may include administration of an immunomodulatory therapy described herein (e.g., a checkpoint molecule inhibitor therapy described herein).

[0188] In one aspect, the immunomodulatory therapy comprises an activator of a costimulatory molecule, such as one or more activators described herein (e.g., an agonist of the GITR molecule described herein). In other aspects, the immunomodulatory therapy comprises an inhibitor of an immune checkpoint molecule, such as one or more inhibitors of the checkpoint inhibitors described herein (e.g., one or more inhibitors of PD-L1, PD-1, TIM-3, or CTLA-4 described herein). In one aspect, the immunomodulatory therapy comprises the anti-PD-L1 antibody molecule described herein. In other aspects, the immunomodulatory therapy comprises a combination of an activator of a costimulatory molecule and an inhibitor of a checkpoint inhibitor.

[0189] In one aspect, the level and / or distribution of CD8, PD-L1, or other checkpoint inhibitors is determined in vivo, e.g., non-invasively (e.g., by detecting an antibody to a T cell marker detectably labeled using a suitable imaging technique, e.g., a positron emission tomography (PET) scan). For example, target antibody-PET or immuno-PET (e.g., anti-CD8 PET or anti-PD-L1 PET) can be used to detect the level and / or distribution (e.g., tumor localization) of target CD8 or PD-L1 expressing cells in vivo. Techniques for antibody imaging (e.g., antibody-PET imaging) are known in the art and are described, for example, in Lamberts, L. E. et al. (2015) J. Clin. Oncol. 33 (DOI: 10.1200 / JCO.2014.57.8278); Tavare, R. et al. (2014) PNAS 111(3):1108-1113; and Boerman and Oyen (2011) The Journal of Nuclear Medicine 52 (8):1171-72, which are incorporated herein by reference. In other aspects, the level of CD8, PD-L1, or other checkpoint inhibitors is determined in a sample obtained from the subject (e.g., a tumor biopsy) (e.g., using immunohistochemical techniques).

[0190] As a useful aspect, an increased number of CD8-expressing cells, or a change in their location from the tumor periphery to the tumor interior, can indicate an improved outcome, such as an increased responsiveness of a subject to immunomodulatory therapy. In certain aspects, a positive immunologically activated state (e.g., an increase in activated T cells) can occur in response to a previous treatment of cancer, e.g., radiation therapy, chemotherapy, oncolytic virus, bispecific T cell engagers, biologic or targeted therapy (e.g., the anti-cancer therapies described herein) of one or more. In one aspect, a subject showing a positive immunologically activated state in response to a previous treatment is a better candidate for immunomodulatory therapy. In one aspect, a subject having a B-Raf mutation (e.g., the B-Raf mutations described herein) is treated with a B-Raf inhibitor (e.g., the B-Raf inhibitors described herein). Since the subject can have an increase in CD8-expressing T cells after treatment with a B-Raf inhibitor, it can be a better candidate for immunomodulatory therapy.

[0191] In another aspect useful for further explanation, the expression of PD-L1 or other checkpoint inhibitors (e.g., compared to a reference) can be used to determine or analyze the responsiveness of a subject to a cancer therapy. In one aspect, a PET imaging agent can indicate that a subject has an existing expression of PD-L1 or other checkpoint molecule. The expression of these molecules in tumors can be useful for patient stratification for companion diagnostic purposes. If a subject does not have an existing detectable immune response and detectable checkpoint molecule expression to an immunomodulatory pharmacotherapy, such a subject may be an inadequate candidate for the therapy. PET imaging can also show the heterogeneity of the expression and indicate the tumors (PET positive) that are the actual tumors / lesions most suitable for response; this can be beneficial as sampling of a few target lesions can be misleading; for example, due to sampling errors of biopsy techniques and a few samples to determine whether a subject is responsive or not. Using PET imaging, it is possible to detect the majority of the tumors, thereby obtaining a more accurate determination of tumor burden efficacy and a more thorough understanding of how the therapy functions.

[0192] In another aspect, the invention features a diagnostic or therapeutic kit comprising the antibody molecules and instructions described herein.

[0193] All publications, patent applications, patents, and other references mentioned herein are hereby incorporated by reference in their entirety.

[0194] Other features, objects, and advantages of the invention will be apparent from the specification and drawings, as well as from the claims.

Brief Description of the Drawings

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Modes for Carrying Out the Invention

[0217] Brief Explanation of the Tables Table 1 provides an overview of the amino acid and nucleotide sequences for mouse, chimeric, and humanized anti-PD-L1 antibody molecules. The antibody molecules include mouse mAb BAP058, chimeric mAb BAP058-chi, and humanized mAbs BAP058-hum01 to BAP058-hum17 and BAP058-clone-K to BAP058-clone-O. The amino acid and nucleotide sequences of the heavy and light chain CDRs, the amino acid and nucleotide sequences of the heavy and light chain variable regions, and the amino acid and nucleotide sequences of the heavy and light chains are shown in this table.

[0218] Table 2 shows the amino acid and nucleotide sequences of the heavy and light chain framework regions of humanized mAbs BAP058-hum01 to BAP058-hum17 and BAP058-clone-K to BAP058-clone-O.

[0219] Table 3 shows the constant region amino acid sequences of human IgG heavy chain and human kappa light chain.

[0220] Table 4 provides an overview of the yield, titer, monomer content, and endotoxin level for selected humanized BAP058 mAbs expressed in CHO cells.

[0221] Table 5 shows the charge isomers detected by Novex IEF analysis for selected humanized BAP058 mAbs expressed in CHO cells.

[0222] Table 6 provides an overview of selected therapeutic agents that can be administered in combination with anti-PD-1 antibody molecules and other immunomodulatory agents described herein (e.g., one or more of activators of costimulatory molecules and / or inhibitors of immune checkpoint molecules). Table 6 shows, from left to right, the compound name, compound structure, and patent publication document disclosing the compound of the second therapeutic agent.

[0223] Table 7 provides an exemplary list of therapeutic agents by antigen presentation combinations (category A), effector cell combinations (category B), and anti-tumor immunosuppressive combinations (category C).

[0224] Table 8 shows the cross-species binding of exemplary anti-PD-L1 antibodies evaluated by Biacore.

[0225] The immune system has the ability to recognize and eliminate tumor cells; however, tumors can use multiple strategies to evade immunity. Blockade of immune checkpoints is one approach to activate or reactivate therapeutic anti-tumor immunity. Programmed death ligand 1 (PD-L1) has been described as a ligand for the immune inhibitory receptor programmed death 1 (PD-1). Binding of PD-L1 to PD-1 results in inhibition of T cell receptor-mediated lymphocyte proliferation and cytokine secretion [Freeman et al. (2000) J Exp Med 192:1027-34]. Thus, blockade of PD-L1 can result in enhancement of anti-tumor immunity.

[0226] Several cell types express PD-L1. For example, PD-L1 is expressed in activated T cells, dendritic cells (DCs), natural killer (NK) cells, macrophages, B cells, monocytes, and vascular endothelial cells. PD-L1 is expressed in many cancers including human lung, ovarian, and colon cancers as well as various myelomas [Iwai et al. (2002) PNAS 99:12293-7; Ohigashi et al. (2005) Clin Cancer Res 11:2947-53; Okazaki et al. (2007) Intern. Immun. 19:813-24; Thompson et al. (2006) Cancer Res. 66:3381-5]. PD-L1 expression strongly correlates with poor prognosis in various types of cancers including kidney, ovarian, bladder, breast, gastric, and pancreatic cancers.

[0227] Many tumor-infiltrating T lymphocytes express PD-1 predominantly compared with T lymphocytes in normal tissues and peripheral blood T lymphocytes. This indicates that upregulation of PD-1 in tumor-reactive T cells may contribute to defective antitumor immune responses [Ahmadzadeh et al. (2009) Blood 114:1537-44]. Thus, PD-L1 signaling mediated by PD-L1-expressing tumor cells that interact with PD-1-expressing T cells can lead to attenuation of T cell activation and avoidance of immune surveillance [Sharpe et al. (2002) Nat Rev Immunol. 2:116-26; Keir et al. (2008) Annu Rev Immunol. 26:677-704]. PD-1 blockade can inhibit hematogenous dissemination of poorly immunogenic tumor cells by enhancing recruitment of effector T cells [Iwai et al. (2005) Int. Immunol. 17:133-144].

[0228] Anti-PD-L1 can enhance T cell immunity, for example, by blocking both inhibitory interactions of PD-1 and B7-1 with PD-L1. Anti-PD-1 can also enable immune regulation by PD-L2 / PD-1. Both PD-1 and B7-1 are expressed in T cells, B cells, DCs and macrophages, which gives rise to the possibility of bidirectional interactions between B7-1 and PD-L1 in these cell types. PD-L1 in non-hematopoietic cells can interact with B7-1 and PD-1 in T cells.

[0229] Accordingly, the present invention provides, at least in part, antibody molecules (e.g., humanized antibody molecules) that bind to programmed death ligand 1 (PD-L1) with high affinity and specificity. In certain embodiments, humanized antibodies to PD-L1 are disclosed that exhibit surprisingly low immunogenicity. For example, a humanized anti-PD-L1 antibody can have a risk score of less than 650, 600, 550, or 500 according to a T cell epitope assay. In other embodiments, for example, as shown in FIGS. 5 and 7, selected combinations of framework regions have been shown to have distinct production efficiencies and binding characteristics.

[0230] Additional aspects of the invention include nucleic acid molecules encoding the antibody molecules, expression vectors, host cells, and methods for making the antibody molecules. Immunoconjugates, multispecific or bispecific molecules, and pharmaceutical compositions comprising the antibody molecules are also provided. The anti-PD-L1 antibody molecules disclosed herein can be used to treat, prevent, and / or diagnose cancerous or malignant disorders [e.g., solid and soft tissue tumors; melanoma, e.g., advanced melanoma; hepatocellular carcinoma; pancreatic cancer; renal cell carcinoma (RCC), e.g., metastatic RCC or clear cell RCC; glioma or glioblastoma; multiple myeloma; colorectal cancer; and lung cancer, e.g., non-small cell cancer], and infectious diseases [e.g., hepatitis, e.g., hepatitis C (e.g., chronic viral hepatitis); infectious disorders such as sepsis]. Thus, methods for detecting PD-L1, as well as methods for treating various disorders, including cancer and infectious diseases, using the anti-PD-L1 antibody molecules are disclosed herein.

[0231] In addition, methods and compositions are disclosed herein that include combinations of two or more therapeutic agents selected from one, two, or all of the following categories (i)-(iii): (i) agents that enhance antigen presentation (e.g., tumor antigen presentation) (e.g., by enhancing one or more of dendritic cell activation or maturation, antigen uptake, or antigen processing); (ii) agents that enhance effector cell responses (e.g., immune effector cell responses, e.g., B cell and / or T cell activation and / or recruitment, e.g., in lymph nodes); or (iii) agents that reduce tumor immunosuppression (e.g., by increasing T cell infiltration and tumor cell death). In certain embodiments, the combination includes a PD-L1 inhibitor (e.g., an anti-PD-L1 antibody molecule as described herein). Without wishing to be bound by theory, it is believed that a therapeutic approach that enhances anti-tumor immunity may function more effectively if the immune response is optimized by multiple targets at different stages of the immune response. Each of these stages is depicted in schematic form in FIG. 20. For example, an approach that results in activation of dendritic cells in combination with an approach that enhances cellular and humoral immunity can result in a more effective and / or prolonged therapeutic response.

[0232] The term "programmed death ligand 1" or "PD-L1" includes isoforms, mammals, e.g., human PD-L1, species homologs of human PD-1, and analogs that include at least one common epitope with PD-L1. The amino acid sequences of PD-L1, e.g., human PD-1, are known in the art and are described, e.g., in Dong et al. (1999) Nat Med. 5(12):1365-9; Freeman et al. (2000) J Exp Med. 192(7):1027-34).

[0233] Additional terms are defined hereinafter and throughout this application. As used herein, singular recitations refer to one or more (e.g., at least one) of the grammatical object of the article.

[0234] As used herein, the term "or" is used to mean "and / or" and is used interchangeably therewith, unless the context clearly indicates otherwise.

[0235] "About" and "approximately" generally mean an acceptable degree of error for the measured quantity, taking into account the nature or precision of the measurement. Exemplary degrees of error are within 20 percent (%) of a given value or range of values, typically within 10%, more typically within 5%.

[0236] "Combination" or "in combination with" is not intended to imply that the therapies or agents must be administered simultaneously and / or formulated for co-delivery, but such delivery methods are within the scope described herein. The therapeutic agents in a combination can be administered concurrently, prior to or subsequent to one or more other additional therapies or agents. The therapeutic agents or protocols can be administered in any order. Generally, each agent will be administered at the dosage and / or time schedule determined for that agent. It will further be recognized that the additional therapeutic agents utilized in a combination may be administered together in a single composition or separately in different compositions. Generally, it is expected that the additional therapeutic agents utilized in a combination will be utilized at levels that do not exceed those at which they are individually utilized. In certain embodiments, the levels utilized in a combination will be lower than those utilized individually.

[0237] In certain embodiments, the additional therapeutic agent is administered at a therapeutic dose or at a dose lower than the therapeutic dose. In certain embodiments, the concentration of the second therapeutic agent required to achieve inhibition, e.g., growth inhibition, is lower when the second therapeutic agent is administered in combination with a first therapeutic agent, e.g., an anti-PD-L1 antibody molecule, than when the second therapeutic agent is administered individually. In certain embodiments, the concentration of the first therapeutic agent required to achieve inhibition, e.g., growth inhibition, is lower when the first therapeutic agent is administered in combination with a second therapeutic agent than when the first therapeutic agent is administered individually. In certain embodiments, in combination therapy, the concentration of the second therapeutic agent required to achieve inhibition, e.g., growth inhibition, is lower than the therapeutic dose of the second therapeutic agent as monotherapy, e.g., 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80% or 80-90% lower. In certain embodiments, in combination therapy, the concentration of the first therapeutic agent required to achieve inhibition, e.g., growth inhibition, is lower than the therapeutic dose of the first therapeutic agent as monotherapy, e.g., 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80% or 80-90% lower.

[0238] The terms "inhibition", "inhibitor" or "antagonist" include a decrease in a particular parameter, e.g., activity, of a given molecule, e.g., an immune checkpoint inhibitor. For example, inhibition of at least 5%, 10%, 20%, 30%, 40% or more of the activity, e.g., PD-1 or PD-L1 activity, is included by this term. Thus, inhibition need not be 100%.

[0239] The terms "activation", "activator" or "agonist" include an increase in a particular parameter, e.g., activity, of a given molecule, e.g., a co-stimulatory molecule. For example, an increase in co-stimulatory activity of at least 5%, 10%, 25%, 50%, 75% or more is included by this term.

[0240] The term "anticancer effect" refers to a biological effect that can be manifested by various means, such as, for example, a decrease in tumor volume, a decrease in the number of cancer cells, a decrease in the number of metastases, an increase in the average life span, a decrease in cancer cell proliferation, a decrease in cancer cell survival, or a remission of various physiological symptoms related to the cancer condition, but is not limited thereto. The "anticancer effect" can also be manifested first by the ability of peptides, polynucleotides, cells, and antibodies in the prevention of cancer development.

[0241] The term "antitumor effect" refers to a biological effect that can be manifested by various means, such as, for example, a decrease in tumor volume, a decrease in the number of tumor cells, a decrease in tumor cell proliferation, or a decrease in tumor cell survival, but is not limited thereto.

[0242] The term "cancer" refers to a disease characterized by the rapid and uncontrolled growth of abnormal cells. Cancer cells can spread locally or to other parts of the body via the bloodstream and lymphatic system. Examples of various cancers are described herein and include, but are not limited to, breast cancer, prostate cancer, ovarian cancer, cervical cancer, skin cancer, pancreatic cancer, colorectal cancer, kidney cancer, liver cancer, brain cancer, lymphoma, leukemia, lung cancer, and others. The terms "tumor" and "cancer" are used interchangeably herein. For example, both terms include solid and liquid tumors, such as diffuse or circulating tumors. As used herein, the term "cancer" or "tumor" includes malignant and tumor cancers as well as premalignant.

[0243] The term "antigen-presenting cell" or "APC" refers to an accessory immune system cell (such as, for example, B cells, dendritic cells, and others) that presents an exogenous antigen complexed with a major histocompatibility antigen complex (MHC) on its surface. T cells can recognize such a complex using their T cell receptor (TCR). APCs process the antigen and present it to T cells.

[0244] The term "costimulatory molecule" refers to a cognate binding partner in T cells, which specifically binds to a costimulatory ligand and thereby mediates a costimulatory response by T cells, including but not limited to proliferation, etc. A costimulatory molecule is a cell surface molecule other than an antigen receptor or its ligand required for an efficient immune response. Examples of costimulatory molecules include, but are not limited to, MHC class I molecules, TNF receptor proteins, immunoglobulin-like proteins, cytokine receptors, integrins, signaling lymphocyte activation molecule (SLAM protein), activating NK cell receptors, BTLA, Toll ligand receptor, OX40, CD2, CD7, CD27, CD28, CD30, CD40, CDS, ICAM-1, LFA-1 (CD11a / CD18), 4-1BB (CD137), B7-H3, CDS, ICAM-1, ICOS (CD278), GITR, BAFFR, LIGHT, HVEM (LIGHTR), KIRDS2, SLAMF7, NKp80 (KLRF1), NKp44, NKp30, NKp46, CD19, CD4, CD8α, CD8β, IL2Rβ, IL2Rγ, IL7Rα, ITGA4, VLA1, CD49a, ITGA4, IA4, CD49D, ITGA6, VLA-6, CD49f, ITGAD, CD11d, ITGAE, CD103, ITGAL, CD11a, LFA-1, ITGAM, CD11b, ITGAX, CD11c, ITGB1, CD29, ITGB2, CD18, LFA-1, ITGB7, NKG2D, NKG2C, TNFR2, TRANCE / RANKL, DNAM1 (CD226), SLAMF4 (CD244, 2B4), CD84, CD96 (Tactile), CEACAM1, CRTAM, Ly9 (CD229), CD160 (BY55), PSGL1, CD100 (SEMA4D), CD69, SLAMF6 (NTB-A, Ly108), SLAM (SLAMF1, CD150, IPO-3), BLAME (SLAMF8), SELPLG (CD162), LTBR, LAT, GADS, SLP-76, PAG / Cbp, CD19a, and CD83, and ligands that specifically bind to them.

[0245] The term "immune effector cell" or "effector cell", as used herein, refers to a cell involved in promoting an immune response, such as an immune effector response. Examples of immune effector cells include T cells, such as alpha / beta T cells and gamma / delta T cells, B cells, natural killer (NK) cells, natural killer T (NKT) cells, mast cells, and myeloid-derived phagocytic cells.

[0246] The term "immune effector" or "effector" "function" or "response", as used herein, refers to the function or response of an immune effector cell that enhances or promotes, for example, an immune attack on a target cell. For example, an immune effector function or response refers to the property of T or NK cells that promotes the death of a target cell or the inhibition of its growth or proliferation. In the case of T cells, primary stimulation and co-stimulation are examples of immune effector functions or responses.

[0247] The term "effector function" refers to a specialized function of a cell. For example, the effector function of a T cell can be a helper activity that includes cytolytic activity or cytokine secretion.

[0248] The term "immune activation state" refers to the likelihood that a subject has with respect to a response to a treatment method, such as immunomodulatory therapy. In an aspect, the immune activation state can be a positive state or a negative state. In an aspect, a positive immune activation state means that the subject has a probability of more than 50% (for example, a probability of more than 50%, 60%, 70%, 80%, 90% or more) of responding to a treatment method, such as immunomodulatory therapy. In an aspect, a negative immune activation state means that the subject has a probability of more than 50% (for example, a probability of more than 50%, 60%, 70%, 80%, 90% or more) of not responding to a treatment method, such as immunomodulatory therapy.

[0249] As used herein, the terms "treating," "treatment," and "being treated" refer to a decrease or alleviation in the progression, severity, and / or duration of a disorder, such as a proliferative disorder, or alleviation of one or more symptoms (preferably one or more distinguishable symptoms) of a disorder, resulting from the administration of one or more therapies. In a particular aspect, the terms "treating," "treatment," and "being treated" refer to alleviation of at least one measurable physical parameter of a proliferative disorder, such as tumor growth, which need not necessarily be distinguishable by the patient. In another aspect, the terms "treating," "treatment," and "being treated" refer to inhibition of the progression of a proliferative disorder, for example, physically, such as by stabilization of a physical parameter, or physiologically, or both, such as by stabilization of a distinguishable symptom. In another aspect, the terms "treating," "treatment," and "being treated" refer to a decrease or stabilization in tumor size or the number of cancerous cells.

[0250] The compositions and methods of the present invention include polypeptides and nucleic acids having a specified sequence, or a sequence that is substantially identical or similar thereto, such as a sequence that is at least 85%, 90%, 95% or more identical to the specified sequence. In the context of amino acid sequences, the term "substantially identical" is used herein to refer to a first amino acid sequence that contains a sufficient or minimal number of amino acid residues that are either i) identical to the aligned amino acid residues in a second amino acid sequence, or ii) conservative substitutions thereof, such that the first and second amino acid sequences can have a common structural domain and / or a common functional activity. For example, an amino acid sequence containing a common structural domain having at least about 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to a reference sequence, such as a sequence presented herein.

[0251] In the context of nucleotide sequences, the term "substantially identical" is used herein to refer to a first nucleic acid sequence that contains a sufficient or minimum number of nucleotides that are identical to the aligned nucleotides in a second nucleic acid sequence such that the first and second nucleotide sequences encode a polypeptide having common functional activity or encode a common structural polypeptide domain or common functional polypeptide activity. For example, a nucleotide sequence having at least about 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to a reference sequence, e.g., a sequence presented herein.

[0252] The term "functional variant" refers to a polypeptide having an amino acid sequence that is substantially identical to a sequence of natural origin or that is encoded by a substantially identical nucleotide sequence and that can have one or more activities of the sequence of natural origin.

[0253] Calculation of homology or sequence identity between sequences (these terms are used interchangeably herein) is performed as follows.

[0254] To determine the percent identity of two amino acid sequences or two nucleic acid sequences, the sequences are aligned for optimal comparison purposes (e.g., gaps can be introduced into one or both of the first and second amino acid or nucleic acid sequences for optimal alignment and non-homologous sequences can be ignored for comparison purposes). In a preferred embodiment, the length of the reference sequence aligned for comparison purposes is at least 30%, preferably at least 40%, more preferably at least 50%, 60%, even more preferably at least 70%, 80%, 90%, 100% of the length of the reference sequence. Subsequently, the amino acid residues or nucleotides at the corresponding amino acid positions or nucleotide positions are compared. If a position in the first sequence is occupied by the same amino acid residue or nucleotide as the corresponding position in the second sequence, the two molecules are identical at that position (as used herein, amino acid or nucleic acid "identity" is equivalent to amino acid or nucleic acid "homology").

[0255] The percent identity between two arrays is a function of the number of identical positions shared by both arrays, taking into account the number of gaps that need to be introduced for optimal alignment of the two arrays and the length of each gap.

[0256] Comparison of arrays and determination of the percent identity between two arrays can be achieved using mathematical algorithms. In a preferred embodiment, the percent identity between two amino acid sequences is determined using either the Blossum62 matrix or the PAM250 matrix, as well as gap weights of 16, 14, 12, 10, 8, 6, or 4 and length weights of 1, 2, 3, 4, 5, or 6, using the Needleman and Wunsch [(1970) J. Mol. Biol. 48:444-453] algorithm incorporated into the GAP program in the GCG software package (available at http: / / www.gcg.com). In yet another preferred embodiment, the percent identity between two nucleotide sequences is determined using the NWSgapdna.CMP matrix as well as gap weights of 40, 50, 60, 70, or 80 and length weights of 1, 2, 3, 4, 5, or 6, using the GAP program in the GCG software package (available at http: / / www.gcg.com). A particularly preferred set of parameters (and the set to be used unless otherwise specified) is the Blossum 62 scoring matrix, a gap penalty of 12, a gap extension penalty of 4, and a frameshift gap penalty of 5.

[0257] The percent identity between two amino acid or nucleotide sequences can be determined using the algorithm of E. Meyers and W. Miller [(1989) CABIOS, 4:11-17] incorporated into the ALIGN program (version 2.0), using the PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4.

[0258] The nucleic acid and protein sequences described in this specification can be used, for example, as "query sequences" for performing searches against public databases to identify other family members or related sequences. Such searches can be performed using the NBLAST and XBLAST programs (version 2.0) of Altschul, et al. (1990) J. Mol. Biol. 215:403-10. A BLAST nucleotide search can be performed with the NBLAST program, score = 100, wordlength = 12, to obtain nucleotide sequences homologous to the nucleic acid molecules of the present invention. A BLAST protein search can be performed with the XBLAST program, score = 50, wordlength = 3, to obtain amino acid sequences homologous to the protein molecules of the present invention. To obtain gapped alignments for comparison purposes, Gapped BLAST can be utilized as described in Altschul et al., (1997) Nucleic Acids Res. 25:3389-3402. When using the BLAST and Gapped BLAST programs, the default parameters of each program (e.g., XBLAST and NBLAST) can be used. See http: / / www.ncbi.nlm.nih.gov.

[0259] As used herein, the term "hybridizes under low stringency, medium stringency, high stringency or ultra-high stringency conditions" describes the conditions for hybridization and washing. Guidance for performing the hybridization reactions can be found in Current Protocols in Molecular Biology, John Wiley & Sons, N.Y. (1989), 6.3.1-6.3.6, which is incorporated herein by reference. Both aqueous and non-aqueous methods are described in that reference and either can be used. The specific hybridization conditions referred to herein are as follows: 1) Low stringency hybridization conditions are 6× sodium chloride / sodium citrate (SSC), approximately 45° C., followed by two washes in 0.2× SSC, 0.1% SDS, at at least 50° C. (the wash temperature can be increased up to 55° C. under low stringency conditions); 2) Medium stringency hybridization conditions are 6× SSC, approximately 45° C., followed by one or more washes in 0.2× SSC, 0.1% SDS, at 60° C.; 3) High stringency hybridization conditions are 6× SSC, approximately 45° C., followed by one or more washes in 0.2× SSC, 0.1% SDS, at 65° C.; and preferably 4) Ultra-high stringency hybridization conditions are 0.5 M sodium phosphate, 7% SDS, 65° C., followed by one or more washes in 0.2× SSC, 1% SDS, at 65° C. Ultra-high stringency conditions (4) are the preferred conditions and are to be used unless otherwise specified.

[0260] It is understood that the molecules of the invention can have additional conservative or non-essential amino acid substitutions that do not substantially affect their function.

[0261] The term "amino acid" is intended to encompass any molecule, whether natural or synthetic, that can be included in a polymer of amino acids of natural origin and that contains both amino functionality and acid functionality. Exemplary amino acids include amino acids of natural origin; their analogs, derivatives, and homologs; amino acid analogs having variant side chains; and any stereoisomers of any of the foregoing. As used herein, the term "amino acid" includes both D- or L-optical isomers and peptidomimetics.

[0262] "Conservative amino acid substitutions" are substitutions in which an amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains are defined in the art. Such families include amino acids having basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), beta-branched side chains (e.g., threonine, valine, isoleucine), and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine).

[0263] The terms "polypeptide," "peptide," and "protein" (if single-chain) are used interchangeably herein to refer to polymers of amino acids of any length. The polymer may be linear or branched, may contain modified amino acids, and may be interrupted by non-amino acids. The term also encompasses modified amino acid polymers; for example, any other manipulations such as disulfide bond formation, glycosylation, lipid addition, acetylation, phosphorylation, or conjugation with a labeling component. Polypeptides can be isolated from natural sources, produced by recombinant techniques from eukaryotic or prokaryotic hosts, or be the product of synthetic procedures.

[0264] The terms "nucleic acid", "nucleic acid sequence", "nucleotide sequence" or "polynucleotide sequence" and "polynucleotide" are used interchangeably. These refer to polymeric forms of nucleotides of any length, which are either deoxyribonucleotides or ribonucleotides or analogs thereof. A polynucleotide can be either single-stranded or double-stranded, and if single-stranded, can be either the coding strand or the non-coding (antisense) strand. A polynucleotide can include modified nucleotides such as methylated nucleotides and nucleotide analogs. The sequence of nucleotides can be interrupted by non-nucleotide components. A polynucleotide can be further modified after polymerization, such as by conjugation with a labeling component. A nucleic acid can be a recombinant polynucleotide, or a polynucleotide of genomic, cDNA, semi-synthetic or synthetic origin, which is not naturally occurring or is linked to another polynucleotide in a non-natural arrangement.

[0265] The term "isolated" as used herein refers to a material that has been removed from its original or native environment (e.g., the natural environment if it is of natural origin). For example, a polynucleotide or polypeptide of natural origin that exists in a living animal is not isolated, but the same polynucleotide or polypeptide that has been separated by human intervention from some or all of the coexisting materials in the natural system is isolated. Such a polynucleotide can be part of a vector, and / or such a polynucleotide or polypeptide can be part of a composition, and such a vector or composition is still isolated in that it is not part of the environment in which it is naturally found.

[0266] Various aspects of the present invention are described in more detail hereinafter. Additional definitions are presented throughout the specification.

[0267] Antibody molecule In one aspect, the antibody molecule binds to a mammalian, e.g., human PD-L1. For example, the antibody molecule specifically binds to an epitope in PD-L1, e.g., a linear or conformational epitope (e.g., an epitope described herein).

[0268] As used herein, the term "antibody molecule" refers to a protein, e.g., an immunoglobulin chain or a fragment thereof, that includes at least one immunoglobulin variable domain sequence. The term "antibody molecule" includes, for example, monoclonal antibodies (including full-length antibodies having an immunoglobulin Fc region). In one aspect, the antibody molecule includes a full-length antibody or a full-length immunoglobulin chain. In one aspect, the antibody molecule includes an antigen-binding or functional fragment of a full-length antibody or a full-length immunoglobulin chain.

[0269] In one aspect, the antibody molecule is a monospecific antibody molecule that binds to a single epitope. For example, a monospecific antibody molecule having a plurality of immunoglobulin variable domain sequences that each bind to the same epitope.

[0270] In one aspect, the antibody molecule is a multispecific antibody molecule, which, for example, includes a plurality of immunoglobulin variable domain sequences, wherein a first immunoglobulin variable domain sequence of the plurality has binding specificity for a first epitope and a second immunoglobulin variable domain sequence of the plurality has binding specificity for a second epitope. In one aspect, the first and second epitopes are present on the same antigen, e.g., the same protein (or a subunit of a multimeric protein). In one aspect, the first and second epitopes overlap. In one aspect, the first and second epitopes do not overlap. In one aspect, the first and second epitopes are present on different antigens, e.g., different proteins (or different subunits of a multimeric protein). In one aspect, the multispecific antibody molecule includes a third, fourth, or fifth immunoglobulin variable domain. In one aspect, the multispecific antibody molecule is a bispecific antibody molecule, a trispecific antibody molecule, or a tetravalent antibody molecule.

[0271] In an aspect, the multispecific antibody molecule is a bispecific antibody molecule. The bispecific antibody has specificity for no more than two antigens. The bispecific antibody molecule is characterized by a first immunoglobulin variable domain sequence having binding specificity for a first epitope and a second immunoglobulin variable domain sequence having binding specificity for a second epitope. In an aspect, the first and second epitopes are present on the same antigen, for example, the same protein (or a subunit of a multimeric protein). In an aspect, the first and second epitopes overlap. In an aspect, the first and second epitopes do not overlap. In an aspect, the first and second epitopes are present on different antigens, for example, different proteins (or different subunits of a multimeric protein). In an aspect, the bispecific antibody molecule comprises a heavy chain variable domain sequence and a light chain variable domain sequence having binding specificity for a first epitope, and a heavy chain variable domain sequence and a light chain variable domain sequence having binding specificity for a second epitope. In an aspect, the bispecific antibody molecule comprises a half antibody having binding specificity for a first epitope and a half antibody having binding specificity for a second epitope. In an aspect, the bispecific antibody molecule comprises a half antibody or a fragment thereof having binding specificity for a first epitope, and a half antibody or a fragment thereof having binding specificity for a second epitope. In an aspect, the bispecific antibody molecule comprises a scFv or a fragment thereof having binding specificity for a first epitope, and a scFv or a fragment thereof having binding specificity for a second epitope. In an aspect, the first epitope is located on PD-L1, and the second epitope is located on TIM-3, LAG-3, CEACAM (e.g., CEACAM-1, CEACAM-3 and / or CEACAM-5), PD-1 or PD-L2.

[0272] In some embodiments, the antibody molecule includes diabodies and single-chain molecules, as well as antigen-binding fragments of antibodies (e.g., Fab, F(ab’)2, and Fv). For example, the antibody molecule can include a heavy (H) chain variable domain sequence (abbreviated herein as VH) and a light (L) chain variable domain sequence (abbreviated herein as VL). In some embodiments, the antibody molecule consists of or comprises one heavy chain and one light chain (referred to herein as a half-antibody). In another example, the antibody molecule includes two heavy (H) chain variable domain sequences and two light (L) chain variable domain sequences, thereby forming two antigen-binding sites, and can be produced by modification of a whole antibody or can be de novo synthesized using recombinant DNA technology, such as Fab, Fab’, F(ab’)2, Fc, Fd, Fd’, Fv, single-chain antibody (e.g., scFv), single variable domain antibody, diabody (Dab) (bivalent and bispecific), and chimeric (e.g., humanized) antibodies. These functional antibody fragments retain the ability to selectively bind to their respective antigens or receptors. Antibodies and antibody fragments can be derived from any class of antibodies, including but not limited to IgG, IgA, IgM, IgD, and IgE, and can be derived from any subclass of antibodies (e.g., IgG1, IgG2, IgG3, and IgG4). Preparations of antibody molecules can be monoclonal or polyclonal. Antibody molecules can also be human, humanized, CDR-grafted, or in vitro-generated antibodies. The antibody can have a heavy chain constant region selected, for example, from IgG1, IgG2, IgG3, or IgG4. The antibody can also have a light chain selected, for example, from kappa or lambda. The term “immunoglobulin” (Ig) is used interchangeably herein with the term “antibody”.

[0273] Examples of antigen-binding fragments of antibody molecules include: (i) Fab fragments; monovalent fragments consisting of the VL, VH, CL, and CH1 domains; (ii) F(ab’)2 fragments; divalent fragments containing two Fab fragments linked by disulfide bridges in the hinge region; (iii) Fd fragments consisting of the VH and CH1 domains; (iv) Fv fragments consisting of the VL and VH domains of a single arm of an antibody; (v) diabody (dAb) fragments consisting of the VH domain; (vi) camelid or camelized variable domains; (vii) single-chain Fv (scFv), see, e.g., Bird et al. (1988) Science 242:423-426; and Huston et al. (1988) Proc. Natl. Acad. Sci. USA 85:5879-5883); (viii) single-domain antibodies. These antibody fragments can be obtained using conventional techniques known to those skilled in the art, and the fragments are screened for utility in the same manner as intact antibodies.

[0274] The term “antibody” includes intact molecules as well as its functional fragments. The constant region of an antibody can be modified, e.g., mutated, to alter the properties of the antibody, such as to increase or decrease one or more of Fc receptor binding, antibody glycosylation, the number of cysteine residues, effector cell function, or complement function.

[0275] An antibody molecule can also be a single-domain antibody. A single-domain antibody can include an antibody whose complementarity-determining region is part of a single-domain polypeptide. Examples include, but are not limited to, heavy-chain antibodies, antibodies that are naturally lacking in light chains, single-domain antibodies derived from conventional four-chain antibodies, genetically engineered antibodies, and single-domain scaffolds other than antibody-derived ones. A single-domain antibody can be any or any future single-domain antibody in the art. A single-domain antibody can be derived from any species including, but not limited to, mouse, human, camel, llama, fish, shark, goat, rabbit, and cow. In another aspect of the invention, the single-domain antibody is a single-domain antibody of natural origin known as a heavy-chain antibody lacking a light chain. Such single-domain antibodies are disclosed, for example, in WO9404678. For clarity, this variable domain derived from a heavy-chain antibody that is naturally lacking in a light chain is known herein as VHH or nanobody and is distinguished from the conventional VH of a four-chain immunoglobulin. Such VHH molecules can be derived from antibodies produced in Camelidae species such as camel, llama, dromedary, alpaca, and guanaco. Other species other than Camelidae can also produce heavy-chain antibodies lacking a light chain; such VHHs are within the scope of the invention.

[0276] The VH and VL regions can be subdivided into regions of high-frequency variability named "complementary-determining regions" (CDRs) interspersed with more conserved regions named "framework regions" (FR or FW).

[0277] The framework regions and the CDR ranges are defined precisely by a number of methods [see Kabat, E. A., et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No. 91-3242; Chothia, C. et al. (1987) J. Mol. Biol. 196:901-917; and the AbM definitions used by Oxford Molecular's AbM antibody modeling software]. Generally, see, for example, Protein Sequence and Structure Analysis of Antibody Variable Domains.:Antibody Engineering Lab Manual (Ed.: Duebel, S. and Kontermann, R., Springer-Verlag, Heidelberg).

[0278] The terms "complementary determining region" and "CDR" as used herein refer to the sequences of amino acids within the antibody variable regions that confer antigen specificity and binding affinity. Generally, there are three CDRs in each heavy chain variable region (HCDR1, HCDR2, HCDR3) and three CDRs in each light chain variable region (LCDR1, LCDR2, LCDR3).

[0279] The exact amino acid sequence boundaries of a given CDR can be determined using any of a number of well-known schemes, such as those described in Kabat et al. (1991), “Sequences of Proteins of Immunological Interest,” 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD (the “Kabat” numbering scheme), Al-Lazikani et al., (1997) JMB 273,927-948 (the “Chothia” numbering scheme). As used herein, a CDR defined according to the “Chothia” numbering scheme may also be referred to as a “hypervariable loop.”

[0280] For example, in Kabat, the CDR amino acid residues in the heavy chain variable domain (VH) are numbered 31-35 (HCDR1), 50-66 (HCDR2), and 99-109 (HCDR3); and the CDR amino acid residues in the light chain variable domain (VL) are numbered 24-34 (LCDR1), 50-56 (LCDR2), and 89-97 (LCDR3). In Chothia, the CDR amino acids in VH are numbered 26-32 (HCDR1), 52-57 (HCDR2), and 99-109 (HCDR3); and the amino acid residues in VL are numbered 26-32 (LCDR1), 50-52 (LCDR2), and 91-96 (LCDR3). By combining the CDR definitions of both Kabat and Chothia, the CDRs consist of amino acid residues 26-35 (HCDR1), 50-66 (HCDR2), and 99-109 (HCDR3) in human VH, and amino acid residues 24-34 (LCDR1), 50-56 (LCDR2), and 89-97 (LCDR3) in human VL.

[0281] Generally, unless otherwise indicated, an anti-PD-L1 antibody molecule can include, for example, any combination of one or more of the Kabat CDRs and / or Chothia hypervariable loops described in Table 1. In certain embodiments, the following definitions are used for the anti-PD-L1 antibody molecules described in Table 1: HCDR1 according to the combined CDR definition of both Kabat and Chothia, and HCCDR2-3 and LCCDR1-3 according to the Kabat CDR definition. In every definition, each VH and VL typically includes three CDRs and four FRs arranged in the following order from the amino terminus to the carboxy terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.

[0282] As used herein, the term “immunoglobulin variable domain sequence” refers to an amino acid sequence capable of forming the structure of an immunoglobulin variable domain. For example, the sequence can include all or a portion of the amino acid sequence of a naturally-occurring variable domain. For example, the sequence can or cannot include one, two or more N- or C-terminal amino acids, or can include other modifications compatible with protein structure formation.

[0283] The term “antigen-binding site” refers to the portion of an antibody molecule that contains determinants that form an interface that binds to a PD-L1 polypeptide or an epitope thereof. With respect to a protein (or protein mimetic), the antigen-binding site typically includes one or more loops (of at least four amino acids or amino acid mimetics) that form an interface that binds to the PD-L1 polypeptide. Typically, the antigen-binding site of an antibody molecule includes at least one or two CDRs and / or hypervariable loops, or more typically, at least three, four, five or six CDRs and / or hypervariable loops.

[0284] The term "monoclonal antibody" or "monoclonal antibody composition" as used herein refers to a preparation of antibody molecules of a single molecular composition. Monoclonal antibody compositions exhibit a single binding specificity and affinity for a particular epitope. Monoclonal antibodies can be produced by hybridoma technology or by methods that do not use hybridoma technology (e.g., recombinant methods).

[0285] A "human-effective" protein is one that does not induce a neutralizing antibody response, e.g., a human anti-mouse antibody (HAMA) response. HAMA can be a problem in a number of situations, e.g., when antibody molecules are administered repeatedly, e.g., in the treatment of chronic or recurrent disease states. The HAMA response can potentially render repeated antibody administration ineffective due to increased antibody clearance from the serum [see, e.g., Saleh et al., Cancer Immunol. Immunother., 32:180-190 (1990)] and due to potential allergic reactions [see, e.g., LoBuglio et al., Hybridoma, 5:5117-5123 (1986)].

[0286] Antibody molecules can be polyclonal or monoclonal antibodies. In other embodiments, the antibodies can be produced recombinantly, e.g., by phage display or combinatorial methods.

[0287] Phage display and combinatorial methods for generating antibodies are known in the art [e.g., Ladner et al. U.S. Patent No. 5,223,409; Kang et al. International Publication WO92 / 18619; Dower et al. International Publication WO91 / 17271; Winter et al. International Publication WO92 / 20791; Markland et al. International Publication WO92 / 15679; Breitling et al. International Publication WO93 / 01288; McCafferty et al. International Publication WO92 / 01047; Garrard et al. International Publication WO92 / 09690; Ladner et al. International Publication WO90 / 02809; Fuchs et al. (1991) Bio / Technology 9:1370-1372; Hay et al. (1992) Hum Antibod Hybridomas 3:81-85; Huse et al. (1989) Science 246:1275-1281; Griffiths et al. (1993) EMBO J 12:725-734; Hawkins et al. (1992) J Mol Biol 226:889-896; Clackson et al. (1991) Nature 352:624-628; Gram et al. (1992) PNAS 89:3576-3580; Garrad et al. (1991) Bio / Technology 9:1373-1377; Hoogenboom et al. (1991) Nuc Acid Res 19:4133-4137; and Barbas et al. (1991) PNAS 88:7978-7982, all of which are hereby incorporated by reference in their entirety].

[0288] In one aspect, the antibody is a fully human antibody (e.g., an antibody made in a mouse genetically engineered to produce an antibody from human immunoglobulin sequences) or a non-human antibody, e.g., a rodent (mouse or rat), goat, primate (e.g., monkey), camelid antibody. Preferably, the non-human antibody is a rodent (mouse or rat antibody). Methods for producing rodent antibodies are known in the art.

[0289] Human monoclonal antibodies can be made using transgenic mice that carry human immunoglobulin genes rather than mouse systems. Spleen cells from such transgenic mice immunized with the antigen of interest are used to produce hybridomas that secrete human mAbs having specific affinity for epitopes derived from human proteins (see, e.g., Wood et al. International Application WO91 / 00906, Kucherlapati et al. PCT Publication WO91 / 10741; Lonberg et al. International Application WO92 / 03918; Kay et al. International Application 92 / 03917; Lonberg, N. et al. 1994 Nature 368:856-859; Green, L.L. et al. 1994 Nature Genet. 7:13-21; Morrison, S.L. et al. 1994 Proc. Natl. Acad. Sci. USA 81:6851-6855; Bruggeman et al. 1993 Year Immunol 7:33-40; Tuaillon et al. 1993 PNAS 90:3720-3724; Bruggeman et al. 1991 Eur J Immunol 21:1323-1326).

[0290] An antibody can be one in which the variable region or a portion thereof, such as a CDR, is an antibody made in a non-human organism, such as a rat or a mouse. Chimeric, CDR-grafted, and humanized antibodies are within the scope of the present invention. Antibodies made in a non-human organism, such as a rat or a mouse, and subsequently modified in the variable framework or constant region to, for example, reduce antigenicity in a human, are within the scope of the present invention.

[0291] Chimeric antibodies can be produced by recombinant DNA techniques known in the art [see Robinson et al., International Patent Publication PCT / US86 / 02269; Akira, et al., European Patent Application No. 184,187; Taniguchi, M., European Patent Application No. 171,496; Morrison et al., European Patent Application No. 173,494; Neuberger et al., International Application WO86 / 01533; Cabilly et al., U.S. Patent No. 4,816,567; Cabilly et al., European Patent Application No. 125,023; Better et al. (1988 Science 240:1041-1043); Liu et al. (1987) PNAS 84:3439-3443; Liu et al., 1987, J. Immunol. 139:3521-3526; Sun et al. (1987) PNAS 84:214-218; Nishimura et al., 1987, Canc. Res. 47:999-1005; Wood et al. (1985) Nature 314:446-449; and Shaw et al., 1988, J. Natl Cancer Inst. 80:1553-1559].

[0292] Humanized or CDR-grafted antibodies will have at least 1 or 2, but generally all 3 recipient CDRs [of the heavy and / or light immunoglobulin chains] replaced by donor CDRs. The antibody can be replaced by at least a portion of the non-human CDRs, or only a small part of the CDRs can be replaced by non-human CDRs. It is only necessary to replace the number of CDRs required for the binding of the humanized antibody to PD-L1. Preferably, the donor will be a rodent antibody, such as a rat or mouse antibody, and the recipient will be a human framework or human consensus framework. Typically, the immunoglobulin providing the CDRs is called the "donor", and the immunoglobulin providing the framework is called the "acceptor". In one aspect, the donor immunoglobulin is non-human (e.g., rodent). The acceptor framework is a naturally occurring (e.g., human) framework or consensus framework, or a sequence that is about 85% or more, preferably 90%, 95%, 99% or more identical thereto.

[0293] As used herein, the term "consensus sequence" refers to a sequence formed from the amino acids (or nucleotides) that occur most frequently in a family of related sequences [see, e.g., Winnaker, From Genes to Clones (Verlagsgesellschaft, Weinheim, Germany 1987)]. In a family of proteins, each position in the consensus sequence is occupied by the amino acid that occurs most frequently at that position in the family. If two amino acids occur equally frequently, either may be included in the consensus sequence. "Consensus framework" refers to the framework region in a consensus immunoglobulin sequence.

[0294] Antibodies can be humanized by methods known in the art (see, e.g., Morrison, S. L., 1985, Science 229:1202-1207, Oi et al., 1986, BioTechniques 4:214 and US5,585,089, US5,693,761 and US5,693,762 by Queen et al., which are hereby incorporated by reference in their entirety).

[0295] Humanized or CDR-grafted antibodies can be produced by CDR grafting or CDR substitution, whereby one, two or all CDRs of an immunoglobulin chain can be replaced. See, e.g., U.S. Pat. No. 5,225,539; Jones et al. 1986 Nature 321:552-525; Verhoeyan et al. 1988 Science 239:1534; Beidler et al. 1988 J. Immunol. 141:4053-4060; Winter, U.S. Pat. No. 5,225,539, the entire contents of which are hereby incorporated by reference. Winter described a CDR grafting method that can be used to prepare the humanized antibodies of the present invention [British Patent Application GB2188638(A), filed Mar. 26, 1987; Winter, U.S. Pat. No. 5,225,539], the contents of which are hereby incorporated by reference in their entirety).

[0296] Humanized antibodies with specific amino acid substitutions, deletions or additions are also within the scope of the present invention. The criteria for selecting amino acids from the donor are described in US5,585,089, e.g., paragraphs 12-16 of US5,585,089, the entire contents of which are hereby incorporated by reference. Other techniques for humanizing antibodies are described in Padlan et al., EP519596(A1), published Dec. 23, 1992).

[0297] The antibody molecule can be a single-chain antibody. The single-chain antibody (scFv) may be genetically engineered [see, for example, Colcher, D. et al. (1999) Ann N Y Acad Sci 880:263-80; and Reiter, Y. (1996) Clin Cancer Res 2:245-52]. The single-chain antibody can dimerize or multimerize to form a multivalent antibody having specificity for different epitopes of the same target protein.

[0298] In still other embodiments, the antibody molecule is selected from, for example, the heavy chain constant regions of IgG1, IgG2, IgG3, IgG4, IgM, IgA1, IgA2, IgD, and IgE; in particular, it has a heavy chain constant region selected from, for example, the (e.g., human) heavy chain constant regions of IgG1, IgG2, IgG3, and IgG4. In another embodiment, the antibody molecule has a light chain constant region selected from, for example, the (e.g., human) kappa or lambda light chain constant regions. The constant region can be altered, e.g., mutated, to modify the properties of the antibody (e.g., to increase or decrease one or more of Fc receptor binding, antibody glycosylation, the number of cysteine residues, effector cell function, and / or complement function). In one embodiment, the antibody has effector function and can fix complement. In other embodiments, the antibody does not recruit effector cells or fix complement. In another embodiment, the antibody has a reduced or no ability to bind to Fc receptors. For example, this can be an isotype or subtype, fragment, or other mutant that does not support binding to Fc receptors, e.g., having a mutagenized or deleted Fc receptor binding region.

[0299] Methods for altering the constant region of an antibody are known in the art. Antibodies having altered functions, such as altered affinity for effector ligands such as FcR or the C1 component of complement in cells, can be produced by replacing at least one amino acid residue in the constant portion of the antibody with a different residue [see, e.g., EP388,151 (A1), U.S. Patent No. 5,624,821, and U.S. Patent No. 5,648,260, which are hereby incorporated by reference in their entirety]. Similar types of alterations that would reduce or eliminate these functions can be described when applied to mouse or other species immunoglobulins.

[0300] Antibody molecules can be derivatized or conjugated to another functional molecule (e.g., another peptide or protein). As used herein, a “derivatized” antibody molecule is an antibody molecule that has been modified. Methods of derivatization include, but are not limited to, the addition of a fluorescent moiety, a radioactive nucleotide, a toxin, an enzyme, or an affinity ligand such as biotin. Accordingly, the antibody molecules of the invention are intended to include antibodies described herein in derivatized and otherwise modified forms, including immunoadhesion molecules. For example, an antibody molecule can be functionally linked (by chemical coupling, genetic fusion, non-covalent binding, or otherwise) to one or more other molecular entities, such as another antibody (e.g., a bispecific antibody or diabody), a detectable agent, a cytotoxic agent, a pharmaceutical agent, and / or another molecule, such as a protein or peptide that can mediate association of the antibody or antibody portion with streptavidin core region or polyhistidine tag, etc.

[0301] One type of derivatized antibody molecule is produced by crosslinking two or more antibodies (of the same or different types, e.g., to produce a bispecific antibody). Suitable crosslinking agents include heterobifunctional (e.g., m-maleimidobenzoyl-N-hydroxysuccinimide ester) or homobifunctional (e.g., disuccinimidyl suberate) crosslinking agents having two separate reactive groups spaced by an appropriate spacer. Such linkers are available from Pierce Chemical Company, Rockford, Ill.

[0302] Useful detectable agents that can derivatize (or label) the antibody molecules of the present invention include fluorescent compounds, various enzymes, families of hapten molecules, luminescent materials, bioluminescent materials, fluorescent-emitting metal atoms such as europium (Eu) and other lanthanides, and radioactive materials (described later). Exemplary fluorescent detectable agents include fluorescein, fluorescein isothiocyanate, rhodamine, 5-dimethylamine-1-naphthalenesulfonyl chloride, phycoerythrin, and others. Antibodies can also be derivatized with detectable enzymes such as alkaline phosphatase, horseradish peroxidase, β-galactosidase, acetylcholinesterase, glucose oxidase, and others. When an antibody is derivatized with a detectable enzyme, it is detected by adding additional reagents used by the enzyme for the production of a detectable reaction product. For example, when horseradish peroxidase, a detectable agent, is present, the addition of hydrogen peroxide and diaminobenzidine results in a colored reaction product that is detectable. Antibody molecules can also be derivatized with families of hapten molecules (e.g., streptavidin / biotin and avidin / biotin). For example, an antibody can be derivatized with biotin and detected by indirect measurement of avidin or streptavidin binding. Examples of suitable fluorescent materials include umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride, or phycoerythrin; an example of a luminescent material is luminol; and examples of bioluminescent materials include luciferase, luciferin, and aequorin.

[0303] The labeled antibody molecules can be used diagnostically and / or experimentally in a number of contexts, such as (i) to isolate a given antigen by standard techniques such as affinity chromatography or immunoprecipitation; (ii) to detect a given antigen (e.g., in a cell lysate or cell supernatant) to assess the abundance and expression pattern of a protein; (iii) to monitor protein levels in tissues as part of a clinical assay procedure, e.g., to determine the effectiveness of a given treatment regimen.

[0304] The antibody molecules can be conjugated to another molecular entity, typically a label or a therapeutic (e.g., cytotoxic or cytostatic) agent or moiety. Radioisotopes can be used in diagnostic or therapeutic applications. Examples of radioisotopes that can bind to an anti-PSMA antibody include, but are not limited to, α-, β- or γ-emitters, or β- and γ-emitters. Such radioisotopes include iodine ( 131 I or 125 I), yttrium ( 90 Y), lutetium ( 177 Lu), actinium ( 225 Ac), praseodymium, astatine ( 211 At), rhenium ( 186 Re), bismuth ( 212 Bi or 213 Bi), indium ( 111 In), technetium ( 99 mTc), phosphorus ( 32 P), rhodium ( 188 Rh), sulfur ( 35 S), carbon ( 14 C), tritium ( 3 H), chromium ( 51 Cr), chlorine ( 36 Cl), cobalt ( 57 Co or 58 Co), iron ( 59 Fe), selenium ( 75 Se) or gallium ( 67Examples include, but are not limited to, (Ga). Radioactive isotopes useful as therapeutic agents include yttrium ( 90 Y), lutetium ( 177 Lu), actinium ( 225 Ac), praseodymium, astatine ( 211 At), rhenium ( 186 Re), bismuth ( 212 Bi or 213 Bi) and rhodium ( 188 Rh). For example, radioactive isotopes useful as labels for diagnostic use include iodine ( 131 I or 125 I), indium ( 111 In), technetium ( 99 mTc), phosphorus ( 32 P), carbon ( 14 C) and tritium ( 3 H), or one or more of the therapeutic isotopes listed above.

[0305] The present invention provides radiolabeled antibody molecules and methods for labeling them. In one aspect, a method for labeling an antibody molecule is disclosed. The method includes contacting the antibody molecule with a chelating agent, thereby producing a conjugated antibody. The conjugated antibody is radiolabeled with a radioactive isotope, such as, for example, 111 indium, 90 yttrium and 177 lutetium, thereby producing a labeled antibody molecule.

[0306] As noted above, the antibody molecule can be conjugated to a therapeutic agent. Radioactive isotopes with therapeutic activity have already been mentioned. Examples of other therapeutic agents include taxol, cytochalasin B, gramicidin D, ethidium bromide, emetine, mitomycin, etoposide, teniposide, vincristine, vinblastine, colchicine, doxorubicin, daunorubicin, dihydroxyanthracinedione, mitoxantrone, mithramycin, actinomycin D, 1-dehydrotestosterone, glucocorticoids, procaine, tetracaine, lidocaine, propranolol, puromycin, maytansinoids such as maytansinol (see U.S. Patent No. 5,208,020), CC-1065 (see U.S. Patents Nos. 5,475,092, 5,585,499, and 5,846,545), and analogs or homologs thereof. Therapeutic agents include antimetabolites [e.g., methotrexate, 6-mercaptopurine, 6-thioguanine, cytarabine, 5-fluorouracil dacarbazine], alkylating agents [e.g., mechlorethamine, thiotepa chlorambucil, CC-1065, melphalan, carmustine (BSNU) and lomustine (CCNU), cyclophosphamide, busulfan, dibromomannitol, streptozotocin, mitomycin C and cis-dichlorodiamine platinum (II) (DDP) cisplatin], anthracyclines [e.g., daunorubicin (formerly daunomycin) and doxorubicin], antibiotics [e.g., dactinomycin (formerly actinomycin), bleomycin, mithramycin and anthramycin (AMC)] and antimitotic agents [e.g., vincristine, vinblastine, taxol and maytansinoids], but are not limited thereto.

[0307] In one aspect, the present invention features a method of preparing a target binding molecule that specifically binds to a PD-L1 receptor. For example, the target binding molecule is an antibody molecule. The method includes preparing a target protein that is at least a portion of a non-human protein and is homologous to the corresponding portion of a human target protein (at least 70, 75, 80, 85, 87, 90, 92, 94, 95, 96, 97, 98% identical), but includes a portion that differs by at least one amino acid (e.g., at least 1, 2, 3, 4, 5, 6, 7, 8, or 9 amino acids); obtaining an antibody molecule that specifically binds to the antigen; and evaluating the effectiveness of a binding agent in modulating the activity of the target protein. The method can further include administering a binding agent (e.g., an antibody molecule) or derivative (e.g., a humanized antibody molecule) to a human subject.

[0308] In certain embodiments, the antibody molecule is a multispecific (e.g., bispecific or trispecific) antibody molecule. Protocols for making bispecific or heterodimeric antibody molecules are known in the art; for example, but not limited to: for example, the "knob and hole" approach described in US5731168; for example, electrostatic steering Fc pairing described in WO09 / 089004, WO06 / 106905 and WO2010 / 129304; for example, strand exchange engineered domain (SEED) heterodimer formation described in WO07 / 110205; for example, Fab arm exchange described in WO08 / 119353, WO2011 / 131746 and WO2013 / 060867; for example, double antibody conjugates by cross-linking antibodies to create a bispecific structure using a heterobifunctional reagent having, for example, amine reactive groups and sulfhydryl reactive groups as described in US4433059; for example, bispecific antibody determinants made by recombining half-antibodies (heavy-light chain pairs or Fabs) from different antibodies by cycles of reduction and oxidation of disulfide bonds between two heavy chains as described in US4444878; for example, trifunctional antibodies such as three Fab' fragments cross-linked by sulfhydryl reactive groups as described in US5273743; for example, biosynthetic binding proteins such as pairs of scFvs cross-linked by C-terminal tails, preferably by disulfide or amine-reactive chemical cross-linking as described in US5534254; for example, bifunctional antibodies such as Fab fragments having different binding specificities dimerized by leucine zippers with constant domains replaced (e.g., c-fos and c-jun) as described in US5591828; for example, bispecific and oligovalent monovalent and oligovalent receptors such as the VH-CH1 regions of two antibodies (two Fab fragments) linked by a polypeptide spacer between the CH1 region of one antibody and the VH region of the other antibody typically by a related light chain as described in US5635602; for example, bispecific DNA-antibody conjugates such as cross-linking of an antibody or Fab fragment by a double-stranded piece of DNA as described in US5635602; for example,The expression constructs described in US5637481, such as bispecific fusion proteins containing two scFvs and a complete constant region with a hydrophilic helical peptide linker therebetween; multivalent and multispecific binding proteins described in, for example, US5837242, such as polypeptides having a first domain with a binding region of an Ig heavy chain variable region and a second domain with a binding region of an Ig light chain variable region, generally called diabodies [higher order structures for making bispecific, trispecific or tetraspecific molecules are also included]; minibody constructs having linked VL and VH chains further connected by a peptide spacer to an antibody hinge region and CH3 region that can dimerize to form a bispecific / polyvalent molecule, described in, for example, US5837821; VH and VL domains linked by a short peptide linker (e.g., 5 or 10 amino acids) in any orientation or without a linker at all that can dimerize to form a bispecific diabody; trimers and tetramers described in, for example, US5844094; strings of VH domains (or VL domains in a family member) connected by peptide bonds having a cross-linkable chemical group at the C-terminus that further associate with VL domains to form a series of Fv (or scFv), described in, for example, US5864019; and single-chain binding polypeptides having both VH and VL domains linked by a peptide linker that are combined into a multivalent structure by non-covalent or chemical cross-linking, using, for example, both scFV or diabody-type formats, to form homo- and hetero-bivalent, trivalent and tetravalent structures. Additional exemplary multispecific and bispecific molecules, and methods for making them, are described, for example, in US5910573, US5932448, US5959083, US5989830, US6005079, US6239259, US6294353, US6333396, US6476198, US6511663, US6670453, US6743896, US6809185, US6833441, US7129330, US7183076,It is found in US7521056, US7527787, US7534866, US7612181, US2002004587A1, US2002076406A1, US2002103345A1, US2003207346A1, US2003211078A1, US2004219643A1, US2004220388A1, US2004242847A1, US2005003403A1, US2005004352A1, US2005069552A1, US2005079170A1, US2005100543A1, US2005136049A1, US2005136051A1, US2005163782A1, US2005266425A1, US2006083747A1, US2006120960A1, US2006204493A1, US2006263367A1, US2007004909A1, US2007087381A1, US2007128150A1, US2007141049A1, US2007154901A1, US2007274985A1, US2008050370A1, US2008069820A1, US2008152645A1, US2008171855A1, US2008241884A1, US2008254512A1, US2008260738A1, US2009130106A1, US2009148905A1, US2009155275A1, US2009162359A1, US2009162360A1, US2009175851A1, US2009175867A1, US2009232811A1, US2009234105A1, US2009263392A1, US2009274649A1, EP346087A2, WO0006605A2, WO02072635A2, WO04081051A1, WO06020258A2, WO2007044887A2, WO2007095338A2, WO2007137760A2, WO2008119353A1, WO2009021754A2, WO2009068630A1, WO9103493A1, WO9323537A1, WO9409131A1, WO9412625A2, WO9509917A1, WO9637621A2, WO9964460A1. The content of the applications referred to above is hereby incorporated by reference in its entirety.

[0309] In other embodiments, an anti-PD-L1 antibody molecule (e.g., a monospecific, bispecific or multispecific antibody molecule) is covalently linked, e.g., fused, to another partner, e.g., a protein, e.g., one or more cytokines, e.g., as a fusion molecule, e.g., a fusion protein. In other embodiments, the fusion molecule comprises one or more proteins, e.g., one or more cytokines. In certain embodiments, the cytokine is an interleukin (IL) selected from one, two or more of IL-1, IL-2, IL-12, IL-15 or IL-21. In certain embodiments, the bispecific antibody molecule has a first binding specificity for a first target (e.g., PD-L1) and a second binding specificity for a second target (e.g., LAG-3 or TIM-3) and may be linked to an interleukin (e.g., IL-12) domain, e.g., full-length IL-12 or a portion thereof.

[0310] "Fusion protein" and "fusion polypeptide" refer to a polypeptide having at least two moieties linked together by a covalent bond, each of which moieties is a polypeptide having different properties. The properties can be biological properties such as activity in vitro or in vivo. The properties can also be simple chemical or physical properties such as binding to a target molecule, catalysis of a reaction, etc. The two moieties can be linked directly by a single peptide bond or via a peptide linker, but are in-frame with each other.

[0311] The present invention provides an isolated nucleic acid molecule encoding the above-described antibody molecule, its vector and host cell. Nucleic acid molecules include, but are not limited to, RNA, genomic DNA and cDNA.

[0312] Exemplary anti-PD-L1 antibody molecules In certain embodiments, the anti-PD-L1 antibody molecule is (i) A heavy chain variable region (VH) comprising a VHCDR1 amino acid sequence selected from SEQ ID NO: 1, SEQ ID NO: 4, or SEQ ID NO: 195, the VHCDR2 amino acid sequence of SEQ ID NO: 2, and the VHCDR3 amino acid sequence of SEQ ID NO: 3, and (ii) A light chain variable region (VL) comprising the VLCDR1 amino acid sequence of SEQ ID NO: 9, the VLCDR2 amino acid sequence of SEQ ID NO: 10, and the VLCDR3 amino acid sequence of SEQ ID NO: 11 comprising.

[0313] In other embodiments, the anti-PD-L1 antibody molecule is (i) A heavy chain variable region (VH) comprising a VHCDR1 amino acid sequence selected from SEQ ID NO: 1, SEQ ID NO: 4, or SEQ ID NO: 195, the VHCDR2 amino acid sequence of SEQ ID NO: 5, and the VHCDR3 amino acid sequence of SEQ ID NO: 3, and (ii) A light chain variable region (VL) comprising the VLCDR1 amino acid sequence of SEQ ID NO: 12, the VLCDR2 amino acid sequence of SEQ ID NO: 13, and the VLCDR3 amino acid sequence of SEQ ID NO: 14 comprising.

[0314] In the embodiments of the aforementioned antibody molecules, VHCDR1 comprises the amino acid sequence of SEQ ID NO: 1. In other embodiments, VHCDR1 comprises the amino acid sequence of SEQ ID NO: 4. In still other embodiments, VHCDR1 comprises the amino acid sequence of SEQ ID NO: 195.

[0315] In embodiments, the aforementioned antibody molecule has a heavy chain variable region comprising at least one framework (FW) region comprising an amino acid sequence of any one of SEQ ID NO: 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150, 152, or 154, or an amino acid sequence that is at least 90% identical thereto, or an amino acid sequence having 2 or fewer amino acid substitutions, insertions, or deletions as compared to an amino acid sequence of any one of SEQ ID NO: 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150, 152, or 154.

[0316] In other embodiments, the antibody molecule described above has a heavy chain variable region comprising at least one framework region comprising an amino acid sequence of any one of SEQ ID NOs: 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150, 152 or 154.

[0317] In still other embodiments, the antibody molecule described above has a heavy chain variable region comprising at least 2, 3 or 4 framework regions comprising an amino acid sequence of any one of SEQ ID NOs: 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150, 152 or 154.

[0318] In other embodiments, the antibody molecule described above comprises the VHFW1 amino acid sequence of SEQ ID NOs: 124, 126, 128 or 130, the VHFW2 amino acid sequence of SEQ ID NOs: 132, 134, 136, 138, 140 or 142, and the VHFW3 amino acid sequence of SEQ ID NOs: 144, 146, 148, 150 or 152, and may further comprise the VHFW4 amino acid sequence of SEQ ID NO: 154.

[0319] In other embodiments, the antibody molecule described above has a light chain variable region comprising at least one framework region comprising an amino acid sequence of any one of SEQ ID NOs: 156, 158, 160, 162, 164, 166, 168, 170, 172, 174, 176, 178, 180, 182, 184 or 186, or an amino acid sequence that is at least 90% identical thereto, or an amino acid sequence having 2 or fewer amino acid substitutions, insertions or deletions as compared to an amino acid sequence of any one of SEQ ID NOs: 156, 158, 160, 162, 164, 166, 168, 170, 172, 174, 176, 178, 180, 182, 184 or 186.

[0320] In other embodiments, the aforementioned antibody molecule has a light chain variable region comprising at least one framework region comprising the amino acid sequence of any one of SEQ ID NOs: 156, 158, 160, 162, 164, 166, 168, 170, 172, 174, 176, 178, 180, 182, 184 or 186.

[0321] In other embodiments, the aforementioned antibody molecule has a light chain variable region comprising at least 2, 3 or 4 framework regions comprising the amino acid sequence of any one of SEQ ID NOs: 156, 158, 160, 162, 164, 166, 168, 170, 172, 174, 176, 178, 180, 182, 184 or 186.

[0322] In other embodiments, the aforementioned antibody molecule comprises the VLFW1 amino acid sequence of any one of SEQ ID NOs: 156, 158, 160, 162, 164 or 166, the VLFW2 amino acid sequence of SEQ ID NOs: 168 or 170, and the VLFW3 amino acid sequence of any one of SEQ ID NOs: 172, 174, 176, 178, 180, 182 or 184, and may further comprise the VLFW4 amino acid sequence of SEQ ID NO: 186.

[0323] In other embodiments, the aforementioned antibody comprises a heavy chain variable domain comprising an amino acid sequence that is at least 85% identical to any one of SEQ ID NOs: 18, 30, 38, 46, 50, 54, 62, 70 or 78.

[0324] In other embodiments, the aforementioned antibody molecule comprises a heavy chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 18, 30, 38, 46, 50, 54, 62, 70 or 78.

[0325] In other embodiments, the aforementioned antibody molecule comprises a light chain variable domain comprising an amino acid sequence that is at least 85% identical to any one of SEQ ID NOs: 22, 26, 34, 42, 58, 66, 74, 82 or 86.

[0326] In other embodiments, the aforementioned antibody molecule comprises a light chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 22, 26, 34, 42, 58, 66, 74, 82 or 86.

[0327] In other embodiments, the aforementioned antibody molecule comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 18. In other embodiments, the aforementioned antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 20.

[0328] In other embodiments, the aforementioned antibody molecule comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 30.

[0329] In other embodiments, the aforementioned antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 32. In other embodiments, the aforementioned antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 96. In other embodiments, the aforementioned antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 197.

[0330] In other embodiments, the aforementioned antibody molecule comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 38.

[0331] In other embodiments, the aforementioned antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 40. In other embodiments, the aforementioned antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 91.

[0332] In other embodiments, the aforementioned antibody molecule comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 46. In other embodiments, the aforementioned antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 48.

[0333] In other embodiments, the aforementioned antibody molecule comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 50. In other embodiments, the aforementioned antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 52.

[0334] In other embodiments, the aforementioned antibody molecule comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 54.

[0335] In other embodiments, the aforementioned antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 56. In other embodiments, the aforementioned antibody molecule comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 62.

[0336] In other embodiments, the aforementioned antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 64. In other embodiments, the aforementioned antibody molecule comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 70.

[0337] In other embodiments, the aforementioned antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 72. In other embodiments, the aforementioned antibody molecule comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 78.

[0338] In other embodiments, the aforementioned antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 80. In other embodiments, the aforementioned antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 247. In other embodiments, the aforementioned antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 260.

[0339] In other embodiments, the aforementioned antibody molecule comprises a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 22. In other embodiments, the aforementioned antibody molecule comprises a light chain comprising the amino acid sequence of SEQ ID NO: 24.

[0340] In other embodiments, the aforementioned antibody molecule comprises a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 26. In other embodiments, the aforementioned antibody molecule comprises a light chain comprising the amino acid sequence of SEQ ID NO: 28.

[0341] In other embodiments, the aforementioned antibody molecule comprises a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 34.

[0342] In other embodiments, the aforementioned antibody molecule comprises a light chain comprising the amino acid sequence of SEQ ID NO: 36. In other embodiments, the aforementioned antibody molecule comprises a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 42.

[0343] In other embodiments, the aforementioned antibody molecule comprises a light chain comprising the amino acid sequence of SEQ ID NO: 44. In other embodiments, the aforementioned antibody molecule comprises a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 58.

[0344] In other embodiments, the aforementioned antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 60. In other embodiments, the aforementioned antibody molecule comprises a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 66.

[0345] In other embodiments, the aforementioned antibody molecule comprises a light chain comprising the amino acid sequence of SEQ ID NO: 68. In other embodiments, the aforementioned antibody molecule comprises a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 74.

[0346] In other embodiments, the aforementioned antibody molecule comprises a light chain comprising the amino acid sequence of SEQ ID NO: 76. In other embodiments, the aforementioned antibody molecule comprises a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 82.

[0347] In other embodiments, the aforementioned antibody molecule comprises a light chain comprising the amino acid sequence of SEQ ID NO: 84. In other embodiments, the aforementioned antibody molecule comprises a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 86.

[0348] In other embodiments, the aforementioned antibody molecule comprises a light chain comprising the amino acid sequence of SEQ ID NO: 88. In other embodiments, the aforementioned antibody molecule comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 18 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 22.

[0349] In other embodiments, the aforementioned antibody molecule comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 18 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 26.

[0350] In other embodiments, the aforementioned antibody molecule comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 18 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 86.

[0351] In other embodiments, the aforementioned antibody molecule comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 30 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 34.

[0352] In other embodiments, the aforementioned antibody molecule comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 30 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 66.

[0353] In other embodiments, the aforementioned antibody molecule comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 38 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 42.

[0354] In other embodiments, the aforementioned antibody molecule comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 38 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 74.

[0355] In other embodiments, the aforementioned antibody molecule comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 46 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 42.

[0356] In other embodiments, the aforementioned antibody molecule comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 50 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 42.

[0357] In other embodiments, the aforementioned antibody molecule comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 50 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 22.

[0358] In other embodiments, the aforementioned antibody molecule comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 50 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 86.

[0359] In other embodiments, the aforementioned antibody molecule comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 54 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 58.

[0360] In other embodiments, the aforementioned antibody molecule comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 54 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 86.

[0361] In other embodiments, the aforementioned antibody molecule comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 62 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 66.

[0362] In other embodiments, the aforementioned antibody molecule comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 62 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 86.

[0363] In other embodiments, the aforementioned antibody molecule comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 70 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 66.

[0364] In other embodiments, the aforementioned antibody molecule comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 78 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 82.

[0365] In other embodiments, the aforementioned antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 20 and a light chain comprising the amino acid sequence of SEQ ID NO: 24.

[0366] In other embodiments, the aforementioned antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 20 and a light chain comprising the amino acid sequence of SEQ ID NO: 28.

[0367] In other embodiments, the aforementioned antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 20 and a light chain comprising the amino acid sequence of SEQ ID NO: 88.

[0368] In other embodiments, the aforementioned antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 32 and a light chain comprising the amino acid sequence of SEQ ID NO: 36.

[0369] In other embodiments, the aforementioned antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 32 and a light chain comprising the amino acid sequence of SEQ ID NO: 68.

[0370] In other embodiments, the aforementioned antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 40 and a light chain comprising the amino acid sequence of SEQ ID NO: 44.

[0371] In other embodiments, the aforementioned antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 40 and a light chain comprising the amino acid sequence of SEQ ID NO: 76.

[0372] In other embodiments, the aforementioned antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 48 and a light chain comprising the amino acid sequence of SEQ ID NO: 44.

[0373] In other embodiments, the aforementioned antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 52 and a light chain comprising the amino acid sequence of SEQ ID NO: 44.

[0374] In other embodiments, the aforementioned antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 52 and a light chain comprising the amino acid sequence of SEQ ID NO: 24.

[0375] In other embodiments, the aforementioned antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 52 and a light chain comprising the amino acid sequence of SEQ ID NO: 88.

[0376] In other embodiments, the aforementioned antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 260 and a light chain comprising the amino acid sequence of SEQ ID NO: 44.

[0377] In other embodiments, the aforementioned antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 56 and a light chain comprising the amino acid sequence of SEQ ID NO: 60.

[0378] In other embodiments, the antibody molecule described above comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 56 and a light chain comprising the amino acid sequence of SEQ ID NO: 88.

[0379] In other embodiments, the antibody molecule described above comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 64 and a light chain comprising the amino acid sequence of SEQ ID NO: 68.

[0380] In other embodiments, the antibody molecule described above comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 64 and a light chain comprising the amino acid sequence of SEQ ID NO: 88.

[0381] In other embodiments, the antibody molecule described above comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 72 and a light chain comprising the amino acid sequence of SEQ ID NO: 68.

[0382] In other embodiments, the antibody molecule described above comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 80 and a light chain comprising the amino acid sequence of SEQ ID NO: 84.

[0383] In other embodiments, the antibody molecule described above comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 247 and a light chain comprising the amino acid sequence of SEQ ID NO: 84.

[0384] In other embodiments, the antibody molecule described above comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 197 and a light chain comprising the amino acid sequence of SEQ ID NO: 36.

[0385] In other embodiments, the antibody molecule described above comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 91 and a light chain comprising the amino acid sequence of SEQ ID NO: 44.

[0386] In other embodiments, the antibody described above comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 96 and a light chain comprising the amino acid sequence of SEQ ID NO: 68.

[0387] In other embodiments, the antibody molecule described above is selected from Fab, F(ab’)2, Fv or single-chain Fv fragment (scFv).

[0388] In other embodiments, the antibody molecule described above comprises a heavy chain constant region selected from IgG1, IgG2, IgG3, and IgG4.

[0389] In other embodiments, the antibody molecule described above comprises a light chain constant region selected from a kappa or lambda light chain constant region.

[0390] In other embodiments, the antibody molecule described above comprises a human IgG4 heavy chain constant region having a mutation at position 228 of SEQ ID NO: 188 or 190, and a kappa light chain constant region.

[0391] In other embodiments, the antibody molecule described above comprises a human IgG4 heavy chain constant region having a serine-to-proline mutation at position 228 of SEQ ID NO: 188 or 190, and a kappa light chain constant region.

[0392] In other embodiments, the antibody molecule described above comprises a human IgG1 heavy chain constant region having an asparagine-to-alanine mutation at position 297 of SEQ ID NO: 192, and a kappa light chain constant region.

[0393] In other embodiments, the antibody molecule described above comprises a human IgG1 heavy chain constant region having an aspartic acid-to-alanine mutation at position 265 and a proline-to-alanine mutation at position 329 of SEQ ID NO: 193, and a kappa light chain constant region.

[0394] In other embodiments, the antibody molecule described above comprises a human IgG1 heavy chain constant region having a leucine-to-alanine mutation at position 234 and a leucine-to-alanine mutation at position 235 of SEQ ID NO: 194, and a kappa light chain constant region.

[0395] In other embodiments, the antibody molecule described above can bind to human PD-L1 with a dissociation constant (K D ) of less than about 0.2 nM.

[0396] In one aspect, the aforementioned antibody molecule has a K, as measured by, for example, the Biacore method, of about 2.5 nM, 2 nM, 1.5 nM, 1 nM, 0.5 nM, 0.2 nM, 0.15 nM, 0.1 nM, 0.05 nM or 0.02 nM, for example, about 0.2 nM to 0.1 nM, for example, about 0.166 nM to 0.176 nM, for example, about 0.171 nM, or for example, about 0.1 nM to 1.5 nM, for example, about 0.25 to 0.46 nM, for example, about 0.137 nM, 0.931 nM or less than 2.14 nM D and binds to human PD-L1.

[0397] In another aspect, the aforementioned antibody molecule has a K, as measured by, for example, the Biacore method, of about 1 nM, 0.8 nM, 0.6 nM, 0.4 nM, 0.2 nM, 0.15 nM, 0.1 nM, 0.05 nM or 0.02 nM, for example, about 0.1 nM to 1 nM, for example, about 0.2 nM to 0.8 nM, for example, about 0.13 nM to 0.11 nM, for example, about 0.124 nM, 0.369 nM, 0.431 nM, less than 0.735 nM D and binds to cynomolgus monkey PD-L1.

[0398] In another aspect, the aforementioned antibody molecule has a K, as measured by, for example, the Biacore method, of about 100 nM, 60 nM, 10 nM, 1 nM, 0.5 nM, 0.2 nM, 0.15 nM, 0.1 nM, 0.05 nM or 0.02 nM, for example, about 0.13 nM to 0.11 nM, for example, about 0.124 nM, 0.04 nM, 0.075 nM or less than 77.4 nM D and binds to mouse PD-L1.

[0399] In another aspect, the aforementioned antibody molecule has a K, as measured by, for example, the Biacore method, of about 15 nM, 10 nM, 5 nM, 1 nM, 0.5 nM, 0.2 nM, 0.15 nM, 0.1 nM, 0.05 nM or 0.02 nM, for example, about 0.1 nM to 3.5 nM, for example, about 0.13 nM to 0.11 nM, for example, about 0.124 nM, 0.04 nM, 0.075 nM, 0.431 nM, 1.36 nM, 6.14 nM or less than 77.4 nM Dand binds to rat PD-L1. In certain embodiments, the antibody molecules described above have a similar K, as measured, for example, by the Biacore method D and binds to both human PD-L1 and cynomolgus monkey PD-L1, for example, within the nM range.

[0400] In one embodiment, the antibody molecules described above have a K of about 0.5 nM, 0.4 nM, 0.3 nM, 0.2 nM, 0.1 nM, 0.075 nM, 0.05 nM, 0.025 nM or 0.01 nM, for example, less than about 0.285 nM, as measured, for example, by FACS analysis D and binds to 300.19 cells expressing human PD-L1 (e.g., 300.19 cells transfected with human PD-L1).

[0401] In one embodiment, the antibody molecules described above have a K of about 1 nM, 0.75 nM, 0.5 nM, 0.25 nM or 0.01 nM, for example, less than about 0.129 nM, as measured, for example, by FACS analysis D and binds to cells expressing cynomolgus monkey PD-L1 (e.g., cells transfected with cynomolgus monkey PD-L1).

[0402] In certain embodiments, the antibody molecules described above are not cross-reactive with mouse or rat PD-L1. In other embodiments, the antibodies described above are cross-reactive with rhesus monkey PD-L1. For example, cross-reactivity can be measured by the Biacore method or a binding assay using cells expressing PD-L1 (e.g., human PD-L1-expressing 300.19 cells). In other embodiments, the antibody molecules described above bind to the extracellular Ig-like domain of PD-L1.

[0403] In other aspects, the antibody molecules described above can reduce the binding of PD-1 or B7-1 to PD-L1 or cells expressing PD-L1. In certain aspects, the antibody molecules described above can reduce (e.g., block) PD-L1 binding to cells expressing PD-L1 (e.g., human PD-L1-expressing 300.19 cells) with an IC50 of about 1.5 nM, 1 nM, 0.8 nM, 0.6 nM, 0.4 nM, 0.2 nM or 0.1 nM, e.g., between about 0.2 nM and about 0.1 nM, e.g., about 0.15 nM or less, e.g., less than about 0.145 nM. In certain aspects, the antibody described above can reduce (e.g., block) B7-1 binding to cells expressing PD-L1 (e.g., human PD-L1-expressing 300.19 cells) with an IC50 of about 2 nM, 1.5 nM, 1 nM, 0.5 nM or 0.2 nM, e.g., between about 0.5 nM and about 0.01 nM or about 0.2 nM or less, e.g., less than about 0.1 nM.

[0404] In other aspects, the antibody molecules described above can enhance antigen-specific T cell responses.

[0405] In certain aspects, the antibody molecules described above increase the expression of IL-2 from cells activated by Staphylococcus enterotoxin B (SEB) (e.g., 25 μg / mL) by at least about 2, 3, 4, 5, 6, 7 or 8-fold, e.g., about 2-3 fold, e.g., about 2-2.6 fold, e.g., about 2.39 fold or e.g., about 2.4-6.4 fold, as compared to the expression of IL-2 when an isotype control (e.g., IgG4) is used, as measured in, e.g., an SEB T cell activation assay using peripheral blood mononuclear cells (PMBC) or a human whole blood ex vivo assay.

[0406] In certain aspects, the antibody molecules described above increase the expression of IFN-γ from T cells activated by SEB (e.g., 3 pg / mL) by at least about 2, 3, 4, 5, 6, 7 or 8-fold, e.g., about 0.5-4.5 fold, e.g., about 2.72 fold or e.g., about 4-7 fold, as compared to the expression of IFN-γ when an isotype control (e.g., IgG4) is used, as measured in, e.g., an IFN-γ activity assay.

[0407] In one aspect, the aforementioned antibody molecule binds to PD-L1 with a Kd slower than, for example, 5×10 -4 、1×10 -4 、5×10 -5 or 1×10 -5 s -1 、for example, about 6.33×10 -5 s -1 measured by, for example, the Biacore method. In one aspect, the aforementioned antibody molecule binds to PD-L1 with an Ka faster than, for example, 1×10 4 、5×10 4 、1×10 5 or 5×10 5 M -1 s -1 、for example, about 3.07×10 4 M -1 s -1 measured by, for example, the Biacore method.

[0408] In an aspect, the anti-PD-L1 antibody molecule is a monospecific antibody molecule or a bispecific antibody molecule. In an aspect, the anti-PD-L1 antibody molecule has a first binding specificity for PD-L1 and a second binding specificity for TIM-3, LAG-3, CEACAM (e.g., CEACAM-1, CEACAM-3 and / or CEACAM-5), PD-1 or PD-L2. In an aspect, the antibody molecule comprises an antigen-binding fragment of an antibody, e.g., a Fab or an antigen-binding fragment of a Fab.

[0409] In another aspect, the present invention provides an isolated nucleic acid molecule encoding any of the aforementioned antibody molecules, a vector thereof, and a host cell.

[0410] An isolated nucleic acid encoding any of the aforementioned antibody molecule's heavy chain variable region or light chain variable region, or both.

[0411] In one aspect, an isolated nucleic acid encoding heavy chain CDR1-3, comprising a nucleotide sequence of SEQ ID NO: 104-108, 113-117 or 205-208.

[0412] In another aspect, an isolated nucleic acid encoding light chain CDR1-3, the nucleic acid comprising the nucleotide sequences of SEQ ID NOs: 109-112, 118-123, 209-214 and 245-246.

[0413] In other aspects, the aforementioned nucleic acid is a nucleotide sequence encoding a heavy chain variable domain, further comprising a nucleotide sequence that is at least 85% identical to any one of SEQ ID NOs: 19, 31, 39, 47, 51, 55, 63, 71, 79, 90, 95, 100, 196 or 201.

[0414] In other aspects, the aforementioned nucleic acid is a nucleotide sequence encoding a heavy chain variable domain, further comprising a nucleotide sequence comprising any one of SEQ ID NOs: 19, 31, 39, 47, 51, 55, 63, 71, 79, 90, 95, 100, 196 or 201.

[0415] In other aspects, the aforementioned nucleic acid is a nucleotide sequence encoding a heavy chain, further comprising a nucleotide sequence that is at least 85% identical to any one of SEQ ID NOs: 21, 33, 41, 49, 53, 57, 65, 73, 81, 92, 97, 101, 198 or 202.

[0416] In other aspects, the aforementioned nucleic acid is a nucleotide sequence encoding a heavy chain, further comprising a nucleotide sequence comprising any one of SEQ ID NOs: 21, 33, 41, 49, 53, 57, 65, 73, 81, 92, 97, 101, 198 or 202.

[0417] In other aspects, the aforementioned nucleic acid is a nucleotide sequence encoding a light chain variable domain, further comprising a nucleotide sequence that is at least 85% identical to any one of SEQ ID NOs: 23, 27, 35, 43, 59, 67, 75, 83, 87, 93, 98, 102, 199 or 203.

[0418] In other aspects, the aforementioned nucleic acid is a nucleotide sequence encoding a light chain variable domain, and further includes a nucleotide sequence containing any one of SEQ ID NO: 23, 27, 35, 43, 59, 67, 75, 83, 87, 93, 98, 102, 199 or 203.

[0419] In other aspects, the aforementioned nucleic acid is a nucleotide sequence encoding a light chain, and further includes a nucleotide sequence that is at least 85% identical to any one of SEQ ID NO: 25, 29, 37, 45, 61, 69, 77, 85, 89, 94, 99, 103, 200 or 204.

[0420] In other aspects, the aforementioned nucleic acid is a nucleotide sequence encoding a light chain, and further includes a nucleotide sequence containing any one of SEQ ID NO: 25, 29, 37, 45, 61, 69, 77, 85, 89, 94, 99, 103, 200 or 204.

[0421] In certain aspects, one or more expression vectors and host cells containing the aforementioned nucleic acid are provided.

[0422] Also provided is a method for producing an antibody molecule or a fragment thereof, which includes culturing the host cells described herein under conditions suitable for gene expression.

[0423] Pharmaceutical Compositions and Kits In another aspect, the present invention provides a composition containing an antibody molecule described herein formulated together with a pharmaceutically acceptable carrier, for example, a pharmaceutically acceptable composition. As used herein, "pharmaceutically acceptable carrier" includes any solvent, dispersion medium, isotonic and absorption delaying agent, and the like that are physiologically compatible. The carrier can be suitable for intravenous, intramuscular, subcutaneous, parenteral, rectal, spinal or epithelial administration (e.g., by injection or infusion).

[0424] The compositions of the present invention can be in various forms. Such forms include, for example, liquid solutions (e.g., solutions for injection and infusion), dispersions or suspensions, liposomes, and suppositories, etc., liquid, semi-solid, and solid dosage forms. The preferred form depends on the intended route of administration and therapeutic application. A typical preferred composition is in the form of a solution for injection or infusion. The preferred route of administration is parenteral (e.g., intravenous, subcutaneous, intraperitoneal, intramuscular). In a preferred embodiment, the antibody is administered by intravenous infusion or injection. In another preferred embodiment, the antibody is administered by intramuscular or subcutaneous injection.

[0425] The terms "parenteral administration" and "administered parenterally" mean, in this specification, routes of administration other than enteral and topical administration, and usually include, without limitation, injection by intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, intratracheal, subcutaneous, subepidermal, intraarticular, subcapsular, subdural, intraspinal, epidural, and intrasternal injection and infusion.

[0426] The therapeutic composition should typically be sterile and stable under the conditions of manufacture and storage. The composition should be formulated as a solution, microemulsion, dispersion, liposome or other ordered structure suitable for high antibody concentrations. Sterile injectable solutions can be prepared by incorporating the required amount of the active compound (i.e., antibody or antibody portion) into a suitable solvent having one or a combination of the ingredients listed above, followed by sterile filtration as required. Generally, dispersions are prepared by incorporating the active compound into a sterile medium containing a base dispersion medium and the required other ingredients derived from the ingredients listed above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and freeze drying, which yield a powder of the active ingredient plus any additional desired ingredients from its previously sterile filtered solution. The proper fluidity of the solution can be maintained, for example, by the use of coatings such as lecithin, by the maintenance of the required particle size in the case of dispersions, and also by the use of surfactants. Prolonged absorption of the injectable composition can be brought about by including in the composition an agent that delays absorption, such as a monostearate salt and gelatin.

[0427] Antibody molecules can be administered by a variety of methods known in the art, but for many therapeutic applications, the preferred route of administration / mode is intravenous injection or infusion. For example, the antibody molecule can be administered by intravenous infusion at a rate greater than 20 mg / min, such as 20 - 40 mg / min, typically 40 mg / min or more, to reach a dose of about 35 - 440 mg / m 2 , typically about 70 - 310 mg / m 2 , more typically about 110 - 130 mg / m 2 . For example, the antibody molecule can be administered at a dose of about 1 - 100 mg / m 2 , typically about 5 - 50 mg / m 2 , about 7 - 25 mg / m 2 , more typically about 10 mg / m 2It can be administered by intravenous infusion at a rate of less than 10 mg / min; typically, at a rate of 5 mg / min or less, so as to reach the dosage. As will be appreciated by those skilled in the art, the route of administration and / or mechanism will vary depending on the desired result. In certain embodiments, the active compound can be prepared with a carrier that will protect the compound from rapid release, such as a controlled release formulation including implants, transdermal patches, and microencapsulated delivery systems. Biodegradable, biocompatible polymers such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid can be used. Many methods for the preparation of such formulations are patented or generally known to those skilled in the art. See, for example, Sustained and Controlled Release Drug Delivery Systems, J. R. Robinson, ed., Marcel Dekker, Inc., New York, 1978.

[0428] In certain embodiments, the antibody molecule can be administered orally, for example, with an inert diluent or an assimilable edible carrier. The compounds (and optionally other ingredients) can also be enclosed in hard or soft shell gelatin capsules, compressed into tablets, or directly incorporated into the subject's diet. For oral therapeutic administration, the compounds can be taken with excipients and used in ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafers, and other forms. To administer the compounds of the invention by a route other than parenteral, it may be necessary to coat the compounds with, or co-administer the compounds with, a material to prevent their inactivation. The therapeutic composition can also be administered by medical devices known in the art.

[0429] The dosage regimen is adjusted to provide an optimal desired response (e.g., a therapeutic response). For example, a single bolus can be administered, several divided doses can be administered over time, or the dosage can be proportionally decreased or increased as indicated by the exigencies of the treatment situation. For ease of administration and for dosage uniformity, it is particularly advantageous to formulate the parenteral composition in dosage unit form. A dosage unit form as used herein refers to physically discrete units suitable as unit dosages for the subject to be treated; each unit contains a predetermined quantity of the active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. The specification for the dosage unit forms of the invention is dictated by and directly dependent on (a) the unique characteristics of the active compound and the particular therapeutic effect to be achieved, as well as (b) the limitations inherent in the art of compounding such active compounds for the treatment of sensitivity in individuals.

[0430] Exemplary non-limiting ranges for a therapeutically or prophylactically effective amount of the antibody molecule are 0.1 to 30 mg / kg, more preferably 1 to 25 mg / kg. The dosage and treatment regimen of the anti-PD-L1 antibody molecule can be determined by one of ordinary skill in the art. In certain embodiments, the anti-PD-L1 antibody molecule is administered by injection (e.g., subcutaneously or intravenously) at a dosage of about 1 to 40 mg / kg, such as 1 to 30 mg / kg, such as about 5 to 25 mg / kg, about 10 to 20 mg / kg, about 1 to 5 mg / kg, 1 to 10 mg / kg, 5 to 15 mg / kg, 10 to 20 mg / kg, 15 to 25 mg / kg or about 3 mg / kg. The dosing schedule can vary, for example, from once a week to once every 2, 3 or 4 weeks. In one embodiment, the anti-PD-L1 antibody molecule is administered at a dosage of about 10 to 20 mg / kg once a week. The antibody molecule is about 35 to 440 mg / m 2 , typically about 70 to 310 mg / m 2 , more typically about 110 to 130 mg / m 2It can be administered by intravenous infusion at a rate exceeding 20 mg / min, for example, 20 - 40 mg / min, typically 40 mg / min or more, so as to reach the dosage of 2 The infusion rate achieves a level of about 3 mg / kg. In other embodiments, the antibody molecule is about 1 - 100 mg / m 2 , for example, about 5 - 50 mg / m 2 , about 7 - 25 mg / m 2 or about 10 mg / m 2 It can be administered by intravenous infusion at a rate less than 10 mg / min, for example, 5 mg / min or less, so as to reach the dosage of 2 , preferably about 5 - 50 mg / m 2 , about 7 - 25 mg / m 2 , more preferably about 10 mg / m 2 It can be administered by intravenous infusion at a rate less than 10 mg / min, preferably 5 mg / min or less, so as to reach the dosage of. Note that the dosage value can vary depending on the type and severity of the condition to be alleviated. For any particular subject, a specific dosage regimen should be adjusted over time according to individual needs and the expert judgment of the person administering or supervising the administration of the composition. Further understand that the dosage ranges set forth herein are merely illustrative and are not intended to limit the scope or practice of the claimed composition.

[0431] The pharmaceutical composition of the present invention can contain a "therapeutically effective amount" or "prophylactically effective amount" of the antibody or antibody portion of the present invention. A "therapeutically effective amount" refers to an amount effective at a dosage and for a period of time necessary to achieve the desired therapeutic result. The therapeutically effective amount of the modified antibody or antibody fragment can vary according to factors such as the medical condition, age, gender and weight of the individual, and the ability of the antibody or antibody portion to induce the desired response in the individual. The therapeutically effective amount is also an amount where the therapeutically beneficial effects outweigh any toxic or detrimental effects of the modified antibody or antibody fragment. A "therapeutically effective dosage" preferably inhibits a measurable parameter, such as tumor growth rate, by at least about 20%, more preferably at least about 40%, even more preferably at least about 60%, and still more preferably at least about 80% compared to an untreated subject. The ability of a measurable parameter, such as a compound to inhibit cancer, can be evaluated in an animal model system that predicts efficacy in human tumors. Alternatively, this property of the composition can be evaluated by testing the ability of the compound to inhibit, and such inhibition in vitro can be evaluated by assays known to those skilled in the art.

[0432] A "prophylactically effective amount" refers to an amount effective at a dosage and for a period of time necessary to achieve the desired prophylactic result. Typically, the prophylactic dosage is used in a subject prior to or at an earlier stage of the disease, so the prophylactically effective amount will be less than the therapeutically effective amount.

[0433] Kits containing the antibody molecules described herein are also within the scope of the present invention. The kit can contain one or more other elements including: instructions for use; other reagents such as labels, therapeutic agents, or agents useful for labeling or chelating or otherwise coupling the antibody to a therapeutic agent, or a radiation protection composition; an apparatus or other material for preparing the antibody for administration; a pharmaceutically acceptable carrier; and an apparatus or other material for administration to a subject.

[0434] Use of anti-PD-L1 antibody molecules The anti-PD-L1 antibody molecules disclosed herein have in vitro and in vivo diagnostic, as well as therapeutic and prophylactic utility. For example, such molecules can be administered to cells in culture, in vitro or ex vivo, or to a subject, e.g., a human subject, to treat, prevent and / or diagnose various disorders such as cancer and infectious diseases.

[0435] Thus, in one aspect, the present invention provides a method of modifying an immune response in a subject, comprising administering to the subject an antibody molecule described herein such that the immune response in the subject is modified. In one embodiment, the immune response is enhanced, stimulated or upregulated. In one embodiment, the antibody molecule enhances the immune response in the subject by blocking PD-L1.

[0436] As used herein, the term "subject" is intended to include humans and non-human animals. In one embodiment, the subject is a human subject, e.g., a human patient having a disorder or condition characterized by abnormal PD-L1 function. The term "non-human animal" includes mammals and non-mammals such as non-human primates. In one embodiment, the subject is a human. In one embodiment, the subject is a human patient in need of enhanced immune response. In one embodiment, the subject is immunocompromised, e.g., the subject has received or is receiving chemotherapy or radiation therapy. Alternatively or in combination therewith, the subject is immunocompromised or at risk thereof as a result of an infection. The methods and compositions described herein are suitable for treating human patients having a disorder that can be treated by increasing a T cell-mediated immune response. For example, the methods and compositions described herein can enhance a number of immune activities. In one embodiment, the subject has an increased number or activity of tumor infiltrating T lymphocytes (TIL). In another embodiment, the subject has increased expression or activity of interferon-gamma (IFN-γ). In yet another embodiment, the subject has decreased PD-L1 expression or activity.

[0437] Therapeutic Use Blocking of PD-L1 by an antibody can enhance the immune response against cancer cells in a patient. PD-L1 is typically not expressed in normal human cells, but is abundant in various human cancers [Dong et al. (2002) Nat Med 8:787-9]. The interaction between PD-1 and PD-L1 results in a decrease in tumor-infiltrating lymphocytes, a decrease in T cell receptor-mediated proliferation, and immune evasion by cancer cells [Dong et al. (2003) J Mol Med 81:281-7; Blank et al. (2005) Cancer Immunol Immunother. 54:307-14); Konishi et al. (2004) Clin. Cancer Res. 10:5094-100]. Immune suppression can be reversed by inhibiting the local interaction of PD-L1 with PD-1, and the effect is additive when the interaction of PD-L2 with PD-1 is also blocked [Iwai et al. (2002) PNAS 99:12293-7; Brown et al. (2003) J. Immunol. 170:1257-66]. An anti-PD-L1 antibody can be used alone to inhibit the growth of cancerous tumors. Alternatively, an anti-PD-L1 antibody can be used in combination with other immunogenic agents, standard cancer treatments or other antibodies, as described herein. Thus, inhibition of PD-L1 can increase the immune response.

[0438] In one aspect, the invention relates to the in vivo treatment of a subject using an anti-PD-L1 antibody molecule such that the growth of a cancerous tumor is inhibited or reduced. An anti-PD-L1 antibody can be used alone to inhibit the growth of cancerous tumors. Alternatively, the anti-PD-L1 antibody can be used in combination with one or more of the following: standard therapeutic treatments (e.g., for cancer or infectious disorders), another antibody or antigen-binding fragment thereof, an immunomodulatory agent (e.g., an activator of a costimulatory molecule or an inhibitor of an inhibitory molecule); a vaccine, e.g., a therapeutic cancer vaccine; or other forms of cellular immunotherapy described hereinafter.

[0439] Thus, in one aspect, the present invention provides a method of inhibiting the growth of tumor cells in a subject, comprising administering to the subject a therapeutically effective amount of an anti-PD-L1 antibody molecule described herein.

[0440] In one aspect, the method is suitable for the treatment of cancer in vivo. To achieve antigen-specific enhancement of immunity, the anti-PD-L1 antibody molecule can be administered together with the antigen of interest. When an antibody against PD-L1 is administered in combination with one or more agents, the combination can be administered in either order or simultaneously.

[0441] Cancer type; seranostic method In another aspect, provided is a method of treating a subject, e.g., reducing or alleviating a proliferative condition or disorder (e.g., cancer) in a subject, e.g., a solid tumor, soft tissue tumor or metastatic lesion. The method comprises administering to the subject one or more of the anti-PD-L1 antibody molecules described herein, either alone or in combination with other agents or treatment modalities.

[0442] As used herein, the term "cancer" means any type of cancerous growth or carcinogenic process, metastatic tissue or malignantly transformed cells, tissues or organs, regardless of histological type or invasive stage. Examples of cancerous disorders include, but are not limited to, solid tumors, hematological cancers, soft tissue tumors and metastatic lesions.

[0443] Examples of solid tumors include malignant lesions of various organ systems, such as those affecting the liver, lung, breast, lymphatic system, gastrointestinal tract (e.g., colon), urogenital system (e.g., kidney, urothelial cells), prostate, and pharynx, such as sarcomas and carcinomas (including adenocarcinomas and squamous cell carcinomas). Adenocarcinomas include malignancies such as most colon cancers, rectal cancers, renal cell carcinomas, liver cancers, non-small cell lung cancers, small intestine cancers, and esophageal cancers. Squamous cell carcinomas include malignancies such as those in the lung, esophagus, skin, head and neck region, oral cavity, anus, and cervix. In one aspect, the cancer is melanoma, e.g., melanoma in a progressive stage. The methods and compositions of the present invention can also be used to treat or prevent metastatic lesions of the cancers described above.

[0444] Exemplary cancers whose growth can be inhibited using the antibody molecules disclosed herein typically include cancers responsive to immunotherapy. Non-limiting examples of cancers preferred for treatment include lymphoma (e.g., diffuse large B-cell lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma), breast cancer [e.g., metastatic breast cancer], lung cancer [e.g., non-small cell lung cancer (NSCLC), e.g., stage IV or recurrent non-small cell lung cancer, NSCLC adenocarcinoma or NSCLC squamous cell carcinoma], myeloma (e.g., multiple myeloma), leukemia (e.g., chronic myelogenous leukemia), skin cancer [e.g., melanoma (e.g., stage III or IV melanoma) or Merkel cell carcinoma], head and neck cancer [e.g., head and neck squamous cell carcinoma (HNSCC)], myelodysplastic syndrome, bladder cancer (e.g., transitional cell carcinoma), kidney cancer (e.g., renal cell carcinoma, e.g., clear cell renal carcinoma, e.g., advanced or metastatic clear cell renal carcinoma), and colon cancer. Moreover, the antibody molecules described herein can be used to treat refractory or recurrent malignant lesions.

[0445] Examples of other cancers that can be treated include bone cancer, pancreatic cancer, skin cancer, head or neck cancer, cutaneous or intraocular malignant melanoma, uterine cancer, ovarian cancer, rectal cancer, anal cancer, gastroesophageal, stomach cancer, testicular cancer, uterine cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, Merkel cell carcinoma, Hodgkin lymphoma, non-Hodgkin lymphoma, esophageal cancer, small intestine cancer, endocrine system cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, chronic or acute leukemia including chronic lymphocytic leukemia, pediatric solid tumors, lymphocytic lymphoma, bladder cancer, multiple myeloma, myelodysplastic syndrome, kidney or ureter cancer, renal pelvis cancer, central nervous system (CNS) neoplasms, primary CNS lymphoma, tumor angiogenesis, spinal cord axis tumors, brainstem glioma, pituitary adenoma, Kaposi sarcoma, epidermoid cancer, squamous cell carcinoma, T cell lymphoma, environmentally induced cancers including cancers induced by asbestos (e.g., mesothelioma), and combinations of the foregoing cancers.

[0446] Treatment of metastatic cancers, e.g., metastatic cancers that express PD-L1 [Iwai et al. (2005) Int. Immunol. 17:133-144], can be effected using the antibody molecules described herein. In certain embodiments, the cancer expresses elevated levels of PD-L1, IFNγ and / or CD8.

[0447] PD-L1 signaling can contribute to increased Bim expression in CD8+ T cells. Treatment of cancers in patients that express high levels of Bim (e.g., elevated Bim levels in PD-1+ CD8+ T cells compared to PD-1-CD8+ T cells), e.g., advanced melanoma, can be effected using the antibody molecules described herein.

[0448] Animal models that can be used to test the efficacy of anti-PD-L1 antibodies in the monotherapy or combination therapy of cancer include, for example, the CT26 colon cancer model [Sakuishi et al. (2010) J Exp Med. 207(10): 2187-2194] and the 5T33 myeloma model [Manning et al. (1992) Br J Cancer. 66(6): 1088-1093].

[0449] In certain embodiments, the cancer expresses elevated levels of PD-L1, IFNγ, and / or CD8.

[0450] Without wishing to be bound by theory, in certain embodiments, when a patient has cancer that highly expresses PD-L1 and / or the cancer is infiltrated by anti-tumor immune cells, such as TILs, the patient is more likely to respond to treatment with an immunomodulatory agent (which may be combined with one or more of the agents described herein). Anti-tumor immune cells can be positive for CD8, PD-L1, and / or IFN-γ; thus, the levels of CD8, PD-L1, and / or IFN-γ can function as readout information for the level of TILs in the microenvironment. In certain embodiments, the cancer microenvironment is referred to as triple positive for PD-L1 / CD8 / IFN-γ.

[0451] Accordingly, in certain aspects, the present application provides a method of administering to a patient a therapeutically effective amount of an anti-PD-L1 antibody molecule that determines whether a tumor sample is positive for one or more of PD-L1, CD8, and IFN-γ, and, if the tumor sample is positive for one or more of the markers, e.g., two or all three, may be combined with one or more other immunomodulatory agents or anti-cancer agents.

[0452] In the indications listed below, a large proportion of patients are triple positive for PD-L1 / CD8 / IFN-γ: lung cancer (squamous cell); lung cancer (adenocarcinoma); head and neck cancer; gastric cancer; NSCLC; HNSCC; gastric body cancer (e.g., MSIhi and / or EBV+); CRC (e.g., MSIhi); nasopharyngeal cancer (NPC); cervical cancer (e.g., squamous cell); thyroid cancer, e.g., papillary thyroid; melanoma; TN breast cancer; and DLBCL (diffuse large B-cell lymphoma). In breast cancer generally, and also in colorectal cancer generally, a moderate proportion of patients are triple positive for PD-L1 / CD8 / IFN-γ. In the indications listed below, a small proportion of patients are triple positive for PD-L1 / CD8 / IFN-γ: ER+ breast cancer and pancreatic cancer. These findings are further described in Example 4. Regardless of whether a large or small proportion of patients are triple positive for these markers, screening of patients for these markers enables the identification of a proportion of patients with a particularly high likelihood of responding favorably to treatment with a PD-L1 antibody (e.g., a blocking PD-L1 antibody), which may be combined with one or more other immunomodulatory agents (e.g., an anti-TIM-3 antibody molecule, an anti-LAG-3 antibody molecule or an anti-PD-L1 antibody molecule) and / or an anticancer agent, e.g., an anticancer agent listed in Table 6 and disclosed in the publications listed in Table 6.

[0453] In one aspect, cancer samples are classified as triple positive for PD-L1 / CD8 / IFN-γ. This measurement can be roughly broken down into two thresholds: whether individual cells are classified as positive, and whether the sample as a whole is classified as positive. First, the levels of PD-L1, CD8, and / or IFN-γ can be measured within individual cells. In one aspect, cells that are positive for one or more of these markers have higher levels of the markers compared to control cells or reference values. For example, in one aspect, a high level of PD-L1 in a given cell is higher than the level of PD-L1 in the corresponding non-cancerous tissue of the patient. As another example, in one aspect, a high level of CD8 or IFN-γ in a given cell is the level of the protein typically observed in TIL. Second, the percentage of cells in a sample that are positive for PD-L1, CD8, and / or IFN-γ can also be measured. (Individual cells do not need to express all three markers.) In one aspect, triple positive samples have a high percentage of cells that are positive for these markers, e.g., a percentage of cells higher than a reference value or higher than a control sample.

[0454] In other aspects, the overall levels of PD-L1, CD8, and / or IFN-γ in a sample can be measured. In this case, a high level of CD8 or IFN-γ in the sample can be the level of the protein typically observed in tumors infiltrated by TIL. Similarly, a high level of PD-L1 can be the level of the protein typically observed in tumor samples, e.g., in the tumor microenvironment.

[0455] As shown in Example 4 in this specification, the identification of a subset of patients who are triple-positive for PD-L1 / CD8 / IFN-γ reveals a specific subpopulation of patients who may be particularly responsive to PD-L1 antibody therapy. For example, many IM-TN (immunomodulatory, triple-negative) breast cancer patients are triple-positive for PD-L1 / CD8 / IFN-γ. IM-TN breast cancer is described, for example, in Brian D. Lehmann et al., “Identification of human triple-negative breast cancer subtypes and preclinical models for selection of targeted therapies”, J Clin Invest. Jul 1, 2011; 121(7): 2750-2767. Triple-negative breast cancer is breast cancer that does not express estrogen receptor (ER), progesterone receptor (PR), and Her2 / neu. Such cancers are typically difficult to treat because they are not responsive to agents that target ER, PR, and Her2 / neu. Triple-negative breast cancer can be further subdivided into different classes, one of which is immunomodulatory. As described in Lehmann et al., IM-TN breast cancer is enriched for factors involved in one or more of the genes involved in immune cell processes such as immune cell signaling (e.g., TH1 / TH2 pathway, NK cell pathway, B cell receptor signaling pathway, DC pathway, and T cell receptor signaling), cytokine signaling (e.g., cytokine pathway, IL-12 pathway, and IL-7 pathway), antigen processing and presentation, signaling via core immune signaling pathways (e.g., NFKB, TNF, and JAK / STAT signaling), T cell function, immune transcription, interferon (IFN) response, and antigen processing.Thus, in one aspect, the cancer to be treated is positive for, or determined to be so for, one or more markers of IM-TN breast cancer, such as immune cell signaling (e.g., TH1 / TH2 pathway, NK cell pathway, B cell receptor signaling pathway, DC pathway and T cell receptor signaling), cytokine signaling (e.g., cytokine pathway, IL-12 pathway and IL-7 pathway), antigen processing and presentation, signaling via core immune signaling pathways (e.g., NFKB, TNF and JAK / STAT signaling), T cell function, immune transcription, interferon (IFN) response and one or more factors that promote genes involved in antigen processing.

[0456] As another example, herein, a subset of colon cancer patients with high microsatellite instability (MSI) is also shown to be triple positive for PD-L1 / CD8 / IFN-γ. Thus, in one aspect, a PD-L1 antibody, e.g., a PD-L1 antibody described herein (which may be combined with one or more immunomodulatory agents and one or more anti-cancer agents, such as the anti-cancer agents described in Table 6 or the publications of Table 6, e.g., LAG-3 antibody, TIM-3 antibody or PD-1 antibody, etc.) is administered to a patient having or identified as having colon cancer with high MSI, thereby treating the cancer. In one aspect, cells with high MSI are cells having MSI at a higher level than a reference value or control cells, e.g., non-cancerous cells of the same histotype as the cancer.

[0457] As another example, in the present specification, a subset of gastric cancer patients having high MSI and / or being EBV+ is also shown to be triple positive for PD-L1 / CD8 / IFN-γ. Thus, in one aspect, a PD-L1 antibody, for example, a PD-L1 antibody described herein (one or more immunomodulatory agents such as LAG-3 antibody, TIM-3 antibody or PD-1 antibody, and one or more anti-cancer agents, for example, in combination with the anti-cancer agents described in Table 6 or the publications of Table 6) may be administered to patients having or identified as having high MSI and / or EBV+ gastric cancer, thereby treating the cancer. In one aspect, cells having high MSI are cells having a higher level of MSI than a reference value or control cells, for example, non-cancerous cells of the same histological type as the cancer.

[0458] Furthermore, methods of assaying cancer for PD-L1 and subsequently treating the cancer with a PD-L1 antibody are disclosed herein. As described in Example 5 herein, cancer samples can be assayed for PD-L1 protein level or mRNA level. Samples having a level of PD-L1 (protein or mRNA) higher than a reference value or control cells (e.g., non-cancerous cells) can be classified as PD-L1 positive. Thus, in one aspect, a PD-L1 antibody, for example, a PD-L1 antibody described herein (optionally in combination with one or more anti-cancer agents) is administered to patients having or identified as having PD-L1 positive cancer. The cancer can be, for example, non-small cell lung (NSCLC) adenocarcinoma (ACA), NSCLC squamous cell carcinoma (SCC) or hepatocellular carcinoma (HCC).

[0459] In one aspect, the methods herein involve the use of a PD-L1 antibody, such as a monotherapy of a PD-L1 antibody described herein, for treating cancer that is PD-L1 positive (or identified as such). In one aspect, the cancer is colorectal cancer (e.g., high MSI), gastric cancer (e.g., high MSI and / or EBV+), NPC, cervical cancer, breast cancer (e.g., TN breast cancer) and ovarian cancer. In one aspect, the cancer is NSCLC, melanoma or HNSCC. In one aspect, the PD-L1 antibody is administered, for example, at a dose of 1, 3, 10 or 20 mg / kg.

[0460] For example, based on Example 4 herein, it was found that a certain gastric cancer that is triple positive for PD-L1 / CD8 / IFN-γ is also positive for PIK3CA. Thus, in one aspect, the cancer can be treated with an anti-PD-1 antibody molecule (which may be combined with one or more immunomodulatory agents, such as an anti-LAG-3 antibody molecule, an anti-TIM-3 antibody molecule or an anti-PD-1 antibody molecule) and an agent that inhibits PIK3CA. Exemplary agents in this category are described in Stein RC (September 2001). "Prospects for phosphoinositide 3-kinase inhibition as a cancer treatment". Endocrine-related Cancer 8 (3): 237-48 and Marone R, Cmiljanovic V, Giese B, Wymann MP (January 2008...

Claims

1. Use of an isolated antibody molecule capable of binding to human programmed death-ligand 1 (PD-L1) in combination with an AKT inhibitor in the manufacture of a medicament for treating cancer in a subject, wherein the antibody comprises (i) a heavy chain variable region (VH) comprising the VHCDR1 amino acid sequence of SEQ ID NO: 1, the VHCDR2 amino acid sequence of SEQ ID NO: 2, and the VHCDR3 amino acid sequence of SEQ ID NO: 3, and a light chain variable region (VL) comprising the VLCDR1 amino acid sequence of SEQ ID NO: 9, the VLCDR2 amino acid sequence of SEQ ID NO: 10, and the VLCDR3 amino acid sequence of SEQ ID NO: 11; (ii) a VH comprising the VHCDR1 amino acid sequence of SEQ ID NO: 4, the VHCDR2 amino acid sequence of SEQ ID NO: 5, and the VHCDR3 amino acid sequence of SEQ ID NO: 3; and a VL comprising the VLCDR1 amino acid sequence of SEQ ID NO: 12, the VLCDR2 amino acid sequence of SEQ ID NO: 13, and the VLCDR3 amino acid sequence of SEQ ID NO: 14; (iii) a VH comprising the VHCDR1 amino acid sequence of SEQ ID NO: 195, the VHCDR2 amino acid sequence of SEQ ID NO: 2, and the VHCDR3 amino acid sequence of SEQ ID NO: 3; and a VL comprising the VLCDR1 amino acid sequence of SEQ ID NO: 9, the VLCDR2 amino acid sequence of SEQ ID NO: 10, and the VLCDR3 amino acid sequence of SEQ ID NO: 11; or (iv) a VH comprising the VHCDR1 amino acid sequence of SEQ ID NO: 195, the VHCDR2 amino acid sequence of SEQ ID NO: 5, and the VHCDR3 amino acid sequence of SEQ ID NO: 3; and a VL comprising the VLCDR1 amino acid sequence of SEQ ID NO: 12, the VLCDR2 amino acid sequence of SEQ ID NO: 13, and the VLCDR3 amino acid sequence of SEQ ID NO: 14 comprising the use.

2. The heavy chain variable region comprising at least one, two, three, or four framework (FW) regions, wherein the antibody molecule comprises an amino acid sequence of any one of SEQ ID NOs: 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150, 152, or 154, or an amino acid sequence that is at least 90% identical thereto, or an amino acid sequence having two or fewer amino acid substitutions, insertions, or deletions as compared to an amino acid sequence of any one of SEQ ID NOs: 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150, 152, or 154; and / or The light chain variable region comprising at least one, two, three, or four framework regions, wherein the antibody molecule comprises an amino acid sequence of any one of SEQ ID NOs: 156, 158, 160, 162, 164, 166, 168, 170, 172, 174, 176, 178, 180, 182, 184, or 186, or an amino acid sequence that is at least 90% identical thereto, or an amino acid sequence having two or fewer amino acid substitutions, insertions, or deletions as compared to an amino acid sequence of any one of SEQ ID NOs: 156, 158, 160, 162, 164, 166, 168, 170, 172, 174, 176, 178, 180, 182, 184, or 186 The use according to claim 1, having the above.

3. The VH comprising an amino acid sequence of SEQ ID NOs: 18, 30, 38, 46, 50, 54, 62, 70, or 78, or an amino acid sequence that is at least 85% identical to any one of SEQ ID NOs: 18, 30, 38, 46, 50, 54, 62, 70, or 78; and / or The VL comprising an amino acid sequence of SEQ ID NOs: 22, 26, 34, 42, 58, 66, 74, 82, or 86, or an amino acid sequence that is at least 85% identical to any one of SEQ ID NOs: 22, 26, 34, 42, 58, 66, 74, 82, or 86 The use according to claim 1 or 2, comprising the above.

4. The antibody molecule comprises a VH comprising an amino acid sequence selected from SEQ ID NO: 18, SEQ ID NO: 30, SEQ ID NO: 38, SEQ ID NO: 46, SEQ ID NO: 50, SEQ ID NO: 54, SEQ ID NO: 62, SEQ ID NO: 70, or SEQ ID NO: 78; and / or a VL comprising an amino acid sequence selected from SEQ ID NO: 22, SEQ ID NO: 26, SEQ ID NO: 34, SEQ ID NO: 42, SEQ ID NO: 58, SEQ ID NO: 66, SEQ ID NO: 74, SEQ ID NO: 82, or SEQ ID NO: 86 The use according to any one of claims 1 - 3, comprising the same.

5. The antibody molecule comprises a heavy chain comprising an amino acid sequence selected from SEQ ID NO: 20, SEQ ID NO: 32, SEQ ID NO: 40, SEQ ID NO: 48, SEQ ID NO: 52, SEQ ID NO: 56, SEQ ID NO: 64, SEQ ID NO: 72, SEQ ID NO: 80, SEQ ID NO: 91, SEQ ID NO: 96, or SEQ ID NO: 197; and / or a light chain comprising an amino acid sequence selected from SEQ ID NO: 24, SEQ ID NO: 28, SEQ ID NO: 36, SEQ ID NO: 44, SEQ ID NO: 60, SEQ ID NO: 68, SEQ ID NO: 76, SEQ ID NO: 84, or SEQ ID NO: 88, The use according to any one of claims 1 - 4, comprising the same.

6. The antibody molecule is (a) a VH comprising the amino acid sequence of SEQ ID NO: 18 and a VL comprising the amino acid sequence of SEQ ID NO: 22; (b) a VH comprising the amino acid sequence of SEQ ID NO: 18 and a VL comprising the amino acid sequence of SEQ ID NO: 26; (c) a VH comprising the amino acid sequence of SEQ ID NO: 18 and a VL comprising the amino acid sequence of SEQ ID NO: 86; (d) a VH comprising the amino acid sequence of SEQ ID NO: 30 and a VL comprising the amino acid sequence of SEQ ID NO: 34; (e) a VH comprising the amino acid sequence of SEQ ID NO: 30 and a VL comprising the amino acid sequence of SEQ ID NO: 66; (f) a VH comprising the amino acid sequence of SEQ ID NO: 38 and a VL comprising the amino acid sequence of SEQ ID NO: 42; (g) a VH comprising the amino acid sequence of SEQ ID NO: 38 and a VL comprising the amino acid sequence of SEQ ID NO: 74; (h) a VH comprising the amino acid sequence of SEQ ID NO: 46 and a VL comprising the amino acid sequence of SEQ ID NO: 42; (i) VH comprising the amino acid sequence of SEQ ID NO: 50 and VL comprising the amino acid sequence of SEQ ID NO: 42; (j) VH comprising the amino acid sequence of SEQ ID NO: 50 and VL comprising the amino acid sequence of SEQ ID NO: 22; (k) VH comprising the amino acid sequence of SEQ ID NO: 50 and VL comprising the amino acid sequence of SEQ ID NO: 86; (l) VH comprising the amino acid sequence of SEQ ID NO: 54 and VL comprising the amino acid sequence of SEQ ID NO: 58; (m) VH comprising the amino acid sequence of SEQ ID NO: 54 and VL comprising the amino acid sequence of SEQ ID NO: 86; (n) VH comprising the amino acid sequence of SEQ ID NO: 62 and VL comprising the amino acid sequence of SEQ ID NO: 66; (o) VH comprising the amino acid sequence of SEQ ID NO: 62 and VL comprising the amino acid sequence of SEQ ID NO: 86; (p) VH comprising the amino acid sequence of SEQ ID NO: 70 and VL comprising the amino acid sequence of SEQ ID NO: 66; or (q) VH comprising the amino acid sequence of SEQ ID NO: 78 and VL comprising the amino acid sequence of SEQ ID NO: 82 The use according to any one of claims 1 - 3, comprising.

7. The antibody molecule is (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 20 and a light chain comprising the amino acid sequence of SEQ ID NO: 24; (b) a heavy chain comprising the amino acid sequence of SEQ ID NO: 20 and a light chain comprising the amino acid sequence of SEQ ID NO: 28; (c) a heavy chain comprising the amino acid sequence of SEQ ID NO: 20 and a light chain comprising the amino acid sequence of SEQ ID NO: 88; (d) a heavy chain comprising the amino acid sequence of SEQ ID NO: 32 and a light chain comprising the amino acid sequence of SEQ ID NO: 36; (e) a heavy chain comprising the amino acid sequence of SEQ ID NO: 32 and a light chain comprising the amino acid sequence of SEQ ID NO: 68; (f) a heavy chain comprising the amino acid sequence of SEQ ID NO: 40 and a light chain comprising the amino acid sequence of SEQ ID NO: 44; (g) a heavy chain comprising the amino acid sequence of SEQ ID NO: 40 and a light chain comprising the amino acid sequence of SEQ ID NO: 76; (h) A heavy chain comprising the amino acid sequence of SEQ ID NO: 48 and a light chain comprising the amino acid sequence of SEQ ID NO: 44; (i) A heavy chain comprising the amino acid sequence of SEQ ID NO: 52 and a light chain comprising the amino acid sequence of SEQ ID NO: 44; (j) A heavy chain comprising the amino acid sequence of SEQ ID NO: 52 and a light chain comprising the amino acid sequence of SEQ ID NO: 24; (k) A heavy chain comprising the amino acid sequence of SEQ ID NO: 52 and a light chain comprising the amino acid sequence of SEQ ID NO: 88; (l) A heavy chain comprising the amino acid sequence of SEQ ID NO: 56 and a light chain comprising the amino acid sequence of SEQ ID NO: 60; (m) A heavy chain comprising the amino acid sequence of SEQ ID NO: 56 and a light chain comprising the amino acid sequence of SEQ ID NO: 88; (n) A heavy chain comprising the amino acid sequence of SEQ ID NO: 64 and a light chain comprising the amino acid sequence of SEQ ID NO: 68; (o) A heavy chain comprising the amino acid sequence of SEQ ID NO: 64 and a light chain comprising the amino acid sequence of SEQ ID NO: 88; (p) A heavy chain comprising the amino acid sequence of SEQ ID NO: 72 and a light chain comprising the amino acid sequence of SEQ ID NO: 68; (q) A heavy chain comprising the amino acid sequence of SEQ ID NO: 80 and a light chain comprising the amino acid sequence of SEQ ID NO: 84; (r) A heavy chain comprising the amino acid sequence of SEQ ID NO: 197 and a light chain comprising the amino acid sequence of SEQ ID NO: 36; (s) A heavy chain comprising the amino acid sequence of SEQ ID NO: 91 and a light chain comprising the amino acid sequence of SEQ ID NO: 44; (t) A heavy chain comprising the amino acid sequence of SEQ ID NO: 96 and a light chain comprising the amino acid sequence of SEQ ID NO: 68; (u) A heavy chain comprising the amino acid sequence of SEQ ID NO: 247 and a light chain comprising the amino acid sequence of SEQ ID NO: 84; or (v) A heavy chain comprising the amino acid sequence of SEQ ID NO: 260 and a light chain comprising the amino acid sequence of SEQ ID NO: 44 The use according to any one of claims 1 - 3, comprising

8. The antibody molecule is a Fab, F(ab')2, Fv or single-chain Fv fragment (scFv); or The heavy chain constant region of IgG1, IgG2, IgG3, or IgG4, and / or comprising the kappa or lambda light chain constant region , the use according to any one of claims 1 - 7.

9. wherein the antibody molecule comprises (a) a human IgG4 heavy chain constant region having a mutation at position 228 of the human IgG4 heavy chain constant region according to EU numbering, and a kappa light chain constant region; (b) a human IgG4 heavy chain constant region having a serine - to - proline mutation at position 228 of the human IgG4 heavy chain constant region according to EU numbering, and a kappa light chain constant region; (c) a human IgG1 heavy chain constant region having an asparagine - to - alanine mutation at position 297 of the human IgG1 heavy chain constant region according to EU numbering, and a kappa light chain constant region; (d) a human IgG1 heavy chain constant region having an aspartic acid - to - alanine mutation at position 265 of the human IgG1 heavy chain constant region according to EU numbering and a proline - to - alanine mutation at position 329 of the human IgG1 heavy chain constant region according to EU numbering, and a kappa light chain constant region; or (e) a human IgG1 heavy chain constant region having a leucine - to - alanine mutation at position 234 and a leucine - to - alanine mutation at position 235 of the human IgG1 heavy chain constant region according to EU numbering, and a kappa light chain constant region , the use according to claim 8.

10. wherein the antibody molecule can bind to human PD - L1 with (a) a dissociation constant (K Ｄ ) of less than about 0.2 nM; (b) binds to the extracellular domain of PD - L1; (c) can reduce the binding of PD - 1 to PD - L1 and / or both; and / or (d) can enhance the antigen - specific T cell response , the use according to any one of claims 1 - 9.

11. Use according to any one of claims 1-10, wherein the antibody molecule is a humanized antibody molecule and / or a monospecific or bispecific antibody molecule.

12. The antibody molecule has a first binding specificity for PD-L1 and a second binding specificity for TIM-3, LAG-3, CEACAM (e.g., CEACAM-1 and / or CEACAM-5), PD-1 or PD-L2; and / or the antibody molecule comprises an antigen-binding fragment of an antibody, a half antibody, or an antigen-binding fragment of a half antibody. Use according to claim 11.

13. Use of an isolated antibody molecule capable of binding to human programmed death-ligand 1 (PD-L1) in combination with an AKT inhibitor and a pharmaceutical composition comprising a pharmaceutically acceptable carrier, excipient or stabilizer in the manufacture of a medicament for treating cancer in a subject, wherein the antibody (i) comprises a heavy chain variable region (VH) comprising the VHCDR1 amino acid sequence of SEQ ID NO: 1, the VHCDR2 amino acid sequence of SEQ ID NO: 2, and the VHCDR3 amino acid sequence of SEQ ID NO: 3, and a light chain variable region (VL) comprising the VLCDR1 amino acid sequence of SEQ ID NO: 9, the VLCDR2 amino acid sequence of SEQ ID NO: 10, and the VLCDR3 amino acid sequence of SEQ ID NO: 11; (ii) comprises a VH comprising the VHCDR1 amino acid sequence of SEQ ID NO: 4, the VHCDR2 amino acid sequence of SEQ ID NO: 5, and the VHCDR3 amino acid sequence of SEQ ID NO: 3; and a VL comprising the VLCDR1 amino acid sequence of SEQ ID NO: 12, the VLCDR2 amino acid sequence of SEQ ID NO: 13, and the VLCDR3 amino acid sequence of SEQ ID NO: 14; (iii) comprises a VH comprising the VHCDR1 amino acid sequence of SEQ ID NO: 195, the VHCDR2 amino acid sequence of SEQ ID NO: 2, and the VHCDR3 amino acid sequence of SEQ ID NO: 3; and a VL comprising the VLCDR1 amino acid sequence of SEQ ID NO: 9, the VLCDR2 amino acid sequence of SEQ ID NO: 10, and the VLCDR3 amino acid sequence of SEQ ID NO: 11; or (iv) VH comprising the VHCDR1 amino acid sequence of SEQ ID NO: 195, the VHCDR2 amino acid sequence of SEQ ID NO: 5, and the VHCDR3 amino acid sequence of SEQ ID NO: 3; and VL comprising the VLCDR1 amino acid sequence of SEQ ID NO: 12, the VLCDR2 amino acid sequence of SEQ ID NO: 13, and the VLCDR3 amino acid sequence of SEQ ID NO: 14 Use comprising

14. wherein the cancer is selected from solid tumors, lung cancer, skin cancer, kidney cancer, liver cancer, prostate cancer, breast cancer, colorectal cancer, gastric cancer, pancreatic cancer, thyroid cancer, brain cancer, uterine cancer, nasopharyngeal cancer, head and neck cancer, ovarian cancer, endometrial cancer, endocrine cancer, bladder cancer, urothelial cancer, or hematological cancer, or metastatic lesions of cancer Use according to any one of claims 1 - 13.

15. (a) the lung cancer is selected from non - small cell lung cancer (NSCLC), lung adenocarcinoma, squamous cell lung cancer, or small cell lung cancer; (b) the skin cancer is melanoma or Merkel cell carcinoma; (c) the kidney cancer is selected from renal cell carcinoma (RCC), metastatic renal cell carcinoma, or clear cell renal cell carcinoma (CCRCC); (d) the hematological cancer is selected from lymphoma, myeloma, or leukemia; (e) the brain cancer is glioblastoma; (f) the breast cancer is triple - negative breast cancer; (g) the liver cancer is hepatocellular carcinoma; or (h) the cancer is MSI - high (due to high microsatellite instability) cancer, Use according to claim 14.

16. (a) the NSCLC comprises a KRAS mutation; or (b) the melanoma is selected from progressive melanoma, unresectable melanoma, metastatic melanoma, melanoma having a BRAF mutation, melanoma having an NRAS mutation, cutaneous melanoma, or intraocular melanoma; Use according to claim 15.

17. Use according to any one of claims 1 - 16 in a further combination with one or more therapeutic agents or procedures.

18. One or more therapeutic agents or procedures are selected from one or more of chemotherapy, targeted anti - cancer therapy, tumor lysing agents, cytotoxic agents, immune - based therapies, cytokines, surgical procedures, radiation procedures, activators of costimulatory molecules, inhibitors of inhibitory molecules, vaccines or cellular immunotherapies, Use according to claim 17.

19. One or more therapeutic agents or procedures are: (a) an agonist of a costimulatory molecule selected from one or more of GITR, OX40, CD2, CD27, CD8, ICAM - 1, LFA - 1 (CD11a / CD18), ICOS (CD278), 4 - 1BB (CD137), CD30, CD40, BAFFR, HVEM, CD7, LIGHT, NKG2C, SLAMF7, NKp80, CD160, B7 - H3 or CD83 ligand; or (b) an inhibitor of an immune checkpoint molecule selected from one or more of PD - L1, PD - L2, CTLA - 4, TIM - 3, LAG - 3, CEACAM - 1, CEACAM - 5, VISTA, BTLA, TIGIT, LAIR1, CD160, 2B4 or TGF - R, Use according to claim 17.

20. One or more therapeutic agents or procedures are: (a) an inhibitor of PD - 1; (b) an inhibitor of LAG - 3; (c) an inhibitor of TIM - 3; (d) an agonist of GITR; (e) an interleukin; (f) a MEK inhibitor; (g) an FGFR inhibitor, for example, for treating hepatocellular carcinoma; (h) chemotherapy for treating lung cancer; (i) An indoleamine-pyrrole 2,3-dioxygenase (IDO) inhibitor for treating lung cancer; (j) An inhibitor of CTLA-4 for treating lung cancer or melanoma; (k) A cancer vaccine, for example, a dendritic cell renal cell carcinoma (DC-RCC) vaccine; (l) One or more of an immunotherapy-based treatment, a targeted agent, a VEGF tyrosine kinase inhibitor, an RNAi inhibitor, or an inhibitor of a downstream mediator of VEGF signaling for treating kidney cancer; (m) One, two, or all of oxaliplatin, leucovorin, or 5-FU for treating melanoma, colorectal cancer, non-small cell lung cancer, ovarian cancer, breast cancer, prostate cancer, pancreatic cancer, hematological cancer, or renal cell carcinoma; or (n) A tyrosine kinase inhibitor (for example, axitinib) for treating kidney cancer, being, The use according to claim 17.

21. (a) The inhibitor of PD-1 is, for example, an anti-PD-1 antibody molecule for treating thyroid cancer, non-small cell lung cancer, triple-negative breast cancer, endometrial cancer, uterine cancer, or lymphoma, or is selected from nivolumab, pembrolizumab, pidilizumab, AMP-224, or AMP514; (b) The inhibitor of LAG-3 is, for example, an anti-LAG-3 antibody molecule for treating NSCLC, melanoma, RCC, or hematological cancer; (c) The inhibitor of TIM-3 is, for example, an anti-TIM-3 antibody molecule for treating melanoma or renal cell carcinoma; (d) The agonist of GITR is, for example, an anti-GITR antibody molecule or a GITR fusion protein for treating non-small cell lung cancer; (e) The interleukin is, for example, IL-15 for treating solid tumors; (f) the MEK inhibitor is selected from, for example, ARRY-142886, G02442104 (GSK1120212), RDEA436, RDEA119 / BAY 869766, AS703026, G00039805 (AZD-6244 or selumetinib), BIX 02188, BIX 02189, CI-1040 (PD-184352), PD0325901, PD98059, U0126, GDC-0973 (Methanone or [3,4-difluoro-2-[(2-fluoro-4-iodophenyl)amino]phenyl][3-hydroxy-3-(25)-2-piperidinyl-l-azetidinyl]-), G-38963, G02443714 (AS703206), or a pharmaceutically acceptable salt or solvate thereof for treating, for example, triple negative breast cancer, NSCLC, or colorectal cancer; (g) the chemotherapy is platinum doublet chemotherapy; (h) the IDO inhibitor is INCB24360; (i) the inhibitor of CTLA-4 is an anti-CTLA-4 antibody (e.g., ipilimumab) or a soluble ligand of CTLA-4, where the antibody molecule or pharmaceutical composition is further used in combination with a BRAF inhibitor, e.g., vemurafenib or dabrafenib; (j) the immunotherapy-based therapy includes interleukin-2 or interferon-α; (k) the targeted agent is a VEGF inhibitor, e.g., an anti-VEGF antibody; (l) the VEGF tyrosine kinase inhibitor is selected from sunitinib, sorafenib, axitinib, or pazopanib; or (m) the inhibitor of a downstream mediator of VEGF signaling is an inhibitor of the mammalian target of rapamycin (mTOR) of rapamycin, e.g., temsirolimus; The use according to claim 20.

22. One or more therapeutic agents are: 1) Protein kinase C (PKC) inhibitor; 2) Heat shock protein 90 (HSP90) inhibitor; 3) Phosphoinositide 3-kinase (PI3K) and / or mammalian target of rapamycin (mTOR) inhibitor; 4) Cytochrome P450 inhibitor (e.g., CYP17 inhibitor or 17alpha-hydroxylase / C17-20 lyase inhibitor); 5) Iron chelating agent; 6) Aromatase inhibitor; 7) p53 inhibitor, e.g., inhibitor of p53 / Mdm2 interaction; 8) Apoptosis inducer; 9) Angiogenesis inhibitor; 10) Aldosterone synthase inhibitor; 11) Smoothened (SMO) receptor inhibitor; 12) Prolactin receptor (PRLR) inhibitor; 13) Wnt signaling inhibitor; 14) CDK4 / 6 inhibitor; 15) Fibroblast growth factor receptor 2 (FGFR2) / Fibroblast growth factor receptor 4 (FGFR4) inhibitor; 16) Macrophage colony-stimulating factor (M-CSF) inhibitor; 17) Inhibitor of one or more of c-KIT, histamine release, Flt3 (e.g., FLK2 / STK1) or PKC; 18) Inhibitor of one or more of VEGFR-2 (e.g., FLK-1 / KDR), PDGFR beta, c-KIT or Raf kinase C; 19) Somatostatin agonist and / or growth hormone release inhibitor; 20) Anaplastic lymphoma kinase (ALK) inhibitor; 21) Insulin-like growth factor 1 receptor (IGF-1R) inhibitor; 22) P-glycoprotein 1 inhibitor; 23) Vascular endothelial growth factor receptor (VEGFR) inhibitor; 24) BCR-ABL kinase inhibitor; 25) FGFR inhibitor; 26) CYP11B2 inhibitor; 27) HDM2 inhibitor, e.g., inhibitor of HDM2-p53 interaction; 28) Tyrosine kinase inhibitor; 29) c-MET inhibitor; 30) JAK inhibitor; 31) DAC inhibitor; 32) 11beta-hydroxylase inhibitor; 33) IAP inhibitor; 34) PIM kinase inhibitor; 35) Porcupine inhibitor; 36) BRAF, e.g., BRAF V600E or wild-type BRAF inhibitor; 37) HER3 inhibitor; 38) MEK inhibitor; or 39) Lipid kinase inhibitor, The use according to claim 17, which is as described above.

23. The use according to claim 17, wherein the one or more therapeutic agents are the compounds A1-A51 described in Table 6. **Claim 24** The one or more therapeutic agents are from one or more of the following categories (i)-(iii): (i) Agents that enhance tumor antigen presentation selected from one or more of STING agonists, TLR agonists, A2AR antagonists, oncolytic viruses, TIM-3 modulators, vascular endothelial growth factor receptor (VEGFR) inhibitors, c-Met inhibitors, TGFb inhibitors, IDO / TDO inhibitors, vaccines or bispecific or trispecific cell-engaging factors, or combinations thereof, (ii) Agents that enhance effector cell responses selected from one or more of GITR agonists, PD-1 inhibitors, PD-L1 inhibitors, inhibitors of IAP (inhibitors of apoptosis proteins), inhibitors of EGFR (epidermal growth factor receptor), inhibitors of mammalian target of rapamycin (mTOR), IL-15 or variants thereof, CTLA-4 inhibitors, bispecific antibody molecules that bind CD3 and tumor antigens, CD40 agonists, OX40 agonists or CD27 agonists, or combinations thereof, and / or, (iii) Agents that reduce tumor immunosuppression selected from one or more of inhibitors of immune checkpoint molecules selected from one or more of GITR agonists, PD-1, LAG-3, TIM-3 or CTLA-4, CSF-1 / 1R inhibitors, IL-17 inhibitors, IL-1β inhibitors, CXCR2 inhibitors, inhibitors of PI3Kγ or PI3Kδ), (vii) BAFF-R inhibitors, MALT-1 / BTK inhibitors, JAK inhibitors, CRTH2 inhibitors, VEGFR inhibitors, IL-15 or variants thereof, CTLA-4 inhibitors, IDO / TDO inhibitors, A2AR antagonists, TGFb inhibitors or PFKFB3 inhibitors, or combinations thereof The use according to claim 17, selected from one, two or all of the above. **Claim 25** The use according to any one of claims 1-24, wherein the AKT inhibitor is RX-0201 or MK-2206.

26. wherein the subject is: (a) a cancer that expresses PD-L1; (b) a cancer that is positive for one, two, or all of PD-L1, CD8, and IFN-γ; (c) a cancer that is triple positive for PD-L1, CD8, and IFN-γ; or (d) a cancer that is positive for tumor-infiltrating lymphocytes (TIL) and having one or more of or identified as having one or more of the foregoing, the use according to any one of claims 1-25.

27. The use according to any one of claims 1-26, wherein the antibody molecule or the pharmaceutical composition comprising the antibody molecule is administered at a dose of about 1-30 mg / kg, such as a dose of about 1-5 mg / kg.

28. The use according to claim 27, wherein the antibody molecule or the pharmaceutical composition comprising the antibody molecule is administered once a week to once every 2, 3, or 4 weeks.

29. The use according to any one of claims 1-28, wherein the combination is administered sequentially or simultaneously.

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